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Rising risks of climate change and supply chain insecurity highlight the need to develop alternative, greener synthesis routes to common materials currently sourced from petroleum. Biological systems excel at interconverting chemicals with exquisite specificity and speed, using networks of enzymes that perform catalysis at mild conditions. Protein complexes in nature colocalize complementary subunits to perform sophisticated biochemistry, and artificial, spatial organization of enzyme systems into synthetic complexes is an attractive strategy for improving biocatalytic process throughputs in industrial settings. While some sets of modular parts that enable designer protein complex construction exist, there is still a need to develop new components that are widely compatible with different enzymes and that are highly engineerable to impart desired self-assembly properties. Fungal cellulosomes, modular protein machines produced by anaerobic fungi in the guts of herbivores to rapidly free sugars from plant matter, represent an unexplored framework for synthetic protein complex construction. Cellulosomes synergistically incorporate enzymes involved in biomass degradation into discrete complexes via modular protein-protein interactions between enzyme fused dockerin domains and cohesin domains repeated on a central scaffoldin protein. Over 80% of the degradative power anaerobic fungi possess is attributed to cellulosomes, but the mechanistic nature of their activity and their assembly mechanism remain unknown. These knowledge gaps have precluded the development of fungal cellulosomes or their parts as biocatalytic technologies with real world applications. We apply a range of experimental techniques towards addressing how cellulosomes are produced in native anaerobic fungal cultures and characterizing the composition, nanostructure, and biochemical activity of purified, native cellulosomes. Immunofluorescence microscopy with cellulosome-labeling antibodies shows celluloso

The Triassic-Jurassic transition (~201.5Ma) is marked by one of the largest mass extinctions in Earth’s history. This was accompanied by significant perturbations in ocean and atmosphere geochemistry, including the global carbon cycle, as expressed by major fluctuations in carbon isotope ratios. Central Atlantic Magmatic Province (CAMP) volcanism triggered environmental changes and played a key role in this biotic crisis. Biostratigraphic and chronostratigraphic studies link the end-Triassic mass extinction with the early phases of CAMP volcanism, and notable mercury enrichments in geographically distributed marine and continental strata are shown to be coeval with the onset of the extrusive emplacement of CAMP. Sulfuric acid induced atmospheric aerosol clouds from subaerial CAMP volcanism can explain a brief, relatively cool seawater temperature pulse in the mid-paleolatitude Pan-European seaway across the T–J transition. The occurrence of CAMP-induced carbon degassing may explain the overall long-term shift toward much warmer conditions. The effect of CAMP volcanism on seawater87Sr/86Sr values might have been indirect by driving enhanced continental weathering intensity. Changes in ocean-atmosphere geochemistry and associated (causative) effects on paleoclimatic, paleoenvironmental, and paleoceanographic conditions on local, regional, and global scales are however not yet fully constrained.

The contribution to knowledge, and the core of the research, is a tonal foundation based on glissandi using compositional techniques derived from synergy of glissandi, microtonality, and isorhythm. The techniques are performed on specially constructed guitars in 18, 24, 30, and 36 tet (tone equal temperament). Guitar based musical artefacts demonstrating some possible techniques are arranged on two compact discs: CD1 ‘Experimental Miniatures’ and CD2 ‘After Twelve’.

The contribution to knowledge, and the core of the research, is a tonal foundation based on glissandi using compositional techniques derived from synergy of glissandi, microtonality, and isorhythm. The techniques are performed on specially constructed guitars in 18, 24, 30, and 36 tet (tone equal temperament). Guitar based musical artefacts demonstrating some possible techniques are arranged on two compact discs: CD1 ‘Experimental Miniatures’ and CD2 ‘After Twelve’.

The contribution to knowledge, and the core of the research, is a tonal foundation based on glissandi using compositional techniques derived from synergy of glissandi, microtonality, and isorhythm. The techniques are performed on specially constructed guitars in 18, 24, 30, and 36 tet (tone equal temperament). Guitar based musical artefacts demonstrating some possible techniques are arranged on two compact discs: CD1 ‘Experimental Miniatures’ and CD2 ‘After Twelve’.

The contribution to knowledge, and the core of the research, is a tonal foundation based on glissandi using compositional techniques derived from synergy of glissandi, microtonality, and isorhythm. The techniques are performed on specially constructed guitars in 18, 24, 30, and 36 tet (tone equal temperament). Guitar based musical artefacts demonstrating some possible techniques are arranged on two compact discs: CD1 ‘Experimental Miniatures’ and CD2 ‘After Twelve’.

Powder diffraction patterns of standard reference material LaB6 (660a) have been used to determine the energy of a synchrotron beam across the range 7-12keV with typical uncertainties of 0.1eV. Diffraction peaks were recorded on X-ray image plates over a

The design and operation of electrosynthesis cells for generation of samples for X-ray absorption spectroscopy are described. Optimization of continuous-flow methods allows the generation of highly reducing species, which may be combined with spectroscopic validation of the composition of the electrogenerated solution. It is shown that the large sample volume (10mL) of the 110mM (in the absorbing element) solution required for such experiments can be reduced to ∼100L using a strategy in which the in situ electrosynthesis cell is amenable to freeze-quenching and transfer to a beamline cryostat. The working electrode in this case doubles as the X-ray absorption spectroscopy sample cell. The application of these techniques is illustrated by the reduction chemistry of Fe3S2(CO)9, 3Fe2S. Spectra recorded in the near-edge region confirm that quantitative preparation of samples of 3Fe2S, 3Fe2S1- and 3Fe2S2- can be prepared by either approach, but samples of a more reduced form, identified as [Fe3S(CO)9]2-, could only be generated using continuous-flow electrosynthesis techniques. Differences in the structural chemistry of the 3Fe2S-0/1/2- redox series were examined from the perspective of their near-edge spectra and the structures of 3Fe2S1- and 3Fe2S2- forms were deduced by a combination of computational (density functional theory), spectroscopic and X-ray absorption fine-structure analyses. These show that addition of the first electron is predominantly localized in one of the FeFe bonds; cleavage of the FeFe bond by addition of a second electron to the FeFe antibonding orbital is associated with a more substantial rearrangement of the molecule. The reduced compounds have structural similarities to the reduced dithiolate-bridged di-iron hexacarbonyl compounds and this is related to the weak electrocatalytic proton reduction exhibited by Fe3S2(CO)9. The methods described provide a strategy for the collection and analysis of experimental data directed towards structure elu

Recent developments in the understanding of the biosynthesis of the active site of the nitrogenase enzyme, the structure of the iron centre of [Fe]-hydrogenase and the structure and biomimetic chemistry of the [FeFe] hydrogenase H-cluster as deduced by application of X-ray spectroscopy are reviewed. The techniques central to this work include X-ray absorption spectroscopy either in the form of extended X-ray absorption fine structure (EXAFS), X-ray absorption near-edge structure (XANES) and nuclear resonant vibrational spectroscopy (NRVS). Examples of the advances in the understanding of the chemistry of the system through integration of a range of spectroscopic and computational techniques with X-ray spectroscopy are highlighted. The critical role played by ab initio calculation of structural and spectroscopic properties of transition-metal compounds using density functional theory (DFT) is illustrated both by the calculation of nuclear resonance vibrational spectroscopy (NRVS) spectra and the structures and spectra of intermediates through the catalytic reactions of hydrogenase model compounds. Crown

This thesis reports on experimental and modelling work carried out in order to make quantitative estimates on the environmental impacts of home composting. The focus of the work was climate relevant gaseous emissions, and developing and utilising a methodology for quantifying them. Experiments using 220L open bottomed home compost bins, alongside purpose built 200L composting reactors with airflow control were performed. A variety of composting conditions were tested, using different compositions of garden and kitchen wastes. The experiments were monitored for headspace gas composition, including CO2, O2, NH3, N2O, CH4 and volatile organic compounds, as well as temperature, humidity, moisture and solids losses and pH. From the CO2 emission rates calculated from the reactor experiments, theoretical analysis and modelling and airflow pathway tests on home compost bins, it was concluded that molecular diffusion, rather than bulk convective flow, is the dominant gas transfer mechanism from home compost bins. There were no detected emissions of N2O but emissions of NH3 up to 16 g/T feed. Only a few cases of CH4 emission were detected, typically in the first 2-3 days following a feed addition, with the highest single concentration measured at 86 ppm within the headspace. The total anthropogenic greenhouse gas emissions from home composting were estimated as between 3 and 12 Kg CO2E/Tw with almost 90% coming from the lifecycle of the compost bin. This compares with between 20 and 56 Kg CO2E/Tw from centralised facilities, at least more than double that for home composting. Total anthropogenic CO2-equivalent emissions from home composting in the UK in 2008 were estimated to be in the region of 7 thousand tonnes CO2E.

This thesis reports on experimental and modelling work carried out in order to make quantitative estimates on the environmental impacts of home composting. The focus of the work was climate relevant gaseous emissions, and developing and utilising a methodology for quantifying them. Experiments using 220L open bottomed home compost bins, alongside purpose built 200L composting reactors with airflow control were performed. A variety of composting conditions were tested, using different compositions of garden and kitchen wastes. The experiments were monitored for headspace gas composition, including CO2, O2, NH3, N2O, CH4 and volatile organic compounds, as well as temperature, humidity, moisture and solids losses and pH. From the CO2 emission rates calculated from the reactor experiments, theoretical analysis and modelling and airflow pathway tests on home compost bins, it was concluded that molecular diffusion, rather than bulk convective flow, is the dominant gas transfer mechanism from home compost bins. There were no detected emissions of N2O but emissions of NH3 up to 16 g/T feed. Only a few cases of CH4 emission were detected, typically in the first 2-3 days following a feed addition, with the highest single concentration measured at 86 ppm within the headspace. The total anthropogenic greenhouse gas emissions from home composting were estimated as between 3 and 12 Kg CO2E/Tw with almost 90% coming from the lifecycle of the compost bin. This compares with between 20 and 56 Kg CO2E/Tw from centralised facilities, at least more than double that for home composting. Total anthropogenic CO2-equivalent emissions from home composting in the UK in 2008 were estimated to be in the region of 7 thousand tonnes CO2E.

English: This thesis evaluates the Court for Sexual Offences in Bloemfontein at the hand of the perceptions of the victims, their families, offenders and professionals involved with this court. The first article is a theoretical orientation to the relatively unique area of sex courts. The remaining four empirical articles discuss the perceptions of the parties involved with the Court. The findings indicate that the victims of sexual offences, their families and the professionals working in the Court and associated auxiliary services, are generally of the opinion that the establishment of specialist sex courts has gone a long way towards ensuring improved justice for victims and decreasing the incidents of secondary victimization during judicial proceedings. However, the Court still exhibits shortcomings in several areas. Furthermore, doubts are expressed with regard to the impartiality of such specialist courts, sentences for sexual offenders are perceived to be ineffectual and the after-care afforded to the victims and their families appears to be very poor. The perpetrators of sexual offences also perceive the Court for Sexual Offences to be slow and inefficient. They were of the opinion that this particular system of courts is inherently biased. The victim's rights are perceived to be overemphasized and the majority of the offenders questioned felt that it was not possible for an individual accused of a sex crime to receive a fair trial in a specialist sex court., Afrikaans: Die doel van hierdie proefskrif is om die Hof vir Seksuele Misdrywe in Bloemfontein te evalueer en wel aan die hand van die persepsies van die slagoffers, hul families, oortreders en professionele persone wat by die Hof betrokke is. Die eerste artikel is 'n teoretiese oriëntering ten opsigte van die relatief unieke terrein van howe vir seksuele misdrywe. Die oorblywende vier artikels is empiries van aard en fokus spesifiek op die genoemde persepsies. Die bevindings dui daarop dat die slagoffers van seksuele misdrywe, hul families en die professionele persone betrokke oor die algemeen van mening is dat die daarstelling van gespesialiseerde howe vir seksuele misdrywe baie bygedra het om geregtigheid vir slagoffers te verseker en die voorkoms van sekondêre viktimisering tydens die juridiese verrigtinge te verminder. Aan die anderkant gaan die Hof steeds mank aan sekere leemtes. Verder bestaan daar twyfel ten opsigte van die onpartydigheid van sodanige spesialishowe, terwyl strawwe vir seksuele oortreders as oneffektief beskou word en die nasorg van slagoffers en hul families veel te wense oorlaat. Die seksuele oortreders beskou ook die Hof vir Seksuele Misdrywe as stadig en ontoereikend. Hulle was van mening dat hierdie spesialishof inherent bevooroordeeld is. Hul persepsie is dat die slagoffers se regte oorbeklemtoon word en dat dit nie moontlik is vir 'n individu wat van 'n seksuele oortreding aangekla word, om 'n regverdige verhoor in 'n spesialishofvir seksuele misdrywe te kry nie.

English: Adolescence is a time of immense physical, cognitive and emotional development. This transition from childhood to adulthood begins with the onset of puberty. Puberty also coincides with the transition many learners have to make from primary to secondary school. This transition confronts adolescents with new educational environments and challenges. Some adolescent learners experience a great deal of stress during this period and are thus not able to maintain the same level of academic achievement as they did in primary school. Other learners, however, appear to make the transition with very little difficulty. These learners are able to maintain or even improve upon their previous level of academic achievement. These differing experiences during the transitional period may be due to differences in the learners' adjustment. Learners' study habits and attitudes serve as important coping mechanisms within the changing educational environment. Specific attention was also given to the role of the family, peers, school and academic skills during the transition from primary to secondary school. The sample consisted of 137 grade eight learners from four secondary schools in the greater Bloemfontein area. The learners were divided into two groups on the basis of whether their academic achievement had improved or decreased from the end of grade seven to the middle of grade eight. These two groups were then compared with respect to their adjustment and general study orientation. The effects of gender, language and residential status upon adjustment and general study orientation were also investigated. Attention was also given to the interaction between changes in academic achievement and each of the above-mentioned independent variables. The statistical analysis of the data was performed by employing a multivariate analysis of variance (MANOVA). An analysis of variance (ANOVA) or post hoc t-test (Scheffé) was then used to determine the direction of any differences that, Afrikaans: Adolessensie is 'n periode van ingrypende fisiese, kognitiewe en emosionele ontwikkeling. Hierdie oorgang vanaf kinderjare na volwassenheid word deur die aanvangs van puberteit gekenmerk. Puberteit val saam met die oorskakeling wat meeste leerders vanaf primêre na sekondêre skool moet maak. Hierdie oorskakeling gaan met nuwe opvoedkundige omgewings en uitdagings gepaard. Sommige adolessente leerders ondervind 'n groot mate van spanning tydens hierdie periode en is nie in staat om dieselfde vlak van akademiese prestasie te handaaf as wat in die primêre skool die geval was nie. Ander leerders blyk die oorgang weer met betreklik min moeite te oorbrug. Laasgenoemde leerders is in staat om hul vorige vlak van akademiese prestasie te handhaaf of selfs daarop te verbeter. Hierdie uiteenlopende ervarings tydens die oorgangsperiode kan aan verskille in die leerders se algemene aanpassing toegeskryf word. Leerders se studiegewoontes en- houdings dien as belangrike hanteringsmeganismes binne die veranderende opvoedkundige konteks. Spesifieke aandag is ook aan die rol van die gesin, die portuurgroep, die skool en akademiese vaardighede tydens die oorgang van primêre na sekondêre skool geskenk. Die steekproef het uit 137 graad agt leerders uit vier sekondêre skole in die groter Bloemfontein-area bestaan. Die leerders is in twee groepe verdeel op grond van of hul akademiese prestasie verbeter of verswak het vanaf die einde van graad sewe tot die helfte van graad agt. Die twee groepe is toe op grond van hul aanpassing en algemene studie-oriëntasie vergelyk. Voorts is die invloed van geslag, taal en residensiële status op aanpassing en algemene studie-oriëntasie ondersoek. Aandag is ook aan die interaksie tussen veranderings in akademiese prestasie en elk van bogenoemde veranderlikes geskenk. Die statistiese ontleding van data is uitgevoer deur die gebruik van meervoudige variansie-ontleding (MANOVA). 'n Variansie ontleding (ANOVA) of post hoc t-toets (Scheffé) is gebrui, Centre for Science Development (HSRC South Africa)

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This thesis examines aspects of the political career of Robert Wilmot Horton (1784-1841), a junior minister in the Tory governments of the 1820s and an advocate of state-aided emigration to the British colonies. It considers how far Wilmot conforms to existing conceptualisations of 'liberal Toryism', which are summarized in Chapter 1. Chapter 2 finds both ambition and principle in Wilmot's choice of party, while identifying fundamental aspects of his political make-up, in particular his devotion to political economy and his hostility to political radicalism. Chapters 3 to 5 explore his economic thinking. Chapter 3 charts Wilmot's gradual move away from a Malthusian approach to the problem of pauperism, and the resulting changes in his view of the role of emigration as a means of relief. Chapter 4 shows how his specific plan of colonization addressed broader considerations of imperial strategy and economic development. Chapter 5, exploring the wider context of economic debate, reveals Wilmot as an advocate of governmental activism in social policy, a critic of 'economical reform', and a moderate protectionist in the short term. Chapter 6 suggests that Wilmot, and the ministry as a whole, were driven by pragmatic rather than ideological considerations in their approach to the amelioration of slavery. Chapter 7 concludes that Wilmot's advocacy of Catholic Emancipation, on grounds of expediency, conformed to the approach normally ascribed to liberal Tories in principle if not in detail. Chapter 8 finds, in Wilmot's pamphleteering and lecturing, a striking instance of an 'outward turn' in political behaviour; and, in his support for parliamentary reform in 1831, a continuing determination to resist political radicalism. Overall, the thesis argues that Wilmot embraced political economy more in its 'secular' than its 'Christian' guise, but took interventionist positions on economic and social questions which set him apart from his colleagues. These conclusions complicate t

Traditionally, the bishops who held office during the civil war which dominated King Stephen's reign (1135-1154) have been considered weak and ineffective, able neither to bring peace between the two sides or among warring local barons nor to protect their flocks or even themselves from the so-called 'Anarchy'. The explanation for this has been found in the bishops' lack of spiritual calibre. Bishops have also been seen as withdrawing their support from the king and ending their involvement in royal government, partly because of increasing general ecclesiastical desire for separation between Church and State and partly because of specific disputes with Stephen. As a consequence of all this, bishops are allowed little importance in modern histories of Stephen's reign. This thesis shows that modern historiographical consensus is based in flawed interpretive frameworks which have led to misinterpretation of the nature of the episcopate and its importance in Stephen's reign. It offers more valid alternatives and then re-examines, the royal, ecclesiastical and, especially, the local evidence in light of them to show that, in fact, the bishops were crucially important figures in regional politics, religion and society during the civil war. It proves as well, that they could possess considerable spiritual authority and continued to be committed to the king and active in the government of the kingdom throughout the period. Additionally, each of these also has consequences for how the episcopacy and Anglo-Norman history in general are understood. This is, therefore, a reassessment of the bishops of King Stephen's reign.

We have developed a new ligand (H\(_3\)L\(^2\)) based on a DTPA bis-amide backbone which improves upon previous work within the group. The quantum yields of the europium and samarium complexes of H\(_3\)L\(^2\) are greatly enhanced over that of H\(_3\)L\(^1\). The previously observed nanoparticle-based quenching of the lanthanide luminescence lifetime of EuL\(^1\) is suppressed in EuL\(^2\). We have demonstrated the functionalisation of gold and platinum nanoparticles of various sizes with the surface active lanthanide complexes of H\(_3\)L\(^1\) and H\(_3\)L\(^2\). As a proof of principle, we have imaged, tracked and measured velocities of individual EuL\(^2\) functionalised nanoparticles in an aqueous flow, providing flow-rates accurate to micrometer resolution of fluids through a 2 mm diameter tube. We have also observed the microscale mixing of a nanoparticle labelled solution with water on time-scales of milliseconds to seconds. We can study the development and measure the dimensions of features present during mixing and therefore assess the efficiency of that mixing. We have also visualised a turn-on event at the boundary of aqueous solutions of Eu(III) ions and H\(_3\)L\(^2\)-functionalised nanoparticles during mixing, demonstrating a novel optical method for ascertaining the degree of mixing between two solutions and the position and area of that mixing.

Traditionally, the bishops who held office during the civil war which dominated King Stephen's reign (1135-1154) have been considered weak and ineffective, able neither to bring peace between the two sides or among warring local barons nor to protect their flocks or even themselves from the so-called 'Anarchy'. The explanation for this has been found in the bishops' lack of spiritual calibre. Bishops have also been seen as withdrawing their support from the king and ending their involvement in royal government, partly because of increasing general ecclesiastical desire for separation between Church and State and partly because of specific disputes with Stephen. As a consequence of all this, bishops are allowed little importance in modern histories of Stephen's reign. This thesis shows that modern historiographical consensus is based in flawed interpretive frameworks which have led to misinterpretation of the nature of the episcopate and its importance in Stephen's reign. It offers more valid alternatives and then re-examines, the royal, ecclesiastical and, especially, the local evidence in light of them to show that, in fact, the bishops were crucially important figures in regional politics, religion and society during the civil war. It proves as well, that they could possess considerable spiritual authority and continued to be committed to the king and active in the government of the kingdom throughout the period. Additionally, each of these also has consequences for how the episcopacy and Anglo-Norman history in general are understood. This is, therefore, a reassessment of the bishops of King Stephen's reign.

We have developed a new ligand (H\(_3\)L\(^2\)) based on a DTPA bis-amide backbone which improves upon previous work within the group. The quantum yields of the europium and samarium complexes of H\(_3\)L\(^2\) are greatly enhanced over that of H\(_3\)L\(^1\). The previously observed nanoparticle-based quenching of the lanthanide luminescence lifetime of EuL\(^1\) is suppressed in EuL\(^2\). We have demonstrated the functionalisation of gold and platinum nanoparticles of various sizes with the surface active lanthanide complexes of H\(_3\)L\(^1\) and H\(_3\)L\(^2\). As a proof of principle, we have imaged, tracked and measured velocities of individual EuL\(^2\) functionalised nanoparticles in an aqueous flow, providing flow-rates accurate to micrometer resolution of fluids through a 2 mm diameter tube. We have also observed the microscale mixing of a nanoparticle labelled solution with water on time-scales of milliseconds to seconds. We can study the development and measure the dimensions of features present during mixing and therefore assess the efficiency of that mixing. We have also visualised a turn-on event at the boundary of aqueous solutions of Eu(III) ions and H\(_3\)L\(^2\)-functionalised nanoparticles during mixing, demonstrating a novel optical method for ascertaining the degree of mixing between two solutions and the position and area of that mixing.

This thesis examines aspects of the political career of Robert Wilmot Horton (1784-1841), a junior minister in the Tory governments of the 1820s and an advocate of state-aided emigration to the British colonies. It considers how far Wilmot conforms to existing conceptualisations of 'liberal Toryism', which are summarized in Chapter 1. Chapter 2 finds both ambition and principle in Wilmot's choice of party, while identifying fundamental aspects of his political make-up, in particular his devotion to political economy and his hostility to political radicalism. Chapters 3 to 5 explore his economic thinking. Chapter 3 charts Wilmot's gradual move away from a Malthusian approach to the problem of pauperism, and the resulting changes in his view of the role of emigration as a means of relief. Chapter 4 shows how his specific plan of colonization addressed broader considerations of imperial strategy and economic development. Chapter 5, exploring the wider context of economic debate, reveals Wilmot as an advocate of governmental activism in social policy, a critic of 'economical reform', and a moderate protectionist in the short term. Chapter 6 suggests that Wilmot, and the ministry as a whole, were driven by pragmatic rather than ideological considerations in their approach to the amelioration of slavery. Chapter 7 concludes that Wilmot's advocacy of Catholic Emancipation, on grounds of expediency, conformed to the approach normally ascribed to liberal Tories in principle if not in detail. Chapter 8 finds, in Wilmot's pamphleteering and lecturing, a striking instance of an 'outward turn' in political behaviour; and, in his support for parliamentary reform in 1831, a continuing determination to resist political radicalism. Overall, the thesis argues that Wilmot embraced political economy more in its 'secular' than its 'Christian' guise, but took interventionist positions on economic and social questions which set him apart from his colleagues. These conclusions complicate t

Small interfering RNA siRNA promise to be highly effective molecules but susceptible to some of the limitations experienced with antisense oligonucleotide ASO technology biological instability, design of target mRNA accessible sequences and efficient delivery to subcellular sites of action. This thesis has aim to address each of these limitations for siRNA. The biological stability of 5'-end 32P radiolabeled siRNA and ASO were tested in FBS, FBS-containing cell culture media and in A431 cell lysates. The biological degradation profiles of ASO and siRNA in serum were comparable ASO t1/2 19 minutes and siRNA t1/2 19 minutes. The stability of siRNA was higher than ASO in A431 cell lysates ASO tU2 37 minutes and siRNA t1/2 43 minutes. siRNA were designed using innovative scanning ASO array technologies to EGFR mRNA accessible sites. Initially seven siRNAs were designed against different mRNA accessible sites. Based on above a further five siRNAs were designed incorporating termini modifications to alter the thermodynamic stability of the siRNA duplex. These sequences were evaluated for biological activity following optimisation of a delivery strategy. siRNA delivery to A431 cells was optimised using a FACs assay with Oligofectamine and fluorescendy labelled siRNA. Oligofectamine delivery showed significant cell fluorescence associated with labelled siRNA compared with untreated controls P 0.05. The activity, cell proliferation and EGFR protein and mRNA knockdown, of the designed siRNA was investigated in A431 and TamR cells. RT-PCR, Western blot and immunofluorescent microscopy showed inhibitory trends, but ultimately lacked sufficient resolution and reproducibility A431 growth was not inhibited. A proprietary siRNA showed knockdown of EGFR mRNA in TamR cells, but all the custom designed siRNA produced highly variable effects, failing to show significant knockdown, even when various designed siRNA were pooled. ASO and siRNA share common pharmaceutical developmental prob

Small interfering RNA siRNA promise to be highly effective molecules but susceptible to some of the limitations experienced with antisense oligonucleotide ASO technology biological instability, design of target mRNA accessible sequences and efficient delivery to subcellular sites of action. This thesis has aim to address each of these limitations for siRNA. The biological stability of 5'-end 32P radiolabeled siRNA and ASO were tested in FBS, FBS-containing cell culture media and in A431 cell lysates. The biological degradation profiles of ASO and siRNA in serum were comparable ASO t1/2 19 minutes and siRNA t1/2 19 minutes. The stability of siRNA was higher than ASO in A431 cell lysates ASO tU2 37 minutes and siRNA t1/2 43 minutes. siRNA were designed using innovative scanning ASO array technologies to EGFR mRNA accessible sites. Initially seven siRNAs were designed against different mRNA accessible sites. Based on above a further five siRNAs were designed incorporating termini modifications to alter the thermodynamic stability of the siRNA duplex. These sequences were evaluated for biological activity following optimisation of a delivery strategy. siRNA delivery to A431 cells was optimised using a FACs assay with Oligofectamine and fluorescendy labelled siRNA. Oligofectamine delivery showed significant cell fluorescence associated with labelled siRNA compared with untreated controls P 0.05. The activity, cell proliferation and EGFR protein and mRNA knockdown, of the designed siRNA was investigated in A431 and TamR cells. RT-PCR, Western blot and immunofluorescent microscopy showed inhibitory trends, but ultimately lacked sufficient resolution and reproducibility A431 growth was not inhibited. A proprietary siRNA showed knockdown of EGFR mRNA in TamR cells, but all the custom designed siRNA produced highly variable effects, failing to show significant knockdown, even when various designed siRNA were pooled. ASO and siRNA share common pharmaceutical developmental prob

Traditionally, the bishops who held office during the civil war which dominated King Stephen's reign (1135-1154) have been considered weak and ineffective, able neither to bring peace between the two sides or among warring local barons nor to protect their flocks or even themselves from the so-called 'Anarchy'. The explanation for this has been found in the bishops' lack of spiritual calibre. Bishops have also been seen as withdrawing their support from the king and ending their involvement in royal government, partly because of increasing general ecclesiastical desire for separation between Church and State and partly because of specific disputes with Stephen. As a consequence of all this, bishops are allowed little importance in modern histories of Stephen's reign. This thesis shows that modern historiographical consensus is based in flawed interpretive frameworks which have led to misinterpretation of the nature of the episcopate and its importance in Stephen's reign. It offers more valid alternatives and then re-examines, the royal, ecclesiastical and, especially, the local evidence in light of them to show that, in fact, the bishops were crucially important figures in regional politics, religion and society during the civil war. It proves as well, that they could possess considerable spiritual authority and continued to be committed to the king and active in the government of the kingdom throughout the period. Additionally, each of these also has consequences for how the episcopacy and Anglo-Norman history in general are understood. This is, therefore, a reassessment of the bishops of King Stephen's reign.

Traditionally, the bishops who held office during the civil war which dominated King Stephen's reign (1135-1154) have been considered weak and ineffective, able neither to bring peace between the two sides or among warring local barons nor to protect their flocks or even themselves from the so-called 'Anarchy'. The explanation for this has been found in the bishops' lack of spiritual calibre. Bishops have also been seen as withdrawing their support from the king and ending their involvement in royal government, partly because of increasing general ecclesiastical desire for separation between Church and State and partly because of specific disputes with Stephen. As a consequence of all this, bishops are allowed little importance in modern histories of Stephen's reign. This thesis shows that modern historiographical consensus is based in flawed interpretive frameworks which have led to misinterpretation of the nature of the episcopate and its importance in Stephen's reign. It offers more valid alternatives and then re-examines, the royal, ecclesiastical and, especially, the local evidence in light of them to show that, in fact, the bishops were crucially important figures in regional politics, religion and society during the civil war. It proves as well, that they could possess considerable spiritual authority and continued to be committed to the king and active in the government of the kingdom throughout the period. Additionally, each of these also has consequences for how the episcopacy and Anglo-Norman history in general are understood. This is, therefore, a reassessment of the bishops of King Stephen's reign.

Traditionally, the bishops who held office during the civil war which dominated King Stephen's reign (1135-1154) have been considered weak and ineffective, able neither to bring peace between the two sides or among warring local barons nor to protect their flocks or even themselves from the so-called 'Anarchy'. The explanation for this has been found in the bishops' lack of spiritual calibre. Bishops have also been seen as withdrawing their support from the king and ending their involvement in royal government, partly because of increasing general ecclesiastical desire for separation between Church and State and partly because of specific disputes with Stephen. As a consequence of all this, bishops are allowed little importance in modern histories of Stephen's reign. This thesis shows that modern historiographical consensus is based in flawed interpretive frameworks which have led to misinterpretation of the nature of the episcopate and its importance in Stephen's reign. It offers more valid alternatives and then re-examines, the royal, ecclesiastical and, especially, the local evidence in light of them to show that, in fact, the bishops were crucially important figures in regional politics, religion and society during the civil war. It proves as well, that they could possess considerable spiritual authority and continued to be committed to the king and active in the government of the kingdom throughout the period. Additionally, each of these also has consequences for how the episcopacy and Anglo-Norman history in general are understood. This is, therefore, a reassessment of the bishops of King Stephen's reign.

Traditionally, the bishops who held office during the civil war which dominated King Stephen's reign (1135-1154) have been considered weak and ineffective, able neither to bring peace between the two sides or among warring local barons nor to protect their flocks or even themselves from the so-called 'Anarchy'. The explanation for this has been found in the bishops' lack of spiritual calibre. Bishops have also been seen as withdrawing their support from the king and ending their involvement in royal government, partly because of increasing general ecclesiastical desire for separation between Church and State and partly because of specific disputes with Stephen. As a consequence of all this, bishops are allowed little importance in modern histories of Stephen's reign. This thesis shows that modern historiographical consensus is based in flawed interpretive frameworks which have led to misinterpretation of the nature of the episcopate and its importance in Stephen's reign. It offers more valid alternatives and then re-examines, the royal, ecclesiastical and, especially, the local evidence in light of them to show that, in fact, the bishops were crucially important figures in regional politics, religion and society during the civil war. It proves as well, that they could possess considerable spiritual authority and continued to be committed to the king and active in the government of the kingdom throughout the period. Additionally, each of these also has consequences for how the episcopacy and Anglo-Norman history in general are understood. This is, therefore, a reassessment of the bishops of King Stephen's reign.