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No description available.

The Planctomycetes are a deep branching phylum of the domain Bacteria that incorporate a diverse group of organisms possessing a number of unusual and distinct characteristics. These features include budding reproduction, the planctomycetecharacteristic crateriform structures on their cell surface, a cell wall that lacks peptidoglycan, internal compartmentalisation and unique molecular features of their rRNA genes. This study chose to investigate a number of aspects of planctomycete cell biology and diversity to further our knowledge of this unique group. In a study of the diversity of ribonuclease P (RNase P) RNA, one molecule of relevance to cell biology and compartmentalisation in planctomycetes, RNase P RNA genes were sequenced for species from all genera of planctomycetes for which a pure culture exists. Secondary structures for RNase P RNA of these strains were deduced, taking to 26 the number of planctomycete RNase P RNA structures. Nucleotide positions were identified in which some planctomycetes possess a less common form, including one thought to be otherwise conserved within all Bacteria and Archaea. Phylogenetic analysis of RNase P RNA genes was relatively consistent with that of 16S rRNA genes with the exception that clustering of Gemmata and anammox sequences occurred, possibly due to either long-branch attraction or lateral gene transfer. Analysis of RNase P RNA secondary structures revealed unusual features of planctomycetes relative to all other bacteria, including an additional helix within the P13 helix of ‘Candidatus Brocadia anammoxidans’, ‘Candidatus Kuenenia stuttgartiensis’ and all Gemmata sequences. The longest P12 helix of any bacteria type A RNase P RNA was found in a Gemmatalike isolate. The short tandem repeats in P12 helices of two Gemmata-like isolates are possibly analogous to short tandem repetitive repeat sequences of some cyanobacteria RNase P RNA. In experiments using Gemmata obscuriglobus as a model for planctomycete cell biology and compartmentalisation functions, electron microscope-level in situ hybridisation (EMISH), and subsequent statistical analysis, was developed to localise 16S rRNA, 23S rRNA and RNase P RNA to particular regions within Gemmata obscuriglobus, the first instance of EMISH being applied in this way to bacteria. Statistical analysis localised 16S rRNA to both nuclear body and to riboplasm outside this region but it was absent from paryphoplasm. While co-localisation of both 16S rRNA and 23S rRNA molecules, which might indicate assembled ribosomes, was rarely observed, 23S rRNA, like 16S rRNA, was distributed in both riboplasm-containing areas of the cell. While statistical analysis revealed minor DNA within riboplasm outside the nuclear body, the majority was localised to that body. These results suggest at least some uncoupling of translation from transcription involving ribosomes in the riboplasm. RNase P RNA was localised both to the nuclear body and to the riboplasm outside this region, suggesting that pre-tRNA processing occurs both within nuclear body, where RNA transcripts are presumably generated, and outside nuclear body, separated from the origin of these transcripts. This is also consistent with the hypothesis that processed tRNA is required in the riboplasm outside the nuclear body, due to occurrence of some uncoupled translation. In research on planctomycetes not yet examined with respect to cell plan or structure, 16S rRNA gene sequencing of isolate ATCC 35122 confirmed its very close relationship to the type strain of Pirellula staleyi and its membership of the phylum Planctomycetes. Morphological characteristics, including polar crateriform structures and the occurrence of a unique internal, single membrane-bounded compartment enclosing nucleoid and ribosome-like particles, the pirellulosome, and a polar cap region, are also consistent with its membership of the planctomycetes and of genus Pirellula. Cells often displayed pointed, hump-like protrusions opposite each other on the cell, constituting prosthecae. Also re-examined using a number of methods were uncultured species Planctomyces bekefii and Pl. guttaeformis. Samples could be enriched for Pl. bekefii via either addition of ferric citrate or ampicillin. An application of a novel approach, laser microdissection and pressure catapulting, was also used physically to enrich P. bekefii rosettes. Fluorescent in situ hybridisation provided the first molecular evidence of Pl. bekefii and Pl. guttaeformis as Planctomycetes. Also confirming Planctomycetes membership of Pl. bekefii was the presence of a cytoplasm divided into two regions by an intracytoplasmic membrane, consistent with membership to the genus Planctomyces. Two new planctomycete-like organisms, MBLW1 and MBLW2, were isolated in this study and possessed a Gemmata-like cell plan. 16S rRNA gene sequencing confirmed these isolates belonged to the Gemmata clade within phylum Planctomycetes, though they may comprise a separate but closely related genus. Via EMISH, both ATCC 35122 and MBLW1 were hybridised with a planctomycete-specific probe, consistent with membership to the planctomycetes. Statistical analysis showed that 16S rRNA was present in both regions of the riboplasm of MBLW1, identical to the distribution observed G. obscuriglobus. This is another example of possible uncoupled translation within a member of the planctomycetes and within organisms in the Gemmata clade of planctomycetes.

The Planctomycetes are a deep branching phylum of the domain Bacteria that incorporate a diverse group of organisms possessing a number of unusual and distinct characteristics. These features include budding reproduction, the planctomycetecharacteristic crateriform structures on their cell surface, a cell wall that lacks peptidoglycan, internal compartmentalisation and unique molecular features of their rRNA genes. This study chose to investigate a number of aspects of planctomycete cell biology and diversity to further our knowledge of this unique group. In a study of the diversity of ribonuclease P (RNase P) RNA, one molecule of relevance to cell biology and compartmentalisation in planctomycetes, RNase P RNA genes were sequenced for species from all genera of planctomycetes for which a pure culture exists. Secondary structures for RNase P RNA of these strains were deduced, taking to 26 the number of planctomycete RNase P RNA structures. Nucleotide positions were identified in which some planctomycetes possess a less common form, including one thought to be otherwise conserved within all Bacteria and Archaea. Phylogenetic analysis of RNase P RNA genes was relatively consistent with that of 16S rRNA genes with the exception that clustering of Gemmata and anammox sequences occurred, possibly due to either long-branch attraction or lateral gene transfer. Analysis of RNase P RNA secondary structures revealed unusual features of planctomycetes relative to all other bacteria, including an additional helix within the P13 helix of ‘Candidatus Brocadia anammoxidans’, ‘Candidatus Kuenenia stuttgartiensis’ and all Gemmata sequences. The longest P12 helix of any bacteria type A RNase P RNA was found in a Gemmatalike isolate. The short tandem repeats in P12 helices of two Gemmata-like isolates are possibly analogous to short tandem repetitive repeat sequences of some cyanobacteria RNase P RNA. In experiments using Gemmata obscuriglobus as a model for planctomycete cell biology and compartmentalisation functions, electron microscope-level in situ hybridisation (EMISH), and subsequent statistical analysis, was developed to localise 16S rRNA, 23S rRNA and RNase P RNA to particular regions within Gemmata obscuriglobus, the first instance of EMISH being applied in this way to bacteria. Statistical analysis localised 16S rRNA to both nuclear body and to riboplasm outside this region but it was absent from paryphoplasm. While co-localisation of both 16S rRNA and 23S rRNA molecules, which might indicate assembled ribosomes, was rarely observed, 23S rRNA, like 16S rRNA, was distributed in both riboplasm-containing areas of the cell. While statistical analysis revealed minor DNA within riboplasm outside the nuclear body, the majority was localised to that body. These results suggest at least some uncoupling of translation from transcription involving ribosomes in the riboplasm. RNase P RNA was localised both to the nuclear body and to the riboplasm outside this region, suggesting that pre-tRNA processing occurs both within nuclear body, where RNA transcripts are presumably generated, and outside nuclear body, separated from the origin of these transcripts. This is also consistent with the hypothesis that processed tRNA is required in the riboplasm outside the nuclear body, due to occurrence of some uncoupled translation. In research on planctomycetes not yet examined with respect to cell plan or structure, 16S rRNA gene sequencing of isolate ATCC 35122 confirmed its very close relationship to the type strain of Pirellula staleyi and its membership of the phylum Planctomycetes. Morphological characteristics, including polar crateriform structures and the occurrence of a unique internal, single membrane-bounded compartment enclosing nucleoid and ribosome-like particles, the pirellulosome, and a polar cap region, are also consistent with its membership of the planctomycetes and of genus Pirellula. Cells often displayed pointed, hump-like protrusions opposite each other on the cell, constituting prosthecae. Also re-examined using a number of methods were uncultured species Planctomyces bekefii and Pl. guttaeformis. Samples could be enriched for Pl. bekefii via either addition of ferric citrate or ampicillin. An application of a novel approach, laser microdissection and pressure catapulting, was also used physically to enrich P. bekefii rosettes. Fluorescent in situ hybridisation provided the first molecular evidence of Pl. bekefii and Pl. guttaeformis as Planctomycetes. Also confirming Planctomycetes membership of Pl. bekefii was the presence of a cytoplasm divided into two regions by an intracytoplasmic membrane, consistent with membership to the genus Planctomyces. Two new planctomycete-like organisms, MBLW1 and MBLW2, were isolated in this study and possessed a Gemmata-like cell plan. 16S rRNA gene sequencing confirmed these isolates belonged to the Gemmata clade within phylum Planctomycetes, though they may comprise a separate but closely related genus. Via EMISH, both ATCC 35122 and MBLW1 were hybridised with a planctomycete-specific probe, consistent with membership to the planctomycetes. Statistical analysis showed that 16S rRNA was present in both regions of the riboplasm of MBLW1, identical to the distribution observed G. obscuriglobus. This is another example of possible uncoupled translation within a member of the planctomycetes and within organisms in the Gemmata clade of planctomycetes.

This research project proposes that the monstrous encounter in art, film and story can signify the change from the understood to the unknown self, using the historical context and literary elements of Little Red Riding Hood as a framework. Within this framework three visual artists, Kiki Smith, Jazmina Cininas and Matthew Barney are investigated to demonstrate how the monstrous encounter signifies the change from the understood to the unknown self. In order to use Little Red Riding Hood as a framework the historical context of the tale has been broken down into three periods, identified as the original, the bourgeois and the contemporary. In each period it is shown how the monstrous encounter signifies the change from the understood to the unknown self. The research project draws on the work of Jack Zipes, The Trials and Tribulations of Little Red Riding Hood (1993) and Fairy Tales and the Art of Subversion (1983); Jon Elster, The Multiple Self (1985); Barbara Creed, The Monstrous Feminine (1993) and Phallic Panic (2005); and Julia Kristeva, Powers of Horror (1982). These key references are used to define important ideas and strengthen terminology specific to the project, such as self, monster and monstrous encounter, abjection and transformation. As a result of using Little Red Riding Hood as a framework the majority of the research looks at the monstrous encounter almost exclusively in the form of lycanthropy, which lends itself most easily to concepts of metamorphoses and the div ided self. The werewolf enjoys enormous popularity in many avenues of contemporary culture and there exists countless references to this particular genre of monstrous encounter. However, this is not a project about werewolves but an investigation of the monstrous encounter, whatever form it takes. Whether it is an internal event like the work of Matthew Barney or an external process as in Cininas's use of Angela Carter's contemporary Red Riding Hood, the monstrous encounter in art, film and story can be demonstrated to signify the change from the understood to the unknown self.

The Planctomycetes are a deep branching phylum of the domain Bacteria that incorporate a diverse group of organisms possessing a number of unusual and distinct characteristics. These features include budding reproduction, the planctomycetecharacteristic crateriform structures on their cell surface, a cell wall that lacks peptidoglycan, internal compartmentalisation and unique molecular features of their rRNA genes. This study chose to investigate a number of aspects of planctomycete cell biology and diversity to further our knowledge of this unique group. In a study of the diversity of ribonuclease P (RNase P) RNA, one molecule of relevance to cell biology and compartmentalisation in planctomycetes, RNase P RNA genes were sequenced for species from all genera of planctomycetes for which a pure culture exists. Secondary structures for RNase P RNA of these strains were deduced, taking to 26 the number of planctomycete RNase P RNA structures. Nucleotide positions were identified in which some planctomycetes possess a less common form, including one thought to be otherwise conserved within all Bacteria and Archaea. Phylogenetic analysis of RNase P RNA genes was relatively consistent with that of 16S rRNA genes with the exception that clustering of Gemmata and anammox sequences occurred, possibly due to either long-branch attraction or lateral gene transfer. Analysis of RNase P RNA secondary structures revealed unusual features of planctomycetes relative to all other bacteria, including an additional helix within the P13 helix of ‘Candidatus Brocadia anammoxidans’, ‘Candidatus Kuenenia stuttgartiensis’ and all Gemmata sequences. The longest P12 helix of any bacteria type A RNase P RNA was found in a Gemmatalike isolate. The short tandem repeats in P12 helices of two Gemmata-like isolates are possibly analogous to short tandem repetitive repeat sequences of some cyanobacteria RNase P RNA. In experiments using Gemmata obscuriglobus as a model for planctomycete cell biology and compartmentalisation functions, electron microscope-level in situ hybridisation (EMISH), and subsequent statistical analysis, was developed to localise 16S rRNA, 23S rRNA and RNase P RNA to particular regions within Gemmata obscuriglobus, the first instance of EMISH being applied in this way to bacteria. Statistical analysis localised 16S rRNA to both nuclear body and to riboplasm outside this region but it was absent from paryphoplasm. While co-localisation of both 16S rRNA and 23S rRNA molecules, which might indicate assembled ribosomes, was rarely observed, 23S rRNA, like 16S rRNA, was distributed in both riboplasm-containing areas of the cell. While statistical analysis revealed minor DNA within riboplasm outside the nuclear body, the majority was localised to that body. These results suggest at least some uncoupling of translation from transcription involving ribosomes in the riboplasm. RNase P RNA was localised both to the nuclear body and to the riboplasm outside this region, suggesting that pre-tRNA processing occurs both within nuclear body, where RNA transcripts are presumably generated, and outside nuclear body, separated from the origin of these transcripts. This is also consistent with the hypothesis that processed tRNA is required in the riboplasm outside the nuclear body, due to occurrence of some uncoupled translation. In research on planctomycetes not yet examined with respect to cell plan or structure, 16S rRNA gene sequencing of isolate ATCC 35122 confirmed its very close relationship to the type strain of Pirellula staleyi and its membership of the phylum Planctomycetes. Morphological characteristics, including polar crateriform structures and the occurrence of a unique internal, single membrane-bounded compartment enclosing nucleoid and ribosome-like particles, the pirellulosome, and a polar cap region, are also consistent with its membership of the planctomycetes and of genus Pirellula. Cells often displayed pointed, hump-like protrusions opposite each other on the cell, constituting prosthecae. Also re-examined using a number of methods were uncultured species Planctomyces bekefii and Pl. guttaeformis. Samples could be enriched for Pl. bekefii via either addition of ferric citrate or ampicillin. An application of a novel approach, laser microdissection and pressure catapulting, was also used physically to enrich P. bekefii rosettes. Fluorescent in situ hybridisation provided the first molecular evidence of Pl. bekefii and Pl. guttaeformis as Planctomycetes. Also confirming Planctomycetes membership of Pl. bekefii was the presence of a cytoplasm divided into two regions by an intracytoplasmic membrane, consistent with membership to the genus Planctomyces. Two new planctomycete-like organisms, MBLW1 and MBLW2, were isolated in this study and possessed a Gemmata-like cell plan. 16S rRNA gene sequencing confirmed these isolates belonged to the Gemmata clade within phylum Planctomycetes, though they may comprise a separate but closely related genus. Via EMISH, both ATCC 35122 and MBLW1 were hybridised with a planctomycete-specific probe, consistent with membership to the planctomycetes. Statistical analysis showed that 16S rRNA was present in both regions of the riboplasm of MBLW1, identical to the distribution observed G. obscuriglobus. This is another example of possible uncoupled translation within a member of the planctomycetes and within organisms in the Gemmata clade of planctomycetes.

BRCA1 is a breast cancer susceptibility gene that is down-regulated in the majority of cases of sporadic breast cancer. Accordingly, there is considerable interest in the mechanisms that regulate normal expression of BRCA1, with a view to elucidating how this could be disrupted in breast cancer. In tumours with reduced BRCA1 protein expression, there can be a concomitant reduction in mRNA level to variable degrees, or no change in mRNA level, suggesting that disruption of multiple different regulatory processes may contribute to BRCA1 down-regulation. Despite this, efforts to date have chiefly focussed on transcriptional and epigenetic regulation of the gene, whilst post-transcriptional processes that regulate the dynamics of the BRCA1 transcript, such as decay, localisation and translation efficiency, are poorly understood. Post-transcriptional regulatory pathways are critical for sustaining normal cellular physiology, as evidenced by many examples where disruption of these processes results in disease, including cancer. Regulation of gene expression at this level is often mediated by RNA-binding proteins that recognise specific cis-acting sequence motifs in the untranslated regions (UTRs) of certain messenger RNAs, and recruit, or shield them from macromolecular complexes involved in RNA metabolism, such as the translation apparatus, exosome, and subcellular transport particles. This thesis is centred on investigating post-transcriptional regulation of BRCA1. Others have shown that expression of the transcript and protein is regulated throughout the mammalian cell division cycle. Results presented in this thesis suggest that changes in mRNA stability may contribute to cell cycle-dependent expression of BRCA1, and therefore that post-transcriptional regulation of BRCA1 is a biologically-relevant phenomenon. To begin to address the molecular mechanisms involved in regulation of BRCA1 mRNA decay, and possibly other post-transcriptional regulatory processes, the 3’UTR of BRCA1, which had not been previously characterised, was analysed for functional regulatory motifs using a combination of bioinformatics, reporter assays and RNA-proteinbinding analysis. An evolutionarily-conserved 3’UTR subsequence was identified which contains sequence elements capable of regulating reporter activity, and forming complexes with multiple proteins from human epithelial cell lines. Some of these elements have been previously characterised in the context of other genes, including a Hu-antigen R (HuR)-binding motif, adenosine-uridine (AU)- rich sequences and a differentiation control element (DICE). Experiments were also conducted to determine the identities of the RNA-binding proteins detected using an RNA probe containing the 3’UTR elements. A preliminary screen of a small group of RNA-binding proteins with previously-characterised roles in 3’UTR-mediated post-transcriptional gene regulation identified HuR as a negative regulator of BRCA1 protein expression. Interestingly, HuR is over-expressed in breast cancer. Evidence presented in this thesis suggests that the mechanism of HuR-mediated down-regulation of BRCA1 involves direct binding of HuR to the BRCA1 3’UTR, and no changes to mRNA stability or abundance. Finally, proteomics-based analysis of protein extracts enriched with BRCA1 3’UTR RNA-binding proteins yielded several interesting candidates with previously-reported RNA-binding and/or post-transcriptional regulatory activities, including Far upstream element-binding protein 1 (FBP1), Glyceraldehyde-3-phosphate dehydrogenase (GAPD) and Heat-shock protein 27 (HSP27). This thesis addresses a clear deficiency in the literature concerning regulation of BRCA1, and contributes to our general understanding of the molecular mechanisms controlling gene expression in mammalian cells. Additionally, the finding that RNA-binding proteins that are over-expressed in breast cancer can negatively regulate BRCA1 expression constitutes important groundwork for identifying potential novel breast cancer therapeutics in the future.

For hundreds of years leadership of religious organisations has been dominated by males, despite the acknowledgement that much religious work and support of the institutions has been done by women. Historically, leadership carried the notion of masculinity and the belief that men were born with certain leadership traits and therefore make better leaders than women. Nevertheless, current thinking contests this view and argues that leadership can be taught and learned and it is possible to develop leader traits in any individual, regardless of gender. This research sets out to contribute in promoting women’s leadership in faith schools. The research employs a qualitative method of data collection, and adopts the critical realist and feminist theorising standpoint. This research presents results of case studies involving the Catholic, Anglican and Muslim faiths. Through in-depth interviews, an exploration on the views of religious authorities, school authorities and female teachers, regarding the role, and position, and experiences of female teachers within faith schools is presented. The research reveals the reality and complexity of barriers encountered by female teachers from a cultural, social, institutional and religious perspective that hinder women’s career advancement. These case studies provide strong first-hand evidence that is hoped to influence both practice and policy. Through the interaction and involvement of head teachers, school governors and religious authorities concerned with the management of the schools, the research aspires to support a process of enlightenment — particularly to individuals who influence decision making processes — to implement strategies that will allow equal representation among the genders in leadership positions within education in faith schools.

This study aims to design and develop effervescent proliposomes that could disintegrate in water and liberate liposomes, and to investigate the potential suitability of liposomes generated for aerosolization to target paranasal sinuses. Novel effervescent proliposomes prepared with Soya phosphatidylcholine (SPC) and Dipalmitoylphosphatidylcholine (DPPC) successfully generated stable liposomes with an improved disintegration time of less than 5 min. Differences in lipid composition were found to influence liposome size and drug entrapment of the hydrophobic drug Beclometasone dipropionate (BDP). Mannitol-based formulations developed with DPPC:Chol (1:1) produced liposomes of 7.54±0.15 µm with a drug entrapment efficiency of 82.15±8.29%. Addition of the mucoadhesives alginic acid or chitosan to effervescent proliposomes made with SPC was found to hamper BDP entrapment in liposomes. Effervescent proliposomes produced SPC:Chol liposomes that also proved beneficial for entrapment of the hydrophilic drug Xylometazoline hydrochloride (XH). The Pari Sinus (pulsating aerosol technology) and Pari Sprint (non-pulsating technology) nebulizers were used for liposome delivery to a nasal cast. Choice of carrier did not affect the liposome’s ability to withstand shearing. A novel system of a Sar-Gel® (water indicating paste) coated clear nasal cast fixed to a two-stage impinger system was set up to analyze drug deposition within the nasal cast cavity. Sinus drug deposition with effervescent mannitol, DPPC:Chol formulation was observed to be highest at 48.45±2.75 cm2 with pulsation compared to deposition of 35.52±11.11 cm2 without pulsation. Drug distribution studies indicated that the Pari Sinus deposited 10.47±2.9% drug, while the Pari Sprint deposited only 4.6±1.4%. The degree of drug loss was higher with conventional liposomes in the Pari Sinus nebulizer, indicating that the degree of bilayers disruption depended on formulation.

Through an analysis of critical reviews and other commentaries on the annual Turner Prize shortlist exhibitions, I examine a philosophical problem which has put into question the rational basis for evaluation in art criticism: the lack of any agreed criteria for the evaluation of artworks. This problem has been most often addressed within philosophical aesthetics through two contrasting approaches: the attempt to formulate evaluative criteria, and the denial that such criteria are either possible or necessary. My response to this meta-critical issue is an interdisciplinary study, in the form of an analysis of published commentaries on the Turner Prize, that examines theories of critical evaluation against an empirical investigation of actual critical practice. The Turner Prize has a number of advantages as a case study. Extensive media coverage of the competition means that it is possible to study a wide range of sources intended for the art-going public, that contain a large body of examples of comparative critical evaluation, and as an annual event it offers the opportunity for both synchronic and diachronic analyses. Moreover, the regular presence of artists whose work has been characterised as ‘conceptual, ensures that many of the commentaries focus on an area of art that presents a particular challenge to aesthetic theory and critical practice. In order to develop a critique of criteria based approaches, the contrasting approaches to art criticism taken by Noel Carroll and Frank Sibley are explored within an analysis of the critical reasons given to justify evaluations of Turner Prize exhibits. Suggestions are offered for ways of developing alternative approaches, drawing upon theories of the aesthetic developed by Suzanne Langer and Kendall Walton.