No description available.

Osteoarthritis (OA) is characterised by cartilage degradation, inflammation and pain within synovial joints. OA is a major cause of morbidity in the elderly human population and in companion animals such as horses. Changes in expression and activity of pro-inflammatory cytokines, chemokines and catabolic mediators contribute towards OA progression, which can be studied using in vitro culture models and proteomic approaches. This project studied the secretome from an in vitro model of equine articular cartilage, aiming to develop understanding of cartilage biology and degradative processes. These studies also aimed to identify protein markers relevant to this explant model for screening anti-inflammatory properties of novel therapeutics. To evaluate responses to OA associated pro-inflammatory IL-1β and the non-steroidal anti-inflammatory drug (NSAID), carprofen, time courses of protein release were established in the explant model. The cartilage secretome contained cartilage extracellular matrix (ECM), non-ECM and intracellular proteins, all of which were identified by high-throughput mass spectrometry (MS). Semi-quantitative differences in protein release were reported between untreated control and IL-1β stimulated cartilage by MS. The release of glycosaminoglycans (GAGs) initiated by IL-1β was delayed when carprofen was present. The proteomic sample preparation method was adapted to deplete high abundance proteins that can hinder the detection of low level proteins in high-throughput MS analysis. Three depletion approaches were applied: CPC precipitation, concanavalin A lectin chromatography and Proteominer™ technology. These approaches provided additional identifications of the non-ECM secreted proteins MMP-10 and IL-9, and of additional intracellular proteins. Further optimization of these methods could further enhance the detection of low level proteins. Proteins identified by MS analysis of the cartilage secretome were assessed using quantitative western blotting analysis. Carprofen significantly reduced IL-1β stimulated release of MMP-1, MMP-3, MMP-13 and a fibronectin degradation product. Levels of clusterin were reduced by IL-1β and carprofen treatments. These specific proteins were shown to be markers of IL-1β stimulated inflammation and degradative processes, which can be significantly reduced by an anti-inflammatory such as carprofen. This thesis describes the use of proteomics with other approaches to study the effects of IL-1β and carprofen on release of several important structural, metabolic and inflammatory related components from cartilage. Carprofen was beneficial in decreasing certain aspects of inflammation and degradation, including significantly reducing release of MMPs and their catabolic products (fibronectin and GAGs) from the ECM. The equine explant model can be further studied with high-throughput MS to assess responses to various stimuli and detect released proteins. In conclusion, anti-degradative effects and MMP inhibition can be specifically monitored within this in vitro equine cartilage model, to screen efficacy of therapeutics and putative anti-inflammatories to relieve OA.

The characteristic curd phenotype of the Brassica cauliflower consists of proliferating, arrested inflorescence and floral meristems. Recent analysis of the similar phenotype in the ap1-1/cal-1 mutant of the related crucifer Arabidopsis thaliana has led to speculation that the orthologous genes from Brassica oleracea L. may be responsible for this characteristic trait. Application of molecular genetic analysis to this hypothesis allows the presentation of a genetic model based on specific, mapped loci of BoCAL and BoAP 1. This model accounts for differences in the stage of arrest between the heading phenotypes of cauliflower (B. oleracea var. botrytis L.) and Calabrese broccoli (B. oleracea var. italica Plenck), and is also predictive in accounting for intermediate stages of arrest similar to those observed in Sicilian Purple types. Further molecular genetic analysis characterised three independent loci of the floral meristem identity gene BoAP 1. Integration of this data into the genetic model proposed for curd development, suggests a combination of point mutations and expression thresholds of several copies of the key meristem identity genes BoCAL and BoAP 1 respectively may account for the development of curd tissue in the Brassica cauliflower. The association of alleles of the BoCAL-a gene with the curding phenotypes of B. oleracea was also demonstrated through a survey of over 200 crop accessions. This reveals strong correlations between specific BoCAL-a alleles and discrete inflorescence morphologies, and allows the presentation of a possible scenario for the domestication of cauliflowers. Molecular genetic analysis of BoCAL-a utilising monosomic addition lines has also demonstrated the potential for integration of the genetic and cytogenetic maps of B. oleracea. Such analysis may have significant utility for physical characterisation of replicated loci in B. oleracea, prior to the development of a strong physical map. Further examination of inflorescence morphologies amongst the heading brassicas revealed a shared trait, termed Fused Inflorescence. Preliminary investigation suggests this trait may be under the control of multiple loci, providing a possible indication of the delineation between heading and sprouting B. oleracea crops.

The overheated atmosphere of English education in contemporary Korean society is popularly represented as English Fever. The phenomenon of English Fever dates back at least to 1945 when there was a socio-political transition from Japanese colonial rule to a newly emerged U.S. army military government rule. This dissertation attempts to elucidate historically accumulated aspects of English education in Korea from a critical macro-level approach (Pennycook, 2010; Phillipson, 1992). In doing so, a theoretical framework for the dissertation is based on language ideologies (Kroskrity, 2000; Schieffelin, Woolard & Kroskrity, 1998; Van Dijk, 1995), which are produced, mediated, transformed, and reproduced by interacting with ever-changing socio-historical, political, economic, and structural contexts of Korea. Language ideologies of English influence and contribute to what is at stake in English education in Korea, depending on what is primarily required in each of three major socio-historical periods in modern Korean: nation building (1945-1960), modernization (1961-1980), and globalization (1990-present). This dissertation elaborates how language ideologies about the English language are dedicated to serving as indexicality, reifying dominance, and prevailing in social institutions (Fairclough, 2001; Gal, 1989; Hill, 2008; Irvine, 1989; Kroskrity, 2000), giving rise to a variety of forms of social power in Korean context. Analysis of the relationship between language ideologies of English and English proficiency in Korea suggests that the English language is ideologized to exert profound influence on Korean social structures, although it hardly functions as a tool of communication within Korean society. In other words, the ubiquitous phenomenon of English Fever in Korea may have little correlation with an actual guarantee of English proficiency. Rather English Fever can be attributed to the accumulated and deeply held English language ideologies permeated into Korean society.

Display of disagreement in a public space under the form of strikes, rallies and not only, is not the sole form of protest. Popular culture can easily be used to send messages of discontent. The paper focuses on popular music by looking at one of the most representative music charts in the world: The Billboard Chart. By screening the Year End Billboard Rock Chart for a period of 5 years the paper tries to identify songs that can be labelled as protest songs and see what they are protesting against, what themes they address, what are their characteristics and how are the messages transmitted in both textual and visual narratives, in order to draw a picture of the contemporary protest song that is present in a popular chart. The theoretical framework of the paper discusses popular culture, the classical image of the protest song, the creational process of music within the music industry and its politic and economic sides. After a first screening of the charts with the help of content analysis, by using the concept of narrative, the paper examines the stories presented in the lyrics and, where possible, the videos made for the songs. The findings of the paper show that even if not respecting the theoretical characteristics of the classical protest song, The Year End Billboard Rock Chart has several songs with strong political messages either in lyrics or videos or in both at the same time.

The majority of UK oilseed rape (Brassica napus L.) is September-sown on heavy clay soils where straw has been incorporated following the cereal harvest in August. A series of germination, emergence and field experiments was conducted to assess the effects of variation in seed quality on establishment and to evaluate the improvements possible by mother crop nitrogen management and pre-sowing seed treatments on commercial and farm-saved seedlots. In germination experiments designed to examme the performance of commercial seed lots at temperatures ranging between 5 and 25°C significant differences were recorded in the speed and unifonnity of germination, particularly at 10 and 15°C, which are comparable to UK field temperatures in late-August to September. The analysis of Apex variety seeds grown from nitrogen-managed mother crops in 1996 and 1998 showed a negative correlation between their nitrogen and oil percentage, which was significantly affected by both the amount (kg ha -1) and timing (vegetative growth period or flowering period) of nitrogen application. The highest nitrogen seeds were produced from mother crops that had received medium (160 kg ha -1) amounts of nitrogen fertiliser during the flowering period. Seeds that were harvested from pods taken from the lower < 1.5 m) section of the crop canopy also had a significantly higher nitrogen and significantly lower oil percentage than those taken from the upper (> 1.5 m) section of the canopy. In germination and emergence experiments the highest nitrogen (3.46 to 3.61 %) seeds germinated significantly faster than the seeds of lowest nitrogen (2.30 to 2.95%) content but they did not emerge as well as low nitrogen seeds from depth. Selecting small (< 2 mm diameter) seeds over large (> 2 mm diameter) seeds significantly improved the rate of geTI11ination and emergence and the final percentage emergence at 10 mm sowing depth although the final percentage emergence at 20 mm sowing depth was significantly greater from the large seeds. Hydrating seeds in water for 18 hours at 15 grad. C before drying them back in the laboratory at 20 grad. C significantly improved the speed of germination and the speed and final percentage emergence at 10 mm sowing depth compared with control treatments provided that the radicle had not emerged before drying back; small seeds derived the most benefit from hydration. Seed heat treatment at temperatures of 80 grad. C significantly delayed the onset of germination and emergence but significantly hastened field establishment. The effects of seed nitrogen percentage, seed size and heat treatment on seedling emergence and subsequent plant growth and development were examined in the field between October 1999 and July 2000. Growth analyses, which were performed in February (growth phase), May (flowering) and July (pre-harvest), showed that the high nitrogen, large seeded and heat-treated populations had a significantly lower rate of plant loss than the low nitrogen, small seeded and control populations. Under field conditions, the higher growth rates and growth parameters of the seedlings produced from the high nitrogen and! or large seeds were not always significant nor were they consistently maintained until harvest. Large seed size and heat treatment significantly increased the number of established plants per m2 and significantly increased the initial plant size. The final yield was not significantly affected by seed nitrogen percentage, seed size or seed heat treatment.

The ability to perform nucleic acid-based detection of plant pathogens away from conventional laboratory facilities has the potential to be beneficial in situations where results are required very rapidly or where resources and access to laboratory equipment are limited. Methods for use in such situations must combine sensitivity and specificity with rapid and simple workflows. The aim of this project was to investigate aspects of on-site testing for plant pathogens by developing detection methods for a range of target species. Detection methods based on loop-mediated isothermal amplification (LAMP) exhibit characteristics which make them potentially suitable for on-site testing. LAMP-based methods were developed for detection of plant pathogens with three potential non-laboratory testing scenarios in mind: testing during plant health inspection (assays for Phytophthora ramorum, P. kernoviae and Guignardia citricarpa); testing to assess inoculum levels in the processing of plant products (an assay for Botrytis cinerea); and testing in under-resourced settings (assays for Cassava brown streak virus and Ugandan cassava brown streak virus). In developing these detection methods, attempts were made to address some of the specific requirements of potential end-users of the tests in each case. For testing in the context of inspection, a particular emphasis was placed on the need for simple, rapid methods for nucleic acid extraction. As well as investigating the use of rapid extraction methods in conjunction with LAMP, work was also carried out to investigate how on-site nucleic acid extraction using lateral flow devices could be integrated with current field and laboratory testing for P. ramorum.

No description available.

The potyvirus Turnip mosaic virus (TuMV) is a major constraint on the cultivation of a wide range of plant species worldwide. It causes significant economic losses in brassica species such as Chinese cabbage (Brassica rapa), which is one the most important vegetable crops in the world. The B. rapa line RLR22 has broad-spectrum resistance to TuMV, which is undefeated. Many recessive resistances against plant viruses in the Potyvirus genus are based on mutations in plant eukaryotic translation initiation factor 4E (eIF4E), or its isoform eIF(iso)4E . B. rapa has three eIF4E genes and three eIF(iso)4E genes. Segregation following a cross between RLR22 and the TuMV-susceptible R-o-18 line of the closely related B. rapa ssp. trilocularis revealed the resistance was due to a recessive gene, retr01 that was epistatic to a dominant gene, ConTR01. My research revealed that retr01 is BraA.eIF(iso)4E.a and that ConTR01 is probably BraA.eIF(iso)4E.c. It also showed that the highly sought after broad-spectrum resistance to TuMV is due to a novel, recessive, natural mechanism, based on the mis-splicing of BraA.eIF(iso)4E.a in B. rapa. This results in a range of eIF(iso)4E splice variants, the most common of which retained the whole of intron 1 and appears to be non-functional for the virus. As the susceptible parent in the original cross, R-o-18, was a different sub-species to RLR22 (B. rapa var. pekinensis, Chinese cabbage), the genetic inheritance of resistance was also investigated in crosses with Chinese cabbage lines; F2 segregation ratios were consistent with those predicted for the single recessive gene (retr01 ). Yeast two-hybrid interactions between the viral protein genome-linked (VPg) of TuMV and eIF(iso)4E from B. rapa seem to be TuMV isolate-specific. Aphid transmission experiments to investigate the complementation of an eIF(iso)4E Arabidopsis thaliana knockout line with B. rapa BraA.eIF(iso)4E.a confirmed the earlier results from mechanical inoculation of these plants. The inability of TuMV to access multiple copies of eIF(iso)4E in Chinese cabbage and the broad-spectrum of the resistance, suggest it may prove to be durable.

Lameness in sheep is an important welfare concern causing financial loss through lost performance. This thesis increases epidemiological understanding of sheep lameness from the farmers’ perspective, using interdisciplinary approaches. Previous work indicated that routine foot trimming (RFT), used by >75% of farmers to control lameness, correlated with higher lameness prevalences. A within farm, clinical trial using stratified random sampling examined the effect of RFT versus no RFT on 173 ewes. RFT was not beneficial and over-trimming was detrimental. Thirty-five farms were visited to assess accuracy of farmers’ estimated lameness prevalence in their sheep flocks compared with a researcher’s observations. Farmers’ estimates were consistently, closely and significantly correlated with the researcher’s estimates. Successful knowledge transfer and research impact requires understanding lameness management from farmers’ perspectives. Qualitative interviews with 17 farmers examining attitudes towards lameness management were used to inform design of a questionnaire, sent to 1000 randomly selected farmers, to quantify farmers’ attitudes towards lameness. Farmers considered interdigital-dermatitis and footrot distinct. Barriers to prompt treatment (e.g. lack of time/labour) accounted for most observed variance, with non-financial motivators rated higher than financial motivators. Ineffective flock record keeping, environmental subsidies and market price fluctuation may reduce financial motivation. Consistent with the RFT trial, there was no difference (p>0.5) in lameness prevalence by RFT frequency/absence. Despite RFT trial evidence given, farmers were reluctant to stop RFT; with reluctance less from those with higher lameness prevalences, large, commercial flocks, infrequently using parenteral antibacterial treatments or not treating mildly lame sheep. This thesis provides evidence that farmers’ lameness estimates are sufficiently accurate and can be used in research. Routine foot trimming appears to be of no benefit to reduce lameness but farmers require further evidence to be convinced of this. Further farmer focused research into RFT, barrier cause and effect, and whole flock managements is required.