Aspartoacylase-LacZ Knockin Mice: An Engineered Model of Canavan Disease

Canavan Disease (CD) is a recessive leukodystrophy caused by loss of function mutations in the gene encoding aspartoacylase (ASPA), an oligodendrocyte-enriched enzyme that hydrolyses N-acetylaspartate (NAA) to acetate and aspartate. The neurological phenotypes of different rodent models of CD vary considerably. Here we report on a novel targeted aspa mouse mutant expressing the bacterial β-Galactosidase (lacZ) gene under the control of the aspa regulatory elements. X-Gal staining in known ASPA expression domains confirms the integrity of the modified locus in heterozygous aspa lacZ-knockin (aspalacZ/+) mice. In addition, abundant ASPA expression was detected in Schwann cells. Homozygous (aspalacZ/lacZ) mutants are ASPA-deficient, show CD-like histopathology and moderate neurological impairment with behavioural deficits that are more pronounced in aspalacZ/lacZ males than females. Non-invasive ultrahigh field proton magnetic resonance spectroscopy revealed increased levels of NAA, myo-inositol and taurine in the aspalacZ/lacZ brain. Spongy degeneration was prominent in hippocampus, thalamus, brain stem, and cerebellum, whereas white matter of optic nerve and corpus callosum was spared. Intracellular vacuolisation in astrocytes coincides with axonal swellings in cerebellum and brain stem of aspalacZ/lacZ mutants indicating that astroglia may act as an osmolyte buffer in the aspa-deficient CNS. In summary, the aspalacZ mouse is an accurate model of CD and an important tool to identify novel aspects of its complex pathology

Correlations of differentially expressed gap junction connexins cx26, cx30, cx32, cx43 and cx46 with breast cancer progression and prognosis.

BACKGROUND AND AIMS: Connexins and their cell membrane channels contribute to the control of cell proliferation and compartmental functions in breast glands and their deregulation is linked to breast carcinogenesis. Our aim was to correlate connexin expression with tumor progression and prognosis in primary breast cancers. MATERIALS AND METHODS: Meta-analysis of connexin isotype expression data of 1809 and 1899 breast cancers from the Affymetrix and Illumina array platforms, respectively, was performed. Expressed connexins were also monitored at the protein level in tissue microarrays of 127 patients equally representing all tumor grades, using immunofluorescence and multilayer, multichannel digital microscopy. Prognostic correlations were plotted in Kaplan-Meier curves and tested using the log-rank test and cox-regression analysis in univariate and multivariate models. RESULTS: The expression of GJA1/Cx43, GJA3/Cx46 and GJB2/Cx26 and, for the first time, GJA6/Cx30 and GJB1/Cx32 was revealed both in normal human mammary glands and breast carcinomas. Within their subfamilies these connexins can form homo- and heterocellular epithelial channels. In cancer, the array datasets cross-validated each other's prognostic results. In line with the significant correlations found at mRNA level, elevated Cx43 protein levels were linked with significantly improved breast cancer outcome, offering Cx43 protein detection as an independent prognostic marker stronger than vascular invasion or necrosis. As a contrary, elevated Cx30 mRNA and protein levels were associated with a reduced disease outcome offering Cx30 protein detection as an independent prognostic marker outperforming mitotic index and necrosis. Elevated versus low Cx43 protein levels allowed the stratification of grade 2 tumors into good and poor relapse free survival subgroups, respectively. Also, elevated versus low Cx30 levels stratified grade 3 patients into poor and good overall survival subgroups, respectively. CONCLUSION: Differential expression of Cx43 and Cx30 may serve as potential positive and negative prognostic markers, respectively, for a clinically relevant stratification of breast cancers

Organisational lobbying activities at the EU: An exploratory analysis of the energy and power generation sector

As a result of the growth of the EU over the years, not only in its physical size and country membership but also in terms of its legislative power, there has been a need for organisations to address the manner in which they represent themselves and their interests at the EU level. For some time now, it has not been possible for large organisations from various industries to simply engage via their home national administrations with the institutions of the EU in terms of seeking to influence the legislation that is now being produced by these supranational bodies. In parallel, the nature and scope of the legislative agenda of the EU has also grown, which has made the EU legislative environment of interest for organisations in terms of both their market and nonmarket environment The growing ability for the EU to influence the setting in which organisations operate and thus have an effect on the overall competitiveness within the business context, of not only individual organisations but industries too, meant that organisations have become more interested in engaging with the EU institutions. The form this engagement takes is via lobbying activities towards the various institutions. This thesis first seeks to explore the aspects of lobbying activities at the EU level for actors from the Energy & Power Generation Sector. It consequently looks at the dynamics between the representation for industry actors via associations or federations, national or international, over more individualistic efforts. In doing so, the strategic aspects to lobbying activities are also explored in terms of the Resource Based View of the firm. In order to address these issues the industry selection is first justified and then a qualitative foundation is laid, this is in terms of seeking primary data for the research via face-to-face interviews with appropriate interview targets. The primary data was ultimately collected from 26 high level face-to-face interviews with actors from the industry side as well as from with in the institutions as well. The findings of the thesis are that activities via associations and federations are very much subject to the consensus position that can be reached within them and the overall effectiveness for their members is subject to the position reached. However, in terms of interaction with the European Commission, associations and federations allow also for smaller actors to make contact in the drafting stage of legislation by the EC. Timing is also a significant issue in terms of the legislative process when it comes to effective lobbying activities towards the institutions. Moreover, the ability to lobby in a transversal manner is an ultimate goal. In terms of achieving this goal of transversal lobbying for industry based actors, this is only possible through the contribution of the appropriate resources and capabilities. If these resources and capabilities are already present and configured in a particular way, then undertaking lobbying in a transversal manner is achievable. In cases 3 " where they are not, then they either need to be sought so that lobbying activity can be increased in the non market environment or alternative routes need to be followed to interact with and lobby the institutions (such as an increased reliance on lobbying activities via associations or federations). 4EThOS - Electronic Theses Online ServiceGBUnited Kingdo