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Markenaufbau von Destinationen ist eine kostenintensive und langfristige Angelegenheit. In den vergangenen Jahren stieg das Interesse von Kommunen, Regionen und Ländern die Markenbildung zu verstärken. Gleichzeitig wird aber auch immer mehr der Ruf lauter ein Instrument zu schaffen, das Markenbewertung für Destinationen vornimmt. Vor allem in der Konsumgüter und Investitionsgüterindustrie finden sich zahlreiche Ansätze zur Markenbewertung, im Destinationsmanagement gibt es das bisher kaum. Es gibt drei verschiedene Arten von Verfahren, die meist von Unternehmensberatungen oder Wissenschaftlern entwickelt wurden: Messung des Markenwertes, Messung der Markenstärke und hybride Verfahren. Für Destinationen machen rein quantitative Verfahren wenig Sinn, da es beim Thema "Reise" auch auf eine Vielzahl von qualitativen Merkmalen ankommt. Von daher wurde im Rahmen dieser Dissertation entschieden, sich primär auf ein qualitatives Verfahren zu konzentrieren und verschiedene Modelle der Markenbewertung zu überprüfen. Das Resultat der Überprüfung zeigte, dass es notwendig war verschiedene Modelle zu kombinieren, um damit die Markenstärke einer Destination zu messen. Als empirisches Beispiel wurde hier Spanien ausgewählt, da dies als ein erfolgreiches Beispiel für Branding in der Fachliteratur gesehen wird. Dieses neu entwickelte Verfahren wurde an der Destination Spanien überprüft im Rahmen einer Projektstudie mit Studierenden der Universität Trier. Das Ergebnis zeigte, dass der Ansatz eines qualitativen Verfahrens richtig war, allerdings könnte es in ein hybrides Verfahren überführt werden. Kooperationspartner für die Dissertation war Turespana, Berlin.
The glucocorticoid (GC) cortisol, main mediator of the hypothalamic-pituitary-adrenal axis, has many implications in metabolism, stress response and the immune system. GC function is mediated mainly via the glucocorticoid receptor (GR) which binds as a transcription factor to glucocorticoid response elements (GREs). GCs are strong immunosuppressants and used to treat inflammatory and autoimmune diseases. Long-term usage can lead to several irreversible side effects which make improved understanding indispensable and warrant the adaptation of current drugs. Several large scale gene expression studies have been performed to gain insight into GC signalling. Nevertheless, studies at the proteomic level have not yet been made. The effects of cortisol on monocytes and macrophages were studied in the THP-1 cell line using 2D fluorescence difference gel electrophoresis (2D DIGE) combined with MALDI-TOF mass spectrometry. More than 50 cortisol-modulated proteins were identified which belonged to five functional groups: cytoskeleton, chaperones, immune response, metabolism, and transcription/translation. Multiple GREs were found in the promoters of their corresponding genes (+10 kb/-0.2 kb promoter regions including all alternative promoters available within the Database for Transcription Start Sites (DBTSS)). High quality GREs were observed mainly in cortisol modulated genes, corroborating the proteomics results. Differential regulation of selected immune response related proteins were confirmed by qPCR and immuno-blotting. All immune response related proteins (MX1, IFIT3, SYWC, STAT3, PMSE2, PRS7) which were induced by LPS were suppressed by cortisol and belong mainly to classical interferon target genes. Mx1 has been selected for detailed expression analysis since new isoforms have been identified by proteomics. FKBP51, known to be induced by cortisol, was identified as the strongest differentially expressed protein and contained the highest number of strict GREs. Genomic analysis of five alternative FKBP5 promoter regions suggested GC inducibility of all transcripts. 2D DIGE combined with 2D immunoblotting revealed the existence of several previously unknown FKBP51 isoforms, possibly resulting from these transcripts. Additionally multiple post-translational modifications were found, which could lead to different subcellular localization in monocytes and macrophages as seen by confocal microscopy. Similar results were obtained for the different cellular subsets of human peripheral blood mononuclear cells (PBMCs). FKBP51 was found to be constitutively phosphorylated with up to 8 phosphosites in CD19+ B lymphocytes. Differential Co-immunoprecipitation for cytoplasm and nucleus allowed us to identify new potential interaction partners. Nuclear FKBP51 was found to interact with myosin 9, whereas cytosolic FKBP51 with TRIM21 (synonym: Ro52, Sjögren`s syndrome antigen). The GR has been found to interact with THOC4 and YB1, two proteins implicated in mRNA processing and transcriptional regulation. We also applied proteomics to study rapid non-genomic effects of acute stress in a rat model. The nuclear proteome of the thymus was investigated after 15 min restraint stress and compared to the non-stressed control. Most of the identified proteins were transcriptional regulators found to be enriched in the nucleus probably to assist gene expression in an appropriate manner. The proteomic approach allowed us to further understand the cortisol mediated response in monocytes/macrophages. We identified several new target proteins, but we also found new protein variants and post-translational modifications which need further investigation. Detailed study of FKBP51 and GR indicated a complex regulation network which opened a new field of research. We identified new variants of the anti-viral response protein MX1, displaying differential expression and phosphorylation in the cellular compartments. Further, proteomics allowed us to follow the very early effects of acute stress, which happen prior to gene expression. The nuclear thymocyte proteome of restraint stressed rats revealed an active preparation for subsequent gene expression. Proteomics was successfully applied to study differential protein expression, to identify new protein variants and phosphorylation events as well as to follow translocation. New aspects for future research in the field of cortisol-mediated immune modulation have been added.