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habilitationsschriftDie Untersuchung von genregulatorischen Mechanismen im Endothelin- und Renin- Angiotensin-System stellt den Fokus dieser Habilitationsschrift dar, wobei (a) Protein-DNA-Interaktionen auf Promotorebene sowie Beeinflussung dieser durch Signaltransduktionswege, (b) Promotorpolymorphismen und (c) epigenetische Aspekte analysiert wurden. Wir konnten den Promotor des humanen Prionenproteins sowie drei isoformspezifische Promotoren des humanen Endothelin-Konvertierungsenzyms-1 (ECE-1) klonieren und funktionell - beispielsweise bezüglich transkriptioneller Startpunkte und cis-Elementen -charakterisieren. Dabei konnte unter anderem demonstriert werden, dass der im Rahmen der Herzentwicklung essentielle Transkriptionsfaktor Nkx2-5 die ECE- 1a-, -1b-und -1c-Isoformen positiv reguliert, wobei Protein-DNA-Interaktionen, Promotoraktivitäten und die mRNA-Ebene analysiert wurden. Damit konnten wir erstmalig eine direkte molekulare Verbindung zwischen dem Endothelinsystem und Nkx2-5 aufzeigen, welcher eine wahrscheinliche Bedeutung in der Pathogenese angeborener Herzfehler zukommt. Bezüglich des ECE-1b-Promotors konnten wir zwei single nucleotide polymorphisms (SNPs) identifizieren und beim Menschen die signifikante Assoziation dieser mit essentieller arterieller Hypertonie aufzeigen. Darüber hinaus konnte gezeigt werden, dass einer dieser Polymorphismen die Bindungsaffinität und die Transaktivierungsaktivität des Transkriptionsfaktors E2F-2 verändert, womit erstmalig eine Verbindung zwischen essentieller Hypertonie und Zellzyklusregulation demonstriert werden konnte. Im humanen ECE-1c-Promotor wurde unter anderem der Methylierungsstatus sowie der funktionelle Effekt einer in vitro-Methylierung analysiert. Weiterhin konnte die ausgeprägte funktionelle Relevanz eines [CpG]m-[CA]n-Mikrosatellitenpolymorphismus nachgewiesen werden. Bezüglich des Renin-Angiotensin-Systems (RAS) konnte das zuvor unbekannte Protein ATBP (AT2R binding protein) durch uns kloniert werden, welches an den zytoplasmatischen Bereich des Angiotensin-AT2-Rezeptors (AT2R) bindet und antiproliferative Eigenschaften dieses Rezeptors vermittelt. Kürzlich konnten wir als erste Arbeitsgruppe die Signaltransduktion des Renin-/ Prorenin-Rezeptors (RER) basierend auf Protein-Protein- und Protein-DNA-Interaktionen aufschlüsseln und nachweisen, dass der Transkriptionsfaktor PLZF ein direktes Adapterprotein des RER darstellt, nach Rezeptoraktivierung in den Zellkern transloziert und dort zielgenabhängig als Aktivator aber auch Repressor fungieren kann. Schließlich konnten wir eine neue Formel - die sog. GED (Gene Expression’s CT Difference)-Formel - zur Auswertung von real-time PCR-Daten mathematisch herleiten und experimentell validieren.Analysis of gene regulatory mechanisms within the endothelin and renin- angiotensin system represents the focus of this habilitation. These mechanisms can be grouped into (a) signal transduction pathways and their downstream protein-DNA interactions on the promoter level, (b) promoter polymorphisms and (c) epigenetic aspects. Our group was able to clone and functionally characterise - e.g. regarding transcriptional start sites and cis-elements - the promoter of the human prion protein as well as three isoform-specific promoters of the human endothelin-converting enzyme-1 (ECE-1) gene. We could establish the first molecular link between the transcription factor Nkx2-5, which is essential for cardiac development, and the endothelin system, an interaction which is likely involved in the pathogenesis of congenital heart defects. Furthermore, two single nucleotide polymorphisms (SNPs) were identified in the human ECE-1b promoter region which could be associated with human arterial hypertension. In addition, we were able to demonstrate that one of these SNPs alters the affinity for and the transactivation potential of the transcription factor E2F-2, thereby linking essential hypertension and cell cylce regulation. In the human ECE-1c promoter the basal methylation status and functional effects of an in vitro methylation were analysed. Additionally, the strong regulatory impact of a polymorphic [CpG]m-[CA]n microsatellite repeat within the human ECE-1c promoter was revealed. Regarding the renin- angiotensin system (RAS) a previously unknown adapter protein of the angiotensin AT2 receptor (AT2R), named ATBP (AT2R binding protein), was discovered mediating for example the antiproliferative effects of this receptor. Recently, our group deciphered the signal transduction cascade of the renin/ prorenin receptor (RER) in which the transcription factor PLZF acts as a direct adapter protein of the RER. Finally, a novel formula, called GED (Gene Expression’s CT Difference), for the analysis of real-time PCR data was mathematically derived and experimentally validated

The Plan-a-Day Approach to Measuring Planning Ability in Patients with Schizophrenia

Deficits in executive functioning are closely related to the level of everyday functioning in patients with schizophrenia. However, many existing neuropsychological measures are limited in their ability to predict functional outcome. To contribute towards closing this gap, we developed a computer-based test of planning ability ("Plan-a-Day”) that requires participants to create daily activity schedules in a simulated work setting. Eighty patients diagnosed with schizophrenia were tested with Plan-a-Day and a battery of cognitive ability tests. Plan-a-Day showed satisfactory psychometric properties in terms of consistency, reliability, and construct validity. Compared to other neuropsychological tests used in this study, it also demonstrated incremental validity with regard to the Global Assessment of Functioning. The Plan-a-Day approach, therefore, seems to represent a valid alternative for measuring planning ability in patients with executive function deficits, occupying a middle ground between traditional neuropsychological tests and real-life assessments. (JINS, 2011, 17, 327-335

Midregional proadrenomedullin and its change predicts recurrent major coronary events and heart failure in stable coronary heart disease patients: The LIPID study

Background: Biomarkers may contribute to risk stratification in coronary heart disease (CHD). We examined whether plasma midregional proadrenomedullin (MR-proADM) concentration at baseline and its change over one year predicts long-term outcomes in stable CHD patients. Methods: The LIPID study randomised patients 3–36 months after an acute coronary syndrome with total cholesterol 4.0–7.0 mmol/L (155–271 mg/dL), to placebo or pravastatin 40 mg. Follow-up was 6.0 years. MR-proADM plasma concentrations at baseline and one year later were determined in 7863 and 6658 patients, respectively. These were categorised into quartiles to perform Cox regression analysis, adjusting for baseline parameters. Results: Baseline MR-proADM concentrations predicted major CHD events (non-fatal myocardial infarction or CHD death; hazard ratio (HR) 1.52, 1.26–1.84 for Q4–Q1), CHD death (HR 2.21, 1.67–2.92), heart failure (HR 2.30, 1.78–2.97) and all-cause mortality (HR 1.82, 1.49–2.23). Associations were still significant after adjustment for baseline B-type natriuretic peptide (BNP) concentration. Increase in MR-proADM after one yearwas associated with increased risk of subsequent CHD events (HR 1.34, 1.08–1.66), non-fatalmyocardial infarction (HR 1.50, 1.12–2.03), heart failure (HR 1.78, 1.37–2.30) and all-causemortality (HR 1.31, 1.04–1.64). Associations with heart failure and all-causemortality remained significant after adjusting for baseline and change in BNP concentration. Change in MR-proADM moderately improved risk reclassification for major CHD events (net reclassification improvement (NRI) 3.48%) but strongly improved risk reclassification for heart failure (NRI 5.60%). Conclusions: Baseline and change in MR-proADM concentrations over one year are associated with risk of major clinical events, even after adjustment for BNP concentrations.National Heart Foundation of Australi

Characterization of the c-specific promoter of the gene encoding human endothelin-converting enzyme-1 (ECE-1)

AbstractHuman ECE-1 is expressed in four isoforms with different tissue distribution and its mRNA and protein levels are altered under certain pathophysiological conditions. To investigate the transcriptional regulation of ECE-1, we studied the regulatory region of ECE-1c, the major ECE-1 isoform. A genomic clone comprising the complete human ECE-1 gene including the putative ECE-1c-specific promoter was obtained. Up to 968 bp upstream of the putative c-specific translation initiation start codon and several serial deletion mutants were subcloned into a reporter vector and transfected into endothelial (BAEC, EA.hy926, ECV304) and epithelial (MDA MB435S, MCF7) cells, showing very strong promoter activity in comparison to the SV40 promoter and to the previously described ECE-1a and 1b promoters. Transfection of serial deletion mutants indicated two positive regulatory regions within the promoter (−142/−240 and −240/490) likely involved in binding GATA and ETS transcription factors. RNase protection assay (RPA) and 5′-RACE revealed multiple transcriptional start sites located at about −110, −140 and −350 bp. Site-directed mutagenesis demonstrated a crucial role for the E2F cis-element for basal ECE-1c promoter activity. Additionally, we found a correlation between isoform-specific ECE-1 mRNA levels and corresponding ECE-1a, 1b, 1c promoter activities

Predictors for Improvement of Problem-Solving during Cognitive Remediation for Patients with Schizophrenia

Cognitive remediation is a promising pathway for ameliorating cognitive impairment of patients with schizophrenia. Here, we investigate predictors of improvement in problem-solving ability for two different types of cognitive remediation - specific problem-solving training and training of basic cognition. For this purpose we conducted a re-analysis of a randomized controlled trial comparing these two training approaches. The main outcome measure was improvement in problem-solving performance. Correlational analyses were used to assess the contribution of clinical, cognitive and training-related predictors. In the problem-solving training group, impaired pre-training planning ability was associated with stronger improvement. In contrast, in the basic cognition training group antipsychotic medication dose emerged as a negative predictor. These results demonstrate that predictors for successful cognitive remediation depend on the specific intervention. Furthermore, our results suggest that at least in the planning domain patients with impaired performance benefit particularly from a specific intervention. (JINS, 2014, 20, 1-6

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journal homepage: www.elsevier.com/locate/schres Planning impairments in schizophrenia: Specificity, task independence an

Development of a GEM-TPC prototype

The use of GEM foils for the amplification stage of a TPC instead of a con- ventional MWPC allows one to bypass the necessity of gating, as the backdrift is suppressed thanks to the asymmetric field configuration. This way, a novel continuously running TPC, which represents one option for the PANDA central tracker, can be realized. A medium sized prototype with a diameter of 300 mm and a length of 600 mm will be tested inside the FOPI spectrometer at GSI using a carbon or lithium beam at intermediate energies (E = 1-3AGeV). This detector test under realistic experimental conditions should allow us to verify the spatial resolution for single tracks and the reconstruction capability for displaced vertexes. A series of physics measurement implying pion beams is scheduled with the FOPI spectrometer together with the GEM-TPC as well.Comment: 5 pages, 4 figures, Proceedings for 11th ICATTP conference in como (italy