Ubiquitin-dependent regulation of the WNT cargo protein EVI/WLS

Cellular communication by WNT signalling is crucial for growth, patterning, and tissue homeo-stasis of metazoan animals and has been associated with various human diseases, such as cancer. The different branches of this signalling cascade are induced after the secretion of WNT ligands by the WNT cargo protein evenness interrupted/Wntless (EVI/WLS). The availa-bility and stability of many proteins involved in the WNT signalling pathways is regulated by post-translational mechanisms. In the absence of WNTs, EVI/WLS is modified with ubiquitin and subjected to endoplasmic reticulum (ER)-associated degradation (ERAD) by the pro-teasome. ERAD is well known to remove misfolded proteins from the ER but can also affect cellular signalling by degrading mature proteins associated with secretory routes. However, the latter type of regulation is not well studied in mammals. In addition, the mechanisms leading to the recognition, ubiquitination, and proteasomal targeting of EVI/WLS remain largely elusive. To gain insights into how ERAD and ubiquitination components regulate EVI/WLS, I performed a RNAi-based screen on EVI/WLS protein stability and used biochemical and cell biological methods in human cells with diverse genetic backgrounds. I discovered that the ER-membrane associated proteins ERLIN2, FAF2, and UBXN4 are novel components of the EVI/WLS ‘destruction complex’. Mechanistically, ERLIN2 links EVI/WLS to the ubiquitination machinery, while FAF2 and UBXN4 interact with EVI/WLS and VCP, potentially to mediate its extraction from the ER membrane. Surprisingly, I also found that EVI/WLS is ubiquitinated and degraded in cells irrespective of their WNT activity. This K11-, K48-, and K63-linked ubiquitina-tion is mediated by the E2 ubiquitin conjugating enzymes UBE2J2, UBE2K, and UBE2N and leads not only to the regulation of EVI/WLS protein levels, but also influences WNT secre-tion. Analysing the functional impact of EVI/WLS abundance revealed that EVI/WLS protein levels and the secretion of WNT11 influence the invasive capacity of malignant melanoma cells. This suggests that the adaptive regulation of EVI/WLS can be important for the phenotypic manifestation and presumably progression of human malignancies. In summary, my data shows an unanticipated complex ubiquitination pattern of EVI/WLS and three novel interaction partners, thus providing important details on the post-translational modification and fate of an endogenous ERAD substrate in mammalian cells. The abundance of EVI/WLS is essential for context-dependent WNT ligand secretion and thus gov-erns the malignancy of several tumours, among them melanoma. Targeting EVI/WLS protein levels via its post-translational regulations could be used to treat WNT-dependent diseases

Boot Camp-Programme als Behandlungsmodell für aggressive Kinder und Jugendliche

In der vorliegenden Diplomarbeit werden so genannte Boot Camp-Programme einer kritischen Erörterung unterzogen. Dazu werden die theoretischen Grundlagen und Zielsetzungen von Boot Camp-Programmen dargelegt und mit ausgesuchten psychoanalytisch-pädagogischen Verwahrlosungstheorien bzw. Konzepten zur Arbeit mit aggressiven/delinquenten Kindern und Jugendlichen kontrastiert.This thesis is a critical discussion of so-called boot camp-programs. It looks at the theoretical fundamentals and the goal of boot camp programs; these are debated and contrasted with well-chosen psychoanalytic-pedagogical delinquent theories as well as concepts for working with aggressive/delinquent children and adolescents

Pax6 interactions with chromatin and identification of its novel direct target genes in lens and forebrain.

Pax6 encodes a specific DNA-binding transcription factor that regulates the development of multiple organs, including the eye, brain and pancreas. Previous studies have shown that Pax6 regulates the entire process of ocular lens development. In the developing forebrain, Pax6 is expressed in ventricular zone precursor cells and in specific populations of neurons; absence of Pax6 results in disrupted cell proliferation and cell fate specification in telencephalon. In the pancreas, Pax6 is essential for the differentiation of α-, β- and δ-islet cells. To elucidate molecular roles of Pax6, chromatin immunoprecipitation experiments combined with high-density oligonucleotide array hybridizations (ChIP-chip) were performed using three distinct sources of chromatin (lens, forebrain and β-cells). ChIP-chip studies, performed as biological triplicates, identified a total of 5,260 promoters occupied by Pax6. 1,001 (133) of these promoter regions were shared between at least two (three) distinct chromatin sources, respectively. In lens chromatin, 2,335 promoters were bound by Pax6. RNA expression profiling from Pax6⁺/⁻ lenses combined with in vivo Pax6-binding data yielded 76 putative Pax6-direct targets, including the Gaa, Isl1, Kif1b, Mtmr2, Pcsk1n, and Snca genes. RNA and ChIP data were validated for all these genes. In lens cells, reporter assays established Kib1b and Snca as Pax6 activated and repressed genes, respectively. In situ hybridization revealed reduced expression of these genes in E14 cerebral cortex. Moreover, we examined differentially expressed transcripts between E9.5 wild type and Pax6⁻/⁻ lens placodes that suggested Efnb2, Fat4, Has2, Nav1, and Trpm3 as novel Pax6-direct targets. Collectively, the present studies, through the identification of Pax6-direct target genes, provide novel insights into the molecular mechanisms of Pax6 gene control during mouse embryonic development. In addition, the present data demonstrate that Pax6 interacts preferentially with promoter regions in a tissue-specific fashion. Nevertheless, nearly 20% of the regions identified are accessible to Pax6 in multiple tissues

Geothermal Potential of the Brenner Base Tunnel—Initial Evaluations

Increasing demands on mobility and transport, but limited space above ground, lead to new traffic routes being built, even more underground in the form of tunnels. In addition to improving the traffic situation, tunnels offer the possibility of contributing to climate-friendly heating by indirectly serving as geothermal power plants. In this study, the geothermal potential of the future longest railway tunnel in the world, the Brenner Base Tunnel, was evaluated. At the Brenner Base Tunnel, warm water naturally flows from the apex of the tunnel towards the city of Innsbruck, Austria. In order to estimate its geothermal potential, hydrological data of discharge rates and temperatures were investigated and analyzed. The investigations indicated the highest geothermal potential in the summertime, while the lowest occurs during winter. It could be shown that these variations were a result of cooling during discharge through areas of low overburden (mid mountain range), where the tunnel atmosphere is increasingly influenced by the air temperatures outside the tunnel. Nevertheless, the calculations showed that there will be a usable potential after completion of the tunnel

Awareness of the role of cardiovascular risk factors and their prevention from the perspective of Tricity adolescents

Introduction. Arterial hypertension (HT) affects 10.4 million adult Poles, and the blood pressure (BP) control rate is only 26%. Beyond any doubt, high blood pressure results in cardiovascular (CV) target organ damage, which markedly influences national healthcare programs. Childhood introduced cardiovascular prophylaxis offers opportunities to decrease incident HT and delay or even eliminate its consequences. Therefore we have decided to study the level of knowledge on HT and CV disease (CVD) among random 1st and 2nd grade high school students in Tricity, Poland. Material and methods. Questionnaire-based study was conducted voluntarily in two selected high schools of the Tricity agglomeration, Poland, in 2000 and repeated in 2016. All participants were presented with a 38-item questionnaire on cardiovascular risk factors knowledge as well as students’ health habits. Results. Studied group consisted of 615 students (57.6% females) at the age of 16, mean BMI 20.89 ± 2.95 kg/m2 (78% BMI norm). The evaluation of the several habits in two time-points was as follows: additional salt intake was reported by 38.3% vs. 35.1% adolescents in 2000 and 2016, respectively; p = 0.43, dining in fast food restaurants at least once a week: 13.9% vs. 44.8%; 2000/2016; p < 0.01; hours per week spent on physical activity: 5.6 ± 4.4 vs. 5.5 } 4.5; p = 0.85; students who never drank alcohol: 19.7% vs. 31.1%; p < 0.01; cigarettes ever smoking: 39.6% vs. 21.2%; p < 0.01; regular smokers: 10.2% vs. 4.1%; p < 0.05. The awareness of CV-preventive measures such as body weight reduction was presented by 88.1% vs. 93.9% students in 2000 vs. 2016; p = 0.01; regular physical activity 92.6% vs. 97.1%; P = 0.01, limiting smoking and alcohol consumption 84.2% vs. 91.2%; p = 0.01; salt reduction: 62.6% vs. 82.3%; p < 0.001. Interestingly, in 2016 the majority of students identified myocardial infarction as a consequence of AH 92.4%, which was markedly less evident in case of stroke and kidney disease (46.2%, and 28%, respectively). Conclusion. Although the awareness of negative consequences of HT and other CV-risk factors has grown over the past 15 years, the implementation of this knowledge is insufficient among high-school adolescents.

Spatio-temporal patterns of Pyrenean exhumation revealed by inverse thermo-kinematic modeling of a large thermochronologic data set

Large thermochronologic data sets enable orogen-scale investigations into spatio-temporal patterns of erosion and deformation. We present the results of a thermo-kinematic modeling study that examines large-scale controls on spatio-temporal variations in exhumation as recorded by multiple low-temperature thermochronometers in the Pyrenees mountains (France/Spain). Using 264 compiled cooling ages spanning ∼200 km of the orogen, a recent model for its topographic evolution, and the thermo-kinematic modeling code Pecube, we evaluated two models for Axial Zone (AZ) exhumation: (1) thrust sheet–controlled (north-south) exhumation, and (2) along-strike (east-west) variable exhumation. We also measured the degree to which spatially variable post-orogenic erosion influenced the cooling ages. We found the best fit for a model of along-strike variable exhumation. In the eastern AZ, rock uplift rates peak at ≥1 mm/yr between 40 and 30 Ma, whereas in the western AZ, they peak between 30 and 20 Ma. The amount of post-orogenic (2.5 km in the west. The data reveal a pattern of exhumation that is primarily controlled by structural inheritance, with ancillary patterns reflecting growth and erosion of the antiformal stack and post-orogenic surface processes.publishedVersio

Cytoplasmic LXR expression is an independent marker of poor prognosis for patients with early stage primary breast cancer

International audiencePurpose The aim of this study was to investigate the expression of liver X receptors α/β (LXR) in primary breast cancer (BC) tissues and to analyze its correlations with clinicopathological parameters including patient survival. Methods In a well-characterized cohort of 305 primary BC, subcellular distribution of LXR was evaluated by immunohistochemistry. Correlations with clinicopathological characteristics as well as with patient outcome were analyzed. Results LXR was frequently localized in both nuclei and cytoplasms of BC cells, with stronger staining in nuclei. Total and nuclear LXR expression was positively correlated with ER and PR status. Overall survival analysis demonstrated that cytoplasmic LXR was significantly correlated with poor survival and appeared as an independent marker of poor prognosis, in stage I but not in stage II–III tumors Conclusion Altogether, these data suggest that cytoplasmic LXR could be defined as a prognostic marker in early stage primary BC

Ginkgo biloba Extract EGb 761 Improves Vestibular Compensation and Modulates Cerebral Vestibular Networks in the Rat

Unilateral inner ear damage is followed by behavioral recovery due to central vestibular compensation. The dose-dependent therapeutic effect of Ginkgo biloba extract EGb 761 on vestibular compensation was investigated by behavioral testing and serial cerebral [18F]-Fluoro-desoxyglucose ([18F]-FDG)-μPET in a rat model of unilateral labyrinthectomy (UL). Five groups of 8 animals each were treated with EGb 761-supplemented food at doses of 75, 37.5 or 18.75 mg/kg body weight 6 weeks prior and 15 days post UL (groups A,B,C), control food prior and EGb 761-supplemented food (75 mg/kg) for 15 days post UL (group D), or control food throughout (group E). Plasma levels of EGb 761 components bilobalide, ginkgolide A and B were analyzed prior and 15 days post UL. Behavioral testing included clinical scoring of nystagmus, postural asymmetry, head roll tilt, body rotation during sensory perturbation and instrumental registration of mobility in an open field before and 1, 2, 3, 5, 7, 15 days after UL. Whole-brain [18F]-FDG-μPET was recorded before and 1, 3, 7, 15 days after UL. The EGb 761 group A (75 mg/kg prior/post UL) showed a significant reduction of nystagmus scores (day 3 post UL), of postural asymmetry (1, 3, 7 days post UL), and an increased mobility in the open field (day 7 post UL) as compared to controls (group E). Application of EGb 761 at doses of 37.5 and 18.75 mg/kg prior/post UL (groups B,C) resulted in faster recovery of postural asymmetry, but did not influence mobility relative to controls. Locomotor velocity increased with higher plasma levels of ginkgolide A and B. [18F]-FDG-μPET revealed a significant decrease of the regional cerebral glucose metabolism (rCGM) in the vestibular nuclei and cerebellum and an increase in the hippocampal formation with higher plasma levels of ginkgolides and bilobalide 1 and 3 days post UL. Decrease of rCGM in the vestibular nucleus area and increase in the hippocampal formation with higher plasma levels persisted until day 15 post UL. In conclusion, Ginkgo biloba extract EGb 761 improves vestibulo-ocular motor, vestibulo-spinal compensation, and mobility after UL. This rat study supports the translational approach to investigate EGb 761 at higher dosages for acceleration of vestibular compensation in acute vestibular loss