A novel MCF-10A line allowing conditional oncogene expression in 3D culture

Introduction Non-transformed mammary epithelial cell lines such as MCF-10A recapitulate epithelial morphogenesis in three-dimensional (3D) tissue culture by forming acinar structures. They represent an important tool to characterize the biological properties of oncogenes and to model early carcinogenic events. So far, however, these approaches were restricted to cells with constitutive oncogene expression prior to the set-up of 3D cultures. Although very informative, this experimental setting has precluded the analysis of effects caused by sudden oncoprotein expression or withdrawal in established epithelial cultures. Here, we report the establishment and use of a stable MCF-10A cell line (MCF-10Atet) fitted with a novel and improved doxycycline (dox)-regulated expression system allowing the conditional expression of any transgene. Methods MCF-10Atet cells were generated by stable transfection with pWHE644, a vector expressing a second generation tetracycline-regulated transactivator and a novel transcriptional silencer. In order to test the properties of this new repressor/activator switch, MCF-10Atet cells were transfected with a second plasmid, pTET-HABRAF-IRES-GFP, which responds to dox treatment with the production of a bi-cistronic transcript encoding hemagglutinin-tagged B-Raf and green fluorescent protein (GFP). This improved conditional expression system was then characterized in detail in terms of its response to various dox concentrations and exposure times. The plasticity of the phenotype provoked by oncogenic B-RafV600E in MCF-10Atet cells was analyzed in 3D cultures by dox exposure and subsequent wash-out. Results MCF-10Atet cells represent a tightly controlled, conditional gene expression system. Using B-RafV600E as a model oncoprotein, we show that its sudden expression in established 3D cultures results in the loss of acinar organization, the induction of an invasive phenotype and hallmarks of epithelial-to-mesenchymal transition (EMT). Importantly, we show for the first time that this severe transformed phenotype can be reversed by dox wash-out and concomitant termination of oncogene expression. Conclusions Taken together, we have generated a stable MCF-10A subline allowing tight dox-controlled and reversible expression of any transgene without the need to modify its product by introducing artificial dimerization or ligand-binding domains. This system will be very valuable to address phenomena such as EMT, oncogene addiction, oncogene-induced senescence and drug resistance

The Hilbert basis method for D-flat directions and the superpotential

We discuss, using the Hilbert basis method, how to efficiently construct a complete basis for D-flat directions in supersymmetric Abelian and non-Abelian gauge theories. We extend the method to discrete (R and non-R) symmetries. This facilitates the construction of a basis of all superpotential terms in a theory with given symmetries.Comment: 11 pages; a related mathematica code can be found at http://einrichtungen.ph.tum.de/T30e/codes/NonAbelianHilbert

Comparing Evapotranspiration from Eddy Covariance Measurements, Water Budgets, Remote Sensing, and Land Surface Models over Canada

This study compares six evapotranspiration ET products for Canada’s landmass, namely, eddy covariance EC measurements; surface water budget ET; remote sensing ET from MODIS; and land surface model (LSM) ET from the Community Land Model (CLM), the Ecological Assimilation of Land and Climate Observations (EALCO) model, and the Variable Infiltration Capacity model (VIC). The ET climatology over the Canadian landmass is characterized and the advantages and limitations of the datasets are discussed. The EC measurements have limited spatial coverage, making it difficult for model validations at the national scale. Water budget ET has the largest uncertainty because of data quality issues with precipitation in mountainous regions and in the north. MODIS ET shows relatively large uncertainty in cold seasons and sparsely vegetated regions. The LSM products cover the entire landmass and exhibit small differences in ET among them. Annual ET from the LSMs ranges from small negative values to over 600 mm across the landmass, with a countrywide average of 256 ± 15 mm. Seasonally, the countrywide average monthly ET varies from a low of about 3 mm in four winter months (November–February) to 67 ± 7 mm in July. The ET uncertainty is scale dependent. Larger regions tend to have smaller uncertainties because of the offset of positive and negative biases within the region. More observation networks and better quality controls are critical to improving ET estimates. Future techniques should also consider a hybrid approach that integrates strengths of the various ET products to help reduce uncertainties in ET estimation

Linking scales and disciplines : an interdisciplinary cross-scale approach to supporting climate-relevant ecosystem management

CITATION: Berger, C. et al. 2019. Linking scales and disciplines : an interdisciplinary cross-scale approach to supporting climate-relevant ecosystem management. Climatic Change, 156:139–150, doi:10.1007/s10584-019-02544-0.The original publication is available at https://www.springer.com/journal/10584Southern Africa is particularly sensitive to climate change, due to both ecological and socioeconomic factors, with rural land users among the most vulnerable groups. The provision of information to support climate-relevant decision-making requires an understanding of the projected impacts of change and complex feedbacks within the local ecosystems, as well as local demands on ecosystem services. In this paper, we address the limitation of current approaches for developing management relevant socio-ecological information on the projected impacts of climate change and human activities.We emphasise the need for linking disciplines and approaches by expounding the methodology followed in our two consecutive projects. These projects combine disciplines and levels of measurements from the leaf level (ecophysiology) to the local landscape level (flux measurements) and from the local household level (socio-economic surveys) to the regional level (remote sensing), feeding into a variety of models at multiple scales. Interdisciplinary, multi-scaled, and integrated socio-ecological approaches, as proposed here, are needed to compliment reductionist and linear, scalespecific approaches. Decision support systems are used to integrate and communicate the data and models to the local decision-makers.https://link.springer.com/article/10.1007/s10584-019-02544-0Publisher's versio

Restricting Glycolysis Preserves T Cell Effector Functions and Augments Checkpoint Therapy

Tumor-derived lactic acid inhibits T and natural killer (NK) cell function and, thereby, tumor immunosurveillance. Here, we report that melanoma patients with high expression of glycolysis-related genes show a worse progression free survival upon anti-PD1 treatment. The non-steroidal anti-inflammatory drug (NSAID) diclofenac lowers lactate secretion of tumor cells and improves anti-PD1-induced T cell killing in vitro. Surprisingly, diclofenac, but not other NSAIDs, turns out to be a potent inhibitor of the lactate transporters monocarboxylate transporter 1 and 4 and diminishes lactate efflux. Notably, T cell activation, viability, and effector functions are preserved under diclofenac treatment and in a low glucose environment in vitro. Diclofenac, but not aspirin, delays tumor growth and improves the efficacy of checkpoint therapy in vivo. Moreover, genetic suppression of glycolysis in tumor cells strongly improves checkpoint therapy. These findings support the rationale for targeting glycolysis in patients with high glycolytic tumors together with checkpoint inhibitors in clinical trials

2020 Proceedings of the 3rd International Conference on Trauma Surgery Technology in Giessen

The 3 rd event of the Giessen International Conference on Trauma Surgery Technology on October, the 17th 2020 was hosted on Zoom in accordance with the worldwide corona situation. Dr Mieczakowski, Dr Yu, and Wolfram drafted in 2018 from Jan’s apartment in Bremen the manuscript which was submitted to and approved for funding by the Deutsche Forschungsgemeinschaft (DFG). At that time, we had no idea what substantial changes the conferencing concept would require. This is why we would like to thank again Michele. She first planned this year’s event after the 2019 date and then in the spring of 2020 had to replan for the new situation

Einfluss von Temperament-, Charakter- und Persönlichkeitsvariablen sowie des Dopamin-D2-Rezeptor (DRD2) rs6276 Genotyps auf die Alkoholtrinkmenge

In der vorliegenden Untersuchung wurde bei Probanden aus dem Subprojekt „Psychosoziale Gesundheit in Familien“ der bevölkerungsrepräsentativen Basisstudie „Study of Health in Pomerania, SHIP“ sowohl der Zusammenhang von Temperament, Charakter und Persönlichkeit, als auch des DRD2 rs6276 Genotyps mit der Alkoholtrinkmenge analysiert. Die Untersuchung erfolgte zum einen in einer Gesamtstichprobe, welche sowohl Probanden mit als auch ohne Alkoholdiagnose enthielt und zum anderen in einer Normalstichprobe, welche nur Probanden ohne Alkoholdiagnose enthielt. Die aus der beschriebenen Literatur abgeleiteten Hypothesen bezüglich erhöhter Trinkmengen bei hohem Neugierverhalten und Neurotizismus, sowie niedriger Belohnungsabhängigkeit und Selbstlenkungsfähigkeit konnte in der Gesamtstichprobe signifikant bestätigt werden. Probanden mit niedrigem Beharrungsvermögen zeigten zudem im Trend erhöhte Trinkmengen. Die Hypothese, dass geringe Werte in Schadensvermeidung mit hohen Trinkmengen assoziiert sind, konnte in der Gesamtstichprobe nicht bestätigt werden. Über die aufgestellten Hypothesen hinaus waren auch niedrige Werte in Kooperativität assoziiert mit erhöhten Trinkmengen. Die Auswertung der Normalstichprobe zeigte wenige mit der Gesamtstichprobe übereinstimmende Ergebnisse. Lediglich niedrige Werte für Kooperativität waren in beiden Stichproben mit hohen Konsummengen assoziiert. Die Hypothesen, dass erhöhte Trinkmengen, sowie kürzere Abstinenzphasen mit dem DRD2 rs6276 A/A-Genotyp assoziiert sind, konnten in der Gesamtstichprobe (mit DNA) bei den männlichen Probanden bestätigt werden. Die Interaktion von Beharrungsvermögen bzw. Selbstlenkungsfähigkeit (sowie im Trend auch Kooperativität) und dem DRD2 rs6276 Genotyp war assoziiert mit der Trinkmenge. Bei den weiblichen Probanden konnte dies nicht beobachtet werden. Die Ergebnisse der vorliegenden Arbeit zeigen, dass gewisse Merkmalsausprägungen von Temperament, Charakter und Persönlichkeit, sowie der Genotyp des DRD2 rs6276 in Abhängigkeit vom Geschlecht, mit der Trinkmenge assoziiert sind. Niedrige Werte in Kooperativität könnten im Hinblick auf die identischen Ergebnisse in der Gesamt- und Normalstichprobe ein Prädiktor für erhöhte Trinkmengen und somit vielleicht für die Entwicklung einer Alkoholabhängigkeit sein. Es erscheint durchaus plausibel, dass Probanden mit niedrigen Werten in Kooperativität (entspricht: intolerant, teilnahmslos, egoistisch, nachtragend, gewissenlos) höhere Trinkmengen aufweisen. So wäre es denkbar, dass diese den aus der mangelnden Kooperativität entstehenden interpersonellen Stress über Alkohol kompensieren. Niedrige Werte in Kooperativität sind zudem ein Charakteristikum zahlreicher Persönlichkeitsstörungen, welche wiederum mit erhöhtem Substanzkonsum assoziiert sind. Zur Überprüfung dieser Hypothese wäre eine longitudinal angelegte Studie indiziert.The influence of temperament-, character- and personality-variables and also the genotype of the DRD2-rs6276-receptor on the alcohol consumption was researched in a mixed sample. The operationalisation was based on the TCI by Cloninger, the NEO-FFI by Costa und McCrae and the SSAGA by Hesselbrock. We saw high values in novelty seeking, neuroticism together with low values in reward dependence, cooperative and self directedness associated with high alcohol consumption. Additional we saw the AA-genotyp of the DRD2-rs6276-receptor in opposite to the AG/GG-genotyp associated with high alcohol consumption in the male sample. Furthermore we saw interactions between the genotype and the temperament-, character- and personality-variables in association to the alcohol consumption

Becoming a professional soccer player in Bavaria : analysis and com-parisons of different career facilities for soccer talents in Bavaria

In dieser Arbeit werden die verschiedenen Ausbildungsmöglichkeiten aufgezeigt, die ein Fußballtalent in Bayern zur Auswahl hat, um dem Traum Profifußball ein Stück näher zu kommen. Die verschiedenen Möglichkeiten werden aufgeführt, vorgestellt und verglichen. Die gängigste Variante ist der Weg durch ein Nachwuchsleistungszentrum eines Profivereins. Jedoch gibt es hierzu gute Alternativen, welche in dieser Arbeit gegenübergestellt werden. Es werden die einzelnen Teilbereiche in der Arbeit eines Profi-NLZ und der eines BFV-NLZ genauer vorgestellt und verglichen. Am Ende der Arbeit wird eine Zusammenfassung erstellt, in der gezeigt wird, welche positiven und negativen Aspekte der jeweilige Ausbildungsweg mit sich bringt

Profiling Foreign Policy Leaders in Their Own Language : New Insights into the Stability and Formation of Leadership Traits

Leaders matter in international politics. One of the main tools for assessing at-a-distance psychological characteristics of political leaders is Leadership Trait Analysis. To facilitate empirical studies, a Leadership Trait Analysis coding scheme for automated text analysis was developed to replace hand-coding. However, the coding scheme has been available only for English-language texts. To broaden research opportunities, this article presents a novel Leadership Trait Analysis coding scheme for the German language. This coding scheme allows engaging in empirical analysis based on original German language sources, thereby shedding new or different light on German foreign policy. At the same time, it contributes to moving automated content analysis beyond the English language more generally