Multiplex Profiling of Cellular Invasion in 3D Cell Culture Models.

To-date, most invasion or migration assays use a modified Boyden chamber-like design to assess migration as single-cell or scratch assays on coated or uncoated planar plastic surfaces. Here, we describe a 96-well microplate-based, high-content, three-dimensional cell culture assay capable of assessing invasion dynamics and molecular signatures thereof. On applying our invasion assay, we were able to demonstrate significant effects on the invasion capacity of fibroblast cell lines, as well as primary lung fibroblasts. Administration of epidermal growth factor resulted in a substantial increase of cellular invasion, thus making this technique suitable for high-throughput pharmacological screening of novel compounds regulating invasive and migratory pathways of primary cells. Our assay also correlates cellular invasiveness to molecular events. Thus, we argue of having developed a powerful and versatile toolbox for an extensive profiling of invasive cells in a 96-well format. This will have a major impact on research in disease areas like fibrosis, metastatic cancers, or chronic inflammatory states

Gli1 mediates lung cancer cell proliferation and sonic hedgehog-dependent mesenchymal cell activation.

Non-Small-Cell-Lung-Cancer (NSCLC) represents approximately 85% of all lung cancers and remains poorly understood. While signaling pathways operative during organ development, including Sonic Hedgehog (Shh) and associated Gli transcription factors (Gli1-3), have recently been found to be reactivated in NSCLC, their functional role remains unclear. Here, we hypothesized that Shh/Gli1-3 could mediate NSCLC autonomous proliferation and epithelial/stromal signaling in the tumoral tissue. In this context, we have investigated the activity of Shh/Gli1-3 signaling in NSCLC in both, cancer and stromal cells. We report here that inhibition of Shh signaling induces a significant decrease in the proliferation of NSCLC cells. This effect is mediated by Gli1 and Gli2, but not Gli3, through regulation of cyclin D1 and cyclin D2 expression. While exogenous Shh was unable to induce signaling in either A549 lung adenocarcinoma or H520 lung squamous carcinoma cells, both cells were found to secrete Shh ligand, which induced fibroblast proliferation, survival, migration, invasion, and collagen synthesis. Furthermore, Shh secreted by NSCLC mediates the production of proangiogenic and metastatic factors in lung fibroblasts. Our results thus provide evidence that Shh plays an important role in mediating epithelial/mesenchymal crosstalk in NSCLC. While autonomous Gli activity controls NSCLC proliferation, increased Shh expression by NSCLC is associated with fibroblast activation in tumor-associated stroma. Our study highlights the relevance of studying stromal-associated cells in the context of NSCLC regarding new prognosis and therapeutic options

Wege aus der Schulden- und Vertrauenskrise in der Europäischen Wirtschafts- und Währungsunion

Die Europäische Wirtschafts- und Währungsunion (EWWU) befindet sich gegenwärtig in einer Schulden- und Vertrauenskrise. Die europäischen Institutionen haben darauf mit einer Reihe von Maßnahmen reagiert: Ein Finanzstabilisierungsmechanismus wurde geschaffen, und die Europäische Zentralbank hat damit begonnen, die Anleihen von denjenigen Mitgliedsstaaten des Euro-Währungsgebietes aufzukaufen, die auf den Finanzmärkten gar nicht mehr oder nur zu relativ hohen Zinsen Finanzmittel aufnehmen können. Zwar können diese Maßnahmen kurzfristig geeignet sein, die Lage zu stabilisieren; langfristig sind sie jedoch problematisch. So wird das Überschuldungsproblem Griechenlands nicht dauerhaft gelöst und die Krisenanfälligkeit sowohl des Finanzsystems als auch der Mitgliedsstaaten selbst wird nicht gemindert. Die durch die ergriffenen Maßnahmen gewonnene Zeit muss unbedingt zur Stärkung der Institutionen im Euro-Währungsgebiet genutzt werden. Eine graduelle Modifikation des Stabilitäts- und Wachstumspaktes oder die Schaffung neuer politischer Institutionen, zum Beispiel einer europäischen Wirtschaftsregierung, wird dies nicht leisten können. Vielmehr bedarf es der Einsicht, dass Krisen Bestandteil marktwirtschaftlich organisierter Volkswirtschaften sind und dass vorab vereinbarte Regeln für den Umgang mit ihnen festgelegt werden müssen. Dazu zählt vor allem eine Insolvenzordnung für Banken und auch für Staaten, um systemische Risiken zu reduzieren.

Strongly inhomogeneous distribution of spectral properties of silicon-vacancy color centers in nanodiamonds

The silicon-vacancy (SiV) color center in diamond is a solid-state single photon emitter and spin quantum bit suited as a component in quantum devices. Here, we show that the SiV center in nanodiamond exhibits a strongly inhomogeneous distribution with regard to the center wavelengths and linewidths of the zero-phonon-line (ZPL) emission at room temperature. We find that the SiV centers separate in two clusters: one group exhibits ZPLs with center wavelengths within a narrow range of approximatly 730 nm to 742 nm and broad linewidths between 5 nm and 17 nm, whereas the second group comprises a very broad distribution of center wavelengths between 715 nm and 835 nm, but narrow linewidths from below 1 nm up to 4 nm. Supported by ab initio Kohn-Sham density functional theory calculations we show that the ZPL shifts of the first group are consistently explained by strain in the diamond lattice. Further, we suggest, that the second group showing the strongly inhomogeneous distribution of center wavelengths might be comprised of modified SiV centers. Whereas single photon emission is demonstrated for SiV centers of both clusters, we show that emitters from different clusters show different spectroscopic features such as variations of the phonon sideband spectra and different blinking dynamics

Extracorporeal Cardiac Shock Wave Therapy Ameliorates Clinical Symptoms and Improves Regional Myocardial Blood Flow in a Patient with Severe Coronary Artery Disease and Refractory Angina

Different therapeutic options are being used for chronic coronary artery disease (CAD). We report about a 51-year-old female with CAD and refractory angina pectoris despite maximally tolerated medical therapy and after both percutaneous coronary intervention (PCI) and coronary artery bypass grafting (CABG). The patient received cardiac shock wave therapy (CSWT) over a period of 6 month. There was no arrhythmia during or after treatment; enzyme levels were normal at all times. PET imaging showed a substantial improvement of myocardial stress perfusion. Since the patient reported that she now was fully capable to deal with her everyday life, further treatment options were postponed. Our case report suggests that ultrasound-guided CSWT is able to improve symptoms and perfusion in ischemic myocardium

Operation strategies of battery energy storage systems for preventive and curative congestion management in transmission grids

Anticipating and relieving congestions is an ongoing challenge for transmission system operators. Distributed grid-scale battery energy storage systems enable operators to shift power flows and remedy congestion through virtual power lines and grid boosters. This paper includes battery energy storage systems in a combined preventive and curative congestion management optimization. First, it analyzes the impact of the two operational strategies in a case study of the German transmission grid. Furthermore, it outlines curative ad-hoc measures to overcome uncertainties during operational planning and real-time operation. The simulation results indicate that battery energy storage systems further increase the use of curative measures and reduce congestion management costs

Robustness of ARS leptogenesis in scalar extensions

Extensions of the Standard Model (SM) with sterile neutrinos are well motivated from the observed oscillations of the light neutrinos and they have shown to successfully explain the Baryon Asymmetry of the Universe (BAU) through, for instance, the so-called ARS leptogenesis. Sterile neutrinos can be added in minimal ways to the SM, but many theories exist where sterile neutrinos are not the only new fields. Such theories often include scalar bosons, which brings about the possibility of further interactions between the sterile neutrinos and the SM. In this paper we consider an extension of the SM with two sterile neutrinos and one scalar singlet particle and investigate the effect that an additional, thermalised, scalar has on the ARS leptogenesis mechanism. We show that in general the created asymmetry is reduced due to additional sterile neutrino production from scalar decays. When sterile neutrinos and scalars are discovered in the laboratory, our results will provide information on the applicability of the ARS leptogenesis mechanism.Comment: 16 pages plus references, 6 figures, 2 table

Pasteurella multocida toxin- induced osteoclastogenesis requires mTOR activation

Background: Pasteurella multocida toxin (PMT) is a potent inducer of osteoclast formation. Pigs suffering from an infection with toxigenic Pasteurella multocida strains develop atrophic rhinitis characterised by a loss of turbinate bones and conchae. However, on the molecular level the process of bone loss remains largely uncharacterised. Results: Recently it was found that PMT activates the serine/threonine kinase mammalian target of rapamycin (mTOR) in fibroblasts. Using RAW264.7 macrophages, we investigated the role of the mTOR complex 1 (mTORC1) in PMT-mediated osteoclast formation. PMT induces the differentiation of RAW264.7 macrophages into multinucleated, tartrate resistant acid phosphatase (TRAP) positive osteoclasts that are capable to resorb bone. In the presence of the mTORC1 inhibitor rapamycin, PMT was significantly less able to induce the formation of TRAP-positive osteoclasts. Accordingly, the resulting resorption of bone was strongly reduced. A major target of mTOR is the 70 kDa ribosomal protein S6 kinase 1 (p70 S6K1). Activated p70 S6K1 decreases the expression of programmed cell death protein 4 (PDCD4), a negative transcriptional regulator of osteoclastogenesis, at the protein and gene level. Ultimately this results in the activation of c-Jun, a component of the activator protein 1 (AP-1) complex, which is a major transcription factor for the induction of osteoclast-specific genes. We now demonstrate that c-Jun and its downstream target, the osteoclast-specific bone degrading protease cathepsin K, are upregulated upon PMT treatment in an mTOR-dependent manner. Conclusions: Activation of mTOR signalling plays a central role in the formation of osteoclasts through the bacterial toxin PMT. On the molecular level, PMT-induced activation of mTOR leads to down regulation of PDCD4, a known repressor of AP-1 complex, culminating in the activation of c-Jun, an essential transcription factor for triggering osteoclastogenesis