iRFP is a real time marker for transformation based assays in high content screening

Anchorage independent growth is one of the hallmarks of oncogenic transformation. Here we show that infrared fluorescent protein (iRFP) based assays allow accurate and unbiased determination of colony formation and anchorage independent growth over time. This protocol is particularly compatible with high throughput systems, in contrast to traditional methods which are often labor-intensive, subjective to bias and do not allow further analysis using the same cells. Transformation in a single layer soft agar assay could be documented as early as 2 to 3 days in a 96 well format, which can be easily combined with standard transfection, infection and compound screening setups to allow for high throughput screening to identify therapeutic targets

New model of calculating the energy transfer efficiency for the spherical theta-pinch device

Ion-beam-plasma-interaction plays an important role in the field of Warm Dense Matter (WDM) and Inertial Confinement Fusion (ICF). A spherical theta pinch is proposed to act as a plasma target in various applications including a plasma stripper cell. One key parameter for such applications is the free electron density. A linear dependency of this density to the amount of energy transferred into the plasma from an energy storage was found by C. Teske. Since the amount of stored energy is known, the energy transfer efficiency is a reliable parameter for the design of a spherical theta pinch device. The traditional two models of energy transfer efficiency are based on assumptions which comprise the risk of systematical errors. To obtain precise results, this paper proposes a new model without the necessity of any assumption to calculate the energy transfer efficiency for an inductively coupled plasma device. Further, a comparison of these three different models is given at a fixed operation voltage for the full range of working gas pressures. Due to the inappropriate assumptions included in the traditional models, one owns a tendency to overestimate the energy transfer efficiency whereas the other leads to an underestimation. Applying our new model to a wide spread set of operation voltages and gas pressures, an overall picture of the energy transfer efficiency results

Development of an inducible mouse model of iRFP713 to track recombinase activity and tumour development in vivo

While the use of bioluminescent proteins for molecular imaging is a powerful technology to further our understanding of complex processes, fluorescent labeling with visible light fluorescent proteins such as GFP and RFP suffers from poor tissue penetration and high background autofluorescence. To overcome these limitations, we generated an inducible knock-in mouse model of iRFP713. This model was used to assess Cre activity in a Rosa Cre-ER background and quantify Cre activity upon different tamoxifen treatments in several organs. We also show that iRFP can be readily detected in 3D organoid cultures, FACS analysis and in vivo tumour models. Taken together we demonstrate that iRFP713 is a progressive step in in vivo imaging and analysis that widens the optical imaging window to the near-infrared spectrum, thereby allowing deeper tissue penetration, quicker image acquisition without the need to inject substrates and a better signal to background ratio in genetically engineered mouse models (GEMMs)

Mapping and explaining the productivity of Pinus radiata in New Zealand

Mapping Pinus radiata productivity for New Zealand not only provides useful information for forest owners, industry stakeholders and policy managers, but also enables current and future plantations to be visualised, quantified, and planned. Using an extensive set of permanent sample plots, split into fitting (n = 1,146) and validation (n = 618) datasets, models of P. radiata 300 Index (an index of volume mean annual increment) and Site Index (an index of height growth) were developed using a regression kriging technique. Spatial predictions were accurate and accounted for 61% and 70% of the variance for 300 Index and Site Index, respectively. Productivity predicted from these surfaces for the entire plantation estate averaged 27.4 m³ ha⁻¹ yr⁻¹ for the 300 Index and 30.4 m for Site Index. Surfaces showed wide regional variation in this productivity, which was attributable mainly to variation in air temperature and root-zone water storage from site to site

Ubiquitination and proteasomal degradation of ATG12 regulates its proapoptotic activity

During macroautophagy, conjugation of ATG12 to ATG5 is essential for LC3 lipidation and autophagosome formation. Additionally, ATG12 has ATG5-independent functions in diverse processes including mitochondrial fusion and mitochondrial-dependent apoptosis. In this study, we investigated the regulation of free ATG12. In stark contrast to the stable ATG12–ATG5 conjugate, we find that free ATG12 is highly unstable and rapidly degraded in a proteasome-dependent manner. Surprisingly, ATG12, itself a ubiquitin-like protein, is directly ubiquitinated and this promotes its proteasomal degradation. As a functional consequence of its turnover, accumulation of free ATG12 contributes to proteasome inhibitor-mediated apoptosis, a finding that may be clinically important given the use of proteasome inhibitors as anticancer agents. Collectively, our results reveal a novel interconnection between autophagy, proteasome activity, and cell death mediated by the ubiquitin-like properties of ATG12

Die Wirkung von FBL-Übungen während der Arbeitszeit

Treten bei vorwiegend sitzenden Tätigkeiten während der Arbeitszeit, z.B. an Bildschirmarbeitsplätzen, über kurz oder lang körperliche Beschwerden auf und kann man diese im Rahmen der betrieblichen Gesundheitsförderung beeinflussen? Diese Frage wurde im Rahmen einer Studienarbeit , die während der Physiotherapieausbildung an der staatlichen Schule für Physiotherapie des Universitätsklinikums Münster durchgeführt wurde, untersucht. Den Probanden wurden Übungen aus der Funktionellen Bewegungslehre gezeigt, die Auswertung ergab keine unmittelbare Schmerzlinderung, aber eine Steigerung des körperlichen Wohlbefindens

High energy Coulomb-scattered electrons for relativistic particle beam diagnostics

A new system used for monitoring energetic Coulomb-scattered electrons as the main diagnostic for accurately aligning the electron and ion beams in the new Relativistic Heavy Ion Collider (RHIC) electron lenses is described in detail. The theory of electron scattering from relativistic ions is developed and applied to the design and implementation of the system used to achieve and maintain the alignment. Commissioning with gold and 3He beams is then described as well as the successful utilization of the new system during the 2015 RHIC polarized proton run. Systematic errors of the new method are then estimated. Finally, some possible future applications of Coulomb-scattered electrons for beam diagnostics are briefly discussed.Comment: 16 pages, 23 figure