Probing the Y2 Receptor on Transmembrane, Intra- and Extra-Cellular Sites for EPR Measurements

The function of G protein-coupled receptors is intrinsically linked to their conformational dynamics. In conjugation with site-directed spin labeling, electron paramagnetic resonance (EPR) spectroscopy provides powerful tools to study the highly dynamic conformational states of these proteins. Here, we explored positions for nitroxide spin labeling coupled to single cysteines, introduced at transmembrane, intra- and extra-cellular sites of the human neuropeptide Y2 receptor. Receptor mutants were functionally analyzed in cell culture system, expressed in Escherichia coli fermentation with yields of up to 10 mg of purified protein per liter expression medium and functionally reconstituted into a lipid bicelle environment. Successful spin labeling was confirmed by a fluorescence assay and continuous wave EPR measurements. EPR spectra revealed mobile and immobile populations, indicating multiple dynamic conformational states of the receptor. We found that the singly mutated positions by MTSL ((1-oxyl-2,2,5,5-tetramethyl-2,5-dihydro-1H-pyrrol-3-yl) methyl methanesulfonothioate) have a water exposed immobilized conformation as their main conformation, while in case of the IDSL (bis(1-oxyl-2,2,5,5-tetramethyl-3-imidazolin-4-yl) disulfide) labeled positions, the main conformation are mainly of hydrophobic nature. Further, double cysteine mutants were generated and examined for potential applications of distance measurements by double electron–electron resonance (DEER) pulsed EPR technique on the receptor

Reduced Acquisition Time [18F]GE-180 PET Scanning Protocol Replaces Gold-Standard Dynamic Acquisition in a Mouse Ischemic Stroke Model

Aim Understanding neuroinflammation after acute ischemic stroke is a crucial step on the way to an individualized post-stroke treatment. Microglia activation, an essential part of neuroinflammation, can be assessed using [ 18 F]GE-180 18 kDa translocator protein positron emission tomography (TSPO-PET). However, the commonly used 60–90 min post-injection (p.i.) time window was not yet proven to be suitable for post-stroke neuroinflammation assessment. In this study, we compare semi-quantitative estimates derived from late time frames to quantitative estimates calculated using a full 0–90 min dynamic scan in a mouse photothrombotic stroke (PT) model. Materials and Methods Six mice after PT and six sham mice were included in the study. For a half of the mice, we acquired four serial 0–90 min scans per mouse (analysis cohort) and calculated standardized uptake value ratios (SUVRs; cerebellar reference) for the PT volume of interest (VOI) in five late 10 min time frames as well as distribution volume ratios (DVRs) for the same VOI. We compared late static 10 min SUVRs and the 60–90 min time frame of the analysis cohort to the corresponding DVRs by linear fitting. The other half of the animals received a static 60–90 min scan and was used as a validation cohort. We extrapolated DVRs by using the static 60–90 min p.i. time window, which were compared to the DVRs of the analysis cohort. Results We found high linear correlations between SUVRs and DVRs in the analysis cohort for all studied 10 min time frames, while the fits of the 60–70, 70–80, and 80–90 min p.i. time frames were the ones closest to the line of identity. For the 60–90 min time window, we observed an excellent linear correlation between SUVR and DVR regardless of the phenotype (PT vs . sham). The extrapolated DVRs of the validation cohort were not significantly different from the DVRs of the analysis group. Conclusion Simplified quantification by a reference tissue ratio of the late 60–90 min p.i. [ 18 F]GE-180 PET image can replace full quantification of a dynamic scan for assessment of microglial activation in the mouse PT model

Reflexive Grundbildung bis zum Ende der Schulpflicht. Konzepte und Prozeduren im Fach

In diesem Band wird ein Ansatz vorgestellt, nach dem der Fächerkanon einer allgemeinbildenden Sekundarstufe I nicht als unverbundene Addition historisch gewachsener, kontingenter und austauschbarer Fächer gesehen werden soll, sondern als sinnvoller Denk- und Handlungszusammenhang. Der fachliche Kern der Fächer, bestehend aus Konzepten und Prozeduren, wird dabei in den Mittelpunkt gerückt. Jedes Fach kann einen unersetzbaren Beitrag zu reflexiver Grundbildung im Sinne eines tiefenstrukturell angelegten Orientierungswissens liefern. Dies wird an vielen Beispielen aufgezeigt. Die Beziehungen der Fächer zueinander ergeben ein Fächerstrukturkonzept. Dieses soll dazu dienen, die angestrebte fachspezifische Kompetenzentwicklung der Schülerinnen und Schüler im Kontext einer Schul- und Unterrichtsentwicklung im Fächerdialog ebenso zu unterstützen wie den Aufbau von nachhaltigen fachlichen Wissensstrukturen. (DIPF/Orig.

Pain measurement in rheumatic and musculoskeletal diseases: where to go from here? Report from a Special Interest Group at OMERACT 2018

Objective: Establishing a research agenda on standardizing pain measurement in clinical trials in rheumatic and musculoskeletal diseases (RMDs). Methods: Discussion during a meeting at OMERACT 2018, prepared by a systematic review of existing core outcome sets and a patient online survey. Results: Several key questions were debated: is pain a symptom or a disease? are pain core (sub)domains consistent across RMDs? how to account for pain mechanistic descriptors (e.g. central sensitization) in pain measurement? Conclusion: Characterizing and assessing the spectrum of pain experience across RMDs in a standardized fashion is the future objective of the OMERACT pain working group

Deutschland, Frankreich und Europa: Perspektiven

"Mit der Osterweiterung und der Baustelle des Konvents zur Zukunft Europas durchlebt die Europäische Union derzeit eine Phase des Wandels, die auch eine Prüfung darstellt. Frankreich und Deutschland müssen jetzt mehr denn je ihre Kräfte vereinen, damit dieser Wandel erfolgreich vollzogen werden kann, Europa seine inneren Konflikte überwindet und seine Rolle als globaler Akteur stärkt. In diesem Jahr feiern die beiden Länder den 40. Jahrestag der Unterzeichnung des Elysée-Vertrages. In den Monaten zuvor haben sie wieder wichtige Kompromisse geschlossen und gemeinsame Initiativen ins Leben gerufen, um in den wesentlichen europäischen Fragen voranzukommen. Auf diesem Weg sollten sie weiter vorangehen, um wieder die unverzichtbare, treibende Kraft eines auf 25 oder 27 Mitgliedstaaten erweiterten Europa zu werden. Vor diesem Hintergrund haben Ifri (Institut français des relations internationales) und DGAP, die seit vielen Jahren durch eine enge Zusammenarbeit miteinander verbunden sind, beschlossen, eine langfristig angelegte gemeinsame Analyse der Rolle des deutsch-französischen Tandems im erweiterten Europa in Gang zu bringen." (Autorenreferat