MERLIN: a novel BRET-based proximity biosensor for studying mitochondria–ER contact sites

The contacts between the ER and mitochondria play a key role in cellular functions such as the exchange of lipids and calcium between both organelles, as well as in apoptosis and autophagy signaling. The molecular architecture and spatiotemporal regulation of these distinct contact regions remain obscure and there is a need for new tools that enable tackling these questions. Here, we present a new bioluminescence resonance energy transfer-based biosensor for the quantitative analysis of distances between the ER and mitochondria that we call MERLIN (Mitochondria-ER Length Indicator Nanosensor). The main advantages of MERLIN compared with available alternatives are that it does not rely on the formation of artificial physical links between the two organelles, which could lead to artifacts, and that it allows to study contact site reversibility and dynamics. We show the applicability of MERLIN by characterizing the role of the mitochondrial dynamics machinery on the contacts of this organelle with the ER.We thank Peter McCormick for helpful advice and discussion and Carolin Stegmuller, Sabine Schafer, Iris Koch, Maria Zarani, Astrid Schauss, Christian Jungst, Felix Babatz, and Marina Nikolova for technical support. This work has been partially supported by the Deutsche Forschungsgemeinschaft (FOR2036 GA1641/2-1 and GA1641/2-2) and the European Research Council (StG 309966)

Wirbelschichtvergasung von Holz und Biomasse fuer Blockheizkraftwerke Abschlussbericht

A wood and biomass gasification process in a circulating fluidized bed with dry gas conditioning and catalytic cracking of tar was tested in a pilot plant with a capacity of about 400 kW. Calorific values of about 5,500 kJ/m"3 in the dry state were achieved for wood chip gasification with air. Average tar concentrations were in the order of 3000-5000 mg/Nm"3 for naphthalene and PAH, which could be reduced to less than 50 mg/Nm"3 with the aid of reformation catalysts. The product gas was combusted in a cogeneration plant. Continuous tests were carried out for up to 150 h, and the resulting electric power was fed into the public grid. Gross electric efficiencies were 32% as referred to wood gas and 27% as referred to primary fuel (biomass).Gegenstand des Vorhabens war die Fortsetzung der Erprobung eines Verfahrens zur Vergasung von Holz/Biomasse in der zirkulierenden Wirbelschicht mit trockener Gaskonditionierung und katalytischer Teerspaltung fuer die Nutzung des Gases in einem Gasmotor an einer Pilotanlage mit ca. 400 kW Feuerungswaermeleistung. Bei der Vergasung von Holzspaenen mit Luft wurden Heizwerte bis ca. 5.500 kJ/m"3 (i.N., tr.) erzielt. Durchschnittliche Teergehalte nach ZWS-Vergaser lagen in der Groessenordnung 3000-5000 mg/Nm"3 fuer Naphthalin + PAK. Diese konnten durch Einsatz von Reformierkatalysatoren im Dauerbetrieb auf <50 mg/Nm"3 gesenkt werden. Mit dem abgekuehlten und entstaubten Holzgas wurde ein Motoren-Blockheizkraftwerk (BHKW) betrieben. Es wurden unterbrechungsfreie Versuche von bis zu 150 h Dauer durchgefuehrt. Versuchszeitbegrenzend waren logistische Gruende. Der erzeute Strom wurde ins Netz eingespeist. Die erzielten elektrischen Bruttowirkungsgrade betrugen 32% bezogen auf Holzgas und 27% bezogen auf Primaerbrennstoff (Biomasse). (orig.)Available from TIB Hannover: F02B112+a / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekSIGLEBundesministerium fuer Ernaehrung, Landwirtschaft und Forsten, Berlin (Germany)DEGerman

Evaluation of the fatigue performance and degradability of resorbable PLDLLA-TMC osteofixations

The fatigue performance of explanted in-situ degraded osteofixations/osteosyntheses, fabricated from poly (70L-lactide-co-24DL-lactide-6-trimethylane-carbonate or PLDLLA-TMC) copolymer was compared to that of virgin products. The fatigue test was performed on 21 explants retrieved from 12 women and 6 men; 16-46 years by a custom-designed three-point bend apparatus using a staircase method and a specified failure criterion (an increase of the deflec-tion of the specimen > 1 mm) with run-out designated as “no failure” after 150,000 loading cycles. While all the virgin products showed run-out at 38N, all of the specimens fabricated from explants failed at this load level. For the explant specimens, although there was a trend of decreased failure load with increased in-situ time, this decrease was pronounced after 4 months in-situ, however, not yet statistically significant, while a 6-month in-situ explant had significantly less fail-ure load. Three and four month in-situ explants had highly significant differences in failure load between measurements close and distant to the osteotomy line: p=0.0017 (the region of maximum load in-situ). In the virgin products, there were only traces of melt joining and cooling, left from a stage in the manufacturing process. For the implants retrieved after 4.5 months in-situ, the fracture surfaces showed signs of degradation of the implants, possibly caused by hydrolysis, and for those retrieved after 9 months in-situ, there were cracks and pores. Thus, the morphological results are consistent with those obtained in the fatigue test. The present results suggest that resorbable osteofixations fabricated from PLDLLA-TMC are stable enough to allow loading of the healing bone and degrade reliably

Technische und wirtschaftliche Optimierung der Vergasung von Holz in der zirkulierenden Wirbelschicht Abschlussbericht

The project's objective was the optimization of a novel process for high efficient combined heat and power production from solid biomass. The processed air blown wood gasification in a pilot scale circulating fluidized bed was added by a catalytic tar reformer which would yield a tar-free gas quality suitable for IC-engine operation. Major efforts were taken for technical improvement of the tar reformer, especially concerning temperature control and cleaning devices which is important for keeping constantly a high activity. Pure natural timber did not yield chemical deactivation at the catalyst whereas the gasification of waste wood yielded decreasing activity which could be partly reversed by special measures taken. Further optimization of the process considered a better automation and improvement of the engine's flue gas emissions. Also a detailed economic consideration and evaluation of the entire process has been carried out. As a result the novel process should have economic advantages compared with conventional technology. (orig.)Gegenstand des Vorhabens war die Weiterentwicklung eines Verfahrens zur effizienteren Strom- und Waermegewinnung aus festen Biobrennstoffen. Durch luftgeblasene Vergasung von Holz im Pilotmassstab in einer zirkulierenden Wirbelschicht und anschliessender katalytischer Teerspaltung konnte ein niederkalorisches Brenngas erzeugt werden, welches zum Betrieb eines Motoren-Blockheizkraftwerks geeignet war. Im Rahmen der Verfahrensoptimierung wurde der katalytische Teer-Reformer, insbesondere in Bezug auf Temperaturfuehrung und die zum Aktivitaetserhalt wichtige Abreinigungsvorrichtung, verbessert. Bei der Vergasung von Naturholz wurde keine chemische Desaktivierung festgestellt. Beim Altholzeinsatz wurde ein ueberwiegend reversibler Aktivitaetsverlust verzeichnet und begruendet. Geeignete Gegenmassnahmen wurden untersucht und beschrieben. Weitere Optimierungen betrafen die Anlagensteuerungstechnik im Hinblick auf die Prozessautomatisierung und das Verbesserungspotenzial hinsichtlich Abgasemissionen am Motor. Darueber hinaus wurde eine detaillierte vergleichende betriebswirtschaftliche Bewertung des Verfahrens vorgenommen. Dabei stellte sich das neue technische Verfahren positiver als etablierte Techniken dar. (orig.)SIGLEAvailable from TIB Hannover: F03B777+a / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekBundesministerium fuer Verbraucherschutz, Ernaehrung und Landwirtschaft (BMVEL), Bonn (Germany); Fachagentur Nachwachsende Rohstoffe e.V., Guelzow (Germany)DEGerman