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Single molecule observation in cells and tissue allows the analysis of physiological processes with molecular detail, but it still represents a major methodological challenge. Here we introduce a microscopic technique that combines light sheet optical sectioning microscopy and ultra sensitive high-speed imaging. By this approach it is possible to observe single fluorescent biomolecules in solution, living cells and even tissue with an unprecedented speed and signal-to-noise ratio deep within the sample. Thereby we could directly observe and track small and large tracer molecules in aqueous solution. Furthermore, we demonstrated the feasibility to visualize the dynamics of single tracer molecules and native messenger ribonucleoprotein particles (mRNPs) in salivary gland cell nuclei of Chironomus tentans larvae up to 200 microm within the specimen with an excellent signal quality. Thus single molecule light sheet based fluorescence microscopy allows analyzing molecular diffusion and interactions in complex biological systems.

Abstract: The half-life of the naturally occurring long-lived rare earth isotope 176Lu was determined by a combination of highly sophisticated experimental procedures in order to further improve the reliability and the precision of literature data. The amount of lutetium in the samples was determined by inductively coupled plasma optical emission spectrometry (ICP-OES) using a NIST reference standard. The isotopic ratio N(176Lu)/N(Lu) in the samples was measured by means of inductively coupled plasma high resolution mass spectrometry (ICP-HRMS). The activity divided by the mass of Lu was determined by applying liquid scintillation (LS) counting. The LS counting efficiency of the beta/gamma emitter 176Lu was determined with the CIEMAT/NIST efficiency tracing technique with low uncertainty. The influences of colour quenching and background effects are discussed in this paper. The half-life was found to be 3.640(35)×1010 y. The result is in good agreement with other evaluations and the relative standard uncertainty of 0.95% is among the lowest of previously published data. [Copyright &y& Elsevier]

Abstract: Calcium-41 plays an important role in the long-term evaluation of the safety of final repositories for nuclear waste and is used to study the fine-scale chronology of the formation of the Solar System. Both applications are hindered by insufficient precision and poor consistency of previous determinations of the half-life. This work reports a half-life for 41Ca of (9.94±0.15)×104 years, which was determined with a combination of methods, chosen to provide the best possible precision. The activity was measured by liquid scintillation counting (LSC) exploiting the triple-to-double coincidence ratio method (TDCR); the absolute isotopic composition was determined by thermal ionization mass spectrometry (TIMS) and isotope dilution. Enhanced precision and accuracy of the 41Ca half-life will allow the improvement of safety analyses for final deposit sites of nuclear waste and of dating first solids, and better constrain the stellar environment of the formation of the Solar System. [Copyright &y& Elsevier]

Abstract: Reliable knowledge of the 79Se half-life is crucial, e.g. for the safety assessments of final repositories for nuclear waste. However, a literature survey reveals an inconsistent picture, indicating remarkably difficult experimental conditions. This work reports a new value of 3.27(8)×105 a for the half-life of 79Se. The uncertainty is less than half of the latest published result. The sample was prepared radiochemically pure from a reprocessing solution by exploitation of reductive deposition on Cu, anion exchange chromatography and sublimation. The specific activity was determined by inductively coupled plasma optical emission spectrometry (ICP-OES) and liquid scintillation counting (LSC). Hydride generation multicollector inductively coupled plasma mass spectrometry (HG-MC-ICP-MS) was applied for the establishment of the isotopic composition. [Copyright &y& Elsevier]

Abstract: The half-life of the long-lived isotope 147Sm was determined by means of activity determination using liquid scintillation counting. The amount of samarium was determined by ICP-OES using a NIST reference standard. The isotopic ratio N(147Sm)/N(Sm) of the samples was measured by means of ICP-HRMS. The half-life was found to be 1.070(9)×1011 y. The result is in good agreement with other evaluations and the relative standard uncertainty of 0.8% is lower than in any previous work. [Copyright &y& Elsevier]

Cholangiocarcinoma is non-curable in many cases at establishing the diagnosis. New insights into epidemiology may promote screening of cholangiocarcinoma in a combination of risk scores, but today PSC is the only risk factor with an established screening recommendation., (© Georg Thieme Verlag KG Stuttgart · New York.)

Aim: To evaluate the safety and technical success of endoscopic radiofrequency ablation (RFA) for palliative treatment of malignant hilar bile duct obstruction., Methods: In this study, a recently CE and FDA-approved endoscopic RFA catheter was first tested in an ex vivo pig liver model to study the effect of electrosurgical variables on the extent of the area of induced necrosis. Subsequently, a retrospective analysis was conducted of all patients treated with endoscopic RFA for malignant biliary obstruction at our center between February 2012 and April 2013. All patients received an additional plastic stent implantation into the biliary tree following RFA., Results: In the pig model, ablation time of 60-90 seconds using the bipolar soft coagulation mode at 8-10 watts with an effect of 8 was found to be the most feasible setting. Twelve patients (5 females, 7 males; mean age, 70 years) underwent 19 endoscopic RFA (range, 1-5) sessions. Deployment of RFA was successful in all patients. Systemic chemotherapy was administered in four patients. We observed biliary bleeding 4-6 wk after the intervention in three cases and two of these patients died: in one patient, spontaneous hemobilia occurred, whereas bleeding started during stent extraction in the other. In the third patient, bleeding was stopped by insertion of a non-covered self-expanding metal stent. Another three patients developed cholangitis during follow-up. Seven patients died during follow-up and median survival was 6.4 mo (95%CI: 0.05-12.7) from the time of the first RFA., Conclusion: Endoscopic RFA is an easy to perform and technically highly successful procedure. However, hemobilia possibly associated with RFA occurred in three of our patients. Therefore, larger prospective studies are needed to further evaluate the safety and efficacy of this promising new method.

Aim: To compare clinical success and complications of uncovered self-expanding metal stents (SEMS) vs covered SEMS (cSEMS) in obstruction of the small bowel., Methods: Technical success, complications and outcome of endoscopic SEMS or cSEMS placement in tumor related obstruction of the duodenum or jejunum were retrospectively assessed. The primary end points were rates of stent migration and overgrowth. Secondary end points were the effect of concomitant biliary drainage on migration rate and overall survival. The data was analyzed according to the Strengthening the Reporting of Observational Studies in Epidemiology guidelines., Results: Thirty-two SEMS were implanted in 20 patients. In all patients, endoscopic stent implantation was successful. Stent migration was observed in 9 of 16 cSEMS (56%) in comparison to 0/16 SEMS (0%) implantations (P = 0.002). Stent overgrowth did not significantly differ between the two stent types (SEMS: 3/16, 19%; cSEMS: 2/16, 13%). One cSEMS dislodged and had to be recovered from the jejunum by way of laparotomy. Time until migration between SEMS and cSEMS in patients with and without concomitant biliary stents did not significantly differ (HR = 1.530, 95%CI 0.731-6.306; P = 0.556). The mean follow-up was 57 ± 71 d (range: 1-275 d)., Conclusion: SEMS and cSEMS placement is safe in small bowel tumor obstruction. However, cSEMS is accompanied with a high rate of migration in comparison to uncovered SEMS.

Saccharomyces cerevisiae reduces the beta-keto ester ethyl 2-chloroacetoacetate to the respective chiral cis- and trans-beta-hydroxy esters. In the course of chiral reduction, competing dehalogenation of the xenobiotic substrate to ethyl acetoacetate occurs, in a reaction mediated by cytosolic glutathione (GSH). Mechanistically, the dechlorination is a novel type of glutathione-dependent dehalogenation catalysed by an as yet unidentified glutathione-dependent dehalogenase. The first step consists of a nucleophilic replacement of the chloride substituent by glutathione. In the subsequent enzyme-catalysed step, a second glutathione molecule liberates the dehalogenation product by thiolytic attack at the thioether bridge, resulting in a net transfer of two electrons to the substrate and in the formation of glutathione disulfide (GSSG). Being effective under aerobic conditions and catalysed by a fungus, this reductive dechlorination of an aliphatic substrate is an outstanding example of a novel, glutathione-mediated microbial dehalogenation.