Monomeric Garnet, a far-red fluorescent protein for live-cell STED imaging

The advancement of far-red emitting variants of the green fluorescent protein (GFP) is crucially important for imaging live cells, tissues and organisms. Despite notable efforts, far-red marker proteins still need further optimization to match the performance of their green counterparts. Here we present mGarnet, a robust monomeric marker protein with far-red fluorescence peaking at 670 nm. Thanks to its large extinction coefficient of 95,000 M-1 cm-1, mGarnet can be efficiently excited with 640-nm light on the red edge of its 598-nm excitation band. A large Stokes shift allows essentially the entire fluorescence emission to be collected even with 640-nm excitation, counterbalancing the lower fluorescence quantum yield of mGarnet, 9.1%, that is typical of far-red FPs. We demonstrate an excellent performance as a live-cell fusion marker in STED microscopy, using 640 nm excitation and 780 nm depletion wavelengths

Global analysis of neutrino masses, mixings and phases: entering the era of leptonic CP violation searches

We perform a global analysis of neutrino oscillation data, including high-precision measurements of the neutrino mixing angle theta_13 at reactor experiments, which have confirmed previous indications in favor of theta_13>0. Recent data presented at the Neutrino 2012 Conference are also included. We focus on the correlations between theta_13 and the mixing angle theta_23, as well as between theta_13 and the neutrino CP-violation phase delta. We find interesting indications for theta_23< pi/4 and possible hints for delta ~ pi, with no significant difference between normal and inverted mass hierarchy.Comment: Updated version, including recent data released at the Neutrino 2012 Conference. Some references adde

The Waveform Digitiser of the Double Chooz Experiment: Performance and Quantisation Effects on PhotoMultiplier Tube Signals

We present the waveform digitiser used in the Double Chooz experiment. We describe the hardware and the custom-built firmware specifically developed for the experiment. The performance of the device is tested with regards to digitising low light level signals from photomultiplier tubes and measuring pulse charge. This highlights the role of quantisation effects and leads to some general recommendations on the design and use of waveform digitisers.Comment: 14 pages, 8 figures, accepted for publication in JINS

Superresolution microscopy reveals a dynamic picture of cell polarity maintenance during directional growth

Polar (directional) cell growth, a key cellular mechanism shared among a wide range of species, relies on targeted insertion of new material at specific locations of the plasma membrane. How these cell polarity sites are stably maintained during massive membrane insertion has remained elusive. Conventional live-cell optical microscopy fails to visualize polarity site formation in the crowded cell membrane environment because of its limited resolution. We have used advanced live-cell imaging techniques to directly observe the localization, assembly, and disassembly processes of cell polarity sites with high spatiotemporal resolution in a rapidly growing filamentous fungus, Aspergillus nidulans. We show that the membrane-associated polarity site marker TeaR is transported on microtubules along with secretory vesicles and forms a protein cluster at that point of the apical membrane where the plus end of the microtubule touches. There, a small patch of membrane is added through exocytosis, and the TeaR cluster gets quickly dispersed over the membrane. There is an incessant disassembly and reassembly of polarity sites at the growth zone, and each new polarity site locus is slightly offset from preceding ones. On the basis of our imaging results and computational modeling, we propose a transient polarity model that explains how cell polarity is stably maintained during highly active directional growth

Superresolution microscopy reveals a dynamic picture of cell polarity maintenance during directional growth

Polar (directional) cell growth, a key cellular mechanism shared among a wide range of species, relies on targeted insertion of new material at specific locations of the plasma membrane. How these cell polarity sites are stably maintained during massive membrane insertion has remained elusive. Conventional live-cell optical microscopy fails to visualize polarity site formation in the crowded cell membrane environment because of its limited resolution. We have used advanced live-cell imaging techniques to directly observe the localization, assembly, and disassembly processes of cell polarity sites with high spatiotemporal resolution in a rapidly growing filamentous fungus, Aspergillus nidulans. We show that the membrane-associated polarity site marker TeaR is transported on microtubules along with secretory vesicles and forms a protein cluster at that point of the apical membrane where the plus end of the microtubule touches. There, a small patch of membrane is added through exocytosis, and the TeaR cluster gets quickly dispersed over the membrane. There is an incessant disassembly and reassembly of polarity sites at the growth zone, and each new polarity site locus is slightly offset from preceding ones. On the basis of our imaging results and computational modeling, we propose a transient polarity model that explains how cell polarity is stably maintained during highly active directional growth

Effect of a fluorinated pyrimidine on cachexia and tumour growth in murine cachexia models: relationship with a proteolysis inducing factor

The fluorinated pyrimidine nucleoside, 5′-deoxy-5-fluorouridine (5′-dFUrd) has been shown to effectively attenuate the progress of cachexia in the murine adenocarcinomas MAC16 and colon 26 as well as in the human uterine cervical carcinoma xenograft, Yumoto. Although concomitant inhibition of tumour growth was observed in all three models this was not sufficient to account for the preservation of body weight. An attempt has been made to correlate the anti-cachectic activity of 5′-dFUrd with the presence of a tumour produced proteolysis-inducing factor (PIF), thought to be responsible for the development of cachexia in the MAC16 model. Two variants of colon 26 adenocarcinoma were employed, clone 20 which produces profound cachexia, and clone 5 which produces no change in body weight in recipient animals. Mice bearing the colon 26, clone 20 variant showed evidence for the presence of PIF in tumour, serum and urine, while there was no evidence for the presence of PIF in tumour or body fluids of mice bearing the clone 5 tumours. Treatment of animals bearing the clone 20 variant with 5′-dF Urd led to the disappearance of PIF from the tumour, serum and urine concomitant with the attenuation of the development of cachexia. The human cervical carcinoma, Yumoto, which also induced cachexia in recipiant animals, showed expression of PIF in tumour, serum and urine in control and vehicle-treated mice, but was absent in mice treated with 5′-dFUrd. Thus in these experimental models cachexia appears to be correlated with the presence of PIF. © 2000 Cancer Research Campaig

Lake Morphometry and River Network Controls on Evasion of Terrestrially Sourced Headwater CO₂

Lakes are central components of the inland water system distinct from, yet inextricably connected to, river networks. Currently, existing network-scale biogeochemistry research, although robust, typically treats each of these components separately or reductively. Here, we incorporate lake morphometry into a fully connected stream/lake network for the Connecticut River watershed and model potential evasion of terrestrially sourced headwater CO2 as transported through the network, ignoring in-stream production. We found that approximately 25%–30% of total potential soil CO2 evasion occurs in lakes, and percent evasion is inversely related to streamflow. A lake's ability to evade CO2 is controlled by residence time and size: most lakes with residence time over 7 days or surface area greater than 0.004 km2 evade functionally all terrestrial CO2 entering from upstream, precluding further downstream transport. We conclude that lakes are important for soil CO2 degassing and that this coupled river/lake approach is promising for CO2 studies henceforth