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Robinson, Alan D. ; Millard, Genevieve A. ; Danis, François ; Guirlet, Marielle ; Harris, Neil R. P. ; Lee, Adrian M. ; McIntyre, J. D. ; Pyle, John A. ; Arvelius, Johan ; Dagnesjo, Saga ; Kirkwood, Sheila ; Nilsson, Hampus ; Toohey, Darin W. ; Deshler, Terry ; Goutail, Florence ; Pommereau, Jean-Pierre ; Elkins, James William ; Moore, Fred L. ; Ray, Eric ; Schmidt, Ulrich ; Engel, Andreas ; Müller, Melanie
Balloon-borne measurements of CFC-11 (on flights of the DIRAC in situ gas chromatograph and the DESCARTES grab sampler), ClO and O3 were made during the 1999/2000 winter as part of the SOLVE-THESEO 2000 campaign. Here we present the CFC-11 data from nine flights and compare them first with data from other instruments which flew during the campaign and then with the vertical distributions calculated by the SLIMCAT 3-D CTM. We calculate ozone loss inside the Arctic vortex between late January and early March using the relation between CFC-11 and O3 measured on the flights, the peak ozone loss (1200 ppbv) occurs in the 440–470 K region in early March in reasonable agreement with other published empirical estimates. There is also a good agreement between ozone losses derived from three independent balloon tracer data sets used here. The magnitude and vertical distribution of the loss derived from the measurements is in good agreement with the loss calculated from SLIMCAT over Kiruna for the same days.
Palayret, Matthieu ; Armes, Helen ; Basu, Srinjan ; Watson, Adam T. ; Herbert, Alex ; Lando, David ; Etheridge, Thomas J. ; Endesfelder, Ulrike ; Heilemann, Mike ; Laue, Ernest ; Carr, Antony M. ; Klenerman, David ; Lee, Steven F.
Single-molecule super-resolution microscopy allows imaging of fluorescently-tagged proteins in live cells with a precision well below that of the diffraction limit. Here, we demonstrate 3D sectioning with single-molecule super-resolution microscopy by making use of the fitting information that is usually discarded to reject fluorophores that emit from above or below a virtual-'light-sheet', a thin volume centred on the focal plane of the microscope. We describe an easy-to-use routine (implemented as an open-source ImageJ plug-in) to quickly analyse a calibration sample to define and use such a virtual light-sheet. In addition, the plug-in is easily usable on almost any existing 2D super-resolution instrumentation. This optical sectioning of super-resolution images is achieved by applying well-characterised width and amplitude thresholds to diffraction-limited spots that can be used to tune the thickness of the virtual light-sheet. This allows qualitative and quantitative imaging improvements: by rejecting out-of-focus fluorophores, the super-resolution image gains contrast and local features may be revealed; by retaining only fluorophores close to the focal plane, virtual-'light-sheet' single-molecule localisation microscopy improves the probability that all emitting fluorophores will be detected, fitted and quantitatively evaluated.
Sandoval, Andres ; Stock, Reinhard ; Stelzer, Herbert ; Renfordt, Rainer Arno Ernst ; Harris, John W. ; Brannigan, James P. ; Geaga, Jorge V. ; Rosenberg, L. J. ; Schroeder, Lee S. ; Wolf, Kevin L.
Exclusive pi - and charged-particle production in collisions of Ar+KCl is studied at incident energies from 0.4 to 1.8 GeV/u. Complete disintegration of both nuclei is observed. The correlation between pi - and total charge multiplicity shows no islands of anomalous pion production. For constant numbers of proton participants the pi - multiplicity distributions are Poissons. For central collisions <n pi -> increases smoothly and to first order linearly with the c.m. energy. Disagreement with the firestreak model is found. Pacs numbers: 25.70.Hi, 24.10.Dp
Al-Shahi Salman, Rustam ; Frantzias, Joseph ; Lee, Robert J. ; Lyden, Patrick D. ; Battey, Thomas W. K. ; Ayres, Alison M. ; Goldstein, Joshua Norkin ; Mayer, Stephan A. ; Steiner, Thorsten ; Wang, Xia ; Arima, Hisatomi ; Hasegawa, Hitoshi ; Oishi, Makoto ; Godoy, Daniel Agustín ; Masotti, Luca ; Dowlatshahi, Dar ; Rodríguez-Luna, David ; Molina, Carlos ; Jang, Dong-Kyu ; Davalos, Antonio ; Castillo, José ; Yao, Xiaoying ; Claassen, Jan ; Volbers, Bastian ; Kazui, Seiji ; Okada, Yasushi ; Fujimoto, Shigeru ; Toyoda, Kazunori ; Li, Qi ; Khoury, Jane ; Delgado, Pilar ; Álvarez-Sabín, José ; Hernández-Guillamon, Mar ; Prats-Sánchez, Luis ; Cai, Chunyan ; Kate, Mahesh ; McCourt, Rebecca ; Venkatasubramanian, Chitra ; Diringer, Michael N. ; Ikeda, Yukio ; Worthmann, Hans ; Ziai, Wendy C. ; D’Esterre, Christopher D. ; Aviv, Richard I. ; Raab, Peter ; Murai, Yasuo ; Zazulia, Allyson R. ; Butcher, Kenneth ; Seyedsaadat, Seyed Mohammad ; Grotta, James C. ; Martí-Fàbregas, Joan ; Montaner, Joan ; Broderick, Joseph ; Yamamoto, Haruko ; Staykov, Dimitre ; Connolly, Edward Sander ; Selim, Magdy ; Leira, Rogelio ; Moon, Byung Hoo ; Demchuk, Andrew M. ; Di Napoli, Mario ; Fujii, Yukihiko ; Anderson, Craig S. ; Rosand, Jonathan
Background: Intracerebral haemorrhage growth is associated with poor clinical outcome and is a therapeutic target for improving outcome. We aimed to determine the absolute risk and predictors of intracerebral haemorrhage growth, develop and validate prediction models, and evaluate the added value of CT angiography.
Methods: In a systematic review of OVID MEDLINE—with additional hand-searching of relevant studies' bibliographies— from Jan 1, 1970, to Dec 31, 2015, we identified observational cohorts and randomised trials with repeat scanning protocols that included at least ten patients with acute intracerebral haemorrhage. We sought individual patient-level data from corresponding authors for patients aged 18 years or older with data available from brain imaging initially done 0·5–24 h and repeated fewer than 6 days after symptom onset, who had baseline intracerebral haemorrhage volume of less than 150 mL, and did not undergo acute treatment that might reduce intracerebral haemorrhage volume. We estimated the absolute risk and predictors of the primary outcome of intracerebral haemorrhage growth (defined as >6 mL increase in intracerebral haemorrhage volume on repeat imaging) using multivariable logistic regression models in development and validation cohorts in four subgroups of patients, using a hierarchical approach: patients not taking anticoagulant therapy at intracerebral haemorrhage onset (who constituted the largest subgroup), patients taking anticoagulant therapy at intracerebral haemorrhage onset, patients from cohorts that included at least some patients taking anticoagulant therapy at intracerebral haemorrhage onset, and patients for whom both information about anticoagulant therapy at intracerebral haemorrhage onset and spot sign on acute CT angiography were known.
Findings: Of 4191 studies identified, 77 were eligible for inclusion. Overall, 36 (47%) cohorts provided data on 5435 eligible patients. 5076 of these patients were not taking anticoagulant therapy at symptom onset (median age 67 years, IQR 56–76), of whom 1009 (20%) had intracerebral haemorrhage growth. Multivariable models of patients with data on antiplatelet therapy use, data on anticoagulant therapy use, and assessment of CT angiography spot sign at symptom onset showed that time from symptom onset to baseline imaging (odds ratio 0·50, 95% CI 0·36–0·70; p<0·0001), intracerebral haemorrhage volume on baseline imaging (7·18, 4·46–11·60; p<0·0001), antiplatelet use (1·68, 1·06–2·66; p=0·026), and anticoagulant use (3·48, 1·96–6·16; p<0·0001) were independent predictors of intracerebral haemorrhage growth (C-index 0·78, 95% CI 0·75–0·82). Addition of CT angiography spot sign (odds ratio 4·46, 95% CI 2·95–6·75; p<0·0001) to the model increased the C-index by 0·05 (95% CI 0·03–0·07).
Interpretation: In this large patient-level meta-analysis, models using four or five predictors had acceptable to good discrimination. These models could inform the location and frequency of observations on patients in clinical practice, explain treatment effects in prior randomised trials, and guide the design of future trials.
Funding: UK Medical Research Council and British Heart Foundation.
Gruber-Rouh, Tatjana ; Lee, Clara ; Bolck, Jan ; Naguib, Nagy Naguib Naeem ; Schulz, Boris ; Eichler, Katrin ; Aschenbach, Rene ; Wichmann, Julian ; Vogl, Thomas J. ; Zangos, Stephan
Objective: To investigate the accuracy, efficiency and radiation dose of a novel laser navigation system (LNS) compared to those of free-handed punctures on computed tomography (CT).
Materials and methods: Sixty punctures were performed using a phantom body to compare accuracy, timely effort, and radiation dose of the conventional free-handed procedure to those of the LNS-guided method. An additional 20 LNS-guided interventions were performed on another phantom to confirm accuracy. Ten patients subsequently underwent LNS-guided punctures.
Results: The phantom 1-LNS group showed a target point accuracy of 4.0 ± 2.7 mm (freehand, 6.3 ± 3.6 mm; p = 0.008), entrance point accuracy of 0.8 ± 0.6 mm (freehand, 6.1 ± 4.7 mm), needle angulation accuracy of 1.3 ± 0.9° (freehand, 3.4 ± 3.1°; p < 0.001), intervention time of 7.03 ± 5.18 minutes (freehand, 8.38 ± 4.09 minutes; p = 0.006), and 4.2 ± 3.6 CT images (freehand, 7.9 ± 5.1; p < 0.001). These results show significant improvement in 60 punctures compared to freehand. The phantom 2-LNS group showed a target point accuracy of 3.6 ± 2.5 mm, entrance point accuracy of 1.4 ± 2.0 mm, needle angulation accuracy of 1.0 ± 1.2°, intervention time of 1.44 ± 0.22 minutes, and 3.4 ± 1.7 CT images. The LNS group achieved target point accuracy of 5.0 ± 1.2 mm, entrance point accuracy of 2.0 ± 1.5 mm, needle angulation accuracy of 1.5 ± 0.3°, intervention time of 12.08 ± 3.07 minutes, and used 5.7 ± 1.6 CT-images for the first experience with patients.
Conclusion: Laser navigation system improved accuracy, duration of intervention, and radiation dose of CT-guided interventions.
Hitzel, Juliane ; Lee, Eunjee ; Zhang, Yi ; Bibli, Sofia Iris ; Li, Xiaogang ; Zukunft, Sven ; Pflüger, Beatrice ; Hu, Jiong ; Schürmann, Christoph ; Vasconez, Andrea Estefania ; Oo, James A. ; Kratzer, Adelheid ; Kumar, Sandeep ; Rezende Felipe, Flávia Figueiredo de ; Josipovic, Ivana ; Thomas, Dominique Jeanette ; Giral, Hector ; Schreiber, Yannick ; Geisslinger, Gerd ; Fork, Christian ; Yang, Xia ; Sigala, Fragiska ; Romanoski, Casey E. ; Kroll, Jens ; Jo, Hanjoong ; Landmesser, Ulf ; Lusis, Aldons J. ; Namgaladze, Dmitry ; Fleming, Ingrid ; Leisegang, Matthias ; Zhu, Jianhui ; Brandes, Ralf
Oxidized phospholipids (oxPAPC) induce endothelial dysfunction and atherosclerosis. Here we show that oxPAPC induce a gene network regulating serine-glycine metabolism with the mitochondrial methylenetetrahydrofolate dehydrogenase/cyclohydrolase (MTHFD2) as a causal regulator using integrative network modeling and Bayesian network analysis in human aortic endothelial cells. The cluster is activated in human plaque material and by atherogenic lipoproteins isolated from plasma of patients with coronary artery disease (CAD). Single nucleotide polymorphisms (SNPs) within the MTHFD2-controlled cluster associate with CAD. The MTHFD2-controlled cluster redirects metabolism to glycine synthesis to replenish purine nucleotides. Since endothelial cells secrete purines in response to oxPAPC, the MTHFD2-controlled response maintains endothelial ATP. Accordingly, MTHFD2-dependent glycine synthesis is a prerequisite for angiogenesis. Thus, we propose that endothelial cells undergo MTHFD2-mediated reprogramming toward serine-glycine and mitochondrial one-carbon metabolism to compensate for the loss of ATP in response to oxPAPC during atherosclerosis.
Robinson, Alan D. ; Millard, Genevieve A. ; Danis, François ; Guirlet, Marielle ; Harris, Neil R. P. ; Lee, Adrian M. ; McIntyre, J. D. ; Pyle, John A. ; Arvelius, Johan ; Dagnesjo, Saga ; Kirkwood, Sheila ; Nilsson, Hampus ; Toohey, Darin W. ; Deshler, Terry ; Goutail, Florence ; Pommereau, Jean-Pierre ; Elkins, James William ; Moore, Fred L. ; Ray, Eric ; Schmidt, Ulrich ; Engel, Andreas ; Müller, Melanie
Balloon-borne measurements of CFC11 (from the DIRAC in situ gas chromatograph and the DESCARTES grab sampler), ClO and O3 were made during the 1999/2000 Arctic winter as part of the SOLVE-THESEO 2000 campaign, based in Kiruna (Sweden). Here we present the CFC11 data from nine flights and compare them first with data from other instruments which flew during the campaign and then with the vertical distributions calculated by the SLIMCAT 3D CTM. We calculate ozone loss inside the Arctic vortex between late January and early March using the relation between CFC11 and O3 measured on the flights. The peak ozone loss (~1200ppbv) occurs in the 440-470K region in early March in reasonable agreement with other published empirical estimates. There is also a good agreement between ozone losses derived from three balloon tracer data sets used here. The magnitude and vertical distribution of the loss derived from the measurements is in good agreement with the loss calculated from SLIMCAT over Kiruna for the same days.
Bergs, Amelie ; Schultheis, Christian ; Fischer, Elisabeth ; Tsunoda, Satoshi P. ; Erbguth, Karen ; Husson, Steven J. ; Govorunova, Elena ; Spudich, John Lee ; Nagel, Georg ; Gottschalk, Alexander ; Liewald, Jana F.
In optogenetics, rhodopsins were established as light-driven tools to manipulate neuronal activity. However, during long-term photostimulation using channelrhodopsin (ChR), desensitization can reduce effects. Furthermore, requirement for continuous presence of the chromophore all-trans retinal (ATR) in model systems lacking sufficient endogenous concentrations limits its applicability. We tested known, and engineered and characterized new variants of de- and hyperpolarizing rhodopsins in Caenorhabditis elegans. ChR2 variants combined previously described point mutations that may synergize to enable prolonged stimulation. Following brief light pulses ChR2(C128S;H134R) induced muscle activation for minutes or even for hours (‘Quint’: ChR2(C128S;L132C;H134R;D156A;T159C)), thus featuring longer open state lifetime than previously described variants. Furthermore, stability after ATR removal was increased compared to the step-function opsin ChR2(C128S). The double mutants C128S;H134R and H134R;D156C enabled increased effects during repetitive stimulation. We also tested new hyperpolarizers (ACR1, ACR2, ACR1(C102A), ZipACR). Particularly ACR1 and ACR2 showed strong effects in behavioral assays and very large currents with fast kinetics. In sum, we introduce highly light-sensitive optogenetic tools, bypassing previous shortcomings, and thus constituting new tools that feature high effectiveness and fast kinetics, allowing better repetitive stimulation or investigating prolonged neuronal activity states in C. elegans and, possibly, other systems.
The G2A receptor (GPR132) contributes to oxaliplatin-induced mechanical pain hypersensitivity
(2017)
Hohmann, Stephan ; Angioni, Carlo ; Tunaru, Sorin ; Lee, Seungkyu ; Woolf, Clifford J. ; Offermanns, Stefan ; Geisslinger, Gerd ; Scholich, Klaus ; Sisignano, Marco
Chemotherapy-induced peripheral neuropathic pain (CIPN) is a common and severe debilitating side effect of many widely used cytostatics. However, there is no approved pharmacological treatment for CIPN available. Among other substances, oxaliplatin causes CIPN in up to 80% of treated patients. Here, we report the involvement of the G-protein coupled receptor G2A (GPR132) in oxaliplatin-induced neuropathic pain in mice. We found that mice deficient in the G2A-receptor show decreased mechanical hypersensitivity after oxaliplatin treatment. Lipid ligands of G2A were found in increased concentrations in the sciatic nerve and dorsal root ganglia of oxaliplatin treated mice. Calcium imaging and patch-clamp experiments show that G2A activation sensitizes the ligand-gated ion channel TRPV1 in sensory neurons via activation of PKC. Based on these findings, we conclude that targeting G2A may be a promising approach to reduce oxaliplatin-induced TRPV1-sensitization and the hyperexcitability of sensory neurons and thereby to reduce pain in patients treated with this chemotherapeutic agent.