dataesr/scientific-paragraphs-categorization huggingface dataset:
Acknowledgments This research has been supported by  RecerCaixa foundation  ( 2010ACUP00378  to P. G. , J. G. , and A. L. ), the  Marató de TV3 Foundation  ( 110230  to J. G. , 110231 to A. L. , 110232 to C. G. , and  111531  to P. G. ), the  Catalan government  ( 2010 BP-A00194  to X. R. , 2012  CTP 00033  and  2012 BE1 00597  to X. G. -S. ,  2014SGR-1251  to P. G. , and  2014SGR-00109  to A. L. ), the  Spanish Government  ( CTQ2014-57020-R  and  PCIN-2013-017-C03-01  to A. L. , and  SAF2014-58396-R  and  PCIN-2013-018-C03-02  to J. G. ), and the  ERANET Neuron LIGHTPAINProject  (to A. L. , J. G. , and J. -P. P. );  SynBio MODULIGHTOR ,  Human Brain Project WAVESCALES  and  Ramón Areces  foundation grants (to P. G. ); the  Agence Nationale de la Recherche  ( ANR-16-CE16-0010  to A. L. and C.
Protocol Experimental data were obtained as follows. Rats were placed in a closed chamber of 10 L volume, fitted with a viewing window. The effects on the rats' body temperature of a 3-hour treatment with Xe-O 2 or He-O 2 gas mixtures containing 25% O 2 at various gas inhalation temperatures were investigated (n = 3-4 per group). The gas flow rate was 6 L/min, a condition that allows maintaining carbon dioxide at a concentration below 0. 03%. 16 The temperature at which the gas mixtures were administered was controlled using a heating/cooling system and a temperature probe placed into the closed chamber, as detailed previously. 16 Immediately before and after the 3-hour period of exposure to the above mentioned gas mixtures, the rats' body temperature was measured.
The respiratory activity was recorded by means of a mechanotransducer placed around the thorax, and the electrocardiogram (ECG) by subcutaneous electrodes placed on each side of the thorax. Moreover, the electrocardiogram and swallowing EMG signals were fed to loud speakers for auditory monitoring. Rectal temperature was monitored and maintained around 37 • C with a heating pad. The EMG, ECG and respiration signals were recorded on a computer using an analog-to-digital interface (PowerLab 8SP data acquisition software for Windows, AD Instruments, USA). A stable control sequence involving three 30-s trains of stimulations was performed before ODN injection. The mean values obtained during this sequence were used as control values. Afterwards, stimulations and recordings were maintained until recovery.
STATISTICAL ANALYSIS Comparisons between groups in Figure  6  were performed with repeated ANOVA with Tukey's HSD (Honestly Significant Difference) test used for post-hoc analysis. P-values <0. 05 were considered significant. Comparisons between groups in Figure  7  were carried out with unpaired 2-tailed Student's t-test. Pvalues <0. 05 were considered significant. S. E. M values were derived from at three independent experiments. For swallowing experiment (Figure  8 ), statistical analyses were performed using one way analysis of variance (ANOVA) followed by Fisher's protected least-significant difference post-hoc test. Differences were considered significant when P < 0. 05. Data were expressed as mean ± SEM. StatView for Windows 5. 0. 1; SAS Institute was used for statistical analysis.
ACKNOWLEDGMENTS This work was supported by funding obtained from the  Aix-Marseille University , the  Institut National de la Santé et de la Recherche Médicale (INSERM) , the  Agence Nationale de la Recherche  grant  EZICROM  ( ANR-16-CE14  to JL and JT). We thank the  Aix-Marseille University Microscopy Center CP2M  for access to their confocal microscopy equipment.
Mice were analyzed at least two weeks after tamoxifen injections. Electrophysiology in acute slices. Coronal hippocampal slices were prepared from young adult mice (36-56 day old). Voltage-clamp and current-clamp recordings were obtained from CA1 pyramidal neurons and hippocampal astrocytes. Details on slice preparation, recording conditions and data analysis are provided in Supplementary Information. Imaging in acute slices. Astrocytes were loaded with Calcium indicator Rhod-2 AM (9 µM) at room temperature (24 °C) for 1 h with 0. 02% Pluronic F-127 (In vitrogen) and 0. 6% DMSO (Sigma) in aCSF. Slices were then incubated at least 30 mins in aCSF to wash out the excess of dye before transferring to the imaging chamber. During experiments, slices were perfused with aCSF containing TTX (0. 5 µM) and GBZ (10 µM).
In control experiments performed on wild type mice, we verified that light stimulations of similar intensity and duration that the ones used to activate ChR2 in Cx30-CreERT2:floxed-ChR2-YFP mice did not induce changes in the holding current recorded in CA1 pyramidal neurons (Supplementary Fig. 1e ). Immunohistochemistry. The expression of the ChR2-EYFP transgene in Cx30-CreERT2:ChR2-EYFP was examined by immunolabeling against EYFP and against specific markers of astrocytes (GS, GFAP) and neurons (NeuN). Details are provided in Supplementary Information. Statistics. Paired and interleaved experiments were performed; no sample size calculation, no randomization or blinding were performed. No mice were excluded from analysis. Data were analyzed and plotted using SigmaPlot and GraphPad InStat.
Acknowledgements We thank  Serge Charpak  for the access to the two-photon microscope and for fruitful discussions. We thank  Ravi Rungta  and  Serge Charpak  for critical reading of an earlier version of the manuscript. We thank  Nicole Ropert ,  Martin Oheim  and  Elke Schmidt  for discussions and help at initial steps of this project,  Yannick Goulam  for his help with two-photon imaging,  Manon Omnes  and  Amelyne David  for maintaining the mouse colonies and genotyping the animals. This work was supported by grants from the  Fondation pour la Recherche Médicale  (FRM:  DEQ. 20140329488 ), the  European Union  ( ERA-NET Neuron BrIE ), the  Université Sorbonne Paris Cité.
Cell transfections and RT-PCR analysis PrSC were transfected overnight with 25 nM of control siRNA (targeting Luciferase mRNA) (Eurogentec, Belgium) or a siRNA raised against TRPA1 mRNA. (siTRPA1-1, target sequence 5′-GGUGGGAUGUUAUUCCAUA(dTdT)-3′ or siTRPA1-2: 5′-GAAGGACGCUCUCCACUUA(dTdT)-3′) as previously described  (32)  and used 48h posttransfection. The efficiency of the siRNAs was validated by RT-PCR and by western blot studies (shown for siTRPA1-1 in Fig. 2D ). RT-PCR experiments and HEK293 stably transfected by either hTRPA1 coding sequence (HEK-hTRPA1) or by pcDNA3 vector (HEKvector) were realized as described earlier  (32). The PCR primers (hTRPA1: 5'-AGTGGCAATGTGGAGCAA-3' and 5'-TCTGATCCACTTTGCGTA -3'; β-actin : 5'-CAGAGCAAGAGAGGCATCCT-3' and 5'-GTTGAAGGTCTCAAACATGATC-3' used in this study were designed on the basis of established GenBank sequences and synthesized by Invitrogen (Carlsbad, CA, USA).
Briefly, cells were seeded in 96-well plate (10 4 cells/well), starved overnight in serum-free RPMI medium and incubated 3 hours at 37 °C in 100 µL/well of serum-free RMPI medium containing or not increasing concentrations of rhCG and rhLH. Then 10 µl of culture supernatants were transferred into 384-well and mixed with 10 µl of a mixture of HTRF mix containing an anti-testosterone antibody and testosterone labeled with Terbium and d2 fluorophores, respectively. TR-FRET signals were detected using Mithras LB 943 plate reader (Berthold Technologies GmbH & Co. Wildbad, Germany). cAMP HTRF-based assay. Intracellular cAMP levels were measured using a homogeneous time-resolved fluorescence (HTRF ® ) assay kit (CisBio Bioassays) 10, 11 according to the manufacturer's instructions. Data analysis.
BRET data were represented as "Induced BRET Changes" by subtracting the 540 nm/480 nm ratio of the non-treated cells from the same ratio of cells stimulated with increasing doses of rhCG and rhLH. The % of responses in BRET and steroid measurement were obtained by taking as 100% the maximal response of rhCG at 10 nM in the different assays. All the results were fitted following the appropriate nonlinear regression equations using GraphPad Prism software (San Diego, CA, USA). Statistical analyses were performed with unpaired t-test (for the E max and EC 50 values) in HEK293 and mLTC-1 cells and Two-way ANOVA (for siRNA β-arrestin data in mLTC-1 cells), both using the GraphPad Prism software.
1C ). Specifically, the EC 50 value of rhCG was approximately 6 times lower than that of rhLH (p = 0. 0067; n = 5) (Table  2 ). To further confirm these data and to exclude any artifact due to the transfection of CAMYEL sensor in mLTC-1 cells, we also measured cAMP levels in native cells using HTRF-based assay, which does not require any cell transfection. We found a similar pattern of rhCG-and rhLH-induced cAMP responses (Fig. 1D ). It is worth noting that consistent with HEK293 cell data, rhCG and rhLH promoted similar maximal cAMP responses in mLTC-1 cells despite the differences in their potencies confirming that both are full agonists for cAMP pathway (Fig. 1C  and D ) (Table  2 ). β-arrestin 2 recruitment and activation. β-arrestins are known to play important roles not only in desensitization/internalization of GPCRs but also in their signaling  [12] [13] [14].
4A,C  and E ) and rhLH (Fig. 4B,D  and F ). Progesterone and testosterone were measured in parallel in the same cells. Our data confirm the partial progesterone response mediated by rhLH compared to rhCG as shown in Fig. 3B. However, on testosterone production there was no difference in hormone efficacy but rhCG was more potent that rhLH confirming the data shown in Fig. 3D. Interestingly, we observed that the depletion of both β-arrestin 1 and β-arrestin 2 led to a partial decrease in the progesterone production induced by rhCG (Fig. 4A ) and rhLH (Fig. 4B ) compared to control siRNA-transfected mLTC-1 cells. The statistical analysis demonstrates the significance of β-arrestin depletion for both rhCG (p < 0. 0001 for siRNA β-arrestin 1 and p = 0. 0233 for siRNA β-arrestin 2; n = 3) and rhLH (p = 0.
Data collection Magnetic fields were recorded on a whole-head MEG system with 151 axial gradiometers  (Omega 151, CTF Systems, Port Coquitlam, B. C. , Canada). This system includes seventeen external reference gradiometers and magnetometers that are used to apply a synthetic third-gradient to all MEG signals for ambient field correction. Three small coils were attached to reference landmarks on the participant (left and right preauricular points, plus nasion) in order to check head position at the beginning of each block. For all participants but one, electrical activity was recorded simultaneously with an amagnetic 64 Ag/AgCl unipolar electrode cap and processed by the MEG system. Electrode placement followed the extended International 10-20 system, including a row of low temporo-occipital electrodes (TP9-TP10, P9-P10, O9-O10, Iz).
One male participant's head was too big to fit in the MEG system with the electrode cap on, so EEG was recorded using only 5 pasted electrodes placed at Cz, TP9, P9, TP10 and P10. Therefore this participant was included only in the ERP and ERF peak analyses, but not in the follow-up GFP analysis and source localization and Deviated head views (mean number of trials averaged ± SEM= 72. 9±6. 0 per condition). These data were filtered with a low-pass filter set at 30Hz. The overall means of the ERF and of the ERP across participants (for the 12 participants with complete electrode coverage) were also calculated.
ERP/ERF analysis. We first performed classical measurements of the parameters of early electromagnetic components. We measured the peak amplitude and latency of the P100 (EEG), the M100 (MEG), the N170 (EEG), and the M170 (MEG), using customized Matlab (The MathWorks, Inc. ) scripts. The peak amplitude and latency of the P1 visual evoked potential were measured between 80 and 140 ms on the electrode where the P1 was maximum in the left and right posterior parieto-occipital regions (P9/10, P7/8, P5/6, PO7/8, PO3/4, TP9/10, O9/10, and O1/2, except for the male participant with 5 EEG electrodes only where the P100 could be measured solely on P9/10 and TP9/10), for each participant and experimental condition. Similarly, the peak amplitude and latency of the M100 magnetic component were These amplitude and latency measures were analyzed using ANOVAs with GAZE DIRECTION, HEAD ORIENTATION, and HEMISPHERE as within-subjects factors.
The analysis was performed using the STEN toolbox developed by Jean-François Knebel (http://www. unil. ch/fenl/home/menuinst/infrastructure/software-analysis-tools. html). In order to compensate for multiple comparisons across time points, we used a temporal stability criterion with a threshold of p <. 05 for a minimal duration of at least 10 ms to assess the time intervals of statistically significant effects or interaction. Follow-up planned comparisons using two-tailed Student t-test were performed when significant interactions were found. In addition, non-parametric Wilcoxon signed-rank tests were performed to confirm the significant effects found with parametric statistical tests.
Source localization Source localization was performed with Brainstorm software  (Tadel, Baillet, Mosher, Pantazis, & Leahy, 2011) , which is documented and freely available for download online under the GNU general public license (http://neuroimage. usc. edu/brainstorm). Cortical current source density mapping was obtained using a distributed source model consisting in ~15,000 current dipoles in each participant and condition. Dipole locations were constrained to the cortical mantle of a generic brain model built from the standard brain of the Montreal Neurological Institute using the BrainSuite software package (http://neuroimage. usc. edu). Dipole orientations were unconstrained. We then used the following forward models: Overlapping spheres for MEG  (Huang, Mosher, & Leahy, 1999)  and three-shell sphere (taking into account the different electrical conductivities of brain tissues-i.
Acknowledgement NB was supported by the  Swiss National Foundation Early Postdoc. Mobility fellowship project,  P2GEP1_151945. NG received funding from the  French Agence Nationale de la Recherche (ANR)  under grant number  ANR 12 SAMA 011 04  (" SENSO ") and from the  "Investissements d'avenir" programmes  under grant numbers  ANR-10-IAIHU-06  and  ANR-11-INBS-0006.
Confocal Microscopy Coronal free-floating sections (30 μm-thick) were obtained from paraformaldehyde-perfused mouse brains. Samples were incubated with antibodies against NeuN (1:500; Chemicon #MAB377), DARPP-32 (1:1000; BD #611520), D 1 R (1:500; Frontier Science #af500), D 2 R (1:500; Frontier Science #af750), S100β (1:500; Abcam #ab868) or GFAP-Cy3 (1:1000; Sigma #C 9205), followed by staining with the corresponding Alexa Fluor 488, 594 or 647 antibodies as appropriate (1:1000; Life Technologies). Nuclei were visualized with DAPI. Analysis of marker-protein immunoreactivity in the dorsal striatum was conducted as described  (Bellocchio et al. 2016 ) in a 1-in-10 series per animal (from bregma +1. 5 to -0. 5 coronal coordinates). For DARPP-32, D 1 R and D 2 R, data were calculated as immunoreactive area per total cell nuclei, and expressed as percentage of the control.
For NeuN, as well as for tdTomato and EGFP fluorescence in Drd1a-tdTomato/Drd2-EGFP mice, data were calculated as number of positive cells per total cell nuclei, and expressed as percentage of the control. Confocal fluorescence images were acquired using TCS-SP2 software and a SP2 AOBS microscope (Leica). Images were analyzed with ImageJ software (NIH).
2012). (  2 ) We  (Blazquez et al. 2011)  and others  (Mievis et al. 2011 ) have previously demonstrated a neuroprotective role of CB 1 R in transgenic mouse models of HD. measured in CB1R floxed/floxed mice, so for clarity only data from Glu-CB1R -/-animals are shown. C, Scheme of the experiment. D, DARPP-32, D1R and D2R immunoreactivity in the dorsal striatum, as well as RotaRod performance (time to fall), in Glu-CB1R -/-mice. Representative images are shown. Scale bar, 50 μm. Data of neuronal markers are expressed as relative values of the wtHtt-vehicle group. E, After termination of chronic pharmacological treatments and RotaRod assays, ambulation was determined in Glu-CB 1 R -/-mice 30 min after a single injection of vehicle or SKF-81297 (1 mg/kg, i. p. ). Data are expressed as SKF-81297-induced ambulation over that of vehicle, in cm.
Epifluorescence Images were obtained using an Olympus motorized inverted system microscope IX81, an X-Cite fluorescence illuminator, and an XM10 camera. Pictures were taken at 10x, 20x(air), and/or 40x(oil) magnifications through the three exposure channels for Alexa Fluor 488, Cy3/Alexa Fluor 555, and DAPI. Images were then processed using software CellSense (Olympus, Leinfelden-Echterdingen, Germany), and corrected for contrast and brightness using ImageJ v2. 0. 0  (Schneider et al. , 2012).
Confocal Microscopy Images were generated using a FluoView FV1000 confocal microscope (Olympus) with 20X UPlanSAPO, NA 0. 75 (air) and 40X UPlanFLN, NA 1. 30(oil) objectives. Stack images were taken by laser illumination at 561 nm (Alexa Fluor 555), 488 nm (Alexa Fluor 488), and 405 nm (DAPI). 12-bit raw images were processed with the imaging software Fluoview, version 4. 1. a (Olympus).
Structured Illumination Microscopy (SIM) Images were captured with a commercial inverted SIM microscope (Zeiss ELYRA, Oberkochen, Germany) using an oil-immersion objective (Plan-Apochromat 63 × /1. 4 Oil Dic M27)  (Gustafsson, 2000; Wegel et al. , 2016). Excitation of the fluorophores was performed by laser illumination at 642 nm (Alexa Fluor 647), 561 nm (Alexa Fluor 555), 488 nm (Alexa Fluor 488), and 405 nm (DAPI) and fluorescence light was filtered by appropriate detection filters: LP 655 (Alexa Fluor 647), BP 570-620 + LP 750 (Alexa Fluor 555), BP 495-550 + LP 750 (Alexa Fluor 488), and BP 420-480 + LP 750 (DAPI). Images were recorded with five rotations and five phase steps of the illumination pattern. Recorded data were processed with the ZEN imaging software (Zeiss). They were processed under standard ELYRA settings of the manual mode, selecting the Raw Scale option to keep the original dynamic range and therefore ensure a reliable comparison of the actual sample and the control samples.
6×250 mm, 5 μm particle size) with linear gradient of acetonitrile in aqueous 0. 1% TFA. Peptide structure was confirmed by both 1 H-NMR and mass spectrometry techniques. 1 H-NMR spectra were obtained using a WM-500 spectrometer (Bruker, Germany). Mass-spectrometry data were obtained using an Ultrafex MALDI TOF/TOF (Bruker Daltonics, Germany) instrument.
Anesthetized rats in vivo Rats were anesthetized with 20% urethane (120 mg/kg body wt i. p. ) and artificially ventilated with a KTR-5 animal respirator (Hugo Sacks Electronik) with a volume of 2-3 ml at a rate of 70-75 breaths/min. Further preparation of animals was performed as described earlier  [27]. Arterial blood pressure was recorded with a pressure transducer (Statham p23Db, Oxnard, USA) using a polygraph Biograph-4 (St. Petersburg, Russia). The mean arterial pressure, HR and standard lead II ECG were recorded on a computer using a LabVIEW 7. 1 data acquisition system (National Instruments, USA)  [20]. In control, after 30-min stabilization of hemodynamic parameters (steady state), LAD coronary artery was occluded for 40 min to simulate regional ischemia; the duration of subsequent reperfusion was 1 h.
Determination of myocardial infarct size After staining with Evans Blue, the heart was excised and the LV was frozen. A frozen LV was transversely cut into 1. 5 mm thick slices which were incubated in 0. 1 M sodium phosphate buffer pH 7. 40, containing 1% 2,3,5-triphenyl-tetrazolium chloride (TTC, Sigma, USA) 10 min at 37°C. The slices were fixed in 10% formalin for 5 min. Then they were placed between two transparent glasses and captured on both sides using a scanner at 600 d. p. i. resolution; the saved images were analyzed by computerized planimetry using Imagecal software. The slices were then weighed for determination of LV weight. The AAR was expressed as a percentage of LV weight, myocardial infarction (MI) was expressed as a percentage of the AAR in each group  [20].
Statistical analysis Data are presented as means ± SEM. Results were analyzed by one-way ANOVA followed by Bonferroni multiple range test post-hoc analysis for calculation differences between more than two groups. Comparisons between two groups involved use of the Student's unpaired t-test. All statistical analyses were performed using SigmaPlot version 12 (Systat Software Inc, San Jose, CA). P < 0. 05 was defined as significant. The data and statistical analysis complied with the recommendations on experimental design and analysis in pharmacology  [23].
Locomotor activity rhythm monitoring Locomotor activity experiments were performed using commercial activity monitors (TriKinetics) placed in incubators equipped with standard white, fluorescent low-energy tubes. Young (10 day-old male flies) or older (30 day-old males) males were maintained 5-6 days under LD 12:12 (Light-Dark 12h:12h), and then switched to at least 5-6 days of constant darkness, all on 5% sucrose-agar medium at 25˚C. Data analysis was performed with the FaasX software, as described previously  [86]. For constant darkness experiments, analysis started the second day of constant darkness. Histograms represent the distribution of the activity through 24 h in 30 min bins, averaged for n flies over 4-5 cycles. All behavioral experiments were repeated 2-3 times to verify reproductibility.
Sleep monitoring 3-4 days old male flies were placed individually in Trikinetics glass tubes, video monitored under infra-red illumination during 3 days in LD, then placed for at least 5 days in LL. The images were processed using pySolo video software  [87]  to determine the distance travelled (in pixels) for each minute of the day. Sleep was defined as 5 min of more of immobility  [88]. At least 40 flies in 2-3 replicates were analyzed for each genotype.
Fluorescent microscopy Fly brains were mounted on slides using as antifade reagent either Prolong Gold (Life Technologies), for brain immunostaining, or Vectashield (Vector Laboratories), for ROS measurements. Brains were visualized and images acquired with a Nikon A1R confocal microscope. A minimum of 10-15 brains was scored over at least 2 trials. Laser, filter and gain settings remained constant within each experiment, and channels were scanned sequentially. Confocal Z-stacks were analyzed using ImageJ software: dopaminergic cells were counted for each cluster, and whole brain average intensity levels were measured for ROS detection.
Acknowledgments We thank  M. Wu  and  K. Koh  for fly strains,  F. Rouyer  for fly strains, the rabbit anti-PDF antiserum, the  FaasX program  and personal communications,  I. Rivals  for advice on statistical analysis,  K. Aghedar ,  P. Fremont  and  C. Couraud  for participating in some experiments during their student internships, and  H. Lucchi  and  A. Lombi  for technical help. We also thank  C. Helfrich ,  P. Menegazzi  and  D. Rieger  ( Biozentrum, Wu ¨rzburg University ) for helpful suggestions.
This work was supported by funding from  Fondation de France  ( 2012-00024435 ),  Rotary Club/Federation pour la Recherche sur le Cerveau ,  Labex MemoLife  ( ANR-10-LABX-54 MEMO LIFE ) to SB, from  Sanofi  (contract  LGCR-VA-2014-01 ) to AK for an undergraduate internship, by PhD fellowships from the  Universite Ṕierre et Marie Curie  ( ED3C ) to AV and  ARI  and.