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The lack of selectivity of anticancer drugs limits current chemotherapy. Light-activatable drugs, whose activity can be precisely controlled with external light, could provide a more localized action of the drugs in the tumor, thus decreasing side effects and increasing efficacy. Herein, we introduce a series of photoswitchable azobenzene histone deacetylase inhibitors (HDACis) whose activity can be controlled by external visible light. Initial HDACis isomerized under ultraviolet light and were up to >50-fold more active under illumination than in the dark in enzyme assays. These were then optimized toward compounds responding to more permeable and less harmful green light by introducing o-halogen atoms into the azobenzene. Selected compounds decreased cell viability only under illumination in four different cancer cell lines. Overall, we present photoswitchable HDACis with optimized activation wavelengths, which inhibit enzyme activity and cell viability only upon illumination with visible light, contributing to the still limited toolbox of photoswitchable anticancer drugs., L.J.-C. has received funding from the European Union’s Horizon2020 research and innovation programme under Marie Sklodowska-Curie Grant Agreement 841089. A.L. received funding from Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación 10.13039/501100011033 and ERDF A way of making Europe (Projects I+D+i CTQ2017-89222-R and PID2020-120499RB-I00), and the Catalan government (2017 SGR 1604). The authors thank Dr. Lourdes Muñoz from SimChem (IQAC-CSIC) for analytical and instrumental support. The authors thank Dr. Wiktor Szymanski (University of Groningen, Groningen, The Netherlands) for helpful discussions.

The total synthesis of the natural (8E,15E,22E)-heptatriaconta-8,15,22-trien-2-one and (15E,22E)-heptatriaconta-15,22-dien-2-one is reported. Our synthesis involved a modular and convergent approach, based on the copper-catalyzed addition of Grignard reagents to allylic olefins. The obtention of these methyl ketones is an important asset for the use of these compounds as analytical patterns with application in Earth sciences., We gratefully acknowledge the Geochemistry and Pollution Research Group from the Institute of Environmental Assessment and Water Research (IDAEA-CSIC) and CSIC (201980E011) for financial support.

G-protein-coupled receptors (GPCRs) are a super-family of membrane proteins and major targets for drug development. However, many GPCRs drug candidates suffer from a lack of selectivity. Photopharmacology gives the possibility of modulating GPCR activity with an unprecedented local and temporal precision with the use of light. In this review, we compile and classify the different strategies in photopharmacology for GPCRs, we revise the different methods of analysis and characterization of light-regulated molecules used for GPCRs, and we give perspective of the impact of photopharmacology in research applications and in the development of new drugs.

Artificial control of neuronal activity enables the study of neural circuits and restoration of neural functions. Direct, rapid, and sustained photocontrol of intact neurons could overcome the limitations of established electrical stimulation such as poor selectivity. We have developed fast photoswitchable ligands of glutamate receptors (GluRs) to enable neuronal control in the auditory system. The new photoswitchable ligands induced photocurrents in untransfected neurons upon covalently tethering to endogenous GluRs and activating them reversibly with visible light pulses of a few milliseconds. As a proof of concept of these molecular prostheses, we applied them to the ultrafast synapses of auditory neurons of the cochlea that encode sound and provide auditory input to the brain. This drug-based method afforded the optical stimulation of auditory neurons of adult gerbils at hundreds of hertz without genetic manipulation that would be required for their optogenetic control. This indicates that the new photoswitchable ligands are also applicable to the spatiotemporal control of fast spiking interneurons in the brain., This research received funding from the European Union Research and Innovation Programme Horizon 2020─Human Brain Project SG3 (945539), DEEPER (ICT-36-2020–101016787), Agency for Management of University and Research Grants/Generalitat de Catalunya (CERCA Programme; 2017-SGR-1442 and 2017-SGR-1604 projects), Fonds Européen de Développement Économique et Régional (FEDER) funds, Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación and ERDF-FEDER European Fund (projects CTQ2017-89222-R and PID2020-120499RB-I00), and Fundaluce and “la Caixa” foundations (ID 100010434, agreement LCF/PR/HR19/52160010). The project Clúster Emergent del Cervell Humà (CECH, 001-P-001682) is cofinanced by the European Union Regional Development Fund within the framework of the ERDF Operational Program of Catalonia 2014–2020 with a grant of 50% of total eligible cost. A.G.-C. was supported by the fellowship BES-2014-068169 and Alexander von Humboldt Foundation. J.H. was supported by Agencia Estatal de Investigación (PID2019-106171RB-100/AEI/10.13039/501100011033) and Generalitat de Catalunya (2017 SGR00465). We thank chemical facility PCB staff (Unai Elezcano, Sònia Varon) and IBEC infrastructure staff (Isabel Oliveira, David Izquierdo). We thank Christiane Senger-Freitag, Sandra Gerke, and Sina Langer for technical support, Gerhard Hoch for his expert technical support, and Patricia Räke-Kügler for excellent administrative support. We thank Thomas Mager for providing the patch clamp setup. This work was funded by the European Research Council through the Advanced Grant “OptoHear” to T.M. under the European Union’s Horizon 2020 Research and Innovation program (grant agreement no. 670759), the Fraunhofer and Max-Planck Cooperation Program (NeurOpto grant) to T.M., was further supported by the German Research Foundation through the Priority Program 1926 “Next generation optogenetics” to A.H. and T.M., and the Leibniz Program (to T.M.) and the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy─EXC 2067/1-390729940 to A.H. and T.M. In addition, this research was supported by the Fakultätinternes Forschungförderungsprogramm from the Universitätsmedizin Göttingen, Georg-August-Universität Göttingen to A.H., and Fondation Pour l’Audition (FPA RD-2020-10) to T.M. Molecular graphics and analyses were performed with UCSF Chimera, developed by the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco, with support from NIH P41-GM103311. We thank Dr. Vladimir Belov, Jurgen Bienert, and Jan Seikowski for HPLC and LC–MS measurements of final TCPfast products.

Acute myeloid leukemia (AML) is a heterogeneous hematological cancer characterized by poor prognosis and frequent relapses. Aside from specific mutation-related changes, in AML, the overall function of lysosomes and mitochondria is drastically altered to fulfill the elevated biomass and bioenergetic demands. On the basis of previous results, in silico drug discovery screening was used to identify a new family of lysosome-/mitochondria-targeting compounds. These novel tetracyclic hits, with a cationic amphiphilic structure, specifically eradicate leukemic cells by inducing both mitochondrial damage and apoptosis, and simultaneous lysosomal membrane leakiness. Lysosomal leakiness does not only elicit canonical lysosome-dependent cell death, but also activates the terminal differentiation of AML cells through the Ca2+-TFEB-MYC signaling axis. In addition to being an effective monotherapy, its combination with the chemotherapeutic arsenic trioxide (ATO) used in other types of leukemia is highly synergistic in AML cells, widening the therapeutic window of the treatment. Moreover, the compounds are effective in a wide panel of cancer cell lines and possess adequate pharmacological properties rendering them promising drug candidates for the treatment of AML and other neoplasias., This work was funded by AGAUR-Producte (R.M.R), Caixaimpulse Program (R.M.R), the Josep Carreras International Leukaemia Foundation (R.M.R), l’Obra Social “La Caixa”-Fundació Bancària “La Caixa” (R.M.R), CERCA Programme/Generalitat de Catalunya (IJC) and Fundació Marató TV3 (grant number: 201930-30)

Treballs Finals de Grau de Matemàtiques, Facultat de Matemàtiques, Universitat de Barcelona, Any: 2023, Director: Rosa M. Miró-Roig, [en] The main goal of this writing is to introduce basic concepts of algebraic geometry and commutative algebra to be able to study the relationship between Perazzo hypersurfaces and the Lefschetz properties. We will introduce graded algebras and how to construct one of them from a hypersurface to further check if they satisfy any of the properties. In this paperwork, we have found an upper bound and a low enough value of the Hilbert vector for Perazzo hypersurfaces. The notable result we have obtained is that the Weak Lefschetz property is failed to obtain when the h-vector is maximal, and conversely, it is always obtained when the h-vector is on that low value.

Rhythm, which refers to the sensation of isochrony conveyed by repeating patterns in speech, is key to add emotion and pragmatic meanings to the dialogues of, for example, voice assistants such as Siri and Alexa. Despite its importance, few papers tested new promising rhythm measures due to the technical challenges involved in this research. The present project aims at ameliorating these challenges by creating a database to study prosody cross-linguistically together with a set of scripts that output over 20 rhythm measures. With the help of undergraduate students majoring in different languages, the database will consist of TED talks in 8 languages, their orthographic transcriptions, and annotated speech wave forms (words, syllables, phonemes, and pauses). Speech annotations will be automatically generated by freely available aligners and sound editing programs. This database will be the input to the scripts that output the 20+ rhythm measures. Both tools, the database and scripts, together with a manual will be made available to the research community worldwide in order to promote international dialogue in this field.

The (poly)pharmacology of drug metabolites is seldom comprehensively characterized in drug discovery and development. However, some drug metabolites can reach high plasma concentrations and display relevant in vivo activity, which can be distinct from its parent drug. Here, we use computational and experimental methods to comprehensively characterise the kinase polypharmacology of M324, the major metabolite of the FDA-approved PARP inhibitor rucaparib. We experimentally demonstrate that M324 displays a distinct in vitro kinome profile from its parent drug, characterized by potent in vitro inhibition of GSK3A and PLK2 at clinically-relevant concentrations. These confirmed kinase activities of M324 could have potential implications for the efficacy and safety of rucaparib and therefore warrant further clinical investigation. The study reported here highlights the importance of thoroughly characterizing the activity of significant drug metabolites to better understanding drug responses in the clinic and maximally exploit the current drug arsenal in personalized and precision medicine.

Metabotropic glutamate 5 receptors (mGlu5) are thought to play an important role in mediating emotional information processing. In particular, negative allosteric modulators (NAMs) of mGlu5 have received a lot of attention as potential novel treatments for several neuropsychiatric diseases, including anxiety-related disorders. The aim of this study was to assess the influence of pre- and post-training mGlu5 inactivation in cued fear conditioned mice on neuronal oscillatory activity during fear retrieval. For this study we used the recently developed mGlu5 NAM Alloswicth-1 administered systemically. Injection of Alloswicth-1 before, but not after, fear conditioning resulted in a significant decrease in freezing upon fear retrieval. Mice injected with Alloswicth-1 pre-training were also implanted with recording microelectrodes into both the medial prefrontal cortex (mPFC) and ventral hippocampus (vHPC). The recordings revealed a reduction in theta rhythmic activity (4-12 Hz) in both the mPFC and vHPC during fear retrieval. These results indicate that inhibition of mGlu5 signaling alters local oscillatory activity in principal components of the fear brain network underlying a reduced response to a predicted threat., This research was funded by the Austrian Science Found grant NR W012060-10 to F.F and by the Dr. Helmut Legerlotz-Stiftung GZ76130 to A.R.-P. Funding by MCIN/AEI/10.13039/501100011033 and by ERDF A way of making Europe, European Union (projects I+D+i CTQ2017-89222-R and PID2020-120499RB-I00), by the Catalan government (2017 SGR 1604) to A.L. European Union’s Horizon 2020 research and innovation program under Marie Skłodowska-Curie grant agreement No. 801342 (Tecniospring INDUSTRY, TECSPR19-1-0062) and the Government of Catalonia’s Agency for Business Competitiveness (ACCIÓ) to X.G. Open Access Funding by the Austrian Science Fund (FWF).

Adrenoceptors are G protein-coupled receptors involved in a large variety of physiological processes, also under pathological conditions. This is due in large part to their ubiquitous expression in the body exerting numerous essential functions. Therefore, the possibility to control their activity with high spatial and temporal precision would constitute a valuable research tool. In this study, we present a caged version of the approved non-selective β-adrenoceptor antagonist carvedilol, synthesized by alkylation of its secondary amine with a coumarin derivative. Introducing this photo-removable group abolished carvedilol physiological effects in cell cultures, mouse isolated perfused hearts and living zebrafish larvae. Only after visible light application, carvedilol was released and the different physiological systems were pharmacologically modulated in a similar manner as the control drug. This research provides a new photopharmacological tool for a wide range of research applications that may help in the development of future precise therapies., We thank Maria José Bleda (IQAC-CSIC, Barcelona), Ignacio Pérez (IQAC-CSIC, Barcelona), Yolanda Pérez (IQAC-CSIC, Barcelona) and Carme Serra (SimChemSiMChem, IQAC-CSIC, Barcelona) for technical support. We thank Dr. Kees Jalink (The Netherlands Cancer Institute, Amsterdam, the Netherlands) for providing the plasmids encoding for the Epac-SH188 biosensor. We thank the University of Vic-Central University of Catalonia (UVic-UCC), Dr. Malu Calle and Dr. Marta Otero for the material assignment which helped in some biological assays. This work was supported by ERDF-FEDER European Fund (projects CTQ2017-89222-R) and by the Catalan government (2017SGR 1604) to AL. Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación (PID2020-120499RB-I00) supported XR and AL. XR research was financed by the Spanish Ministry of Economy, Industry and Competitiveness (SAF2015-74132-JIN). MF was supported by the “Agencia Estatal deInvestigación” from the Spanish Ministry of Science and Innovation and the IDAEA-CSIC, a Centre of Excellence Severo Ochoa (CEX2018-000794-S). ARS has a consolidated Miguel Servet contract and was financed by by the Catalan government (2017-SGR-1807). ADC received the support of a fellowship from “la Caixa” Foundation (ID 100010434) under the fellowship codeLCF/BQ/DE18/11670012.

Neuropathic pain is a common health problem, resulting in exacerbated response to noxious and non noxious stimuli, as well as impaired emotional and cognitive responses. Unfortunately, neuropathic pain is also one of the most difficult pain syndromes to manage, highlighting the importance of better understanding of the brain regions and neuromodulatory mechanisms involved in its regulation. Among the many interconnected brain areas which process pain, the amygdala is known to play an important role in the integration of sensory and emotional pain signals. Here, we questioned the ability of a recently identified neuromodulatory mechanisms associated to the metabotropic glutamate receptors mGlu4in the amygdala to modulate neuropathic pain. In a murine model of peripheral mononeuropathy induced by a chronic constriction of the sciatic nerve, we demonstrate that pharmacological activation of amygdala mGlu4receptors efficiently alleviates sensory and depressive-like symptoms in both male and female mice. Moreover, we reveal a differential modulation of those symptoms, activating mGlu4receptors in the controlateral amygdala, relatively to the side of the mononeuropathy, is necessary and sufficient to relieve both sensory and depressive-like symptoms while ipsilateral activation solely reduces depressive-like symptoms. Furthermore, using photopharmacology, a recent strategy allowing a precise spatiotemporal photocontrol of deep brain endogenous targets, we further demonstrate the rapid and reversible action of mGlu4-mediated neuromodulation on neuropathic pain symptoms. Finally, coupling photopharmacology and analgesic conditioned place preference, we show an important pain-reducing effect of mGlu4 activation. Taken together, these data highlight the analgesic potential of enhancing amygdala mGlu4activity to counteract neuropathy in the hope of improving existing treatments.

Beta-adrenoceptors (ß-AR) are prototypical G proteincoupled receptors and important pharmacological targets for many diseases. Indeed, a number of approved drugs target these receptors due to their key role on many physiological functions. Among other examples, we encounter ß1-AR antagonists (ß- Blockers), which constitute the first-line therapy for the treatment of heart diseases, and ß2-AR agonists, which act as bronchodilators for the treatment of breathing pathologies. Considering the relevance of these receptors, achieving a reversible and localised control of their activity would provide a powerful tool, both for its research applications and its clinical potential. In this context, photopharmacology arises as a potent approach. Photopharmacology is an emerging field based on the use of synthetic light-regulated molecules to allow reversible spatiotemporal control of target receptors in native tissues. These ligands have the potential to provide a precise and controllable therapeutic action with increased efficacy and reduced side effects. Moreover, the fine regulation on demand of the receptor activation state is of great interest for their study in non-modified cells, tissues and organisms. The present project provides the first proof of concept for beta-adrenoceptor photopharmacology. We first designed and synthesised libraries of lightregulated compounds in order to regulate ß-AR activity with spatiotemporal precision. Subsequent testing highlighted the successful development of compounds with promising pharmacological properties which can be reversibly and irreversibly controlled by light. The discovered molecules enable a fine control of ß-AR in their native environment that will certainly open the door to innovative research procedures and may inspire future personalized therapies targeting these receptors.

Beta-adrenoceptors (ß-AR) are prototypical G protein-coupled receptors (GPCR) and important pharmacological targets for numerous diseases. Indeed, a number of approved drugs target ß-AR, which are key regulators of many physiological functions. Among other examples, ß1-AR antagonists (known as ß-Blockers) are first-line therapies for the treatment of heart failure, and ß2-AR agonists, which act as bronchodilators, are widely used for the treatment of breathing pathologies. Considering the medical relevance of these receptors, achieving a reversible and localized control of their activity would provide a powerful research and clinical tool. GPCR signaling is currently recognized as a multidimensional process governed by molecular, spatial and temporal components. Uncovering the role of each of these dimensions is crucial to improve our knowledge on cell communication, to understand how different pathways give rise to cellular and physiological effects, and to know how can we interact with biological systems with precision using drugs. Photopharmacology is an emerging field in which light-sensitive molecules are used to control the function of a given target protein in native tissues. The modulation of the target activity is achieved by small, drug-like, photoregulated ligands. By the use of light, both spatial and temporal control of the compound activity can be achieved in unprecedented manners compared to conventional pharmacology. These ligands have the potential to provide highly precise and controllable therapeutic actions that may result in increased efficacies and reduced side effects. Importantly, photopharmacology may allow to gain mechanistic insight on the interplay between the activation time and the receptor location during signaling processes in non-modified cells, tissues and whole organisms. Our research focused on the generation of new molecular tools for beta-adrenoceptors photopharmacology will be presented in this communication. First, several libraries of light-sensitive compounds with the aim to regulate ß-AR activity with spatiotemporal precision were designed and synthesized. Subsequent testing in cell preparations demonstrated the successful development of compounds with promising pharmacological properties, which can be reversibly and irreversibly controlled by light. Among those, several hit compounds were identified as ligands for beta-1 and beta-2 adrenoceptors with low nanomolar activities. These libraries compounds were found to be active enough to become useful photopharmacological tools, so we also performed in vivo experiments to determine their research potential in physiological environments. Indeed, the discovered molecules enabled a fine control of ß-AR in their native environment. We believe that the results of these studies will certainly open the door to innovative research procedures and may inspire future therapies targeting ß-AR.

New technologies based on luminescence have been essential to monitor the organization, signaling, trafficking or ligand binding of G Protein-Coupled Receptors (GPCRs), but they rely on the overexpression of genetically modified receptors. As more and more studies indicate the importance of studying native receptors in their natural environment, it is essential to develop approaches allowing the specific labeling of native receptors. Here we report an innovative ligand directed approach to specifically label residues of native GPCRs upon ligand binding. To this end, we developed a ligand-directed toolbox based on a novel approach that uses molecular modules to build fluorescent ligand-directed probes that can label an archetypical aminergic GPCR (D1R). Our probes can be readily prepared before the labeling reaction from two molecular modules: an activated electrophilic linker which includes a fluorescent dye and a GPCR ligand that may include nucleophilic groups. Thanks to a fast and specific click reaction, the nucleophilic ligand can barely react with the activated linker before it is bound to the native target GPCR and the labeling reaction occurs. Subsequently, the ligand unbinds the GPCR pocket, leaving the receptor fluorescently labeled and fully functional. This novel labeling approach allowed us to label both D1 receptor in transfected cells and native receptors in neuronal cell lines. This approach will pave the way to develop new reagents and assays to monitor endogenous GPCRs distribution, trafficking, activity or binding properties in their native environment., Funding Agence Nationale de la Recherche ANR-17-CE11-0046 Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación and ERDF - A way of making Europe CTQ2017-89222-R Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación and ERDF - A way of making Europe PCI2018-093047 Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación and ERDF - A way of making Europe PID2020-120499RB-I00 Catalan government 2017 SGR 1604 the European Union's Horizon 2020 research and innovation program under Marie Skłodowska-Curie grant agreement No. 801342 (TecniospringINDUSTRY) and the Government of Catalonia's Agency for Business Competitive-ness (ACCIÓ). TECSPR19-1-0062

Metabotropic glutamate 5 receptors (mGlu

Breakthrough cancer pain (BTcP) refers to a sudden and transient exacerbation of pain, which develops in patients treated with opioid analgesics. Fast-onset analgesia is required for the treatment of BTcP. Light-activated drugs offer a novel potential strategy for the rapid control of pain without the typical adverse effects of systemic analgesic drugs. mGlu5 metabotropic glutamate receptor antagonists display potent analgesic activity, and light-induced activation of one of these compounds (JF-NP-26) in the thalamus was found to induce analgesia in models of inflammatory and neuropathic pain. We used an established mouse model of BTcP based on the injection of cancer cells into the femur, followed, 16 days later, by systemic administration of morphine. BTcP was induced by injection of endothelin-1 (ET-1) into the tumor, 20 min after morphine administration. Mice were implanted with optic fibers delivering light in the visible spectrum (405 nm) in the thalamus or prelimbic cortex to locally activate systemically injected JF-NP-26. Light delivery in the thalamus caused rapid and substantial analgesia, and this effect was specific because light delivery in the prelimbic cortex did not relieve BTcP. This finding lays the groundwork for the use of optopharmacology in the treatment of BTcP., This work was supported by the Italian Ministry of Health (Project code: GR-2016-02362046).

Metabotropic glutamate receptors (mGlu) are a family of class C G protein-coupled receptors (GPCRs) with important biological functions and widespread expression. The mechanisms of mGlu activation and the development of allosteric modulators for these dimeric proteins have attracted singular attention including the use of light regulated ligands. Photopharmacology involves the integration of a photoactive moiety into the ligand structure that following specific illumination undergoes a structural rearrangement and changes its biological activity. The use of light-regulated allosteric ligands offers the opportunity to manipulate mGlu signalling with spatiotemporal precision, unattainable with classical pharmacological approaches. In this review, we will discuss some of the innovations that have been made in the allosteric photopharmacology of mGlu receptors to date. We discuss the prospects of these molecular tools in the control of mGluRs and the new perspectives in understanding mGlu mechanisms, pharmacology and (patho)physiology that can ultimately result in innovative drug discovery concepts., The project on which these results are based has received funding from Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación 10.13039/501100011033 and ERDF A way of making Europe (projects I+D+i CTQ2017-89222-R, PCI2018-093047 and PID2020-120499RB-I00), the Neuron-ERANET program (MAGNOLIA project), by the Catalan government (2017 SGR 1604) to AL and XR, and the European Union's Horizon 2020 research and innovation programme under Marie Skłodowska-Curie grant agreement No. 801342 (Tecniospring INDUSTRY, TECSPR19-1-0062) and the Government of Catalonia's Agency for Business Competitiveness (ACCIÓ) to XG-S.

Lepidopteran pests are major factors limiting soybean productivity in South America. In some cases, the control of these species requires the use of foliar insecticides. For a sustainable use of these insecticides, they should be sprayed when insect population sizes reach an economic threshold. Since this estimation requires to determine the consumption of different species, this work aimed to integrate all the main factors, studying the consumption of small-and medium-size larvae of major lepidopteran pests to vegetative and reproductive tissues on Bt and non-Bt soybeans. The damage to vegetative tissues was tested in detached-leaf assays in grow chambers, and to reproductive structures was measured in greenhouse with infestation at early (flowering) and mid reproductive (mid grain filling) stages. Based on the feeding behavior of the species tested, they were cast in four groups: a) A. gemmatalis and C. includens, defoliating only the RR variety with the lowest consumption of foliar area; b) S. eridania, defoliating both RR and IPRO varieties, consuming twice than the species mentioned above; c) H. armigera, defoliating and being the most damaging species to pods in the RR variety; d) S. cosmioides and S. frugiperda, defoliating and damaging pods in both varieties. The species differed in their ability to feed on IPRO varieties, so a different economic threshould could be considered. This clasification contributes to a recommendation of insecticide use sustainable, taking into account the behavior of these species that are major soybeans pests in South America.

The insecticide carbaryl is commonly found in indirectly exposed freshwater ecosystems at low concentrations considered safe for fish communities. In this study, we showed that after only 24 h of exposure to environmental concentrations of carbaryl (0.066-660 ng/L), zebrafish larvae exhibit impairments in essential behaviours. Interestingly, the observed behavioural effects induced by carbaryl were acetylcholinesterase-independent. To elucidate the molecular initiating event that resulted in the observed behavioural effects, in silico predictions were followed by in vitro validation. We identified two target proteins that potentially interacted with carbaryl, the α2B adrenoceptor (ADRA2B) and the serotonin 2B receptor (HTR2B). Using a pharmacological approach, we then tested the hypothesis that carbaryl had antagonistic interactions with both receptors. Similar to yohimbine and SB204741, which are prototypic antagonists of ADRA2B and HTR2B, respectively, carbaryl increased the heart rate of zebrafish larvae. When we compared the behavioural effects of a 24-h exposure to these pharmacological antagonists with those of carbaryl, a high degree of similarity was found. These results strongly suggest that antagonism of both ADRA2B and HTR2B is the molecular initiating event that leads to adverse outcomes in zebrafish larvae that have undergone 24 h of exposure to environmentally relevant levels of carbaryl., This work was supported by “Agencia Estatal de Investigación” from the Spanish Ministry of Science and Innovation (project PID2020-113371RB-C21), IDAEA-CSIC, Severo Ochoa Centre of Excellence (CEX2018-000794-S), which financed M.F. with Severo Ochoa funds. Juliette Bedrossiantz was supported by a PhD grant (PRE2018-083513) co-financied by the Spanish Government and the European Social Fund (ESF). This work was supported by Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación and ERDF-FEDER European Fund (projects CTQ2017-89222-R and PID2020-120499RB-I00) and by the Catalan Government (2017 SGR 1604 and 2017-SGR-1807) to AL. XR research was financed by the Spanish Ministry of Economy, Industry and Competitiveness (SAF2015-74132-JIN). We thank Dr. Kees Jalink (The Netherlands Cancer Institute, Amsterdam, the Netherlands) for providing the plasmids encoding for the Epac-SH188 biosensor and Dr. Karen Martinez (University of Copenhagen, Copenhagen, Denmark) for providing the HEK 293 SNAP-β1AR). The findings of this report are not to be construed as an official Department of the Army position unless so designated by other authorized documents.

Although learning second language phonology is a difficult task, orthographic input may support the learning of difficult sound contrasts through a process known as orthographic facilitation. We extended this research by examining the effects of orthographic input together with individual differences in three different phonological learning processes, namely, the production of, perception of, and memorization of words containing three Marathi phonemic contrasts (i.e., [k-kh], [], and []) by native English speakers. Moreover, because the [] and [] contrasts were particularly challenging in previous auditory training studies (e.g., Polka, 1991), we used cross-modal training in order to enhance learning by pairing auditory perception tasks with visual orthographic information, the amplification of relevant acoustic cues, and proprioceptive descriptions to the articulation of target phonemes. Results showed significant learning from the pre- to the posttest across tasks and contrasts, supporting the effectiveness of cross-modal training. Furthermore, incongruent orthographic input could inhibit perception, and orthographic input generally supported memory for word pronunciations. Moreover, individual differences regarding phonological skills and nonspeech auditory discrimination predicted participants’ success in different phonological learning processes. These results provide a detailed picture of the complexity between different aspects of second language phonological learning and cross-modal training.

Catecholamine-triggered β-adrenoceptor (β-AR) signaling is essential for the correct functioning of the heart. Although both β

SummaryCatecholamine-triggered β-adrenoceptor (β-AR) signaling is essential for the correct functioning of the heart. Although both β1- and β2-AR subtypes are expressed in cardiomyocytes, drugs selectively targeting β1-AR have proven this receptor as the main target for the therapeutic effects of beta blockers in heart. Here, we report a new strategy for the spatiotemporal control of β1-AR activation by means of light-regulated drugs with a high level of β1-/β2-AR selectivity. All reported molecules allow for an efficient real time optical control of receptor function in vitro. Moreover, using confocal microscopy we demonstrate that the binding of our best hit, pAzo-2, can be reversibly photocontrolled. Strikingly, pAzo-2 also enables a dynamic cardiac rhythm management on alive zebrafish larvae using light, thus highlighting the therapeutic and research potential of the developed photoswitches. Overall, this work provides the first proof of precise control of the therapeutic target β1-AR in native environments using light.

In the last two decades, new technologies based on luminescence have been developed to monitor the organization, signalling or ligand binding of G Protein-Coupled Receptors. These technologies rely on the overexpression of genetically modified (and/or fluorescently tagged) receptors of interest. However, there is an increasing interest in developping approaches to conjugate chemical labels to specific residues of native GPCRs, despite of the low reactivity and the high abundance of such residues. Ligand directed approaches, may offer a solution to this problem. Such approaches consist in molecular probes that include a selective ligand moiety and a reactive moiety. Upon ligand binding, the labelling is directed to a nucleophilic amino acid in the vicinity of the binding pocket. However, this requires the use of non-nucleophilic ligand moieties, which is particularly difficult since many GPCR ligands contain amines or other nucleophilic functional groups. In the present work, we developed an innovative ligand-directed toolbox based on a novel approach. The later uses molecular modules to build fluorescent ligand-directed probes to label an archetypical aminergic GPCR (D1R). Our molecular probes can be readily prepared before the labelling reaction from two molecular modules: an activated electrophilic linker which includes a fluorescent dye and a GPCR ligand that may include nucleophilic groups. Thanks to a fast and specific chemical reaction, the nucleophilic ligand can barely react with the activated linker before it is bound to the native target GPCR and the labelling reaction occurs. Subsequently, the ligand will unbind the GPCR pocket, leaving the receptor fluorescently labelled and fully functional. This novel labelling approach allowed us to label endogenous D1 receptor both in transfected cells and primary cultures of neurons and will pave the way to develop new reagents and assays to monitor endogenous GPCRs distribution and activity in their native environment.

Positive allosteric modulators (PAMs) for metabotropic glutamate receptors have been postulated to treat neuropsychiatric diseases. Besides, obtaining a reversible and efficient spatiotemporal control of mGlu activity would be therapeutically advantageous. Photopharmacology may provide a solution on this topic, since it is based on the use of light and photoswitchable ligands to modulate a protein activity. This approach offers new perspectives for drug discovery and promises a better drug action control reducing side effects to unattained levels.

Catecholamine-triggered β-adrenoceptor (β-AR) signaling is essential for the correct functioning of the heart. Although both β1 - and β2 -AR subtypes are expressed in cardiomyocytes, drugs selectively targeting β1 -AR have proven this receptor as the main target for the therapeutic effects of beta blockers in the heart. Here, we report a new strategy for the light-control of β1 -AR activation by means of photoswitchable drugs with a high level of β1 -/β2 -AR selectivity. All reported molecules allow for an efficient real-time optical control of receptor function in vitro. Moreover, using confocal microscopy we demonstrate that the binding of our best hit, pAzo-2, can be reversibly photocontrolled. Strikingly, pAzo-2 also enables a dynamic cardiac rhythm management on living zebrafish larvae using light, thus highlighting the therapeutic and research potential of the developed photoswitches. Overall, this work provides the first proof of precise control of the therapeutic target β1 -AR in native environments using light., We thank Ignacio Pérez (IQAC-CSIC, Barcelona), Yolanda Pérez (IQAC-CSIC, Barcelona), Lourdes Muñoz (SimChem, IQAC-CSIC, Barcelona) and Carme Serra (SimChem, IQAC-CSIC, Barcelona) for technical support. We thank Diana Baxter (Institute for Research in Biomedicine, IRB, Barcelona) for her thorough revision of the language of this manuscript. We thank Dr. Kees Jalink (The Netherlands Cancer Institute, Amsterdam, the Netherlands) for providing the plasmids encoding for the Epac-SH188 biosensor. We thank the University of Vic-Central University of Catalonia (UVic-UCC) and Dr. Marta Otero for the material assignment which helped in some biological assays. We thank Nikos Hatzakis for access to the Olympus IX81 confocal microscope (UCPH, DK). This work was supported by ERDF-FEDER European Fund and Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación (projects CTQ2017-89222-R and PID2020-120499RB-I00) and by the Catalan government (2017 SGR 1604) to A.L. X.R. research was financed by the Spanish Ministry of Economy, Industry and Competitiveness (SAF2015-74132-JIN). D.R. research was supported by “Agencia Estatal de Investigación” from the Spanish Ministry of Science and Innovation (project PID2020-113371RB-C21) and IDAEA-CSIC, Severo Ochoa Centre of Excellence (CEX2018-000794-S), which financed M.F. A.D.C. received the support of a fellowship from “la Caixa” Foundation (ID 100010434) under the fellowship code LCF/BQ/DE18/11670012. K.L.M., A.D. and Y.M. were supported by the Novo Nordisk Foundation (NNF20OC0064565).

Neuropathic pain is a common health problem resulting in exacerbated response to noxious and non noxious stimuli, as well as impaired emotional and cognitive responses. Unfortunately, neuropathic pain is also one of the most difficult pain syndromes to manage, highlighting the importance of better understanding the brain regions and neuromodulatory mechanisms involved in its regulation. Among the many interconnected brain areas which process pain, the amygdala is known to play an important role in the integration of sensory and emotional pain signals. Here we questioned the ability of a recently identified neuromodulatory mechanism associated with the metabotropic glutamate receptors mGlu4 in the amygdala to modulate neuropathic pain. In a murine model of peripheral mononeuropathy, we demonstrate that pharmacological activation of amygdala mGlu4 efficiently alleviates sensory and depressive-like symptoms in both male and female mice. Moreover, we reveal a differential modulation of these symptoms. Activating mGlu4 in the contralateral amygdala relative to the side of the mononeuropathy, is necessary and sufficient to relieve both sensory and depressive-like symptoms, while ipsilateral activation solely reduces depressive-like symptoms. Furthermore, using photopharmacology, a recent strategy allowing precise photocontrol of endogenous proteins, we further demonstrate the dynamic alleviation of neuropathic pain through light-dependent facilitation of mGlu4 by a photoswitchable positive allosteric modulator. Finally, coupling photopharmacology and analgesic conditioned place preference, we show a significant pain-reducing effect of mGlu4 activation. Taken together, these data highlight the analgesic potential of enhancing amygdala mGlu4 activity to counteract neuropathy reinforcing its therapeutic interest for the treatment of pathological pain., This work was supported by the International Emerging Action (IEA) from the Centre National de la Recherche Scientifique (CNRS) to CG and AL and by grants from the French National Research Agency (ANR): ANR-16-CE16–0010 to CG and ANR-17-NEU3–0001 under the frame of Neuron Cofund to CG and AL.

Lepidopteran pests are major factors limiting soybean productivity in South America. In some cases, effective management of these species requires the use of foliar insecticides. For sustainable use of these insecticides, they should only be applied when insect population size exceeds an economic threshold. Since this estimation requires to determine the consumption of different species, this work aimed to integrate all these factors, studying the consumption of small (less than 1 cm long) and medium (1 to 1.5 cm long) size larvae of major lepidopteran pests to vegetative and reproductive tissues on Bt (M7739IPRO variety, containing the event MON87701 which expresses the Cry1Ac protein from Bacillus thuringiensis) and non-Bt (BMX Desafio RR variety) soybeans. The feeding injury to vegetative tissues was tested in detached-leaf assays in grow chambers, and for reproductive structures the study was conducted in greenhouse with infestations at early (flowering) and mid reproductive (mid grain filling) stages. Based on the feeding behavior of the species tested, they were cast in four groups: a) Anticarsia gemmatalis and Chrysodeixis includens, defoliating only the RR variety with the lowest consumption of foliar area; b) Spodoptera eridania, defoliating both RR and IPRO varieties, consuming twice than the species mentioned above; c) Helicoverpa armigera, defoliating and being the most damaging species to pods in the RR variety; and d) S. cosmioides and S. frugiperda, defoliating and damaging pods in both varieties. The species differed in their ability to feed on IPRO varieties, so a different economic threshold should be considered. Consequently, in cases where more than one species are found simultaneously, the species composition should be considered in estimating the economic threshold. Additionally, our findings may contribute to a better decision-making to control insect feeding injury in IPRO varieties, because a slower larval growth provides more time to ensure the need of control with insecticides. In summary, this clasification contributes to an improved recommendation of sustainable insecticide use, taking into account the behavior of each species that are major soybeans pests in South America.

Cystinuria, a rare inherited aminoaciduria condition, is characterized by the hyperexcretion of cystine, ornithine, lysine, and arginine. Its main clinical manifestation is cystine stone formation in the urinary tract, being responsible for 1–2% total and 6–8% pediatric lithiasis. Cystinuria patients suffer from recurrent lithiasic episodes that might end in surgical interventions, progressive renal functional deterioration, and kidney loss. Cystinuria is monitored for the presence of urinary cystine stones by crystalluria, imaging techniques or urinary cystine capacity; all with limited predicting capabilities. We analyzed blood and urine levels of the natural antioxidant L-ergothioneine in a Type B cystinuria mouse model, and urine levels of its metabolic product S-methyl-L-ergothioneine, in both male and female mice at two different ages and with different lithiasic phenotype. Urinary levels of S-methyl-L-ergothioneine showed differences related to age, gender and lithiasic phenotype. Once normalized by L-ergothioneine to account for interindividual differences, the S-methyl-L-ergothioneine to L-ergothioneine urinary ratio discriminated between cystine lithiasic phenotypes. Urine S-methyl-L-ergothioneine to L-ergothioneine ratio could be easily determined in urine and, as being capable of discriminating between cystine lithiasis phenotypes, it could be used as a lithiasis biomarker in cystinuria patient management., This work has been supported by the Spanish Health Institute Carlos III Grants FIS PI13/00121-R-FEDER, PI16/00267-R-FEDER and PI20/00200-R_FEDER (to VN), Generalitat de Catalunya Grant SGR2017-191 (to VN). VN acknowledge the support of the CERCA Program/Generalitat de Catalunya.

Background and Purpose Opioid-based drugs are the gold standard medicines for pain relief. However, tolerance and several side effects (i.e. constipation and dependence) may occur upon chronic opioid administration. Photopharmacology is a promising approach to improve the benefit/risk profiles of these drugs. Thus, opioids can be locally activated with high spatiotemporal resolution, potentially minimizing systemic-mediated adverse effects. Here, we aimed at developing a morphine photo-derivative (photocaged morphine), which can be activated upon light irradiation both in vitro and in vivo. Experimental Approach Light-dependent activity of pc-morphine was assessed in cell-based assays (intracellular calcium accumulation and electrophysiology) and in mice (formalin animal model of pain). In addition, tolerance, constipation and dependence were investigated in vivo using experimental paradigms. Key results In mice, pc-morphine was able to elicit antinociceptive effects, both using external light-irradiation (hind paw) and spinal cord implanted fibre-optics. In addition, remote morphine photoactivation was devoid of common systemic opioid-related undesired effects, namely, constipation, tolerance to the analgesic effects, rewarding effects and naloxone-induced withdrawal. Conclusion and Implications Light-dependent opioid-based drugs may allow effective analgesia without the occurrence of tolerance or the associated and severe opioid-related undesired effects., This work was supported by FEDER/Ministerio de Ciencia, Innovación y Universidades-Agencia Estatal de Investigación (SAF2017-87349-R, SAF2017-87199-R and CTQ2017-89222-R), ISCIII (PIE14/00034), and the Catalan government (2017 SGR 1604 and 2017 SGR 465). We thank Centres de Recerca de Catalunya (CERCA) Programme/Generalitat de Catalunya for IDIBELL institutional support. We thank Esther Castaño and Benjamín Torrejón from the Scientific and Technical Services (SCT) group at the Bellvitge Campus of the University of Barcelona for their technical assistance. We thank Lourdes Muñoz, Carme Serra and Juanlo Catena (SimChem, IQAC-CSIC, Barcelona) for support in the synthesis of compounds. Xavier Gasull and Aida Castellanos (Facultat de Medicina, Universitat de Barcelona) are acknowledged by their help with the obtention of DRG neurons from mouse. Also, we thank María Pilar Pérez Villamor (Laboratorios Dr. Esteve, Barcelona) for the kind gift of the HEK293 cell line permanently expressing μ receptors.

Summary Metabotropic glutamate receptors (mGluRs) are dimeric G-protein-coupled receptors activated by the main excitatory neurotransmitter, L-glutamate. mGluR activation by agonists binding in the venus flytrap domain is regulated by positive (PAM) or negative (NAM) allosteric modulators binding to the 7-transmembrane domain (7TM). We report the cryo-electron microscopy structures of fully inactive and intermediate-active conformations of mGlu5 receptor bound to an antagonist and a NAM or an agonist and a PAM, respectively, as well as the crystal structure of the 7TM bound to a photoswitchable NAM. The agonist induces a large movement between the subunits, bringing the 7TMs together and stabilizing a 7TM conformation structurally similar to the inactive state. Using functional approaches, we demonstrate that the PAM stabilizes a 7TM active conformation independent of the conformational changes induced by agonists, representing an alternative mode of mGlu activation. These findings provide a structural basis for different mGluR activation modes., Graphical abstract, Highlights • Cryo-EM analysis of thermostabilized mGlu5 receptor bound to inhibitors or activators • X-ray structure of trans-Alloswitch-1 bound to thermostable mGlu5 7TMs • Photopharmacology provides insight into allosteric regulation of mGlu5 7TMs • Multiple conformations of mGlu5 receptor activate G protein, Nasrallah et al. present cryo-EM structures of thermostabilized mGlu5 dimer bound to inhibitors and activators as well as an X-ray structure of mGlu5 7TM-bound photoswitchable ligand alloswitch-1. The structural and functional analyses reveal different modes of mGLu5 receptor activation and provide a structural basis for mGlu receptor activation mechanism.

International audience; We present the polarimetric analysis of the white-light images of the corona obtained with the Large-Angle Spectrometric COronagraph LASCO-C3 onboard the Solar and Heliospheric Observatory (SOHO) from 1996 to 2019, leading to the separation of the K- and F-components and the derivation of the electron-density distribution. The analysis makes use of polarized sequences composed of three images obtained through three polarizers oriented at +60°, 0°, and -60°, complemented by a neighboring unpolarized image. However, the degradation of the 0° polarizer noticed in 1999 compelled us to reconstruct the corresponding images from those obtained with the two other polarizers and the unpolarized ones thereafter. The analysis closely follows the method developed for LASCO-C2 (Lamy et al. in Solar Phys. 295, 89, 2020) and implements the formalism of Mueller, albeit with additional difficulties notably the presence of a non-axially symmetric component of stray light. Critical corrections were derived from a SOHO roll sequence and from consistency criteria (e.g. the “tangential” direction of polarization). The quasi-uninterrupted photopolarimetric analysis of the outer corona over two complete Solar Cycles 23 and 24 was successfully achieved and our final results encompass the characterization of its polarization, of its polarized radiance, of the two-dimensional electron density, and of the K-corona. Comparison between the C3 and C2 results in the region where their fields of view overlap shows an overall agreement. The C3 results are further in agreement with those of eclipses and radio-ranging measurements to an elongation of ≈10 R⊙ but tend to diverge further out. Although the coronal polarization out to 20 R⊙ is still highly correlated with the temporal variation of the total magnetic field, this divergence probably results from the increasing polarization of the F-corona with increasing solar elongation.

Including contributions from a team of world-renowned international scholars, this volume is a state-of-the-art survey of second language speech research, showcasing new empirical studies alongside critical reviews of existing influential speech learning models. It presents a revised version of Flege's Speech Learning Model (SLM-r) for the first time, an update on a cornerstone of second language research. Chapters are grouped into five thematic areas: theoretical progress, segmental acquisition, acquiring suprasegmental features, accentedness and acoustic features, and cognitive and psychological variables. Every chapter provides new empirical evidence, offering new insights as well as challenges on aspects of the second language speech acquisition process. Comprehensive in its coverage, this book summarises the state of current research in second language phonology, and aims to shape and inspire future research in the field. It is an essential resource for academic researchers and students of second language acquisition, applied linguistics and phonetics and phonology.

Natural Killer T (NKT) cells play an important role in the immune response and can be activated by glycolipids presented by CD1d protein. We present MCS-0208, an unprecedented arylthioether-phytoceramide able to induce potent invariant NKT (iNKT) cell activation, notably when tested in human iNKT cells. This arylsphingolipid analog has a simple phenyl group containing a single hydroxyl substituent as a surrogate of the sugar ring. The phenylthioether structure contrasts with α-galactosylceramide (1), the prototypical glycolipid used to induce iNKT cell stimulation, where the galactose 2’-OH and 3’-OH substituents are accepted as the minimal footprint and considered critical for NKT T cell receptor (TCR) recognition. A computational study supports the recognition of aromatic compound by the CD1d and TCR proteins as radically new structures for NKT cell stimulation., Financial support by the Ministry of Economy, Industry and Competitiveness (CTQ2014-57020-R and CTQ2011-29549-C02-01 grants) is gratefully acknowledged. Pre-doctoral FPU Spanish research and teaching fellowship from the Ministry of Education, Culture and Sport (FPU2013-01783 for R. Borràs-Tudurí grant and AP2007-01826 for A. Alcaide grant) are also acknowledged.

El Centre Nacional de Certificació Digital (NCDC) del Regne d'Aràbia Saudita (KSA) busca proporcionar una solució de confiança digital basada en la seva infraestructura de clau pública pública (PKI) per millorar la seva cartera de serveis oferint entre altres serveis, e-Signatura, e-Notari, Segells de marca de temps, Autenticació electrònica i altres serveis potencials. L'objectiu d'aquesta solució és proporcionar una infraestructura centralitzada, que redueixi la inversió, el temps i l'esforç d'integració i habilitació de PKI requerits per tercers (empreses governamentals i no governamentals), i aplicar amb eficàcia i coherència els estàndards adequats i les millors pràctiques per millorar la seguretat de les aplicacions participants a KSA. NCDC. També busca promoure l'ús de certificats digitals PKI en diverses aplicacions de KSA. El Centro Nacional de Certificación Digital (NCDC) del Reino de Arabia Saudita (KSA) está buscando proporcionar una solución de Confianza Digital basada en su Infraestructura de Clave Pública (PKI) nacional para mejorar su cartera de servicios ofreciendo, entre otros servicios, firma electrónica, e-Notariado, Sellado de tiempo, Autenticación y otros servicios potenciales. El objetivo de esta solución es proporcionar una infraestructura centralizada, que reducirá la inversión, el tiempo y el esfuerzo de habilitación e integración de PKI requeridos por terceros (empresas gubernamentales y no gubernamentales), y aplicar de manera eficiente y coherente los estándares adecuados y las mejores prácticas para mejorar la seguridad de las aplicaciones participantes en KSA. NCDC. También busca promover el uso de certificados digitales PKI en varias aplicaciones en KSA. The National Centre for Digital Certification (NCDC) of the Kingdom of Saudi Arabia (KSA) is looking to provide a national Public Key Infrastructure (PKI) based Digital Trust solution to enhance its services portfolio on offering among other services, e-Signature, e-Notary, Timestamping, e-Authentication, and other potential services. The objective of this solution is to provide a centralized infrastructure, which will reduce the investment, time and PKI enablement and integration effort required by third parties (government and non-government), and to efficiently and consistently apply appropriate standards and best-practices to improve the security for participating applications in KSA. NCDC.Is also looking to promote the use of PKI digital certificates across various applications in KSA.

Stroke and traumatic injury in brain or spinal cord are often life-threating conditions and major causes of death or permanent disability with high impact in the health care system. There are several stages of intervention to improve the neurological outcome. Acutely, fast interventions aiming to reestablish cerebral blood flow in ischemic stroke, to stop bleeding after brain hemorrhage, and to reduce edema after contusions are amongst mandatory actions. Current studies aim to develop accompanying strategies for brain cell protection based on enhancing endogenous protective mechanism, blocking cell death pathways, or through immunomodulation. After the acute phase, interventions are intended to promote recovery of function using rehabilitation with state-of-the-art technologies enabled by robotics. Other advanced strategies include cell, gene, and immune therapies, and brain function modulation with the aid of smart nanotechnologies. There is great expectation in the fast evolving novel approaches for improvement of neurological deficits in these unpredictable and devastating condition

[EN] Non-glycosidic analogues ofalpha-galactosylceramide of formula (I), wherein A, Z, Ra, Rb and Re have the meaning indicated in the deOH scription, as NKT cell activators, [ES] Análogos no glicosídicos de alfa-galactosilceramida de fórmula (I) donde A, Z, Ra, Rb Re tienen el significado indicado en la descripción como activadores de células NKT, Consejo Superior de Investigaciones Científicas (España), A1 Solicitud de patente con informe sobre el estado de la técnica

We present a photometrically accurate restitution of the K and F coronae from white-light images obtained over 24 Years [1996--2019] by the Large-Angle Spectrometric COronagraph "LASCO-C2" onboard the Solar and Heliospheric Observatory (SOHO). The procedure starts with the data set coming from the polarimetric separation of images of 512 x 512 pixels in which the F-corona and the instrumental stray light are entangled. Disentangling these components proceeds in three stages, each composed of several steps. Stage 1 establishes the distinct variations of the radiance of these components with the Sun--SOHO distance and generate a new data set of median images calculated for each Carrington rotation. Stage 2 achieves the restitution of a set of 36 stray light images reflecting its temporal variation and the periodic rolls of SOHO which started in 2003. Stage 3 achieves the restitution of the F-corona and a time series of daily images is generated. These results allowed us processing the whole set of routine LASCO-C2 images of 1024 x 1024 pixels (approximately 626000 images) and producing calibrated, high resolution images of the K-corona. We extend our past conclusions that the temporal variation of the integrated radiance of the K-corona tracks the solar activity over two solar cycles 23 and 24 and that it is highly correlated with the temporal variation of the total magnetic field. The behaviours of the integrated radiance during the last few years of the declining phases of solar cycles 23 and 24 are remarkably similar, reaching the same floor level and leading to a duration of 11.0 year for the latter cycle, in agreement with the sunspot determination., 1. Added two new Sections 3.5 and 4.2 discussing uncertainties, with new Figures (21 and 23); results unchanged. 2. Added a comparison with the results of Hayes et al. (2001) in Fig. 24

We present an in-depth characterization of the polarimetric channel of the Large-Angle Spectrometric COronagraph/LASCO-C3 onboard SOHO. The polarimetric analysis of the white-light images makes use of polarized sequences composed of three images obtained through three polarizers oriented at +60$^\circ$, 0$^\circ$, and -60$^\circ$, complemented by a neighboring unpolarized image. However, the degradation of the 0$^\circ$ polarizer noticed in 1999 compelled us to reconstruct the corresponding images from the other ones thereafter. The analysis closely follows the method developed for LASCO-C2 (Lamy, et al. Solar Physics 295, 89, 2020 and arXiv:2001.05925) and implements the formalism of Mueller, albeit with additional difficulties notably the presence of a non-axially symmetric component of stray light. Critical corrections were derived from a SOHO roll sequence and from consistency criteria (e.g., the tangential direction of polarization). The quasi-uninterrupted photopolarimetric analysis of the outer corona over two complete Solar Cycles 23 and 24 was successfully achieved and our final results encompass the characterization of its polarization, of its polarized radiance, of the two-dimensional electron density, and of the K-corona. Comparison between the C3 and C2 results where their field of view overlaps shows an overall agreement. The C3 results are further in agreement with those of eclipses and radio ranging measurements to an elongation of about 10 solar radii but tend to diverge further out. Whereas the coronal polarization out to 20 solar radii is still highly correlated with the temporal variation of the total magnetic field, this divergence probably results from the increasing polarization of the F-corona., 61 pages, 45 figures. arXiv admin note: text overlap with arXiv:2001.05925

β

β2-Adrenoceptors (β2-AR) are prototypical G-protein-coupled receptors and important pharmacological targets with relevant roles in physiological processes and diseases. Herein, we introduce Photoazolol-1–3, a series of photoswitchable azobenzene β2-AR antagonists that can be reversibly controlled with light. These new photochromic ligands are designed following the azologization strategy, with a p-acetamido azobenzene substituting the hydrophobic moiety present in many β2-AR antagonists. Using a fluorescence resonance energy transfer (FRET) biosensor-based assay, a variety of photopharmacological properties are identified. Two of the light-regulated molecules show potent β2-AR antagonism and enable a reversible and dynamic control of cellular receptor activity with light. Their photopharmacological properties are opposite, with Photoazolol-1 being more active in the dark and Photoazolol-2 demonstrating higher antagonism upon illumination. In addition, we provide a molecular rationale for the interaction of the different photoisomers with the receptor. Overall, we present innovative tools and a proof of concept for the precise control of β2-AR by means of light., We thank Montserrat Masoliver (UVic-UCC, Vic, Spain), Joan Bertrán (UVic-UCC, Vic, Spain), Jordi Serra (UVic-UCC, Vic, Spain), Lourdes Muñoz (SiMChem, IQAC-CSIC, Barcelona), Maria José Bleda (IQAC-CSIC, Barcelona), Ignacio Pérez (IQAC-CSIC, Barcelona), Yolanda Pérez (IQAC-CSIC, Barcelona) and Carme Serra (SiMChem, IQAC-CSIC, Barcelona) for technical support. We thank Dr. Kees Jalink (The Netherlands Cancer Institute, Amsterdam, the Netherlands) for providing the plasmids encoding for the Epac-SH188 biosensor. We thank the Cisbio Bioassays for their support and technical discussion, as well as for providing plasmids in the preliminary optimization of the biological assays. The project that gave rise to these results received the support of a fellowship from “la Caixa” Foundation (ID 100010434). The fellowship code is (LCF/BQ/DE18/11670012). This work was supported by FEDER/Ministerio de Ciencia, Innovación y Universidades–Agencia Estatal de Investigación (CTQ2017-89222-R), the Catalan government (2017SGR1604), PO FEDER of Catalonia 2014-2020 (project PECT Osona Transformació Social, Ref. 001-P-000382) and the Spanish Ministry of Economy, Industry and Competitiveness (SAF2015-74132-JIN).

International audience; We present an in-depth characterization of the polarimetric channel of the Large-Angle Spectrometric COronagraph/LASCO-C2 onboard the Solar and Heliospheric Observatory (SOHO). The polarimetric analysis of the white-light images makes use of polarized sequences composed of three images obtained though three polarizers oriented at +60∘, 0∘, and −60∘, complemented by a neighboring unpolarized image, and relies on the formalism of Mueller. The Mueller matrix characterizing the C2 instrument was obtained through extensive ground-based calibrations of the optical components and global laboratory tests. Additional critical corrections were derived from in-flight tests relying prominently on roll sequences of SOHO, on the basis of consistency criteria (e.g. the “tangential” direction of polarization), and from several applications, notably the time-dependent tomographic reconstruction of the coronal electron density. This took several years of effort, but resulted in the quasi-uninterrupted photopolarimetric analysis of the corona over two complete Solar Cycles 23 and 24, and the comparison with a variety of eclipse data obtained at different phases of these cycles. Our final results encompass the characterization of the polarization of the white-light corona, of its polarized radiance, of the two-dimensional electron density, and of the K-corona. The agreement with the eclipse data is excellent except for slight discrepancies affecting the innermost part of the C2 field of view. The present work leaves the two unpolarized components, F-corona and stray light, entangled and their complex separation will be dealt with in a follow-on article.

Supplemental material, supplemental_material for Chinese-English Speakers’ Perception of Pitch in Their Non-Tonal Language: Reinterpreting English as a Tonal-Like Language by Marta Ortega-Llebaria and Zhaohong Wu in Language and Speech

Altres ajuts: CERCA Programme/Generalitat de Catalunya Aquest article té una correcció a 10.1039/d0sc90189j The spatiotemporal control of cellular dynamic processes has great fundamental interest but lacks versatile molecular tools. Dynamin is a key protein in endocytosis and an appealing target to manipulate cell trafficking using patterns of light. We have developed the first photoswitchable small-molecule inhibitors of endocytosis (dynazos), by a stepwise design of the photochromic and pharmacological properties of dynasore, a dynamin inhibitor. We have characterized their photochromism with UV-visible and transient absorption spectroscopy and their biological activity using fluorescence microscopies and flow cytometry. Dynazos are water-soluble, cell permeable, and photostable, and enable fast, single-wavelength photoswitchable inhibition of clathrin-mediated endocytosis at micromolar concentration.

G protein-coupled receptors (GPCR), including the metabotrobic glutamate 5 receptor (mGlu5), are important therapeutic targets and the development of allosteric ligands for targeting GPCRs has become a desirable approach toward modulating receptor activity. Traditional pharmacological approaches toward modulating GPCR activity are still limited since precise spatiotemporal control of a ligand is lost as soon as it is administered. Photopharmacology proposes the use of photoswitchable ligands to overcome this limitation, since their activity can be reversibly controlled by light with high precision. As this is still a growing field, our understanding of the molecular mechanisms underlying the light-induced changes of different photoswitchable ligand pharmacology is suboptimal. For this reason, we have studied the mechanisms of action of alloswitch-1 and MCS0331; two freely diffusible, mGlu5 phenylazopyridine photoswitchable negative allosteric modulators. We combined photochemical, cell-based, and in vivo photopharmacological approaches to investigate the effects of trans–cis azobenzene photoisomerization on the functional activity and binding ability of these ligands to the mGlu5 allosteric pocket. From these results, we conclude that photoisomerization can take place inside and outside the ligand binding pocket, and this leads to a reversible loss in affinity, in part, due to changes in dissociation rates from the receptor. Ligand activity for both photoswitchable ligands deviates from high-affinity mGlu5 negative allosteric modulation (in the trans configuration) to reduced affinity for the mGlu5 in their cis configuration. Importantly, this mechanism translates to dynamic and reversible control over pain following local injection and illumination of negative allosteric modulators into a brain region implicated in pain control., We thank Carolina Cera and Teresa Sarrias (SimChem, IQAC-CSIC, Barcelona) and Anna Duran, Roser Borràs and Gloria Somalo (MCS group, IQAC-CSIC, Barcelona) for technical support. We thank Maria José Bleda Hernández (IQAC-CSIC, Barcelona) for statistical support. We thank Jean-Philippe Pin (IGF, Montpellier) for scientific support and contributions to this work. We thank Christine Enjalbal (IBMM, Montpellier), Thierry Durroux (IGF, Montpellier), Jean-Louis Banères (IBMM, Montpellier) and Guillaume Lebon (IGF, Montpellier) for scientific discussion. This research was supported by FEDER/Ministerio de Ciencia, Innovación y Universidades–Agencia Estatal de Investigación (CTQ2017-89222-R and PCI2018-093047), the Catalan government (2017SGR1604), by CSIC (PICS program 08212), by Neuron-ERANET, by grants from the Agence Nationale de la Recherche (ANR-16-CE16-0010 and ANR-17-NEU3-0001 under the frame of Neuron Cofund) and by the Programme International de Collaboration Scientifique of the CNRS (PICS 08212). AEB was supported by the Labex EpiGenMed (program « Investissements d’avenir », ANR-10-LABX-12-01).

We present an in-depth characterization of the polarimetric channel of the Large-Angle Spectrometric COronagraph LASCO-C2 onboard the Solar and Heliospheric Observatory (SoHO). The polarimetric analysis of the white-light images makes use of polarized sequences composed of three images obtained though three polarizers oriented at +60{\deg}, 0{\deg} and -60{\deg}, complemented by a neighboring unpolarized image, and relies on the formalism of Mueller. The Mueller matrix characterizing the C2 instrument was obtained through extensive ground-based calibrations of the optical components and global laboratory tests. Additional critical corrections were derived from in-flight tests relying prominently on roll sequences and on consistency criteria, mainly the tangential direction of polarization. Our final results encompass the characterization of the polarization of the white-light corona, of its polarized radiance, of the two-dimensional electron density, and of the K-corona over two solar cycles. They are in excellent agreement with measurements obtained at several solar eclipses except for slight discrepancies affecting the innermost part of the C2 field-of-view, probably resulting from an imperfect removal of the bright diffraction fringe surrounding the occulter., Comment: 72 pages, 51 figures