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    204915 research outputs found

    A simplified courtship conditioning protocol to test learning and memory in Drosophila

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    In Drosophila, a male that has previously been sexually rejected reduces its courtship behavior when confronted again with an unreceptive female. This reduced courting time reflects a memory formation process. Here, we describe a simplified protocol to perform the courtship conditioning assay for assessing the reduced courting time, using regular lab equipment and handmade tools. Every step of the procedure, from raising flies and training to testing and quantification of this memory-related behavior, can be implemented in any practice laboratory.We would like to thank Javier Gil Castillo for its invaluable help and advices in 3D printing. We also thank the flies from Bloomington Stock Center. We would like to thank BioRender (www.biorender.com) for the open-access platform used to create the graphical abstract. This work was supported by the Spanish Research Agency (Ministerio de Innovacion y Ciencia [MICINN]) under the grant PGC2018-094630-B-100 to F.A.M., cofinanced by the European Regional Development Fund (ERDF) to F.A.M. F.A.M. is a recipient of a RyC-2014-14961 contract. B.G.-M. is a recipient of a FPI-UAM predoctoral fellowship, grant number SFPI/2020/00878. C.G.B. is a recipient of a FPU predoctoral fellowship, grant number FPU19/04449 (MEFP). S.P.-F. is a recipient of a JAE intro fellowship, grant number JAEINT_21_02520 (CSIC)

    Can we manipulate brain connectivity? A systematic review of cortico-cortical paired associative stimulation effects

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    Objective: Cortico-cortical paired associative stimulation (ccPAS) is a form of dual-site transcranial magnetic stimulation (TMS) entailing a series of single-TMS pulses paired at specific interstimulus intervals (ISI) delivered to distant cortical areas. The goal of this article is to systematically review its efficacy in inducing plasticity in humans focusing on stimulation parameters and hypotheses of underlying neurophysiology. Methods: A systematic review of the literature from 2009–2023 was undertaken to identify all articles utilizing ccPAS to study brain plasticity and connectivity. Six electronic databases were searched and included. Results: 32 studies were identified. The studies targeted connections within the same hemisphere or between hemispheres. 28 ccPAS studies were in healthy participants, 1 study in schizophrenia, and 1 in Alzheimer's disease (AD) patients. 2 additional studies used cortico-cortical repetitive paired associative stimulation (cc-rPAS) in generalized anxiety disorder (GAD) patients. Outcome measures include electromyography (EMG), behavioral measures, electroencephalography (EEG), and functional magnetic resonance imaging (fMRI). ccPAS seems to be able to modulate brain connectivity depending on the ISI. Conclusions: ccPAS can be used to modulate corticospinal excitability, brain activity, and behavior. Although the stimulation parameters used across studies reviewed in this paper are varied, ccPAS is a promising approach for basic research and potential clinical applications. Significance: Recent advances in neuroscience have caused a shift of interest from the study of single areas to a more complex approach focusing on networks of areas that orchestrate brain activity. Consequently, the TMS community is also witnessing a change, with a growing interest in targeting multiple brain areas rather than a single locus, as evidenced by an increasing number of papers using ccPAS. In light of this new enthusiasm for brain connectivity, this review summarizes existing literature and stimulation parameters that have proven effective in changing electrophysiological, behavioral, or neuroimaging-derived measures.JCHP and JLP want to thank The Shirley Ryan AbilityLab for support

    First 3-D structural evidence of a native-like intertwined dimer in the acylphosphatase family

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    Acylphosphatase (AcP, EC 3.6.1.7) is a small model protein conformed by a ferredoxin-like fold, profoundly studied to get insights into protein folding and aggregation processes. Numerous studies focused on the aggregation and/or amyloidogenic properties of AcPs suggest the importance of edge-β-strands in the process. In this work, we present the first crystallographic structure of Escherichia coli AcP (EcoAcP), showing notable differences with the only available NMR structure for this enzyme. EcoAcP is crystalised as an intertwined dimer formed by replacing a single C-terminal β-strand between two protomers, suggesting a flexible character of the C-terminal edge of EcoAcP. Despite numerous works where AcP from different sources have been used as a model system for protein aggregation, our domain-swapped EcoAcP structure is the first 3-D structural evidence of native-like aggregated species for any AcP reported to date, providing clues on molecular determinants unleashing aggregation.This work was supported by the Spanish Ministry of Science and Innovation/FEDER funds Grant PID2020-116261GB-I00/AEI/10.13039/501100011033 (JAG). SMR is grateful to the Andalusian Regional Government through the Endocrinology and Metabolism Group (CTS-202) and to University of Granada “Plan Propio de Investigación de la UGR” (PP2022.PP.18). ACA is grateful to Junta de Andalucía (PY20_00149). We are grateful to the European Synchrotron Radiation Facility (ESRF), Grenoble, France, for providing time through proposal MX2353 and MX2454, and the staff at ID30-A3 beamlines for assistance during data collection. We are also grateful to the Spanish Synchrotron Light Facility (ALBA), Barcelona, Spain, for providing time through proposals 2021085252 and 2022086950, and the staff at XALOC beamline for assistance during data collection. We deeply thank Ma Carmen López-Sánchez for technical assistance

    Guilherme Grandi (ed.). Transportes e formações econômicas na América Latina. São Paulo, Editora Annablume, 2016

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    The phosphodiesterase RmcA contributes to the adaptation of Pseudomonas putida to l-Arginine

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    Amino acids are crucial in nitrogen cycling and to shape the metabolism of microorganisms. Among them, arginine is a versatile molecule able to sustain nitrogen, carbon, and even ATP supply and to regulate multicellular behaviors such as biofilm formation. Arginine modulates the intracellular levels of 3′-5′cyclic diguanylic acid (c-di-GMP), a second messenger that controls biofilm formation, maintenance and dispersion. In Pseudomonas putida, KT2440, a versatile microorganism with wide biotechnological applications, modulation of c-di-GMP levels by arginine requires the transcriptional regulator ArgR, but the connections between arginine metabolism and c-di-GMP are not fully characterized. It has been recently demonstrated that arginine can be perceived by the opportunistic human pathogen Pseudomonas aeruginosa through the transducer RmcA protein (Redox regulator of c-di-GMP), which can directly decrease c-di-GMP levels and possibly affect biofilm architecture. A RmcA homolog is present in P. putida, but its function and involvement in arginine perceiving or biofilm life cycle had not been studied. Here, we present a preliminary characterization of the RmcA-dependent response to arginine in P. putida in modulating biofilm formation, c-di-GMP levels, and energy metabolism. This work contributes to further understanding the molecular mechanisms linking biofilm homeostasis and environmental adaptation.This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement number 101004806 MOSBRI, DSB-UROM TNA access provider for Seahorse experiment (to M.E.U.). Financial support from the Sapienza University of Rome (RM120172A7AD98EB to S.R. and AR12117A63EE6037, AR2221816C44C7B3 to C.S.R.), and grant PID2019-109372GB-I00 funded by MCIN/AEI/10.13039/501100011033 (M.E.U. and M.A.M.H.), are also acknowledged

    Seasonal upwelling influence on trophic indices of mesozooplankton in a coastal food web estimated from δ15N in amino acids

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    Mesozooplankton communities in upwelling ecosystems are known to be influenced by seasonal upwelling enhancing primary productivity. However, the extent to which changes in trophic dynamics of mesozooplankton are driven by variations in the baseline of nitrogen or in the trophic position (TP) is poorly understood. We used nitrogen stable isotopes (δN) in bulk and in specific amino acids (CSIA-AA), as well as the taxonomic composition of mesozooplankton samples collected monthly during two full years, to investigate whether the upwelling intensity affects the isotopic N baseline and other trophic indices at the community level in the coastal system off NW Iberian Peninsula. We hypothesized that enhanced phytoplankton productivity during upwelling events would lead to a shorter and more efficient food web. Upwelling induced mesozooplankton herbivory (by increasing the abundance and biomass of omnivorous-herbivorous copepods and other taxa), resulting in a linear decrease of TP (estimated using Glx = glutamic acid + glutamine, which mainly involves metazoan links) from downwelling to upwelling situations. In contrast, TP (estimated using alanine, which also reflects microbial trophic pathways) was highest in low-moderate upwelling and decreased with upwelling strength. Thus, an optimal coupling between metazoan and microbial food webs was reached at low-moderate upwelling intensities when the microbial contribution was ca. 25 %, while this contribution decreased (<15 %) either during downwelling or during strong upwelling. However, the significance of differences in mean values of TP and zooplankton composition was small due to the rapid succession of upwelling and downwelling events. Our results illustrate how a combination of CSIA-AA and community data can provide valuable information on rapid changes in mesozooplankton food web structure in highly dynamic upwelling systems.This research was partially supported by projects ANILE (CTM2009-08396), from Plan Nacional de I+D+i (Spain), and QLOCKS (PID2020-115620RB-100), from MCIN/AEI/10.13039/501100011033 (Spain). R. García-Seoane was supported by a postdoctoral research grant Juan de la Cierva-Formación (FJC2019-040921-I), from MCIN/AEI/10.13039/501100011033 (Spain) and EU NextGenerationEU/PRTR programmes, and I. G. Viana was supported by a postdoctoral research grant Juan de la Cierva-Incorporación (IJC2019-040554-I), from MCIN/AEI/10.13039/501100011033 (Spain)

    Constant 4He Concentration and 190Pt-4He age of Detrital Pt-Alloy Grains from the Santiago River, Ecuador: Potential as a 4He Mineral Reference Material

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    Over the last few decades geo- and thermochronological techniques based on the accumulation of radiogenic He in minerals have gained popularity. Currently there is no mineral-based He reference material that allows routine screening of gas extraction and measurement procedures. The commonly-used mineral reference material for (U-Th)/He thermochronology have variable U and Th contents which result in variable He concentrations. Here we demonstrate that native platinum grains (RS-Pt) from placer deposits in the Santiago River, Ecuador, yield constant He contents and may be used as a reference material. Helium concentrations measured at the Scottish Universities Environmental Research Centre (n = 8) and Institute of Precambrian Geology and Geochronology RAS (n = 9) noble gas laboratories yield mean He concentrations that overlap at the 95% confidence level. The weighted mean He concentration is 215 ± 4 × 10 at g (2s) and mean Pt-He age is 39.6 ± 0.7 Ma (2s). Samples of the Santiago River Pt concentrate are available on request from the SUERC laboratory.This research was supported financially by RSF 22-77-10088 and SUERC

    Unveiling the Origin of Multidomain Structures in Compositionally Modulated Cylindrical Magnetic Nanowires

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    [EN] CoNi/Ni multisegmented cylindrical nanowires were synthesized via an electrochemical route. The wires are 140 nm in diameter, with 1000 nm long Ni segments and CoNi segments between 600 and 1400 nm in length. The magnetic configuration was imaged by XMCD-PEEM in the demagnetized state and at remanence after magnetizing axially and perpendicularly. Ni segments, with cubic crystal symmetry, show an axial magnetic configuration with a small curling component at the surface. In turn, CoNi segments, with hexagonal crystal symmetry and a strong magnetocrystalline anisotropy perpendicular to the nanowires, show a single vortex state in the shorter segments and multivortex or multitransverse magnetic configurations in medium and long segments, respectively. A detailed study by micromagnetic simulations reveals that the magnetic configuration is determined mainly by the coupling between soft Ni and harder CoNi segments. For short CoNi segments, Ni segments are magnetostatically coupled and the chirality of the single vortex formed in CoNi remains the same as that of the curling in neighboring Ni segments. For longer CoNi segments, the remanent state is either the multivortex or multitransverse state depending on whether the previously applied field was parallel or perpendicular to the magnetocrystalline axis. The results point out the relevance of the cylindrical geometry to promote the occurrence of complex magneto-chiral effects and provide key information for the design of cylindrical magnetic nanowires for multiple applications.Support ofSpanish MINECO under Projects MAT2016-76824-C3-1-Rand FIS2016-78591-C3-3-R and the Regional Government of Madrid under Project S2018/NMT-4321 NANOMAGCOST-CM. L.A. and M.F. acknowledge funding through Project RTI2018-095303-B-C53. A.F.R. acknowledges support fromthe Spanish MINECO (Project MAT2015-68772-P). We alsoacknowledge the service from the MiNa Laboratory at IMNand funding from CM (Project SpaceTec, S2013/ICE2822),MINECO (Project CSIC13-4E-1794), and EU (FEDER,FSE)The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acsnano.0c03579Peer reviewe

    Tomato POLLEN DEFICIENT 2 encodes a G-Type lectin receptor kinase required for viable pollen grain formation

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    Pollen development is a crucial biological process indispensable for seed set in flowering plants and for successful crop breeding. However, little is known about the molecular mechanisms regulating pollen development in crop species. This study reports a novel male-sterile tomato mutant, pollen deficient 2 (pod2), characterized by the production of non-viable pollen grains and resulting in the development of small parthenocarpic fruits. A combined strategy of mapping-by-sequencing and RNA interference-mediated gene silencing was used to prove that the pod2 phenotype is caused by the loss of Solanum lycopersicum G-Type lectin receptor kinase II.9 (SlG-LecRK-II.9) activity. In situ hybridization of floral buds showed that POD2/SlG-LecRK-II.9 is specifically expressed in tapetal cells and microspores at the late tetrad stage. Accordingly, abnormalities in meiosis and tapetum programmed cell death in pod2 occurred during microsporogenesis, resulting in the formation of four dysfunctional microspores leading to an aberrant microgametogenesis process. RNA-seq analyses supported the existence of alterations at the final stage of microsporogenesis, since we found tomato deregulated genes whose counterparts in Arabidopsis are essential for the normal progression of male meiosis and cytokinesis. Collectively, our results revealed the essential role of POD2/SlG-LecRK-II.9 in regulating tomato pollen development.This work was supported by research grants PID2019-110833RB-C31, PID2019-110833RB-C32, and PID2020-113324GB-100 funded by the Spanish Ministry of Science and Innovation (MCIN/AEI/10.13039/501100011033), and the Research and Innovation Programme of the European Union Horizon 2020 (BRESOV Project, ID 774244). A PhD fellowship to MGA was funded by the FPU Programme of the Spanish Ministry of Education and Culture (ref. AP2010-4528). RLe was supported by a Junta de Andalucía and FEDER research contract (DOC_01129)

    Involvement of Bcl-2 Family Proteins in Tetraploidization-Related Senescence

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    The B-cell lymphoma 2 (Bcl-2) family of proteins is the main regulator of apoptosis. However, multiple emerging evidence has revealed that Bcl-2 family proteins are also involved in cellular senescence. On the one hand, the different expression of these proteins determines the entry into senescence. On the other hand, entry into senescence modulates the expression of these proteins, generally conferring resistance to apoptosis. With some exceptions, senescent cells are characterized by the upregulation of antiapoptotic proteins and downregulation of proapoptotic proteins. Under physiological conditions, freshly formed tetraploid cells die by apoptosis due to the tetraploidy checkpoint. However, suppression of Bcl-2 associated x protein (Bax), as well as overexpression of Bcl-2, favors the appearance and survival of tetraploid cells. Furthermore, it is noteworthy that our laboratory has shown that the joint absence of Bax and Bcl-2 antagonist/killer (Bak) favors the entry into senescence of tetraploid cells. Certain microtubule inhibitory chemotherapies, such as taxanes and vinca alkaloids, induce the generation of tetraploid cells. Moreover, the combined use of inhibitors of antiapoptotic proteins of the Bcl-2 family with microtubule inhibitors increases their efficacy. In this review, we aim to shed light on the involvement of the Bcl-2 family of proteins in the senescence program activated after tetraploidization and the possibility of using this knowledge to create a new therapeutic strategy targeting cancer cells.L.S. was supported by the Spanish Agencia Estatal de Investigación, AEI (grant number PID2021-126426OB-I00), the Strategic Program from the Institute of Biomedicine and Molecular Genetics (IBGM) of Valladolid (CCVC8485) and the Internationalization Project of the Unit of Excellence IBGM of Valladolid from the Junta de Castilla y León (CL-EI-2021 IBGM), as well as the “Beatriz Galindo senior” Program from the Spanish Ministry of Universities. G.K. was supported by the Ligue contre le Cancer (équipe labellisée), Agence National de la Recherche (ANR)—Projets blancs, AMMICa US23/CNRS UMS3655, Association pour la recherche sur le cancer (ARC), Cancéropôle Ile-de-France, European Research Council Advanced Investigator Grand “ICD-Cancer”, Fondation pour la Recherche Médicale (FRM), a donation by Elior, Equipex Onco-Pheno-Screen, European Joint Programme on Rare Diseases (EJPRD), European Research Council (ICD-Cancer), European Union Horizon 2020 Projects Oncobiome and Crimson, Fondation Carrefour, Institut National du Cancer (INCa), Institut Universitaire de France, LabEx Immuno-Oncology (ANR-18-IDEX-0001), a Cancer Research ASPIRE Award from the Mark Foundation, the RHU Immunolife, Seerave Foundation, SIRIC Stratified Oncology Cell DNA Repair and Tumor Immune Elimination (SOCRATE) and SIRIC Cancer Research and Personalized Medicine (CARPEM). This study contributes to the IdEx Université de Paris ANR-18-IDEX-0001. D.B. and R.P.-R. were supported by the University of Valladolid. L.G.-G. was supported by the “Margarita Salas” Program from the Spanish Ministry of Universities

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