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Reactive oxygen and nitrogen species and redox signals are at the crossroads of plant responses to abiotic and biotic stress and mediate plant-induced resistance/sensitivity, which mainly depends on the specific interaction., Complex signalling pathways are involved in plant protection against single and combined stresses. Plants are able to coordinate genome-wide transcriptional reprogramming and display a unique programme of transcriptional responses to a combination of stresses that differs from the response to single stresses. However, a significant overlap between pathways and some defence genes in the form of shared and general stress-responsive genes appears to be commonly involved in responses to multiple biotic and abiotic stresses. Reactive oxygen and nitrogen species, as well as redox signals, are key molecules involved at the crossroads of the perception of different stress factors and the regulation of both specific and general plant responses to biotic and abiotic stresses. In this review, we focus on crosstalk between plant responses to biotic and abiotic stresses, in addition to possible plant protection against pathogens caused by previous abiotic stress. Bioinformatic analyses of transcriptome data from cadmium- and fungal pathogen-treated plants focusing on redox gene ontology categories were carried out to gain a better understanding of common plant responses to abiotic and biotic stresses. The role of reactive oxygen and nitrogen species in the complex network involved in plant responses to changes in their environment is also discussed.

W.D.V.H. acknowledges support from National Institutes of Health (Grant: R01GM112077). S.B.O. acknowledges support from the Gordon and Betty Moore Foundations and National Science Foundation (Awards: 1715591 and 1901709). J.M.S.R. acknowledges support from Spanish Ministry of Economy and Competitiveness/FEDER Funds (Grants BIO2015-66426-R and RTI2018-097142-B-100) and the Human Frontier Science Program (Grant RGP0041/2017). V.A.R. acknowledges support from FEDER/Junta de Andalucia-Consejeria de Economia y Conocimiento (Grant E.FQM.113.UGR18). We would like to thank the beamline staff of ID30B of the ESRF (European Synchrotron Radiation Facility, Grenoble, France) for their assistance during data collection and the ESRF for the provision of time through proposals MX-2064., TEM-1 β-lactamase degrades β-lactam antibiotics with a strong preference for penicillins. Sequence reconstruction studies indicate that it evolved from ancestral enzymes that degraded a variety of β-lactam antibiotics with moderate efficiency. This generalist to specialist conversion involved more than 100 mutational changes, but conserved fold and catalytic residues, suggesting a role for dynamics in enzyme evolution. Here, we develop a conformational dynamics computational approach to rationally mold a protein flexibility profile on the basis of a hinge-shift mechanism. By deliberately weighting and altering the conformational dynamics of a putative Precambrian β-lactamase, we engineer enzyme specificity that mimics the modern TEM-1 β-lactamase with only 21 amino acid replacements. Our conformational dynamics design thus re-enacts the evolutionary process and provides a rational allosteric approach for manipulating function while conserving the enzyme active site., United States Department of Health & Human Services National Institutes of Health (NIH) - USA R01GM112077, Gordon and Betty Moore Foundations, National Science Foundation (NSF) 1715591 1901709, Spanish Ministry of Economy and Competitiveness/FEDER Funds BIO2015-66426-R RTI2018-097142-B-100, Human Frontier Science Program RGP0041/2017, FEDER/Junta de Andalucia-Consejeria de Economia y Conocimiento E.FQM.113.UGR18

The most frequent variant of multiple sclerosis (MS) is the relapsing–remitting form, characterized by symptomatic phases followed by periods of total/partial recovery. Hence, it is possible that these patients can benefit from endogenous agents that control the inflammatory process and favor spontaneous remyelination. In this context, there is increasing interest in the role of myeloid‐derived suppressor cells (MDSCs) during the clinical course of experimental autoimmune encephalomyelitis (EAE). MDSCs speed up infiltrated T‐cell anergy and apoptosis. In different animal models of MS, a milder disease course is related to higher presence/density of MDSCs in the periphery, and smaller demyelinated lesions in the central nervous system (CNS). These observations lead us to wonder whether MDSCs might not only exert an anti‐inflammatory effect but might also have direct influence on oligodendrocyte precursor cells (OPCs) and remyelination. In the present work, we reveal for the first time the relationship between OPCs and MDSCs in EAE, relationship that is guided by the distance from the inflammatory core. We describe the effects of MDSCs on survival, proliferation, as well as potent promoters of OPC differentiation toward mature phenotypes. We show for the first time that osteopontin is remarkably present in the analyzed secretome of MDSCs. The ablation of this cue from MDSCs‐secretome demonstrates that osteopontin is the main MDSC effector on these oligodendroglial cells. These data highlight a crucial pathogenic interaction between innate immunity and the CNS, opening ways to develop MDSC‐ and/or osteopontin‐based therapies to promote effective myelin preservation and repair in MS patients., Main Points The abundance of myeloid‐derived suppressor cells (MDSCs) correlates with the density of oligodendrocyte precursor cells (OPCs) in experimental autoimmune encephalomyelitis demyelinating lesions.MDSC secretome favors OPC survival, proliferation, and differentiation toward myelinating phenotypes.Osteopontin is crucial in MDSCs secretome to act on OPC biology.

Background Introduction of pneumococcal conjugate vaccines (PCVs) has reduced the disease caused by vaccine serotypes in children, providing herd protection to adults. However, the emergence of nonvaccine serotypes is of great concern worldwide. Methods This study includes national laboratory data from invasive pneumococcal disease (IPD) cases that affected pediatric and adult populations during 2009–2019. The impact of implementing different vaccine strategies for immunocompetent adults by comparing Spanish regions that used the 13-valent PCV (PCV13) vs regions that used the 23-valent pneumococcal polysaccharide vaccine (PPV23) was also analyzed for 2017−2019. Results The overall reductions in IPD cases by PCV13 serotypes in children and adults were 88% and 59%, respectively, during 2009–2019, with a constant increase in serotype 8 in adults since 2015. IPD cases by additional serotypes covered by PPV23 increased from 20% in 2009 to 52% in 2019. In children, serotype 24F was the most frequent in 2019, whereas serotypes 3 and 8 accounted for 36% of IPD cases in adults. Introduction of PCV13 or PPV23 in the adult calendar of certain Spanish regions reduced the IPD cases by PCV13 serotypes by up to 25% and 11%, respectively, showing a decrease of serotype 3 when PCV13 was used. Conclusions Use of PCV13 in children has affected the epidemiology, reducing the burden of IPD in children but also in adults by herd protection; however, the increase in serotype 8 in adults is worrisome. Vaccination with PCV13 in adults seems to control IPD cases by PCV13 serotypes including serotype 3.

The genetic diversity of bacterial populations’ nodulating Retama sphaerocarpa grown in the soils of Maamora cork forest (Morocco) was examined. ERIC-PCR fingerprinting of 30 strains distributed them in 2 groups, of which a representative strain from each group was studied by multilocus sequence analysis of the 16S rRNA, atpD, and gyrB genes. The two representative strains RSM25 and RSM32, grouped with “Microvirga tunisiensis”. This is the first description of Retama nodule bacteria as members of the genus Microvirga. A nodC-based phylogeny confirmed that the two representative strains RSM25 and RSM32 are affiliated with symbiovar mediterranense. The 2 strains were capable of nodulating not only R. sphaerocarpa but also R. monosperma, R. dasycarpa and L. luteus, and unable to nodulate Phaseolus vulgaris, Vachellia gummifera, Cicer arietinum, Vigna unguiculata and Glycine max. The inoculation of R. sphaerocarpa with RSM25 or RSM32 produced a 1.22-, and 1.36-fold increase in the dry weight of the plants compared to those grown in the presence of 0.05% KNO. The 2 strains used monosaccharides and disaccharides as the sole C source, but fructose, glucose, galactose, arabinose and starch were not used. They were unable to grow on glycine as a N source. Phosphate solubilization and siderophore production was not detected, but IAA or IAA-related compounds were produced. The alkaline pH of the sampling site soil in the Maamora forest where Retama grows could explain why Microvirga was the dominant species in the root nodules of the plants., The authors want to thank the Hassan II Academy of Sciences for funding this research in the frame of the BIOMICS project. Financial support was also obtained from the ERDF-cofinanced project AGL2017-85676R from Ministerio de Economia, Industria y Competitividad (Spain).

Tumor stiffening is a hallmark of malignancy that actively drives tumor progression and aggressiveness. Recent research has shed light onto several molecular underpinnings of this biomechanical process, which has a reciprocal crosstalk between tumor cells, stromal fibroblasts, and extracellular matrix remodeling at its core. This dynamic communication shapes the tumor microenvironment; significantly determines disease features including therapeutic resistance, relapse, or metastasis; and potentially holds the key for novel antitumor strategies. Caveolae and their components emerge as integrators of different aspects of cell function, mechanotransduction, and ECM-cell interaction. Here, we review our current knowledge on the several pivotal roles of the essential caveolar component caveolin-1 in this multidirectional biomechanical crosstalk and highlight standing questions in the field. Research in our laboratory has been supported by grants from the Spanish Ministry of Economy, Industry and Competitiveness (MINECO; SAF2011-25047, SAF2014-51876-R, SAF2017-83130-R, IGP-SO grant MINSEV1512-07-2016, CSD2009-0016 and BFU2016-81912-REDC), and the Worldwide Cancer Research Foundation (no. 15-0404) to M.A.d.P. M.A.d.P is also a member of the Tec4Bio consortium (ref. P2018/NMT4443; “Actividades de I+D entre Grupos de Investigación en Tecnologías,” Comunidad Autónoma de Madrid/FEDER, Spain) and is co-recipient of grants from Fundació LaMarató de TV3 (674/C/2013 and 201936).M.A.d.P’s group received funding from the European Union Horizon 2020 research and innovation programme under Marie Sklodowska-Curie grant agreement no. 641639 (BIOPOL ETN), of which V.J-J. was ESR trainee. M.S-A. was recipient of a CNIC IPP fellowship (COFUND programme 2014). The CNIC is supported by the Instituto de Salud Carlos III (ISCIII), the Ministerio de Ciencia, Innovación y Universidades (MCNU), and the Pro CNIC Foundation and is a Severo Ochoa Center of Excellence (SEV-2015-0505). Sí

Objective: Pulmonary arterial hypertension (PAH), one of the major complications of systemic sclerosis (SSc), is a rare disease with unknown etiopathogenesis and noncurative treatments. As mice deficient in P-selectin glycoprotein ligand 1 (PSGL-1) develop a spontaneous SSc-like syndrome, we undertook this study to analyze whether they develop PAH and to examine the molecular mechanisms involved. Methods: Doppler echocardiography was used to estimate pulmonary pressure, immunohistochemistry was used to assess vascular remodeling, and myography of dissected pulmonary artery rings was used to analyze vascular reactivity. Angiotensin II (Ang II) levels were quantified by enzyme-linked immunosorbent assay, and Western blotting was used to measure Ang II type 1 receptor (ATR), ATR, endothelial cell nitric oxide synthase (eNOS), and phosphorylated eNOS expression in lung lysates. Flow cytometry allowed us to determine cytokine production by immune cells and NO production by endothelial cells. In all cases, there were 4–8 mice per experimental group. Results: PSGL-1 mice showed lung vessel wall remodeling and a reduced mean ± SD expression of pulmonary ATR (expression ratio [relative to β-actin] in female mice age >18 months: wild-type mice 0.799 ± 0.508 versus knockout mice 0.346 ± 0.229). With aging, female PSGL-1 mice had impaired up-regulation of estrogen receptor α (ERα) and developed lung vascular endothelial dysfunction coinciding with an increase in mean ± SEM pulmonary Ang II levels (wild-type 48.70 ± 5.13 pg/gm lung tissue versus knockout 78.02 ± 28.09 pg/gm lung tissue) and a decrease in eNOS phosphorylation, leading to reduced endothelial NO production. These events led to a reduction in the pulmonary artery acceleration time:ejection time ratio in 33% of aged female PSGL-1 mice, indicating pulmonary hypertension. Importantly, we found expanded populations of interferon-γ–producing PSGL-1 T cells and B cells and a reduced presence of regulatory T cells. Conclusion: The absence of PSGL-1 induces a reduction in Treg cells, NO production, and ERα expression and causes an increase in Ang II in the lungs of female mice, favoring the development of PAH., Spanish Ministry of Economy and Competitiveness (grants SAF2015-69396-R, SAF2011-28150, and SAF2014-55399) and the Spanish Ministry of Health and Instituto de Salud Carlos III (cofinanced by Fondos FEDER; grants AC17-00027, FIS-PI14-01698, FIS-PI17-01819, and FIS-PI12-01578, Red de Investigación de Enfermedades Reumáticas [RIER] RD12/0009/0017). The Centro Nacional de Investigaciones Cardiovasculares (CNIC) is supported by the Ministerio de Economía, Industria y Competitividad, and the Pro CNIC Foundation and is a Severo Ochoa Center of Excellence (SEV-2015-0505))

Cells remodel their structure in response to mechanical strain. However, how mechanical forces are translated into biochemical signals that coordinate the structural changes observed at the plasma membrane (PM) and the underlying cytoskeleton during mechanoadaptation is unclear. Here, we show that PM mechanoadaptation is controlled by a tension-sensing pathway composed of c-Abl tyrosine kinase and membrane curvature regulator FBP17. FBP17 is recruited to caveolae to induce the formation of caveolar rosettes. FBP17 deficient cells have reduced rosette density, lack PM tension buffering capacity under osmotic shock, and cannot adapt to mechanical strain. Mechanistically, tension is transduced to the FBP17 F-BAR domain by direct phosphorylation mediated by c-Abl, a mechanosensitive molecule. This modification inhibits FBP17 membrane bending activity and releases FBP17-controlled inhibition of mDia1-dependent stress fibers, favoring membrane adaptation to increased tension. This mechanoprotective mechanism adapts the cell to changes in mechanical tension by coupling PM and actin cytoskeleton remodeling., Mechanical forces are sensed by cells and can alter plasma membrane properties, but biochemical changes underlying this are not clear. Here the authors show tension is sensed by c-Abl and FBP17, which couples changes in mechanical tension to remodelling of the plasma membrane and actin cytoskeleton.

Anthropogenic disturbance is a major cause of the biodiversity crisis. Nevertheless, the role of anthropogenic substrate vibrations in disrupting animal behavior is poorly understood. Amphibians comprise the terrestrial vertebrates most sensitive to vibrations, and since communication is crucial to their survival and reproduction, they are a suitable model for investigating this timely subject. Playback tests were used to assess the effects of substrate vibrations produced by two sources of anthropogenic activity– road traffic and wind turbines– on the calling activity of a naïve population of terrestrial toads. In their natural habitat, a buried tactile sound transducer was used to emit simulated traffic and wind turbine vibrations, and changes in the toads’ acoustic responses were analyzed by measuring parameters important for reproductive success: call rate, call duration and dominant frequency. Our results showed a significant call rate reduction by males of Alytes obstetricans in response to both seismic sources, whereas other parameters remained stable. Since females of several species prefer males with higher call rates, our results suggest that anthropogenically derived substrate-borne vibrations could reduce individual reproductive success. Our study demonstrates a clear negative effect of anthropogenic vibrations on anuran communication, and the urgent need for further investigation in this area., This study was funded by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (VG: #380320/2018-5; CB. 401076/2014-8; 312968/2013-2) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (www.capes.gov.br/CAPES VC.3471301; 88881.135522/2016-01). Partial funding for this project was generously provided by PPG-BAN UFRGS in VZC and MBM lab; by Daniel Hurley for support of student research in the PMN lab and by Ministerio de Economía, Industria y Competitividad (CGL2017-88764-R, MINECO/AEI/FEDER, Spain) in the DL lab.

Over the years, human adenovirus (HAdV) has progressively been recognized as a significant viral pathogen. Traditionally associated with self-limited respiratory, gastrointestinal, and conjunctival infections, mainly in immunocompromised patients, HAdV is currently considered to be a pathogen presenting significant morbidity and mortality in both immunosuppressed and otherwise healthy individuals. Currently available therapeutic options are limited because of their lack of effectivity and related side effects. In this context, there is an urgent need to develop effective anti-HAdV drugs with suitable therapeutic indexes. In this work, we identified new serinol-derived benzoic acid esters as novel scaffolds for the inhibition of HAdV infections. A set of 38 compounds were designed and synthesized, and their antiviral activity and cytotoxicity were evaluated. Four compounds (13, 14, 27, and 32) inhibited HAdV infection at low micromolar concentrations (2.82–5.35 μM). Their half maximal inhibitory concentration (IC50) values were lower compared to that of cidofovir, the current drug of choice. All compounds significantly reduced the HAdV DNA replication process, while they did not block any step of the viral entry. Our results showed that compounds 13, 14, and 32 seem to be targeting the expression of the E1A early gene. Moreover, all four derivatives demonstrated a significant inhibition of human cytomegalovirus (HCMV) DNA replication. This new scaffold may represent a potential tool useful for the development of effective anti-HAdV drugs., This work has been supported by Ministerio de Ciencia, Innovación y Universidades, Plan Estatal 2017-2020 Retos-Proyectos I+D+i (PID2019-104767RB-I00), Ministerio de Economı́a y Competitividad, Plan Estatal 2013-2016 Retos-Proyectos I + D + i (CTQ2016-78580-C2-2-R) and by Instituto de Salud Carlos III, Subdirección General de Redes y Centros de Investigación Cooperativa, Ministerio de Economia, Industria y Competitividad, Spanish Network for Research in Infectious Diseases (REIPI RD16/0016/0009), cofinanced by European Development Regional Fund “A way to achieve Europe”, the Instituto de Salud Carlos III, Proyectos de Investigación en Salud (PI17/01055; PI18/01191) and Proyectos de Desarrollo Tecnológico en Salud (DTS17/00130), the Spanish Adenovirus Network (AdenoNet, BIO2015/68990-REDT), and the program “Nicolás Monardes” (C-0059-2018), Servicio Andaluz de Salud, Junta de Andalucía. M.V.-H. also thanks Ministerio de Economı́a y Competitividad, Plan Estatal 2013-2016 Excelencia I+D+i (CTQ2016-78703-P).

Analysing biological images coming from the microscope is challenging; not only is it complex to acquire the images, but also the three-dimensional shapes found on them. Thus, using automatic approaches that could learn and embrace that variance would be highly interesting for the field. Here, we use an evolutionary algorithm to obtain the 3D cell shape of curve epithelial tissues. Our approach is based on the application of a 3D segmentation algorithm called LimeSeg, which is a segmentation software that uses a particle-based active contour method. This program needs the fine-tuning of some hyperparameters that could present a long number of combinations, with the selection of the best parametrisation being highly time-consuming. Our evolutionary algorithm automatically selects the best possible parametrisation with which it can perform an accurate and non-supervised segmentation of 3D curved epithelial tissues. This way, we combine the segmentation potential of LimeSeg and optimise the parameters selection by adding automatisation. This methodology has been applied to three datasets of confocal images from Drosophila melanogaster, where a good convergence has been observed in the evaluation of the solutions. Our experimental results confirm the proper performing of the algorithm, whose segmented images have been compared to those manually obtained for the same tissues., B.P. research is funded by FEDER/Ministerio de Ciencia e Innovación—Agencia Estatal de Investigación/Proyecto TIN2017-88209-C2 and by the Andalusian Regional Government under the projects: US-1263341 and P18-RT-2778. P.V.-M. and P.G.-G. work is funded by the Ministry of Economy, Industry and Competitiveness grant BFU2016-74975-P co-funded by FEDER funds, and by the Spanish Ministry of Science and Innovation Ministry of Science through grant PID2019-103900GB-I00.

© 2021 by the authors., The discovery and application of antibiotics in the common clinical practice has undeniably been one of the major medical advances in our times. Their use meant a drastic drop in infectious diseases-related mortality and contributed to prolonging human life expectancy worldwide. Nevertheless, antibiotics are considered by many a double-edged sword. Their extensive use in the past few years has given rise to a global problem: antibiotic resistance. This factor and the increasing evidence that a wide range of antibiotics can damage mammalian mitochondria, have driven a significant sector of the medical and scientific communities to advise against the use of antibiotics for purposes other to treating severe infections. Notwithstanding, a notorious number of recent studies support the use of these drugs to treat very diverse conditions, ranging from cancer to neurodegenerative or mitochondrial diseases. In this context, there is great controversy on whether the risks associated to antibiotics outweigh their promising beneficial features. The aim of this review is to provide insight in the topic, purpose for which the most relevant findings regarding antibiotic therapies have been discussed., This work was supported by FIS PI16/00786 (2016) and FIS PI19/00377 (2019) grants, Ministerio de Sanidad, Spain and Fondo Europeo de Desarrollo Regional (FEDER Unión Europea), Spanish Ministry of Education, Culture and Sport. This activity has been co-financed by the European Regional Development Fund (ERDF) and by the Regional Ministry of Economic Transformation, Industry, Knowledge and Universities of the Junta de Andalucía, within the framework of the ERDF Andalusia operational program 2014–2020 Thematic objective “01—Reinforcement of research, technological develop-ment and innovation” through the reference research project CTS-5725 and PY18-850. We acknowledge the support of “Ayudas para la Formación de Profesorado Universitario” (FPU/MINECO), AEPMI (Asociación de Enfermos de Patología Mitocondrial), ENACH (Asociación de enfermos de Neurodegeneración con Acumulación Cerebral de Hierro), FEDER (Federación Española de Enfermedades Raras), Yo Nemálínica Association, KAT6A Association, Fundación MERCK Salud and Fundación MEHUER/Colegio Oficial de Farmacéuticos de Sevilla.

Departamento de Biotecnología (INIA), The success in the response of a plant to a pest depends on the regulatory networks that connect plant perception and plant response. Meta-analyses of transcriptomic responses are valuable tools to discover novel mechanisms in the plant/herbivore interplay. Considering the quantity and quality of available transcriptomic analyses, Arabidopsis thaliana was selected to test the ability of comprehensive meta-analyses to disentangle plant responses. The analysis of the transcriptomic data showed a general induction of biological processes commonly associated with the response to herbivory, like jasmonate signaling or glucosinolate biosynthesis. However, an uneven induction of many genes belonging to these biological categories was found, which was likely associated with the particularities of each specific Arabidopsis-herbivore interaction. A thorough analysis of the responses to the lepidopteran Pieris rapae and the spider mite Tetranychus urticae highlighted specificities in the perception and signaling pathways associated with the expression of receptors and transcription factors. This information was translated to a variable alteration of secondary metabolic pathways. In conclusion, transcriptomic meta-analysis has been revealed as a potent way to sort out relevant physiological processes in the plant response to herbivores. Translation of these transcriptomic-based analyses to crop species will permit a more appropriate design of biotechnological programs., Research was supported by the Ministerio de Economía, Industria y Competitividad (MEIC, Grants BIO2017-83472-R, RED2018-102407-T, and RyC17MESFB) and by the Convenio Plurianual between Comunidad de Madrid (CM) and Universidad Politécnica de Madrid (UPM) through the Programa de Apoyo a la Realización de Proyectos de I+D para Jóvenes Investigadores (APOYO-jovenes-SUR6Q9-22-Y). RyC grant (RyC2017-21814) from the MEIC and Plan Propio from the UPM financed MES. Programa Propio UPM financed AG., 15 Pág.

43 p.-5 fig., Plants react to environmental challenges by integrating external cues with endogenous signals to optimize survival and reproductive success. However, the mechanisms underlying this integration remain obscure. While stress conditions are known to impact plant development, how developmental transitions influence responses to adverse conditions has not been addressed. Here, we reveal a molecular mechanism of stress response attenuation during the onset of flowering in Arabidopsis (Arabidopsis thaliana). We show that Arabidopsis MORF-RELATED GENE (MRG) proteins, components of the NuA4 histone acetyltransferase complex that bind trimethylated-lysine 36 in histone H3 (H3K36me3), function as a chromatin switch on the floral integrator SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) to coordinate flowering initiation with plant responsiveness to hostile environments. MRG proteins are required to activate SOC1 expression during flowering induction by promoting histone H4 acetylation. In turn, SOC1 represses a broad array of genes that mediate abiotic stress responses. We propose that during the transition from vegetative to reproductive growth, the MRG-SOC1 module constitutes a central hub in a mechanism that tunes down stress responses to enhance the reproductive success and plant fitness at the expense of costly efforts for adaptation to challenging environments., This work was funded by the Ministerio de Economía,Industria y Competitividad, Gobierno de España (MINECO) ~grants BIO2016-77559-R (to J.A.J. and M.P.) and BIO2016-79187-R (to J.S.); the Ministerio de Ciencia e Innovación,Gobierno de España grant PID2019-104899GB-I00 (to J.A.J.and M.P.); a FPU fellowship from the Spanish Ministry of Education (to A.M.); The authors thank the “Severo Ochoa Program for Centres of Excellence in R&D” from the Agencia Estatal de Investigación of Spain (grant SEV-2016-0672(2017-2021) for supporting the scientific services used in this work.

© 2021 Ramírez, Torrecilla-Parra, Pardo-Marqués, de-Frutos, Pérez-García, Tabraue, de la Rosa, Martín-Rodriguez, Díaz-Sarmiento, Nuñez, Orizaola, Través, Camps, Boscá and Castrillo., Macrophages are immune cells that play crucial roles in host defense against pathogens by triggering their exceptional phagocytic and inflammatory functions. Macrophages that reside in healthy tissues also accomplish important tasks to preserve organ homeostasis, including lipid uptake/efflux or apoptotic-cell clearance. Both homeostatic and inflammatory functions of macrophages require the precise stability of lipid-rich microdomains located at the cell membrane for the initiation of downstream signaling cascades. Caveolin-1 (Cav-1) is the main protein responsible for the biogenesis of caveolae and plays an important role in vascular inflammation and atherosclerosis. The Liver X receptors (LXRs) are key transcription factors for cholesterol efflux and inflammatory gene responses in macrophages. Although the role of Cav-1 in cellular cholesterol homeostasis and vascular inflammation has been reported, the connection between LXR transcriptional activity and Cav-1 expression and function in macrophages has not been investigated. Here, using gain and loss of function approaches, we demonstrate that LXR-dependent transcriptional pathways modulate Cav-1 expression and compartmentation within the membrane during macrophage activation. As a result, Cav-1 participates in LXR-dependent cholesterol efflux and the control of inflammatory responses. Together, our data show modulation of the LXR-Cav-1 axis could be exploited to control exacerbated inflammation and cholesterol overload in the macrophage during the pathogenesis of lipid and immune disorders, such as atherosclerosis., We thank MINECO FPI predoctoral fellowship granted to MCO (BES-2015-075339). Experimental work was supported by grants from Ministerio de Ciencia, Investigación y Universidades, y Fondo Europeo de Desarrollo Regional (FEDER) Grant REF: PID2019-104284RB-I00/AEI/10.13039/501100011033 (to AC) and support from Networks of Excellence from MINECO (Nuclear Receptors in Cancer, Metabolism and Inflammation [NuRCaMeIn] SAF2017-90604-REDT to AC. Ministerio de Economía, Industria y Competitividad, Ministerio de Ciencia, Investigación y Universidades, and Agencia Estatal de Investigación (SAF2017-82436-R, RTC2017-6283-1), Centro de Investigación Biomédica en Red en Enfermedades Cardiovasculares (CB16/11/00222), Consorcio de Investigación en Red de la Comunidad de Madrid, S2017/BMD-3686 and Fondo Europeo de Desarrollo Regional (to LB). Ministerio de Ciencia, Investigación y Universidades, and Agencia Estatal de Investigación Proyectos de I+D+i Retos Investigación 2018 (RTI2018-095061-B-I00); TALENTO Grant from Madrid Government, Spain (2017-T1/BMD-5333); Consejería de Ciencia, Universidades e Innovación Comunidad de Madrid, Spain (PEJD-2018-POST/BMD-8900 and PEDJ-2018-AI/BDM-9724) to CMR.

Antibiotic resistance in Streptococcus pneumoniae has increased worldwide, making fluoroquinolones an alternative therapeutic option. Fluoroquinolones inhibit the type II DNA topoisomerases (topoisomerase IV and gyrase). In this study we have evaluated the in vivo activity of seconeolitsine, an inhibitor of topoisomerase I. Levofloxacin (12.5 to 50 mg/kg) or seconeolitsine (5 to 40 mg/kg) were administered every 12 h during two days in mice infected with a serotype 8-resistant strain. At 48 h, a 70% protection was obtained with seconeolitsine (40 mg/kg, p &lt, 0.001). However, survival with levofloxacin was 20%, regardless of the dose. In addition, seconeolitsine decreased bacteremia efficiently. Levofloxacin had higher levels in serum than seconeolitsine (Cmax of 14.7 vs. 1.6, 0.01) and higher values of area under the serum concentration-time curve (AUC0-12h of 17.3 vs. 5, 0.01). However, seconeolitsine showed higher levels of time to peak concentration and elimination half-life. This is consistent with the higher binding of seconeolitsine to plasma proteins (40% and 80% when used at 1 µg/mL and 50 µg/mL, respectively) in comparison to levofloxacin (12% at 5 µg/mL and 33% at 50 µg/mL). Our results suggest that seconeolitsine would be a promising therapeutic alternative against pneumococcal isolates with high fluoroquinolone resistance levels.

[EN] Protein sorting at the trans-Golgi network (TGN) usually requires the assistance of cargo adaptors. However, it remains to be examined how the same complex can mediate both the export and retention of different proteins or how sorting complexes interact among themselves. In Saccharomyces cerevisiae, the exomer complex is involved in the polarized transport of some proteins from the TGN to the plasma membrane (PM). Intriguingly, exomer and its cargos also show a sort of functional relationship with TGN clathrin adaptors that is still unsolved. Here, using a wide range of techniques, including time-lapse and BIFC microscopy, we describe new molecular implications of the exomer complex in protein sorting and address its different layers of functional interaction with clathrin adaptor complexes. Exomer mutants show impaired amino acid uptake because it facilitates not only the polarized delivery of amino acid permeases to the PM but also participates in their endosomal traffic. We propose a model for exomer where it modulates the recruitment of TGN clathrin adaptors directly or indirectly through the Arf1 function. Moreover, we describe an in vivo competitive relationship between the exomer and AP-1 complexes for the model cargo Chs3. These results highlight a broad role for exomer in regulating protein sorting at the TGN that is complementary to its role as cargo adaptor and present a model to understand the complexity of TGN protein sorting., Ministerio de Economía, Industria y Competitividad, Gobierno de España (Ministeri d'Economia, Indústria i Competitivitat), Grant/Award Number: CICYT/FEDER BFU2017-84508-P; Consejería de Educación, Junta de Castilla y León (Ministry of Education, Government of Castile-Leon), Grant/Award Number: SA116G19; Ministerio de Economía, Industria y Competitividad, Gobierno de España (Ministeri d'Economia, Indústria i Competitivitat), Grant/Award Number: RTC-2017-6468-2-AR; Ministerio de Economía, Industria y Competitividad, Gobierno de España (Ministeri d'Economia, Indústria i Competitivitat), Grant/Award Number: BIO2016-77776-P; Foundation for the National Institutes of Health (FNIH), Grant/Award Number: R01 GM092741

9 páginas.-4 figuras.- 45 referencias.- Additional Supporting Information may be found online in theSupporting Information section at the end of the article http://dx.doi.org/10.1111/nph.16177, Efficient water transport from soil to leaves sustains stomatal opening and steady-statephotosynthesis. The aboveground portion of this pathway is well-described, yet the roots andtheir connection with the soil are still poorly understood due to technical limitations. Here we used a novel rehydration technique to investigate changes in the hydraulic pathwaybetween roots and soil and within the plant body as individual olive plants were subjected to arange of water stresses. Whole root hydraulic resistance (including the radial pathway from xylem to the soil–rootinterface) constituted 81% of the whole-plant resistance in unstressed plants, increasing to>95% under a moderate level of water stress. The decline in this whole root hydraulicconductance occurred in parallel with stomatal closure and contributed significantly to thereduction in canopy conductance according to a hydraulic model. Our results demonstrate that losses in root hydraulic conductance, mainly due to a disconnectionfrom the soil during moderate water stress in olive plants, are profound and sufficient to inducestomatal closure before cavitation occurs. Future studies will determine whether this coreregulatory role of root hydraulics exists more generally among diverse plant species, Thisstudy was supported by funds from an Individual Fellowship fromthe European Union’s Horizon 2020 research and innovationprogramme under the Marie Skłodowska-Curie grant agreementno. 751918-AgroPHYS to CMR-D and from the AustralianResearch Council, Discovery Projects, DP170100761 to TJB

Carbapenems are considered the treatment of choice for extended-spectrum β-lactamase (ESBL)- or AmpC β-lactamase-producing Enterobacteriaceae bacteraemia. Data on the effectiveness of non-intravenous carbapenem-sparing antibiotic options are limited. This study compared the 30-day mortality and clinical failure associated with the use of carbapenems versus alternative non-intravenous antibiotics for the definitive treatment of ESBL/AmpC-positive Enterobacteriaceae bacteraemia. This 12-year retrospective study (2004–2015) included all patients with bacteraemia due to ESBL/AmpC-producing Enterobacteriaceae at a Spanish hospital. Given the lack of randomisation of initial therapies, a propensity score for receiving carbapenems was calculated. There were 1115 patients with a first episode of bacteraemia due to Escherichia coli or Klebsiella pneumoniae, of which 123 (11.0%) were ESBL/AmpC-positive. There were 101 eligible patients: 59 in the carbapenem group and 42 in the alternative treatment group (trimethoprim/sulfamethoxazole 59.5%, quinolones 21.4%). The most frequent sources of infection were urinary (63%) and biliary (15%). Compared with the carbapenem group, patients treated with an alternative regimen had a shorter hospital stay [median (IQR) 7 (5–10) days vs. 12 (9–18) days; P < 0.001]. Use of an alternative non-intravenous therapy did not increase mortality (OR = 0.27, 95% CI 0.05–1.61; P = 0.15). After controlling for confounding factors with the propensity score, the adjusted OR of carbapenem treatment was 4.95 (95% CI 0.94–26.01; P = 0.059). Alternative non-intravenous carbapenem-sparing antibiotics could have a role in the definitive treatment of ESBL/AmpC-positive Enterobacteriaceae bacteraemia, allowing a reduction in carbapenem use. Use of trimethoprim/sulfamethoxazole in this series showed favourable results., CP, NF-H and JR-B are supported by Plan Nacional de I+D+i 2013?2016 and Instituto de Salud Carlos III, Subdirección General de Redes y Centros de Investigación Cooperativa, Ministerio de Economía, Industria y Competitividad, Spanish Network for Research in Infectious Diseases [REIPI RD16/0016/0001 and 0003], which is co-financed by the European Regional Development Fund ?A way to achieve Europe?, Operative Programme Intelligent Growth 2014?2020.

Multiple sclerosis (MS) is an autoimmune demyelinating disease of the human central nervous system (CNS), mainly affecting young adults. Among the immunomodulatory disease modifying treatments approved up to date to treat MS, IFN-β remains to be one of the most widely prescribed for the Relapsing-Remitting (RR) variant of the disease, although its mechanism of action is still partially understood. RR-MS variant is characterized by phases with increasing neurological symptoms (relapses) followed by periods of total or partial recovery (remissions), which implies the existence of immunomodulatory agents to promote the relapsing-to-remitting transition. Among these agents, it has been described the immunosuppressive role of a heterogeneous population of immature myeloid cells, namely the myeloid-derived suppressor cells (MDSCs) during the clinical course of the experimental autoimmune encephalomyelitis (EAE), the most used MS model to study RRMS. However, it is still unknown how the current MS disease modifying treatments, e.g. IFN- β, affects to MDSCs number or activity. Our present results show that a single injection of IFN-β at the onset of the clinical course reduces the severity of the EAE, enhancing the presence of MDSCs within the smaller demyelinated areas. Moreover, the single dose of IFN-β promotes MDSC immunosuppressive activity both in vivo and in vitro, augmenting T cell apoptosis. Finally, we show that IFN-ß preserves MDSC immaturity, preventing their differentiation to mature and less suppressive myeloid cell subsets. Taking together, all these data add new insights into the mechanism of IFN-β treatment in EAE and point to MDSCs as a putative endogenous mediator of its beneficial role in this animal model of MS., This work was supported by the Spanish Ministerio de Economía, Industria y Competitividad-MINEICO (SAF2012-40023; SAF2016-77575-R; RD12-0032-12; RD16-0015-0019; PI15-00963; PI18/00357, partially financed by F.E.D.E.R.: European Union, “Una Manera de hacer Europa”), the Spanish Research Council/Consejo Superior de Investigaciones Científicas-CSIC (CSIC-2015201023), ADEM-TO (Spain) and ARSEP Foundation (France). DC, RL-G and IM-D were financed by SESCAM; CM-J holds a predoctoral Research Training contract from MINEICO (BES-2013-062630, –associated to SAF2012-40023 and PI15-00963) and is currently hired under SAF2016-77577-R. Dr. Clemente's group was sponsored by Aciturri Aeronáutica SLU, Vesuvius Ibérica LA and Fundación Galletas Coral.

Autism spectrum disorders are early onset neurodevelopmental disorders characterized by deficits in social communication and restricted repetitive behaviors, yet they are quite heterogeneous in terms of their genetic basis and phenotypic manifestations. Recently, de novo pathogenic mutations in DYRK1A, a chromosome 21 gene associated to neuropathological traits of Down syndrome, have been identified in patients presenting a recognizable syndrome included in the autism spectrum. These mutations produce DYRK1A kinases with partial or complete absence of the catalytic domain, or they represent missense mutations located within this domain. Here, we undertook an extensive biochemical characterization of the DYRK1A missense mutations reported to date and show that most of them, but not all, result in enzymatically dead DYRK1A proteins. We also show that haploinsufficient Dyrk1a+/− mutant mice mirror the neurological traits associated with the human pathology, such as defective social interactions, stereotypic behaviors and epileptic activity. These mutant mice present altered proportions of excitatory and inhibitory neocortical neurons and synapses. Moreover, we provide evidence that alterations in the production of cortical excitatory neurons are contributing to these defects. Indeed, by the end of the neurogenic period, the expression of developmental regulated genes involved in neuron differentiation and/or activity is altered. Therefore, our data indicate that altered neocortical neurogenesis could critically affect the formation of cortical circuits, thereby contributing to the neuropathological changes in DYRK1A haploinsufficiency syndrome., This work was supported by grants from the Spanish Ministry of Economia, Industria y Competitividad (MINECO) (BFU2016-81887-REDT, SAF2013-46676-P and SAF2016-77971-R to M.L.A. and BFU2013-44513 and BFU2016-76141 to S.L.), the Secretariat of Universities and Research-Generalitat de Catalunya (2014SGR674), and the Fundación Alicia Koplowitz (Spain). The group of S.L. acknowledges the support of the MINECO Centro de Excelencia Severo Ochoa Programme and of the CERCA Programme (Generalitat de Catalunya). G.S-E. and M.P.S. acknowledge the support of the National Institute of Neurological Disorders and Stroke of the National Institutes of Health(USA) (P01NS097197) and the Spanish Instituto de Salud Carlos III (ISCIII), (PI13/00865, Fondo Europeo de Desarrollo Regional -FEDER “A way of making Europe”, Spain). M.J.B. is supported by the CIBERER, an initiative of the ISCIII. J.A., E.B. and S.N. were supported by MINECO predoctoral fellowships (AP2012-3064 and BES2011-047472) and G.S-E. by a predoctoral fellowship from the Fundación Conchita Rábago, Spain.

White root rot, caused by the soilborne fungus Rosellinia necatrix, is an important constraint to production for a wide range of woody crop plants such as avocado trees. The current methods of detection of white root rot are based on microbial and molecular techniques, and their application at orchard scale is limited. In this study, physiological parameters provided by imaging techniques were analyzed by machine learning methods. Normalized difference vegetation index (NDVI) and normalized canopy temperature (canopy temperature - air temperature) were tested as predictors of disease by several algorithms. Among them, logistic regression analysis (LRA) trained on NDVI data showed the highest sensitivity and lowest rate of false negatives. This algorithm based on NDVI could be a quick and feasible method to detect trees potentially affected by white root rot in avocado orchards., This work was supported by a grant from Consejería de Economía, Innovación, Ciencia y Empleo, Junta de Andalucía (P12-AGR-0370) to M. Barón, Ministerio de Economía, Industria y Competitividad Consejo Superior de Investigaciones Científicas and European Regional Development Fund (RECUPERA 2020/20134R060) to M. Barón, and Agencia de Innovación y Desarrollo de Andalucía (AGL14-52518-C2-1-R). C. Vida was recipient of a Ph.D. fellowship from the FPI program (MINECO).

The nervous system is a target and a source of steroids. Neuroactive steroids are steroids that target neurons and glial cells. They include hormonal steroids originated in the peripheral glands, steroids locally synthesized by the neurons and glial cells (neurosteroids) and synthetic steroids, some of them used in clinical practice. Here we review the mechanisms of synthesis, metabolism and action of neuroactive steroids, including the role of epigenetic modifications and the mitochondria in their sex specific actions. We examine sex differences in neuroactive steroid levels under physiological conditions and their role in the establishment of sex dimorphic structures in the nervous system and sex differences in its function. In addition, particular attention is paid to neuroactive steroids under pathological conditions, analyzing how pathology alters their levels and their role as neuroprotective factors, considering the influence of sex in both cases., This research was supported by grants from MIUR Progetto Eccellenza. We also acknowledge support from Fondazione CARIPLO (Rif.2012-0547), Colciencias (Convocatoria 777-2017, Contract #120377757021), Ministerio de Economía, Industria y Competitividad (MINECO), Spain (grant numbers BFU2014–51836-C2-1-R and BFU2017-82754-R), CIBER de Investigación Biomédica en Red de Fragilidad y Envejecimiento Saludable (CIBERFES), Instituto de Salud Carlos III, Madrid, Spain and Fondos Feder.

The aim of this study was to determine the impact of three different technological processes and in vitro digestion on the profile and content of betalains and inhibition of angiotensin I converting enzyme (ACE) in red beet. Betalains were analysed by the micro-HPLC-TOF-MS/MS method, while ACE inhibition was determined by an in vitro assay. In the tested samples, thirteen betalains were identified, constituting nine betacyanins, two betaxanthins and two betalain precursors. The applied technological processes (boiling, fermentation and microwave-vacuuming treatment) reduced the content of betalain by 42–70% in the obtained products. The contribution of betalains released from red beet products after in vitro digestion was detected within the range of 0.001–0.10%. The applied treatments reduced the inhibition of ACE by the non-digested red beet products, while in vitro digestion caused an increase in ACE inhibition in the case of the digested solid red beet products., This research was supported by the statutory funds of the Department of Chemistry and Biodynamics of Food of the Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences in Olsztyn and by the Ministry of Economy, Industry and Competitiveness (MEIC, grant number AGL2017-83718-R).

[Background] In winter barley plants, vernalization and photoperiod cues have to be integrated to promote flowering. Plant development and expression of different flowering promoter (HvVRN1, HvCO2, PPD-H1, HvFT1, HvFT3) and repressor (HvVRN2, HvCO9 and HvOS2) genes were evaluated in two winter barley varieties under: (1) natural increasing photoperiod, without vernalization, and (2) under short day conditions in three insufficient vernalization treatments. These challenging conditions were chosen to capture non-optimal and natural responses, representative of those experienced in the Mediterranean area., [Results] In absence of vernalization and under increasing photoperiods, HvVRN2 expression increased with day-length, mainly between 12 and 13 h photoperiods in our latitudes. The flowering promoter gene in short days, HvFT3, was only expressed after receiving induction of cold or plant age, which was associated with low transcript levels of HvVRN2 and HvOS2. Under the sub-optimal conditions here described, great differences in development were found between the two winter barley varieties used in the study. Delayed development in ‘Barberousse’ was associated with increased expression levels of HvOS2. Novel variation for HvCO9 and HvOS2 is reported and might explain such differences., [Conclusions] The balance between the expression of flowering promoters and repressor genes regulates the promotion towards flowering or the maintenance of the vegetative state. HvOS2, an ortholog of FLC, appears as a strong candidate to mediate in the vernalization response of barley. Natural variation found would help to exploit the plasticity in development to obtain better-adapted varieties for current and future climate conditions., Study financially supported by the Spanish Ministry of Economy, Industry and Competitiveness (Projects AGL2013–48756-R, including a scholarship granted to AM, and AGL2016–80967-R). The funding body did not play any role in the design of the study, collection, analysis, interpretation of the data or in writing the manuscript. We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI).

Plants respond to the loss of vertical growth re-orientating their affected organs. In trees, this phenomenon has received the scientific attention due to its importance for the forestry industry. Nowadays it is accepted that auxin distribution is involved in the modulation of the tilting response, but how this distribution is controlled is not fully clear. Auxin transporters that determine the spatio-temporal auxin distribution in radiate pine seedlings exposed to 45° of tilting were identified. Additionally, based on indications for an intimate plant hormone crosstalk in this process, IAA and JA contents were evaluated. The experiments revealed that expression of the auxin transporters was down-regulated in the upper half of the tilted stem, while being induced in the lower half. Moreover, transporter-coding genes were first induced at the apical zone of the stem. IAA was consistently redistributed toward the lower half, which is in accordance with the expression profile of the auxin transporters. In contrast, JA was mainly accumulated in the upper half of tilted stems. Finally, lignin content and monomeric composition were analyzed in both sides of stem and along the time course of tilting. As expected, lignin accumulation was higher at the lower half of stem at longer times of tilting. However, the most marked difference was the accumulation of the H-lignin monomer in the lower half, while the G-lignin unit was more dominant in the upper half. Here, we provide detailed insight in the distribution of IAA and JA, affecting the lignin composition during the tilting response in Pinus radiata seedlings., FONDECYT #11121170 supported this work. P. Ramos acknowledges ‘Núcleo Científico Multidisciplinario’ from Universidad de Talca. S. Pollmann is supported by project BFU2014-55575-R by the Spanish Ministry of Economy, Industry and Competitiveness (MINECO). David Caparrós-Ruiz was supported by the Spanish Ministry of Economy and Competitiveness (AGL2014-58126-R and SEV-2015-0533 from the Severo Ochoa Programme for Centres of Excellence in R&D 2016–2019) and by CERCA Programme/Generalitat de Catalunya, and the Suport a Grups de Recerca Programme (SGR2014-1434) from the Autonomous Government of Catalonia.

This work highlights the utility of brush border membrane vesicles (BBMV) from the pig small intestine as a reliable model for gathering information about the reaction mechanisms involved in the human digestion of dietary carbohydrates. Concretely, the elucidation of the transgalactosylation mechanism of pig BBMV to synthesize prebiotic galacto-oligosaccharides (GOS) is provided, unravelling the catalytic activity of mammalian small intestinal β-galactosidase towards the hydrolysis of GOS. This study reveals that pig BBMV preferably synthesizes GOS linked by β-(1 → 3) bonds, since major tri- and disaccharide were produced by the transfer of a galactose unit to the C-3 of the non-reducing moiety of lactose and to the C-3 of glucose, respectively. Therefore, these results point out that dietary GOS having β-(1 → 3) as predominant glycosidic linkages could be more prone to hydrolysis by mammalian intestinal digestive enzymes as compared to those linked by β-(1 → 2), β-(1 → 4), β-(1 ↔ 1) or β-(1 → 6). Given that these data are the first evidence on the transglycosylation activity of mammalian small intestinal glycosidases, findings contained in this work could be crucial for future studies investigating the structure-small intestinal digestibility relationship of a great variety of available prebiotics, as well as for designing tailored fully non-digestible GOS., This work has been financed by the Spanish Ministry of Economy, Industry and Competitiveness (Project AGL2017-84614-C2-1-R). Julio-González thanks the governorship of Bolivar-Colombia and CeiBA Foundation for the scholarship granted in the project “Bolívar Gana con Ciencia”.

Pain is a prevalent complex medical problem, characterized by physically debilitating and mentally destabilizing conditions. Current pain therapeutics mainly include non-steroidal anti-inflammatory drugs and narcotics (opioids), but they exhibit limitations in efficacy, unwanted side effects and the problem of drug abuse. To overcome these issues, the discovery of different molecular players within pain pathways could lead to new opportunities for therapeutic intervention. Among other strategies, peptides could be powerful pharmaceutical agents for effective opioid-free medications for pain treatment. This review is a compendium of representative non-opioid analgesic peptides acting directly or indirectly at different ion channels and receptors distributed in nociceptive pathways. They include peptides targeting Ca, Na and K voltage-gated ion channels, the neuronal nicotinic receptors (nAChR), transient receptor potential channels (TRP), and different non-opioid G-protein coupled receptors (GPCRs), like the calcitonin gen-related peptide (CGRP), cannabinoid, bradykinin and neurotensin receptors, among others. Peptides engineered from protein-protein interactions among pain-related receptors and regulatory proteins also led to new therapeutic approaches for pain management. Following some successful examples, already in the clinics or under clinical trials, the improved understanding of pain mechanisms, and the advances in peptide permeation and/or delivery, could afford new analgesic peptides in the near future., This work was supported by the Spanish Ministry of Economy and Competitiveness [SAF 2015-66275-C2-R]; and Consejo Superior de Investigaciones Científicas, Spain (201580E073, 201880E109).

The study was partially presented as a poster in the 53rd Annual Meeting of the European Association for the Study of Diabetes, Lisbon 2017., The role of insulin-degrading enzyme (IDE), a metalloprotease with high affinity for insulin, in insulin clearance remains poorly understood. OBJECTIVE: This study aimed to clarify whether IDE is a major mediator of insulin clearance, and to define its role in the etiology of hepatic insulin resistance., [Methods] We generated mice with liver-specific deletion of Ide (L-IDE-KO) and assessed insulin clearance and action., [Results] L-IDE-KO mice exhibited higher (~20%) fasting and non-fasting plasma glucose levels, glucose intolerance and insulin resistance. This phenotype was associated with ~30% lower plasma membrane insulin receptor levels in liver, as well as ~55% reduction in insulin-stimulated phosphorylation of the insulin receptor, and its downstream signaling molecules, AKT1 and AKT2 (reduced by ~40%). In addition, FoxO1 was aberrantly distributed in cellular nuclei, in parallel with up-regulation of the gluconeogenic genes Pck1 and G6pc. Surprisingly, L-IDE-KO mice showed similar plasma insulin levels and hepatic insulin clearance as control mice, despite reduced phosphorylation of the carcinoembryonic antigen-related cell adhesion molecule 1, which upon its insulin-stimulated phosphorylation, promotes receptor-mediated insulin uptake to be degraded., [Conclusion] IDE is not a rate-limiting regulator of plasma insulin levels in vivo., This work was supported by grants from the Ministerio de Economía, Industria y Competitividad: SAF2014-58702-C2-1-R and SAF2016-77871-C2-1-R to ICC; SAF2014-58702-C2-2-R and SAF2016-77871-C2-2-R to GP; supported by the EFSD European Research Programme on New Targets for Type 2 Diabetes supported by an educational research grant from MSD to ICC and GP; the National Institutes of Health: R01-DK054254, R01-DK083850 and RO1-HL-112248 to SMN, and R01-GM115617 to MAL; and the American Diabetes Association: Career Development Award 7-11-CD-13 to MAL.

[Objectives] To describe the population pharmacokinetics of fosfomycin for patients with bacteraemic urinary tract infection (BUTI). The analysis identified optimal regimens on the basis of pharmacodynamic targets and assessed the adequacy of Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) susceptibility breakpoints for Escherichia coli., [Methods] Data of 16 patients with BUTI caused by multidrug-resistant E. coli (FOREST clinical trial) received intravenous fosfomycin (4 g every 6 hours) were analysed. A population pharmacokinetic analysis was performed, and Monte Carlo simulations were undertaken using 4 g every 6 hours and 8 g every 8 hours. The probability of pharmacodynamic target attainment was assessed using pharmacodynamic targets for E. coli for static effect, 1-log drop in bacterial burden and resistance suppression., [Results] Sixty-four plasma samples were collected over a single dosing interval (day 2 or 3 after starting fosfomycin treatment). Fosfomycin concentrations were highly variable. Pharmacodynamic target attainment analysis showed mild improvement by increasing fosfomycin dosing (4 g every 6 hours vs. every 8 hours). These dosages showed success for decreasing 1-log bacterial burden in 89% to 96% (EUCAST breakpoints) and 33% to 54% (CLSI breakpoints) of patients, but they were unable to reach bacterial resistance suppression targets., [Conclusions] Fosfomycin concentrations are highly variable—a fact partially explained by renal impairment. The present work supports the use of 4 g every 6 hours as an effective regimen for the treatment of non–critically ill patients with BUTI caused by multidrug-resistant E. coli, as higher dosages might increase toxicity but may not significantly increase efficacy. The current information may suggest that fosfomycin susceptibility breakpoints need to be reappraised., Supported in part by the Ministerio de Economía y Competitividad, Instituto de Salud Carlos III (PI13/01282 and PI16/01824), Spain; and by Plan Nacional de I+D+i 2013-2016 and Instituto de Salud Carlos III, Subdirección General de Redes y Centros de Investigación Cooperativa, Ministerio de Economía, Industria y Competitividad, Spanish Network for Research in Infectious Diseases (REIPI RD16/0015/0010; RD16/0016/0001), cofinanced by European Development Regional Fund ‘A way to achieve Europe,’ Operative Program Intelligent Growth 2014–2020. FDP was supported by a VPPIUS fellowship from the University of Seville. WWH was supported by a National Institute of Health Research Clinician Scientist award (CS/08/08).

[Background]: The Pantosteus plebeius-nebuliferus species-group is a group of freshwater fishes distributed in endo- and exorheic drainage basins in the Mexican Sierra Madre Occidental mountain range system and central North Mexico. The geological history of this region is considered an important factor in explaining the evolutionary history of low vagility animals like freshwaters fishes. The aim of this study was to examine the phylogenetic relationships and describe the evolutionary history of the species-group. We hypothesized that the genetic structure and distribution of the main clades of Pantosteus plebeius-nebuliferus are associated with the geological history of Northern Mexico. To this end, we obtained DNA sequences of mitochondrial and nuclear genes and performed phylogenetic and phylogeographic analyses. Divergence time estimation and ancestral area reconstruction were also carried out to propose a biogeographical hypothesis, and species boundaries within the species-group were also tested., [Results]: We identified four clades within the Pantosteus plebeius-nebuliferus species-group in both markers. Divergence ranged from 5.9% to 9.2% for cytb and 0.1% to 0.9% for GHI. We observed significant genetic structure and no shared haplotypes between clades. We estimated that the clades diverged during the last 5.1 Myr, with a biogeographic scenario suggesting eight vicariant and four dispersal events through the historic range of the species-group. We found that the best species-delimitation model is when four species are assumed, which correspond to the main clades. We identified nine evolutionary significance units (ESUs), pertinent to the conservation of the group, each representing populations present in distinct drainage basins., [Conclusions]: The evolutionary history of the Pantosteus plebeius-nebuliferus species-group is characterized by vicariant post-dispersal processes, linked to geological changes in the Sierra Madre Occidental and central Northern Mexico since the Pliocene. This is congruent with biogeographic patterns described for other co-distributed fish species. We propose a new phylogenetic hypothesis for the species-group, clarifying the taxonomy of this evolutionarily complex group. Our results suggest that the species-group consists of at least four clades with independent evolutionary histories, two of which may represent new undescribed species. Our identification of ESUs provides a basis upon which conservation measures can be developed for the species-group., This work was partial supported by the Coordinación de la Investigación Científica (CIC-UMSNH 2014), the Ministerio de Economía y Competitividad of Spain (grant reference CGL2013-41375-P). Both resources were used to finance fieldwork, laboratory (DNA isolation and amplification of molecular markers) and sequencing service, as well as for the publication of this work. The Consejo Nacional de Ciencia y Tecnología CONACYT with sabbatical grant and the scholarship to ODD (grant number: 260596) and DKCS (number grant: 254270/411621) respectively.

Chia mucilage has been considered as a reliable source of soluble fibre, with the ability to form highly aqueous viscous dispersions at low concentrations; however, its effects in the gastrointestinal tract have never been assessed. Therefore, the present work aims to study the relationship between apparent viscosity of chia mucilage at three concentrations (0.3, 0.5 and 0.8%) and growth and metabolic activity of human gut microbiota by using a dynamic gastrointestinal model, simgi®. Results showed differences in viscosities between mucilage concentrations before chia feeding and in the first stage of the digestion; however, in the descending colon, these differences disappear and were dependent on the shear applied. On a microbiological level, the crude chia mucilage affects the growth of some intestinal bacterial groups, such as Enterococcus spp and Lactobacillus spp. In conclusion, the chia mucilage concentrations do not affect the physical properties of the intestine (viscosity) but could affect the colonic microbial growth., This work was carried out with the financial support of National Fund for Scientific and Technological Development FONDECYT Project 11150307, Chile; the Spanish Ministry of Economy, Industry, and Competitiveness (AGL2015-64522-C2-1), Spain and the Comunidad de Madrid Program (ALIBIRD-CM S2013/ABI-2728), Spain.

The atmosphere-land-ocean dynamics of semivolatile organic compounds in polar regions is poorly understood, also for the abundant and ubiquitous polycyclic aromatic hydrocarbons (PAHs). We report the concentrations and fluxes of PAHs in a polar coastal ecosystem (Livingston Island, Antarctica). From late spring (December 2014) to late summer (February 2015), we sampled air, snow, coastal seawater, plankton, and the fugacity in soils and snow. The concentrations of PAHs in seawater were low but increased during the austral summer. The PAH concentrations in snow were significantly higher than in coastal seawater. Soil-air fugacity ratios showed a net volatilization of PAH when soils were covered with lichens, and close to air-soil equilibrium for bare soils. Concentrations in surface snow were also close to equilibrium with atmospheric PAHs. Conversely, there was a net diffusive deposition of PAHs to coastal seawater during late spring, but a net volatilization from seawater during late summer. Volatilization fluxes were correlated with seawater temperature and salinity, consistent with a key role of snowmelt to the fluxes and dissolved phase concentrations during the austral summer. The comprehensive assessment provided here shows that the fugacity amplification in snow is transferred to soils and coastal seawater supporting PAH concentrations and fluxes., Inorganic nutrient analyses were performed by Mrs. M. I. Abad in coordination with Dr. E. Berdalet (Head of the Service) at the “Nutrient Analysis Service” of the ICM (CSIC). We thank F. Cerqueria and A. Martinez for support with DNA analysis. This work was supported by Spanish Ministry of Science to P.C. through a predoctoral fellowhip, by Fundación BBVA award to M.V.C., European Commission to A.C. through a Marie Curie international outgoing fellowship, and by the Spanish MINECO through projects REMARCA (CTM2012- 34673) and ISOMICS (CTM2015-65691-R). The research group of Global Change and Genomic Biogeochemistry is supported by the Catalan Government (2017SGR800).

This study analyzes the pharmacokinetic variability of piperacillin in non-critically ill patients with Enterobacteriaceae bloodstream infections (EBSI) and explores predicted clinical outcomes and piperacillin-related neurotoxicity under different renal conditions. Hospitalized, non-critically ill patients treated with piperacillin–tazobactam for EBSI were included. Four serum samples per patient were collected and analyzed. A population pharmacokinetic model was developed using the Pmetrics package for R. Monte Carlo simulations of various dosage regimens of 4 g piperacillin, administered q8 h or q12 h by short (0.5 h) or long (4 h) infusion, following the different glomerular filtration rate (GFR) categories used to classify chronic kidney disease (Kidney Disease: Improving Global Outcomes, KDIGO) to determine the probability of target attainment (PTA) using a free drug concentrations above the minimal inhibitory concentration (fT &gt, MIC) of 50% for efficacy and targets for piperacillin-associated neurotoxicity. Twenty-seven patients (102 samples) were included. Extended piperacillin infusions reached a PTA &gt, 90% (50%fT &gt, MIC) within the susceptibility range, although a loading dose did not greatly improve the expected outcome. Long infusions reduced the expected toxicity in patients with severe renal impairment. The study supports the use of extended infusions of piperacillin in non-critically ill patients with EBSI. No benefits of a loading dose were expected in our population. Finally, extended infusions may reduce the risk of toxicity in patients with severe renal impairment.

Maqui-berry is characterised by presenting a high concentration of (poly)phenols, accounting anthocyanins (cyanidin and delphinidin) for over 85% of the total. These coloured flavonoids have demonstrated potential neurological activity, but the evidence of their antinociceptive properties is scarce. In order to cover this gap, different doses (suitable for human administration) of a maqui-berry powder (1.6% anthocyanin), using enteral and parenteral routes of administration, were compared at central and peripheral levels using a nociceptive pain model (formalin test) in mice. Gastric damage analysis as possible adverse effects of analgesic and anti-inflammatory drugs was also explored. Dose-antinociceptive response was confirmed using both routes of administration and in both neurogenic and inflammatory phases of the formalin test, without gastric damage. In conclusion, these preliminary data provide evidence of pharmacological properties of maqui-berry to alleviate nociceptive pain., This work was partially supported by Consejo Nacional de Ciencia y Tecnologıa (CONACYT) grant numbers 226454 and 256448 and Institutional projects numbers INPNC123280.0 and INP-NC17073.0. Also, was partially funded by the Spanish MINECO through Research Project AGL2016-75332-C2-1-R (AEI/FEDER, UE), the Spanish MICIN through Research Project PID2019-104212RD-100, and VA by a FPI [grant number BES-2017-079754

Streptococcus pneumoniae is a pathogen responsible for millions of deaths worldwide. Currently, the available vaccines for the prevention of S. pneumoniae infections are the 23-valent pneumococcal polysaccharide-based vaccine (PPV-23) and the pneumococcal conjugate vaccines (PCV10 and PCV13). These vaccines only cover some pneumococcal serotypes (up to 100 different serotypes have been identified) and are unable to protect against non-vaccine serotypes and non-encapsulated pneumococci. The emergence of antibiotic-resistant non-vaccine serotypes after these vaccines is an increasing threat. Therefore, there is an urgent need to develop new pneumococcal vaccines which could cover a wide range of serotypes. One of the vaccines most characterized as a prophylactic alternative to current PPV-23 or PCVs is a vaccine based on pneumococcal protein antigens. The choline-binding proteins (CBP) are found in all pneumococcal strains, giving them the characteristic to be potential vaccine candidates as they may protect against different serotypes. In this review, we have focused the attention on different CBPs as vaccine candidates because they are involved in the pathogenesis process, confirming their immunogenicity and protection against pneumococcal infection. The review summarizes the major contribution of these proteins to virulence and reinforces the fact that antibodies elicited against many of them may block or interfere with their role in the infection process. This research was funded by Ministerio de Economía, Industria y Competitividad (Grant number SAF2017-83388). Sí

Departamento de Biotecnología, CLECs are a group of molecules of the superfamily of C-type lectin domain containing receptors. Several receptors of this group have been described in humans and mice, as well as in other species. Many of them are expressed in immune cells, and have been shown to be involved in immune response modulation. Several molecules of this family have been proposed as targets for antigen delivery, which is what drew our interest in studying them in the pig. In this species only CLEC7A (also named Dectin-1) had been studied in some detail, and we have characterized two other members, CLEC12A and CLEC12B. Another molecule of interest was CLEC4A, for which we could not find any expression studies in pigs, although up to 5 different mRNA variants had been predicted by bioinformatics analysis of the genomic sequence in databases. Here we present an analysis of the expression of CLEC4A mRNA isoforms in tissues and blood cell subsets by PCR. Cloning and sequencing of cDNA of the different isoforms allowed us to identify all 5 predicted isoforms, in addition to characterizing 5 additional isoforms expressed in alveolar macrophages, of which the form coding for a canonical CLEC4A molecule was apparently the most abundant isoform. Analysis of the sequences of the 10 cDNA cloned allowed us to determine their exon usage and to identify their potential for protein coding. Only 8 of the characterized variants contain a recognizable coding sequence. The sequences of the coded proteins show both proteins with a putative transmembrane segment, and others without this feature, as well as proteins with the complete CLEC domain or with diverse truncated forms. We also prove that most of the cDNA can drive protein expression upon transfection in the CHO cell line. We discuss the relevance that the diversity of products of the porcine CLEC4A gene may have in the regulation of the expression and function of these molecules., This work was supported by grant AGL2015-66187-P from the Ministry of Economy, Industry and Competitiveness of Spain.

18 p.-8 fig.-1 tab., The interaction between the T‐lymphocyte costimulatory molecule ICOS and its ligand (ICOS‐L) is needed for efficient immune responses, but expression levels are tightly controlled, as altered expression of ICOS or ICOS‐L may lead to immunodeficiency, or favor autoimmune diseases and tumor growth. Using cells of mouse B cell lymphoma (M12.C3) and melanoma (B16), or hamster CHO cells transfected with various forms of mouse ICOS‐L, and ICOS+ T cell lines, we show that, within minutes, ICOS induces significant downmodulation of surface ICOS‐L that is largely mediated by endocytosis and trans‐endocytosis. So, after interaction with ICOS+ cells, ICOS‐L was found inside permeabilized cells, or in cell lysates, with significant transfer of ICOS from ICOS+ T cells to ICOS‐L‐expressing cells, and simultaneous loss of surface ICOS by the T cells. Data from cells expressing ICOS‐L mutants show that conserved, functionally important residues in the cytoplasmic domain of mouse ICOS‐L (Arg300, Ser307 and Tyr308), or removal of ICOS‐L cytoplasmic tail have minor effect on its internalization. Internalization was dependent on temperature, and was partially dependent on actin polymerization, the GTPase dynamin, protein kinase C, or the integrity of lipid rafts. In fact, a fraction of ICOS‐L was detected in lipid rafts. On the other hand, proteinase inhibitors had negligible effects on early modulation of ICOS‐L from the cell surface. Our data add a new mechanism of control of ICOS‐L expression to the regulation of ICOS‐dependent responses., Supported by Grants PI13/01809 (to J.M.R.), PI13/02153 and PI16CIII/00012 (to P.P.) from “Acción Estratégica en Salud, Plan Estatal I+D+i”, Ministerio de Economía, Industria y Competitividad(MINECO, Spain) and by the Associazione Italiana Ricerca sul Cancro(Grant IG20714, AIRC, Milan), Fondazione Amici di Jean (Torino), and Fondazione Cariplo (2017-0535) (to U.D.).

24 p.-10 fig.-3 tab., D-amino acid oxidase (DAAO) is an enzyme that catalyzes the oxidation of D-amino acids generating H2O2. The enzymatic chimera formed by DAAO bound to the choline-binding domain of N-acetylmuramoyl-L-alanine amidase (CLytA) induces cytotoxicity in several pancreatic and colorectal carcinoma and glioblastoma cell models. In the current work, we determined whether the effect of CLytA-DAAO immobilized in magnetic nanoparticles, gold nanoparticles, and alginate capsules offered some advantages as compared to the free CLytA-DAAO. Results indicate that the immobilization of CLytA-DAAO in magnetic nanoparticles increases the stability of the enzyme, extending its time of action. Besides, we compared the effect induced by CLytA-DAAO with the direct addition of hydrogen peroxide, demonstrating that the progressive generation of reactive oxygen species by CLytA-DAAO is more effective in inducing cytotoxicity than the direct addition of H2O2. Furthermore, a pilot study has been initiated in biopsies obtained from pancreatic and colorectal carcinoma and glioblastoma patients to evaluate the expression of the main genes involved in resistance to CLytA-DAAO cytotoxicity. Based on our findings, we propose that CLytA-DAAO immobilized in magnetic nanoparticles could be effective in a high percentage of patients and, therefore, be used as an anti-cancer therapy for pancreatic and colorectal carcinoma and glioblastoma., This research was funded by Foundation for the Promotion of Health and Biomedical Research of Valencia Region (FISABIO), grant number UGP-19-063, and co-supported by PRECIPITA crowdfunding platform from Spanish Foundation for Science and Technology (FECYT) and donations from Association of women affected by breast cancer in Elche and the region (AMACMEC) to M.S., Miguel Servet Program from Instituto de Salud Carlos III (CP19/00095) to C.d.J.R. and grants from the Spanish Ministry of Economy, Industry and Competitiveness, grant numbers BIO2013-47684-R and BIO2016-79323-R, and the RETICS-FEDER RICET, RD16/0027/0010, to J.M.S. The CIBER of Enfermedades Respiratorias (CIBERES) is an initiative of the Spanish Instituto de Salud Carlos III and Spanish National Research Council (CSIC Grant 201820I132).

This work was supported by grants from the following institutions: Grant for international postdoctoral stay. Jose Castillejo Program (Grant CAS15/00257), Ministerio de Educación Cultura y Deporte. Gobierno de España. Proyecto de Investigación en Salud, Consejería de Salud y Familias (Grant PI-0139-2018). EU-ERDF-Instituto de Salud Carlos III (Grant PI19/01577). Proyectos dedesarrollo tecnológico, Ministerio de Ciencia e Innovación (Grant DTS19/00125). Ministerio de Economía, Industria y Competitividad (Grant RTC-2019-007329-1).

Stress caused by drought and salinity may compromise growth and productivity of olive (Olea europaea L.) tree crops. Several studies have reported the use of beneficial rhizobacteria to alleviate symptoms produced by these stresses, which is attributed in some cases to the activity of 1-aminocyclopropane-1-carboxylic acid deaminase (ACD). A collection of beneficial olive rhizobacteria was in vitro screened for ACD activity. Pseudomonas sp. PICF6 displayed this phenotype and sequencing of its genome confirmed the presence of an acdS gene. In contrast, the well-known root endophyte and biocontrol agent Pseudomonas simiae PICF7 was defective in ACD activity, even though the presence of an ACD-coding gene was earlier predicted in its genome. In this study, an unidentified deaminase was confirmed instead. Greenhouse experiments with olive ‘Picual’ plants inoculated either with PICF6 or PICF7, or co-inoculated with both strains, and subjected to drought or salt stress were carried out. Several physiological and biochemical parameters increased in stressed plants (i.e., stomatal conductance and flavonoids content), regardless of whether or not they were previously bacterized. Results showed that neither PICF6 (ACD positive) nor PICF7 (ACD negative) lessened the negative effects caused by the abiotic stresses tested, at least under our experimental conditions., This research was funded by the Spanish Ministerio de Economía, Industria y Competitividad/Agencia Estatal de Investigación, grant number AGL2016-75729-C2-1-R, and Junta de Andalucía (Consejería de Economía, Innovación y Ciencia), grant number P12-AGR-0667, both co-financed by the European Regional Development Fund (ERDF).

Acetylation is known to regulate the activity of cytosolic phosphoenolpyruvate carboxykinase (PCK1), a key enzyme in gluconeogenesis, by promoting the reverse reaction of the enzyme (converting phosphoenolpyruvate to oxaloacetate). It is also known that the histone acetyltransferase p300 can induce PCK1 acetylation in cells, but whether that is a direct or indirect function was not known. Here we initially set out to determine whether p300 can acetylate directly PCK1 in vitro. We report that p300 weakly acetylates PCK1, but surprisingly, using several techniques including protein crystallization, mass spectrometry, isothermal titration calorimetry, saturation-transfer difference nuclear magnetic resonance and molecular docking, we found that PCK1 is also able to acetylate itself using acetyl-CoA independently of p300. This reaction yielded an acetylated recombinant PCK1 with a 3-fold decrease in kcat without changes in Km for all substrates. Acetylation stoichiometry was determined for 14 residues, including residues lining the active site. Structural and kinetic analyses determined that site-directed acetylation of K244, located inside the active site, altered this site and rendered the enzyme inactive. In addition, we found that acetyl-CoA binding to the active site is specific and metal dependent. Our findings provide direct evidence for acetyl-CoA binding and chemical reaction with the active site of PCK1 and suggest a newly discovered regulatory mechanism of PCK1 during metabolic stress., P. L. -M. was funded by a PhD fellowship from the ‘‘la Caixa’’ Foundation and received financial support from the Universidad de Zaragoza, Fundación Bancaria Ibercaja, and Fundación CAI (CM 1/16) during his stay at the UW–Madison. We acknowledge Ministerio de Economía, Industria y Competitividad for grant AGL2015-66177 (J. A. C.), the University of Zaragoza for grant UZ-2015-BIO-01 (J. A. C.), and OTRI (University of Zaragoza), Ministerio de Ciencia, Innovación y Universidades for grants CTQ2013-44367-C2-2-P and BFU2016-75633-P (R. H. -G.) and BFU2016-78232-P (A. V. -C.), Gobierno de Aragón for grants E34_R17 and LMP58_18 (R. H. -G.) and E45_17R (J. A. C and A. V. -C.) and FEDER (2014–2020) funds for “Building Europe from Aragón”. R. H. -G. and A. V. -C. thank the ARAID foundation for support. The research leading to these results also received funding from the FP7 (2007–2013) under BioStruct-X (grant agreement N283570 and BIOSTRUCTX_5186). We also thank the ALBA synchrotron radiation source, in particular, the beamline XALOC facility. J. A. and I. D. acknowledge access to UEA Faculty of Science Research Facilities. J. A. acknowledges funding support from the Biotechnology and biological Sciences Research Council (BBSRC; BB/P010660/1) and the Spanish Ministry of Science, Innovation and Universities through grant PID2019-109395GB-100. T. H. acknowledges a BBSRC DTP studentship. Financial support from the Universidad de Sevilla (Acciones Especiales del VI Plan Propio de Investigación y Transferencia) is also acknowledged. This work was also supported by National Institutes of Health (NIH) Grant GM065386 (J. M. D.), NIH National Research Service Award T32 GM007215 (J. B.) and the National Science Foundation Graduate Research Fellowship Program (NSF-GRFP) DGE-1256259 (J. B.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

The Irano-Turanian (IT) floristic region in Asia contains three biodiversity hotspots and has been a source of xerophytic taxa for neighboring regions. Despite its species richness and large territory, the evolutionary history and biogeography of this floristic region is poorly understood. Acanthophyllum, encompassing ca. 70 subshrubby mostly thorny-cushion species, is one of the important components of the steppe and mountain vegetation of this region. In this study, we investigate the tempo and mode of diversification as well as the biogeographic patterns of this genus. The ancestral area analysis suggests that Acanthophyllum originated east of the Zagros Mountains in the Miocene, from where it expanded westwards. A shift in the speciation rate in the late Pliocene (ca. 3.24 Ma) was detected, based on a nrDNA ITS tree, affecting the lineage of the largest section in the genus, Acanthophyllym sect. Oligosperma. Parallelisms with another IT species-rich genera in aspects such as largely coincident ranges and habitats, presence of a thorny-cushion life-form, similar areas reconstructed for their MRCA, and the inference of a shift in speciation in the region suggest common underlying abiotic and biotic factors or even drivers for diversification and speciation in the high elevation dry continental landscape of IT region., This study was supported by the grant CGL2017-88500-P (AEI/FEDER, EU) to GNF from the Spanish Ministry of Economy and Competitivity.

RNA splicing is widely dysregulated in cancer, frequently due to altered expression or activity of splicing factors (SFs). Microexons are extremely small exons (3–27 nucleotides long) that are highly evolutionarily conserved and play critical roles in promoting neuronal differentiation and development. Inclusion of microexons in mRNA transcripts is mediated by the SF Serine/Arginine Repetitive Matrix 4 (SRRM4), whose expression is largely restricted to neural tissues. However, microexons have been largely overlooked in prior analyses of splicing in cancer, as their small size necessitates specialized computational approaches for their detection. Here, we demonstrate that despite having low expression in normal nonneural tissues, SRRM4 is further silenced in tumors, resulting in the suppression of normal microexon inclusion. Remarkably, SRRM4 is the most consistently silenced SF across all tumor types analyzed, implying a general advantage of microexon down-regulation in cancer independent of its tissue of origin. We show that this silencing is favorable for tumor growth, as decreased SRRM4 expression in tumors is correlated with an increase in mitotic gene expression, and up-regulation of SRRM4 in cancer cell lines dose-dependently inhibits proliferation in vitro and in a mouse xenograft model. Further, this proliferation inhibition is accompanied by induction of neural-like expression and splicing patterns in cancer cells, suggesting that SRRM4 expression shifts the cell state away from proliferation and toward differentiation. We therefore conclude that SRRM4 acts as a proliferation brake, and tumors gain a selective advantage by cutting off this brake., This project was funded in part by a grant from the Plan Estatal de Investigación Científica y Técnica y de Innovación to L.S. (PGC2018-101271-B-I00, http://www.ciencia.gob.es). S.A.H. is supported by a Marie Skłodowska-Curie Individual Fellowship from the European Union’s Horizon 2020 research and innovation programme (MSCA-IF-2017-794629, http://ec.europa.eu/). X.H. is supported by a PhD fellowship from the Fundación Ramón Areces (http://www.fundacionareces.es). We acknowledge the support of the Spanish Ministry of Economy and Competitiveness, ‘Centro de Excelencia Severo Ochoa’, the CERCA Programme / Generalitat de Catalunya, and the Spanish Ministry of Economy, Industry and Competitiveness (MEIC) to the EMBL partnership.

In cyanobacteria and most green algae of the eukaryotic green lineage, the copper-protein plastocyanin (Pc) alternatively replaces the heme-protein cytochrome c6 (Cc6) as the soluble electron carrier from cytochrome f (Cf) to photosystem I (PSI). The functional and structural equivalence of ‘green’ Pc and Cc6 has been well established, representing an example of convergent evolution of two unrelated proteins. However, plants only produce Pc, despite having evolved from green algae. On the other hand, Cc6 is the only soluble donor available in most species of the red lineage of photosynthetic organisms, which includes, among others, red algae and diatoms. Interestingly, Pc genes have been identified in oceanic diatoms, probably acquired by horizontal gene transfer from green algae. However, the mechanisms that regulate the expression of a functional Pc in diatoms are still unclear. In the green eukaryotic lineage, the transfer of electrons from Cf to PSI has been characterized in depth. The conclusion is that in the green lineage, this process involves strong electrostatic interactions between partners, which ensure a high affinity and an efficient electron transfer (ET) at the cost of limiting the turnover of the process. In the red lineage, recent kinetic and structural modeling data suggest a different strategy, based on weaker electrostatic interactions between partners, with lower affinity and less efficient ET, but favoring instead the protein exchange and the turnover of the process. Finally, in diatoms the interaction of the acquired green-type Pc with both Cf and PSI may not yet be optimized., The Spanish Ministry of Economy, Industry and Competitiveness (BIO2015-64169-P and BIO2016-79930-R) and the Andalusian Government (PAIDI BIO-022). These grants were partially financed by the EU FEDER Program. C.C. is the recipient of a FPU Program fellowship (FPU16/04040; Spanish Ministry of Education, Culture and Sports). We acknowledge the support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI).

Transposable elements (TEs) are a rich source of genetic variability. Among TEs, miniature inverted-repeat TEs (MITEs) are of particular interest as they are present in high copy numbers in plant genomes and are closely associated with genes. MITEs are deletion derivatives of class II transposons, and can be mobilized by the transposases encoded by the latter through a typical cut-and-paste mechanism. However, MITEs are typically present at much higher copy numbers than class II transposons. We present here an analysis of 103 109 transposon insertion polymorphisms (TIPs) in 738 Oryza sativa genomes representing the main rice population groups. We show that an important fraction of MITE insertions has been fixed in rice concomitantly with its domestication. However, another fraction of MITE insertions is present at low frequencies. We performed MITE TIP-genome-wide association studies (TIP-GWAS) to study the impact of these elements on agronomically important traits and found that these elements uncover more trait associations than single nucleotide polymorphisms (SNPs) on important phenotypes such as grain width. Finally, using SNP-GWAS and TIP-GWAS we provide evidence of the replicative amplification of MITEs., RC is recipient of a Juan de la Cierva-formación contract from the Spanish Ministerio de Ciencia y Innovación. This work was supported by grants from the Spanish Ministerio de Economia, Industria y Competitividad (AGL2016-78992-R/FEDER) and Ministerio de Ciencia y Innovación (PID2019-106374RB-I00/AEI/10.13039/501100011033) to JMC. Computing resources have been provided by the Red Española de Supercomputación at the Pirineus machine (RES activity BCV-2019-2-0006).

LIS1 is one of the principal genes related to Type I lissencephaly, a severe human brain malformation characterized by an abnormal neuronal migration in the cortex during embryonic development. This is clinically associated with epilepsy and cerebral palsy in severe cases, as well as a predisposition to developing mental disorders, in cases with a mild phenotype. Although genetic variations in the LIS1 gene have been associated with the development of schizophrenia, little is known about the underlying neurobiological mechanisms. We have studied how the Lis1 gene might cause deficits associated with the pathophysiology of schizophrenia using the Lis1/sLis1 murine model, which involves the deletion of the first coding exon of the Lis1 gene. Homozygous mice are not viable, but heterozygous animals present abnormal neuronal morphology, cortical dysplasia, and enhanced cortical excitability. We have observed reduced number of cells expressing GABA-synthesizing enzyme glutamic acid decarboxylase 67 (GAD67) in the hippocampus and the anterior cingulate area, as well as fewer parvalbumin-expressing cells in the anterior cingulate cortex in Lis1/sLis1 mutants compared to control mice. The cFOS protein expression (indicative of neuronal activity) in Lis1/sLis1 mice was higher in the medial prefrontal (mPFC), perirhinal (PERI), entorhinal (ENT), ectorhinal (ECT) cortices, and hippocampus compared to control mice. Our results suggest that deleting the first coding exon of the Lis1 gene might cause cortical anomalies associated with the pathophysiology of schizophrenia., This work was supported by the Generalitat Valenciana (Prometeo/2018/041); MINECO/AEI/ERDF, EU, Spanish Ministry of Economy, Industry and Competitiveness, the Spanish State Research Agency and the European Union through the European Regional Development Fund (ERDF) – “Una manera de hacer Europa” (SAF2017-83702-R); and Instituto de Salud Carlos III (“RD16/001/0010”, co-funded by European Regional Development Fund/European Social Fund).

The human Alzheimer’s disease (AD) brain accumulates angiogenic markers but paradoxically, the cerebral microvasculature is reduced around Aß plaques. Here we demonstrate that angiogenesis is started near Aß plaques in both AD mouse models and human AD samples. However, endothelial cells express the molecular signature of non-productive angiogenesis (NPA) and accumulate, around Aß plaques, a tip cell marker and IB4 reactive vascular anomalies with reduced NOTCH activity. Notably, NPA induction by endothelial loss of presenilin, whose mutations cause familial AD and which activity has been shown to decrease with age, produced a similar vascular phenotype in the absence of Aß pathology. We also show that Aß plaque-associated NPA locally disassembles blood vessels, leaving behind vascular scars, and that microglial phagocytosis contributes to the local loss of endothelial cells. These results define the role of NPA and microglia in local blood vessel disassembly and highlight the vascular component of presenilin loss of function in AD., A.E.R.-N. was the recipient of a JdlC-F fellowship from the Spanish Ministry of Economy, Industry, and Competitiveness (MINEICO) (FJCI-2015-23708), M.I.A.-V., N.L.-U., and C.O.-d.S.L. were the recipient of an FPU fellowship from Spanish Ministry of Education, Culture, and Sport (respectively, FPU15/02898, FPU14-02115, and AP2010‐1598), and R.M.-D. was the recipient of a “Sara Borrell” fellowship from ISCIII (CD09/0007). Work was supported by grants to A.P. by the Spanish MINEICO, ISCIII, and FEDER (SAF2012‐33816, SAF2015‐64111‐R, RTI2018-096629-B-100, SAF2017-90794-REDT, and PIE13/0004), by the regional Government of Andalusia (“Proyectos de Excelencia”, P12‐CTS‐2138 and P12‐CTS‐2232) co-funded by CEC and FEDER funds, and by the “Ayuda de Biomedicina 2018”, Fundación Domingo Martínez; J.Vitorica: Instituto de Salud Carlos III (ISCiii) of Spain, co-financed by FEDER funds from European Union (PI18/01556) by La Marató-TV3 Foundation grant 20141431; by CIBERNED (CB06/05/0094); and by Junta de Andalucia Consejería de Economía y Conocimiento through grant US-1262734; A.G.: Instituto de Salud Carlos III (ISCiii) of Spain, co-financed by FEDER funds from European Union, through grant PI18/01557; and by Junta de Andalucia Consejería de Economía y Conocimiento through grants UMA18-FEDERJA-211 and P18-RT-2233 co-financed by Programa Operativo FEDER 2014-2020.

MRCTLH (muskelin/RanBPM/CTLH) is a protein complex found in humans (MRCTLH) that is involved in the regulation of numerous cellular processes, such as gluconeogenesis, cell signaling, development, nuclear extrusion, cell morphology, or stability of different proteins. According to genomic data, all eukaryotes have similar protein complexes. In yeast, a similar protein complex named GID was found to be involved in the regulation of gluconeogenesis. LICC1 is a maize protein whose sequence resembles that of TWA1 in humans and GID8 in yeast, which are central components of the MRCTLH and GID complexes. LICC1 contains three highly conserved protein domains, LisH, CTLH, and CRA, typical of this protein family. Twa1 and gid8 are unique genes in human and yeast genomes. However, three copies of licc1 are present in the maize genome and multiple copies are present in other plant genomes. This result suggests the presence of multiple variants of the MRCTLH/GID complex in plants, which could increase its regulatory capacity. We also demonstrate here that LICC1 has the ability to interact with microtubules, similarly to the human TWA1. This interaction reinforces the idea that the LICC1 protein from maize, and its homologues in plants and, in general, the GID/MRCTLH complex in plants, can perform biological functions similar to those in humans and yeast., This work was supported by grants from the Spanish Government (BIO2001-1721), the Government of Catalonia (Generalitat de Catalunya) (2014SGR-1434) and the Spanish Ministry of Economy, Industry and Competitiveness “Severo Ochoa Programme for Centres of Excellence in R&D” 2016-2019 (SEV2015-0533)”.

16 Pág. Departamento de Reproducción Animal​, Mutations in splicing factors are recurrent somatic alterations identified in myelodysplastic syndromes (MDS) and they frequently coincide with mutations in epigenetic factors. About 25% of patients present concurrent mutations in such pathways, suggesting a cooperative role in the pathogenesis of MDS. We focused on the splicing factor U2AF1 involved in the recognition of the 3' splice site during pre-mRNA splicing. Using a CRISPR/Cas9 system, we created heterozygous mice with a carboxy-terminal truncated U2af1 allele (U2af1mut/+), studied the U2af1mut/+ hematopoietic system, and did not observe any gross differences in both young (12-13 weeks) and old (23 months) U2af1mut/+ mice, except for a reduction in size of approximately 20%. However, hematopoietic stem/progenitor cells lacked reconstitution capacity in transplantation assays and displayed an aberrant RNA splicing by RNA sequencing. We also evaluated U2af1mut/+ in conjunction with Tet2-deficiency. Novel double mutant U2af1mut/+Tet2-/- mice showed increased monogranulocytic precursors. Hematopoietic stem/progenitor cells were also enhanced and presented functional and transcriptomic alterations. Nonetheless, U2af1mut/+Tet2-/- mice did not succumb to MDS disease over a 6-month observation period. Collectively, our data suggest that cooperation between mutant U2af1 and Tet2 loss is not sufficient for MDS initiation in mice., This work is supported by SAF2017-82171-R from the Spanish Ministry of Economy, Industry, and Competitiveness. C.M.-V. is funded by a predoctoral fellowship from Generalitat Valenciana (ACIF/2018/255). C.G.-R. is funded by AECC predoctoral fellowship. A.S.-P. is supported by the Ramón y Cajal Program (RYC2015-17534).