Transcriptome sequencing identifies SPL7-regulated copper acquisition genes FRO4/FRO5 and the copper dependence of iron homeostasis in Arabidopsis

24 Pags., 9 Figs., 2 Tabls., with Supplemental Data (15 Figs., 3 Tabls., 1 Method, 1 Data Set).The transition metal copper (Cu) is essential for all living organisms but is toxic when present in excess. To identify Cu deficiency responses comprehensively, we conducted genome-wide sequencing-based transcript profiling of Arabidopsis thaliana wild-type plants and of a mutant defective in the gene encoding SQUAMOSA PROMOTER BINDING PROTEIN-LIKE7 (SPL7), which acts as a transcriptional regulator of Cu deficiency responses. In response to Cu deficiency, FERRIC REDUCTASE OXIDASE5 (FRO5) and FRO4 transcript levels increased strongly, in an SPL7-dependent manner. Biochemical assays and confocal imaging of a Cu-specific fluorophore showed that high-affinity root Cu uptake requires prior FRO5/FRO4-dependent Cu(II)-specific reduction to Cu(I) and SPL7 function. Plant iron (Fe) deficiency markers were activated in Cu-deficient media, in which reduced growth of the spl7 mutant was partially rescued by Fe supplementation. Cultivation in Cu-deficient media caused a defect in root-to-shoot Fe translocation, which was exacerbated in spl7 and associated with a lack of ferroxidase activity. This is consistent with a possible role for a multicopper oxidase in Arabidopsis Fe homeostasis, as previously described in yeast, humans, and green algae. These insights into root Cu uptake and the interaction between Cu and Fe homeostasis will advance plant nutrition, crop breeding, and biogeochemical research.We acknowledge postdoctoral fellowships to M.B. from the Alexander von Humboldt Foundation and the Spanish Ministry of Science and Innovation; a Deutsche Forschungsgemeinshaft Heisenberg fellowship and funding from the FRONTIERS program at the University of Heidelberg, Germany, and the European Union InP Public Health Impact of Long-Term, Low-Level Mixed Element Exposure in Susceptible Population Strata (FOOD-CT-2006-016253) to U.K.; a grant from the National Science Foundation (IOS-0919739) to E.L.C.; a postdoctoral fellowship from the Spanish Foundation of Science and Technology (MEC-FECYT) to D.C.; National Institutes of Health Grant GM42143 to S.S.M.; and support from the University of California, Los Angeles–Department of Energy Institute for Genomics and Proteomics under Contract DE-FC02-02ER63421 to M.P.Peer reviewe

Detection of Neutralizing Antibodies against SARS-CoV-2 Post-Vaccination in Health Care Workers of a Large Tertiary Hospital in Spain by Using a Rapid Test LFIC and sVNT-ELISA

The presence of neutralizing antibodies (NAbs) against SARS-CoV-2 represent a surrogate marker of immunologic protection in populations at high risk of infection such as healthcare workers caring for hospitalized patients with COVID-19. As recommended by CDC and the European CDC, the use of rapid diagnostic tests during population-based evaluations offers an opportunity to identify individuals with serologic evidence of natural infection or who have undergone vaccination. We carried out a cross-sectional study to assess the presence of neutralizing antibodies against SARS-CoV-2 among medical providers at an intensive care unit of a large referral hospital in Alicante, Spain. In addition, we tested for the presence of neutralizing antibodies compared to serum of uninfected individuals from a Biobank. We were also interested in evaluating the use of a rapid lateral flow immunochromatography (LFIC) test against a surrogate ELISA viral neutralization test (sVNT). This rapid test demonstrated a specificity of 1.000 95% CI (0.91–1.00) and the sensitivity of 0.987 95% CI (0.93–1.00). The negative predictive value was 95%. After six months, this rapid test demonstrated that those immunized with two doses of BioNTech/Pfizer vaccine, maintained optimal levels of neutralizing antibodies. We concluded that all Health Care Workers develop NAbs and the use of this rapid immunochromatographic test represents a potential tool to be used in population-based studies to detect serological antibody responses to vaccination. Vaccination policies could benefit from this tool to assess additional doses of vaccine or boosters among high-risk populations.This work was supported by an advisory contract from the University of Alicante with reference WENZHOU-OJABIO1-21T and no one reports personal fees received from the biotech company

Endocannabinoid regulation of amyloid-induced neuroinflammation

The modulation of endocannabinoid (EC) levels and the activation of cannabinoid receptors are seen as promising therapeutic strategies in a variety of diseases, including Alzheimer’s disease (AD). We aimed to evaluate the effect of the pharmacological and genetic inhibiton of anandamide (AEA)-degrading enzyme in a mouse model of AD (5xFAD). Pharmacological inhibition of the fatty acid amide hydrolase (FAAH) had little impact on the expression of key enzymes and cytokines as well as on the cognitive impairment and plaque deposition and gliosis in 5xFAD mice. CB1 blockade exacerbated inflammation in this transgenic mouse model of AD. The genetic inactivation of FAAH led to increases in the expression of inflammatory cytokines. At the same time, FAAH-null 5xFAD mice exhibited a behavioral improvement in spatial memory that was independent of the level of anxiety and was not CB1-mediated. Finally, mice lacking FAAH showed diminished soluble amyloid levels, neuritic plaques and gliosis. These data reinforce the notion of a role for the endocannabinoid system in neuroinflammation and open new perspectives on the relevance of modulating endocannabinoid levels in the inflammed brain.pre-print687 K

Efecto de la concentración salina y el regimen hídrico sobre la germinación, emergencia y establecimiento de alfalfa (Medicago sativa L.)

Los suelos halo-hidromórficos de la región húmeda y subhúmeda de Argentina suman aproximadamente 20 millones de hectáreas, un amplio porcentaje de estos presentan vegetación nativa de baja producción de biomasa y consumo de agua. Entender el funcionamiento de especies forrajeras perennes como alfalfa en estos sitios es de suma importancia para mejorar la producción de los sistemas allí ubicados. El objetivo de este trabajo fue comprender y cuantificar el efecto de la salinidad y el contenido hídrico edáfico sobre la germinación, emergencia y establecimiento temprano del cultivo de alfalfa. Así se realizaron tres experimentos a diferentes escalas: (E1) experimento en cámara de germinación, con el fin de determinar, bajo condiciones controladas, el efecto de diferentes condiciones de salinidad y disponibilidad hídrica sobre la germinación y emergencia de alfalfa; (E2) un experimento en invernáculo para evaluar el efecto del estrés hídrico (por exceso o por defecto) sobre plantas de alfalfa en la etapa de establecimiento temprano; y (E3) un experimento a campo, con el fin de identificar y jerarquizar indicadores ambientales determinantes de la emergencia, establecimiento y producción del cultivo de alfalfa. Se halló que en ambientes halo-hidromórficos la concentración salina fue la principal variable determinante del crecimiento y supervivencia de la especie durante las etapas tempranas con límites cercanos a 4 dS.m-1. El exceso o déficit hídrico afectó al stand de plantas, reduciendo a la mitad el número de individuos. La plasticidad fenotípica de la alfalfa frente a diferentes tenores salinos, así como al estrés hídrico (por exceso o por defecto) determina que sea una especie para considerar cuando se pretende producir biomasa de alta calidad, distribuida en el tiempo, en este tipo de ambientes.The halo-hydromorphic soils of the humid and sub-humid region of Argentina cover approximately 20 million hectares. A large proportion of them have native vegetation with low biomass production and water consumption. Understanding the functioning of perennial forage species such as alfalfa in these sites is of paramount importance to improve the productivity of the grazing systems located there. The objective of this work is to understand and quantify the effect of salinity and soil water content on the germination, emergence and early establishment of alfalfa. Three experiments were carried out at different scales: (E1) germination chamber experiment, to determine, under controlled conditions, the effect of different salinity and water availability conditions on the germination and emergence of alfalfa; (E2) a greenhouse experiment to assess the effect of water stress (by insufficiency or excess) on alfalfa plants in the early establishment stage; and (E3) a field experiment, to identify and rank environmental indicators that determine the emergence, establishment and production of alfalfa. It was found that, in halo-hydromorphic environments, saline concentration was the main variable affecting alfalfa growth and survival during the early stages, where salinity levels close to 4 dS.m-1. Water stress by excess or water deficit affected plant stands, halving the number of individuals. The phenotypic plasticity of alfalfa against different salinity and water conditions, makes it a species to consider to produce high quality biomass, distributed over time, in this type of environment.EEA Marcos JuárezFil: Bertram, Nicolas. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juárez; ArgentinaFil: Alfonso, C. Asociación Universitaria de Enseñanza Agropecuaria Superior. Becaria INTA-AUDEASFil: Grande, S. Universidad Nacional de Río Cuarto. Facultad de Ciencias Agrarias; ArgentinaFil: Chiacchiera, Sebastián. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juárez; ArgentinaFil: Ohanian, Alfredo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Agrarias; ArgentinaFil: Bonvillani, Julieta. Universidad Nacional de Río Cuarto. Facultad de Ciencias Agrarias; ArgentinaFil: Conde, Maria Belen. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juárez; ArgentinaFil: Angeletti, Fernando Ramón. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juárez; Argentin

Cannabinoid CB2 Receptors Modulate Microglia Function and Amyloid Dynamics in a Mouse Model of Alzheimer's Disease

The distribution and roles of the cannabinoid CB2 receptor in the CNS are still a matter of debate. Recent data suggest that, in addition to its presence in microglial cells, the CB2 receptor may be also expressed at low levels, yet biologically relevant, in other cell types such as neurons. It is accepted that the expression of CB2 receptors in the CNS is low under physiological conditions and is significantly elevated in chronic neuroinflammatory states associated with neurodegenerative diseases such as Alzheimer's disease. By using a novel mouse model (CB2EGFP/f/f), we studied the distribution of cannabinoid CB2 receptors in the 5xFAD mouse model of Alzheimer's disease (by generating 5xFAD/CB2EGFP/f/f mice) and explored the roles of CB2 receptors in microglial function. We used a novel selective and brain penetrant CB2 receptor agonist (RO6866945) as well as mice lacking the CB2 receptor (5xFAD/CB2-/-) for these studies. We found that CB2 receptors are expressed in dystrophic neurite-associated microglia and that their modulation modifies the number and activity of microglial cells as well as the metabolism of the insoluble form of the amyloid peptide. These results support microglial CB2 receptors as potential targets for the development of amyloid-modulating therapies.Funding The present work has been supported by a grant from Ministerio de Ciencia e Innovacion (ref PID2019-108992RB-I00 and ref PID2019-107548RB-I00) to JR and PG, respectively, by the Basque Government (ref IT1230-19) to PG, and the Research and Education Component of the Advancing a Healthier Wisconsin Endowment at the Medical College of Wisconsin to CJH

Spatial evolution of an AMD stream in the Iberian Pyrite Belt: process characterization and control factors on the hydrochemistry

This paper presents hydrochemical data of an AMD stream in the Iberian Pyrite Belt, obtained from its source, in the Poderosa Mine Portal, till its confluence at the Odiel River. The main objective is to establish potential interdependent relations between sulfate and metals’ loads and the following physical-chemical variables: pH, electric conductivity (EC), redox potential (EH), and dissolved oxygen (O2). All the parameters show a global increasing tendency since the tunnel’s exit to the confluence at Odiel River. The TDS and EC are two relevant exceptions. They behave similarly, showing a decreasing trend and a strong inflection that describes a minimum immediately after the discharging point. The spatial analysis combined with statistical tools put in evidence the typical AMD processes and the respective physical-chemical implications. Inputs with distinctive hydrochemical signatures impose relevant modifications in the Poderosa creek waters. This indicates low hydrochemical inertia and high vulnerability to external stimulus.Financial support for this research was provided by DGCICYT National Plan, project CGL2010-21268-C02-01 and the Andalusian Autonomous Government Excellence Projects, Project RNM-6570

Impact of SARS-Cov-2 infection in patients with hypertrophic cardiomyopathy : results of an international multicentre registry

To describe the natural history of SARS-CoV-2 infection in patients with hypertrophic cardiomyopathy (HCM) compared with a control group and to identify predictors of adverse events. Three hundred and five patients [age 56.6 ± 16.9 years old, 191 (62.6%) male patients] with HCM and SARS-Cov-2 infection were enrolled. The control group consisted of 91 131 infected individuals. Endpoints were (i) SARS-CoV-2 related mortality and (ii) severe clinical course [death or intensive care unit (ICU) admission]. New onset of atrial fibrillation, ventricular arrhythmias, shock, stroke, and cardiac arrest were also recorded. Sixty-nine (22.9%) HCM patients were hospitalized for non-ICU level care, and 21 (7.0%) required ICU care. Seventeen (5.6%) died: eight (2.6%) of respiratory failure, four (1.3%) of heart failure, two (0.7%) suddenly, and three (1.0%) due to other SARS-CoV-2-related complications. Covariates associated with mortality in the multivariable were age {odds ratio (OR) per 10 year increase 2.25 [95% confidence interval (CI): 1.12-4.51], P = 0.0229}, baseline New York Heart Association class [OR per one-unit increase 4.01 (95%CI: 1.75-9.20), P = 0.0011], presence of left ventricular outflow tract obstruction [OR 5.59 (95%CI: 1.16-26.92), P = 0.0317], and left ventricular systolic impairment [OR 7.72 (95%CI: 1.20-49.79), P = 0.0316]. Controlling for age and sex and comparing HCM patients with a community-based SARS-CoV-2 cohort, the presence of HCM was associated with a borderline significant increased risk of mortality OR 1.70 (95%CI: 0.98-2.91, P = 0.0600). Over one-fourth of HCM patients infected with SARS-Cov-2 required hospitalization, including 6% in an ICU setting. Age and cardiac features related to HCM, including baseline functional class, left ventricular outflow tract obstruction, and systolic impairment, conveyed increased risk of mortality

The Spanish Infrared Camera onboard the EUSO-BALLOON (CNES) flight on August 24, 2014

The EUSO-Balloon (CNES) campaign was held during Summer 2014 with a launch on August 24. In the gondola, next to the Photo Detector Module (PDM), a completely isolated Infrared camera was allocated. Also, a helicopter which shooted flashers flew below the balloon. We have retrieved the Cloud Top Height (CTH) with the IR camera, and also the optical depth of the nonclear atmosphere have been inferred with two approaches: The first one is with the comparison of the brightness temperature of the cloud and the real temperature obtained after the pertinent corrections. The second one is by measuring the detected signal from the helicopter flashers by the IR Camera, considering the energy of the flashers and the location of the helicopter

Caveolin-1-Enhanced Motility and Focal Adhesion Turnover Require Tyrosine-14 but Not Accumulation to the Rear in Metastatic Cancer Cells

Caveolin-1 is known to promote cell migration, and increased caveolin-1 expression is associated with tumor progression and metastasis. In fibroblasts, caveolin-1 polarization and phosphorylation of tyrosine-14 are essential to promote migration. However, the role of caveolin-1 in migration of metastatic cells remains poorly defined. Here, caveolin-1 participation in metastatic cell migration was evaluated by shRNA targeting of endogenous caveolin-1 in MDA-MB-231 human breast cancer cells and ectopic expression in B16-F10 mouse melanoma cells. Depletion of caveolin-1 in MDA-MB-231 cells reduced, while expression in B16-F10 cells promoted migration, polarization and focal adhesion turnover in a sequence of events that involved phosphorylation of tyrosine-14 and Rac-1 activation. In B16-F10 cells, expression of a non-phosphorylatable tyrosine-14 to phenylalanine mutant failed to recapitulate the effects observed with wild-type caveolin-1. Alternatively, treatment of MDA-MB-231 cells with the Src family kinase inhibitor PP2 reduced caveolin-1 phosphorylation on tyrosine-14 and cell migration. Surprisingly, unlike for fibroblasts, caveolin-1 polarization and re-localization to the trailing edge were not observed in migrating metastatic cells. Thus, expression and phosphorylation, but not polarization of caveolin-1 favor the highly mobile phenotype of metastatic cells