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Cancer immunotherapies strive to overcome tumor-induced immune suppression and activate antitumor immune responses. Although cytotoxic T lymphocytes (CTLs) play a pivotal role in this process, natural killer (NK) cells have also demonstrated remarkable tumor-killing abilities, given their ability to discriminate tumor cells from normal cells and mediate specific antitumoral cytotoxicity. NK cells activation depends on a balance between activation and inhibition signals from several ligands/receptors. Among them, MICA/NKG2D axis is a master regulator of NK activation. MHC class I chain-related polypeptide A (MICA) expression is upregulated by many tumor cell lines and primary tumors and serves as a ligand for the activating NK group 2D (NKG2D) receptor on NK cells and subpopulations of T cells. However, cancer cells can cleave MICA, making it soluble and de-targeting tumor cells from NK cells, leading to tumor immune escape.In this study, we present ICOVIR15KK-MICAMut, an oncolytic adenovirus (OAdv) armed with a transgene encoding a non-cleavable MICA to promote NK-mediated cell-killing capacity and activate the immune response against cancer cells. We first demonstrated the correct MICA overexpression from infected cells. Moreover, our MICA-expressing OAdv promotes higher NK activation and killing capacity than the non-armed virus in vitro. In addition, the armed virus also demonstrated significant antitumor activity in immunodeficient mice in the presence of human PBMCs, indicating the activation of human NK cells. Finally, OAdv-MICA overexpression in immunocompetent tumor-bearing mice elicits tumor-specific immune response resulting in a greater tumor growth control.In summary, this study highlights the significance of NK cells in cancer immunotherapy and presents an innovative approach using a modified oncolytic virus to enhance NK cell activation and antitumor immune response. These findings suggest promising potential for future research and clinical applications.
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Adenoviridae , Subfamilia K de Receptores Similares a Lectina de Células NK , Humanos , Animales , Ratones , Adenoviridae/genética , Subfamilia K de Receptores Similares a Lectina de Células NK/genética , Activación de Linfocitos , Genes MHC Clase I , Escape del TumorRESUMEN
Tests using algae and/or cyanobacteria, invertebrates (crustaceans) and fish form the basic elements of an ecotoxicological assessment in a number of regulations, in particular for classification of a substance as hazardous or not to the aquatic environment according to the Globally Harmonised System of Classification and Labelling of Chemicals (GHS-CLP) (GHS, 2022) and the REACH regulation (Registration, Evaluation, Authorisation and Restriction of Chemicals, EC, 2006). Standardised test guidelines (TGs) of the Organisation for Economic Co-operation and Development (OECD) are available to address the regulatory relevant endpoints of growth inhibition in algae and cyanobacteria (TG 201), acute toxicity to invertebrates (TG 202), and acute toxicity in fish (TG 203). Applying these existing OECD TGs for testing two dimensional (2D) graphene nanoforms may require more attention, additional considerations and/or adaptations of the protocols, because graphene materials are often problematic to test due to their unique attributes. In this review a critical analysis of all existing studies and approaches to testing used has been performed in order to comment on the current state of the science on testing and the overall ecotoxicity of 2D graphene materials. Focusing on the specific tests and available guidance's, a complete evaluation of aquatic toxicity testing for hazard classification of 2D graphene materials, as well as the use of alternative tests in an integrated approach to testing and assessment, has been made. This information is essential to ensure future assessments generate meaningful data that will fulfil regulatory requirements for the safe use of this "wonder" material.
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Grafito , Organización para la Cooperación y el Desarrollo Económico , Animales , Pruebas de Toxicidad/métodos , Peces , InvertebradosRESUMEN
Nanobiomaterials (NBMs) are nanostructured materials for biomedical applications that can reach aquatic organisms. The short and long-term effects of these emerging contaminants are unknown in fish. The RTgill-W1 cell line has been proposed as a model to predict the acute toxicity of chemicals to fish (OECD Test Guideline nº 249). We assessed the applicability of this cell line to study the short and long-term toxicity of 15 NBMs based on hydroxyapatites (HA), lipid (LSNP/LNP), gold, iron oxide, carbon, poly l-Lactide acid (PLLA) fibers with Ag and poly (lactide-co-glycolide) acid. Two more rainbow trout cell lines (RTL-W1, from liver, and RTS-11, from spleen) were exposed, to identify possible sensitivity differences among cells. Exposures to a range of concentrations (0.78-100 µg/mL) lasted for 24 h. Additionally, the RTgill-W1 was used to perform long-term (28 d exposure) and recovery (14 d exposure/14 d recovery) assays. Cells were exposed to the 24 h-IC20 and/or to 100 µg/mL. A triple cytotoxicity assay was conducted. After 24 h, only PLLA Fibers-Ag showed cytotoxicity (IC50 < 100 µg/mL). However, the NBMs in general provoked concentration-dependent effects after long-term exposures, except the LSNPs. A recovery of viability was only observed for AuNPs, AuNRods, Fe3O4PEG-PLGA, MgHA-Collag_Scaffolds, Ti-HA and TiHA-Alg NPs.These results evidenced the need to test the long-term toxicity of NBMs and showed differences in cytotoxicity probably associated to different mechanisms of toxic action. The RTgill-W1 was useful to screen short and long-term toxicities of NBMs and appears as a promiseful model to assess possible toxicity of NBMs in fish.
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Nanopartículas del Metal , Oncorhynchus mykiss , Animales , Oro/metabolismo , Línea Celular , Oncorhynchus mykiss/metabolismo , Carbono/metabolismo , Hidroxiapatitas/metabolismo , LípidosRESUMEN
BACKGROUND AND AIM: Classically acute syndesmosis injuries have been treated using screws. A few years ago more flexible implants appeared evolving to current TighRope® Knotless. The primary aim of this study is to compare clinical and radiographic outcome of both implants. MATERIAL AND METHODS: From April 2019 to September 2020 68 patients diagnosed with acute syndesmosis injury were randomised to use screws or TighRope® Knotless. Syndesmosis reduction was assessed using bilateral CT potsoperatively. Outcomes were clinically and radiologically assessed at three, six, and twelve months after surgery. RESULTS: No significant differences were identified in the AOFAS Scale between groups at three months (83.1 vs. 81.80; P=1.03), nor at six (88.27 vs. 88; P=.26) or at twelve (93.03 vs. 92.10; P=.93). There were also no differences in Olerud-Molander scale at three (65 vs. 61.50; P=3.5), six (82.33 vs. 80.67; P=1.67) and twelve months (92.67 vs. 90; P=2.67). Likewise, there were no differences in rate of postoperative malreduction (no cases in both groups), loss of reduction (three cases in screw group vs. four in TighRope® Knotless group, P=.54) or complications (P=1). CONCLUSIONS: Treatment of acute syndesmosis injuries with screws or the TighRope® Knotless implant is similar in both clinical and radiological results.
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BACKGROUND AND AIM: Classically acute syndesmosis injuries have been treated using screws. A few years ago more flexible implants appeared evolving to current TighRope® Knotless. The primary aim of this study is to compare clinical and radiographic outcome of both implants. MATERIAL AND METHODS: From April 2019 to September 2020 68 patients diagnosed with acute syndesmosis injury were randomized to use screws or TighRope® Knotless. Syndesmosis reduction was assessed using bilateral CT potsoperatively. Outcomes were clinically and radiologically assessed at three, six, and twelve months after surgery. RESULTS: No significant differences were identified in the AOFAS Scale between groups at three months (83.1 vs. 81.80; P=1.03), nor at six (88.27 vs. 88; P=.26) or at twelve (93.03 vs. 92.10; P=,93). There were also no differences in Olerud-Molander scale at three (65 vs. 61.50; P=3.5), six (82.33 vs. 80.67; P=1.67) and twelve months (92.67 vs. 90; P=2.67). Likewise, there were no differences in rate of postoperative malreduction (no cases in both groups), loss of reduction (three cases in screw group vs. four in TighRope® Knotless group, P=.54) or complications (P=1). CONCLUSIONS: Treatment of acute syndesmosis injuries with screws or the TighRope® Knotless implant is similar in both clinical and radiological results.
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Biocidal substances and their environmental relevant metabolites are highly toxic for fish. However, an important scarcity of toxicity data for metabolites is recognised. This article provides new data about the toxicity to fish of these compounds and evaluates the potential use of fish cell lines as screening tools to assess the acute toxicity of these compounds in fish. To this aim, acute toxicity of 7 substances was tested in Oncorhynchus mykiss (OECD TG203) and cytotoxicity of 16 substances was assessed in fish cell lines from two species; Poeciliopsis lucida (PLHC-1) and O. mykiss (RTH-149, RTG-2 and RTgill-W1) performing three cytotoxicity tests: Alamar-Blue, 5-carboxyfluorescein diacetate, acetoxymethyl ester and Neutral Red Uptake. Additionally, in vitro and in vivo data from the LIFE-COMBASE database were included in a dataset finally comprising 33 biocides and 14 metabolites. Hazard data were categorized into 4 toxicity groups, according to the intervals established in Regulation (EC) 1272/2008. Finally, the Spearman correlation test was performed and coincidences between in vitro-in vivo data established. In vitro and in vivo results revealed a high positive correlation, with a complete coincidence for 56.5% of the substances, a 2% of false positives (non-toxic in vivo) and a 13% of false negatives (toxic in vivo) for the 4 toxicity categories. However, when results were grouped in toxic or non-toxic coincidence was obtained for 85% of the substances. In conclusion, although fish denote a greater sensitivity, the use of at least two fish cell lines and three cytotoxicity endpoints appear to be valid approaches for fish acute toxicity screening of biocides and their metabolites.
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Ciprinodontiformes , Desinfectantes , Oncorhynchus mykiss , Pruebas de Toxicidad Aguda , Contaminantes Químicos del Agua , Animales , Línea Celular , Contaminantes Químicos del Agua/toxicidadRESUMEN
Plants have evolved different mechanisms to increase their tolerance to aluminum (Al) toxicity and low pH in the soil. The Zn finger transcription factor SENSITIVE TO PROTON RHIZOTOXICITY1 (STOP1) plays an essential role in the adaptation of plants to Al and low pH stresses. In this work, we isolated the ScSTOP1 gene from rye (Secale cereale L.), which is located on chromosome 3RS. The ectopic expression of ScSTOP1 complements the Arabidopsis stop1 mutation in terms of root growth inhibition due to Al and pH stress, as well as phosphate starvation tolerance, suggesting that rye ScSTOP1 is a functional ortholog of AtSTOP1. A putative STOP1 binding motif was identified in the promoter of a well-known STOP1 target from rye and Arabidopsis and was later corroborated by genomic DAP-seq analyses. Coexpression analyses verified that ScSTOP1 activated the promoter of ScALMT1. We have also identified a putative phosphorylatable serine in STOP1 that is phylogenetically conserved and critical for such activation. Our data indicated that ScSTOP1 also regulated Al and pH tolerance in rye.
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Aluminio/toxicidad , Proteínas de Arabidopsis/metabolismo , Arabidopsis/efectos de los fármacos , Arabidopsis/metabolismo , Transportadores de Anión Orgánico/metabolismo , Secale/metabolismo , Proteínas de Arabidopsis/genética , Cromosomas de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/genética , Concentración de Iones de Hidrógeno , Mutación/genética , Transportadores de Anión Orgánico/genética , Regiones Promotoras Genéticas/efectos de los fármacos , Regiones Promotoras Genéticas/genética , Secale/efectos de los fármacos , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Aiming for the introduction of stability requirements in nanofluids processing, an interface-based three-step method is proposed in this work. It is theory-based design framework for nanofluids that aims for a minimum tension at the solid-liquid interface by adjusting the polar and dispersive components of the base fluid to meet those of disperse nanomaterial. The method was successfully tested in the preparation of aqueous nanofluids containing single-walled carbon nanotubes that resulted to be stable and to provide good thermal properties, i.e. thermal conductivity increases by 79.5% and isobaric specific heat by 8.6% for a 0.087 vol.% load of nanotubes at 70 °C. Besides, a system for these nanofluids was modelled. It was found to be thermodynamically consistent and computationally efficient, providing consistent response to changes in the state variable temperature in a classical Molecular Dynamics environment. From an analysis of the spatial components of the heat flux autocorrelation function, using the equilibrium approach, it was possible to elucidate that heat conduction through the host fluid is enhanced by phonon propagation along nanotubes longitudinal axes. From an analysis of the structural features described by radial distribution functions, it was concluded that additional heat storage arises from the hydrophobic effect.
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BACKGROUND AND OBJECTIVE: Knowing whether a subject is conscious or not is a current challenge with a deep potential clinical impact. Recent theoretical considerations suggest that consciousness is linked to the complexity of distributed interactions within the corticothalamic system. The fractal dimension (FD) is a quantitative parameter that has been extensively used to analyse the complexity of structural and functional patterns of the human brain. In this study we investigate FD to assess whether it can discriminate between consciousness and different states of unconsciousness in healthy individuals. METHODS: We study 69 high-density electroencephalogram (hd-EEG) measurements after transcranial magnetic stimulation (TMS) in 18 healthy subjects progressing from wakefulness to non-rapid eye movement (NREM) sleep and sedation induced by different anaesthetic agents (xenon and propofol). We quantify the integration of thalamocortical networks by calculating the FD of a spatiotemporal voxelization obtained from the locations of all sources that are significantly activated by the perturbation (4DFD). Moreover, we study the temporal evolution of the evoked spatial distributions and compute a measure of the differentiation of the response by means of the Higuchi FD (HFD). Finally, a Fractal Dimension Index (FDI) of perturbational complexity is computed as the product of both quantities: integration FD (4DFD) and differentiation FD (HFD). RESULTS: We found that FDI is significantly lower in sleep and sedation when compared to wakefulness and provides an almost perfect intra-subject discrimination between conscious and unconscious states. CONCLUSIONS: These results support the combination of FD measures of cortical integration and cortical differentiation as a novel paradigm of tracking complex spatiotemporal dynamics in the brain that could provide further insights into the link between complexity and the brain's capacity to sustain consciousness.
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Estado de Conciencia , Electroencefalografía , Estimulación Magnética Transcraneal , Inconsciencia , Adolescente , Adulto , Anestesia , Encéfalo/diagnóstico por imagen , Femenino , Fractales , Voluntarios Sanos , Humanos , Masculino , Propofol , Procesamiento de Señales Asistido por Computador , Sueño , Vigilia , Xenón , Adulto JovenRESUMEN
The toxicity of mycotoxins is well recognized in mammals, but their effects on fish have received less attention. Moreover, in the last years several studies have reported that some mycotoxins may act as endocrine disruptors. The aim of this study was to determine the cytotoxic effects and endocrine activities of three mycotoxins: beauvericin, deoxynivalenol and ochratoxin-A. Cytotoxicity in two fish hepatoma and one mammalian hepatoma cell lines was determined by the AlamarBlue, Neutral Red Uptake and CFDA-AM assays. For the assessment of androgenic, estrogenic and thyroidal agonistic/antagonistic effects three cell lines stably expressing luciferase as reporter gene under the control of hormone receptors were used. Results showed that both fish and mammalian cell lines were very sensitive to the mycotoxins tested. OTA was the least toxic mycotoxin and DON and BEA showed similar acute toxicity. None of the three mycotoxins tested presented agonistic effects at the receptors studied, but all of them showed strong antagonistic effect at the thyroid receptor. BEA showed a weak antagonistic effect at the androgen receptor and OTA produced a biphasic dose-response curve at the estrogen receptor. The data obtained in this work are of high interest for aquaculture industries and for regulators.
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Depsipéptidos/toxicidad , Disruptores Endocrinos/toxicidad , Ocratoxinas/toxicidad , Oncorhynchus mykiss , Tricotecenos/toxicidad , Animales , Línea Celular , Concentración 50 Inhibidora , Ratas , Receptores Androgénicos/efectos de los fármacos , Receptores de Estrógenos/efectos de los fármacosRESUMEN
The current wide use of manufactured nanomaterials (MNs) is leading to the release of nanoparticles (NPs) to water bodies. Aquatic organisms, including fish, are exposed to low concentrations of NPs for long periods of time being necessary to develop laboratory toxicity tests reflecting realistic conditions. Additionally, today there is a demand of in vitro assays respecting the 3Rs principle. Thus, the main aim of this work was to stablish an in vitro tool for the assessment of long-term NPs ecotoxicity. Considering the key role of liver in detoxification, a rainbow trout liver cell line, RTL-W1, was used. CuO NPs were chosen to validate this tool taking into account their important production level. Cells were exposed for 21â¯days to 25 or 100⯵g CuO NPs/ml. Every seven days cells were split and one fourth of them transferred to a new plate with appropriate concentrations of NPs in culture medium. Lower concentrations of CuO NPs did not cause any deleterious effect, whereas higher concentrations led to significant mortality after 14â¯days and to the intracellular accumulation of Cu particles. Identical results were observed in cells exposed to CuSO4 at the same Cu concentrations. Therefore, the observed toxic effects might be mainly due to Cu2+ ions.
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Nanoestructuras/toxicidad , Pruebas de Toxicidad/métodos , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Cobre/toxicidad , Sulfato de Cobre/toxicidad , Oncorhynchus mykissRESUMEN
Endocrine disruptors (EDs) are natural or man-made chemicals that can affect the health of organisms by interfering with their normal hormonal functions. Many of these substances can cause their effects at very low doses and, considering the key role played by the endocrine system on development, organisms in early phases of growth (foetal, childhood, puberty) are especially sensitive to the action of EDs. In addition, when combined, they can show additive, antagonistic and synergistic activities. Taking all this into account it is essential to determine the presence of this kind of compounds in drinking water. Thus the main aim of the present study was to monitor the presence of substances with suspected or known endocrine activity in drinking water of the Madrid Region (MR) (Central Spain) and determine possible estrogenic, androgenic, or thyroidal activities. Water samples were collected at different times from a number of supply points that received water from reservoirs or rivers. The sampling point with the highest concentration of the analysed substances (up to 30 compounds) was DW1 (1203â¯ngâ¯L-1). This sampling point receives water from a drinking water treatment plant (DWTP) that serves the population from the south of the MR with treated water from the Tajuña River. DW2 was the second point with the highest concentration of the analysed substances (1021â¯ngâ¯L-1). DW2 receives water from one of the reservoirs in the north of the MR. The highest daily concentrations detected corresponded to the flame retardant Tris (2-chloroethyl)phosphate (TCEP) (266.55â¯ngâ¯L-1) and to the nonylphenol diethoxylate (188.57 ngâ¯L-1) at points DW1 and DW4, respectively, both of which are supplied with treated river water. None of the water samples exhibited androgenic, oestrogenic, or thyroidal activities in in vitro assays based on cells stably transfected with the receptors of interest and luciferase as reporter gene. These results demonstrate that water quality in the MR is high and does not present a health risk for the population, although the concentrations of some substances justify the need for local authorities to continually monitor the presence of these contaminants in order to implement any corrective measures if necessary.
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Agua Potable/química , Disruptores Endocrinos/análisis , Monitoreo del Ambiente/métodos , Calidad del Agua/normas , Andrógenos/análisis , Agua Potable/efectos adversos , Estrógenos/análisis , Humanos , España , Hormonas Tiroideas/análisis , Contaminantes Químicos del Agua/análisis , Abastecimiento de Agua/normasRESUMEN
Complex tasks require the learning and integration of multiple cognitive, sensory, and psychomotor skills for correct execution. Driving-related skills are developed step by step through the increase of mileage driven and the accumulation of practice in different traffic situations. The acquisition of these skills should be reflected in the brain structure. However, no previous studies have explored brain structural variations associated with driving experience. The aim of the present study was to investigate whether driving frequency, defined as average annual driving mileage, is related to neuroanatomical variations in gray matter (GM) volume and white matter (WM) integrity using voxel-based morphometry (VBM) and DTI-based fractional anisotropy (FA), respectively. We recruited 83 drivers with variable range of annual driving mileage and controlled for age, sex, handedness, IQ, time since the acquisition of driving license, use of motorcycles/mopeds and bicycles, perceived driving skills, and subjective probability of having an accident. Our results showed variations in white matter FA as a function of mileage driven. Driving experience was related to a significant increase of FA in parts of the right hemisphere superior and inferior longitudinal fasciculus, anterior thalamic radiation, forceps majors, inferior fronto-occipital fasciculus, and corticospinal tract. No significant differences were observed in gray matter volumes. FA variations were found in brain regions that have been associated with cognitive, visual, and motor processes necessary for skilled performance in driving. These results suggest that variations in white matter diffusivity can underlie the development of driving skills and safer driving.
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Conducción de Automóvil , Encéfalo/diagnóstico por imagen , Práctica Psicológica , Adulto , Encéfalo/anatomía & histología , Imagen de Difusión Tensora , Femenino , Sustancia Gris/anatomía & histología , Sustancia Gris/diagnóstico por imagen , Humanos , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Tamaño de los Órganos , Sustancia Blanca/anatomía & histología , Sustancia Blanca/diagnóstico por imagen , Adulto JovenRESUMEN
BACKGROUND AND AIMS: The brain reward system is key to understanding adolescent obesity in the current obesogenic environment, rich in highly appetising stimuli, to which adolescents are particularly sensitive. We aimed to examine the association between body fat levels and brain reward system responsivity to general (monetary) rewards in male and female adolescents. METHODS: Sixty-eight adolescents (34 females; mean age (s.d.)= 16.56 (1.35)) were measured for body fat levels with bioelectric impedance, and underwent a functional magnetic resonance imaging (fMRI) scan during the Monetary Incentive Delay (MID) task. The MID task reliably elicits brain activations associated with two fundamental aspects of reward processing: anticipation and feedback. We conducted regression analyses to examine the association between body fat and brain reward system responsivity during reward anticipation and feedback, while controlling for sex, age and socioeconomic status. We also analysed the moderating impact of sex on the relationship between fat levels and brain responsivity measures. Brain imaging analyses were corrected for multiple comparisons, with a cluster-defining threshold of P<0.001, and minimum cluster size of 38 contiguous voxels. RESULTS: Higher body fat levels were associated with lower activation of the primary somatosensory cortex (S1) and the supramarginal gyrus during reward feedback after controlling for key sociodemographic variables. Although we did not find significant associations between body fat and brain activations during reward anticipation, S1/supramarginal gyrus activation during feedback was linked to increased negative prediction error, that is, less reward than expected, in illustrative post hoc analyses. Sex did not significantly moderate the association between body fat and brain activation in the MID task. CONCLUSIONS: In adolescents, higher adiposity is linked to hypo-responsivity of somatosensory regions during general (monetary) reward feedback. Findings suggest that adolescents with excess weight have blunted activation in somatosensory regions involved in reward feedback learning.
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Tejido Adiposo/fisiología , Sobrepeso/fisiopatología , Recompensa , Corteza Somatosensorial/fisiología , Adolescente , Índice de Masa Corporal , Estudios Transversales , Femenino , Humanos , Masculino , Motivación/fisiología , Obesidad/epidemiología , Obesidad/fisiopatología , Sobrepeso/epidemiologíaRESUMEN
Plant roots have the potential capacity to grow almost indefinitely if meristematic and lateral branching is sustained. In a genetic screen we identified an Arabidopsis mutant showing limited root growth (lrg1) due to defects in cell division and elongation in the root meristem. Positional cloning determined that lrg1 affects an alpha-1,2-mannosyltransferase gene, LEW3, involved in protein N-glycosylation. The lrg1 mutation causes a synonymous substitution that alters the correct splicing of the fourth intron in LEW3, causing a mix of wild-type and truncated protein. LRG1 RNA missplicing in roots and short root phenotypes in lrg1 are light-intensity dependent. This mutation disrupts a GC-base pair in a three-base-pair stem with a four-nucleotide loop, which seems to be necessary for correct LEW3 RNA splicing. We found that the lrg1 short root phenotype correlates with high levels of reactive oxygen species and low pH in the apoplast. Proteomic analyses of N-glycosylated proteins identified GLU23/PYK10 and PRX34 as N-glycosylation targets of LRG1 activity. The lrg1 mutation reduces the positive interaction between Arabidopsis and Serendipita indica. A prx34 mutant showed a significant reduction in root growth, which is additive to lrg1. Taken together our work highlights the important role of N-glycosylation in root growth and development.
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Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Basidiomycota/fisiología , Manosiltransferasas/metabolismo , Peroxidasas/metabolismo , beta-Glucosidasa/metabolismo , Arabidopsis/crecimiento & desarrollo , Arabidopsis/fisiología , Arabidopsis/efectos de la radiación , Proteínas de Arabidopsis/genética , División Celular , Glicosilación , Concentración de Iones de Hidrógeno , Intrones/genética , Manosiltransferasas/genética , Meristema/genética , Meristema/crecimiento & desarrollo , Meristema/efectos de la radiación , Mutación , Peroxidasas/genética , Fenotipo , Raíces de Plantas/genética , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/efectos de la radiación , Proteómica , Empalme del ARN , Especies Reactivas de Oxígeno/metabolismo , Plantones/genética , Plantones/crecimiento & desarrollo , Plantones/efectos de la radiación , beta-Glucosidasa/genéticaRESUMEN
Risk assessment of engineered nanomaterials (ENMs) is being hindered by the sheer production volume of these materials. In this regard, the grouping and ranking of ENMs appears as a promising strategy. Here we sought to evaluate the usefulness of in vitro systems based on fish cell lines for ranking a set of ENMs on the basis of their cytotoxicity. We used the topminnow (Poeciliopsis lucida) liver cell line (PLHC-1) and the rainbow trout (Oncorhynchus mykiss) fibroblast-like gonadal cell line (RTG-2). ENMs were obtained from the EU Joint Research Centre repository. The size frequency distribution of ENM suspensions in cell culture media was characterized. Cytotoxicity was evaluated after 24 h of exposure. PLHC-1 cells exhibited higher sensitivity to the ENMs than RTG-2 cells. ZnO-NM was found to exert toxicity mainly by altering lysosome function and metabolic activity, while multi-walled carbon nanotubes (MWCNTs) caused plasma membrane disruption at high concentrations. The hazard ranking for toxicity (ZnO-NM > MWCNT ≥ CeO2-NM = SiO2-NM) was inversely related to the ranking in size detected in culture medium. Our findings reveal the suitability of fish cell lines for establishing hazard rankings of ENMs in the framework of integrated approaches to testing and assessment.
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Ecotoxicología/métodos , Nanotubos de Carbono/toxicidad , Dióxido de Silicio/toxicidad , Pruebas de Toxicidad/métodos , Animales , Línea Celular , Relación Dosis-Respuesta a Droga , Peces , Hepatocitos , Lisosomas/efectos de los fármacos , Medición de Riesgo/métodos , Dióxido de Silicio/farmacocinéticaRESUMEN
Three bacterial isolates were recovered from a disease outbreak with high mortality affecting brill Scophthalmus rhombus (Linnaeus, 1758). Moribund fish showed no external signs of disease, but plentiful haemorrhages were observed in liver. On the basis of phenotypic and genotypic characterization, the isolates were identified as Aliivibrio fischeri. The phenotypic profile of the isolates was basically similar to that of the type strain of this species, although some discrepancies were observed, mainly in the BIOLOG GN profile. The main cellular fatty acids of strain a591 were also consistent with this species. The highest 16S rDNA sequence similarities were recorded with the type strain of A. fischeri (99.07%); other Aliivibrio species showed similarity values below 96%. The highest sequence similarities with gyrB, rpoD and recA genes were also recorded with A. fischeri type strain (99.31, 98.99 and 95.29% similarity, respectively). DNA-DNA hybridization assays confirmed that these isolates belong to A. fischeri; levels of DNA relatedness were 73.5 to 86.2% with isolate a591 (reciprocal values of 86.9 to 99.04%). Finally, a virulence evaluation of the isolates using Senegalese sole fry was also performed; significant mortalities (100% mortality within 5 d) were recorded by intraperitoneal injection, but only with high doses of bacteria (2 × 106 cfu g-1 body weight).
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Aliivibrio fischeri/genética , Brotes de Enfermedades/veterinaria , Enfermedades de los Peces/microbiología , Peces Planos/microbiología , Infecciones por Bacterias Gramnegativas/veterinaria , Animales , Acuicultura , ADN Bacteriano/genética , Infecciones por Bacterias Gramnegativas/microbiología , Filogenia , Reacción en Cadena de la PolimerasaRESUMEN
Four Gram-negative bacterial isolates were recovered from 2 disease outbreaks that occurred in 2013 affecting European sea bass Dicentrarchus labrax fry and sea bream Sparus aurata adults. Main symptoms were erratic swimming, eroded fins and, in the sea bream outbreak, haemorrhages on the body surface; bacteria were always recovered from internal organs, almost in pure culture. On the basis of phenotypic characterization and 16S rRNA gene sequence analysis, the isolates were identified as Lacinutrix venerupis, a bacterium not previously reported as a fish pathogen. The highest 16S rDNA sequence similarities were recorded with the type strain of this species (99.9-100% similarity), while other species showed similarities below 97%, the closest relative being L. mariniflava (96.3% similarity). Phenotypic characterization showed some discrepancies with the L. venerupis type strain (mainly in BIOLOG GN profile); however, DNA-DNA hybridization assays with L. venerupis and L. mariniflava type strains confirmed that these isolates belong to the former species (levels of DNA relatedness were 98-100% and 38-50%, respectively). Finally, a virulence evaluation of the isolates using Senegalese sole Solea senegalensis fry was also performed; significant mortalities (80-100% mortality within 4 d) were recorded after intraperitoneal injection, but only with high doses of bacteria (107colony forming units fish-1). Further studies will be necessary to determine the importance of this species as a fish pathogen.
Asunto(s)
Lubina , Brotes de Enfermedades/veterinaria , Enfermedades de los Peces/microbiología , Infecciones por Flavobacteriaceae/veterinaria , Flavobacteriaceae/aislamiento & purificación , Dorada , Animales , ADN Bacteriano/genética , Flavobacteriaceae/genética , Infecciones por Flavobacteriaceae/microbiología , FilogeniaRESUMEN
OBJECTIVES: To estimate how many of the trauma patients admitted to ICU would be candidates for a secondary prevention programme for trauma related to alcohol or drug use by brief motivational intervention and to define what factors prevent that intervention being performed. METHODS: All 16-70year old trauma patients (n=242) admitted to ICU in 32 non-consecutive months (November 2011 to March 2015) were included in the study, coinciding with the implementation of a screening and brief motivational intervention programme for trauma patients related to substance consumption. The programme includes screening for exposure to substances at admission. Sociodemographic and clinical variables were collected prospectively. RESULTS: The screening for substances was not performed in 38 (15.7%) of all admitted patients. Of the patients screened, 101 (49.5%) were negative. The variables that in greater proportion impeded intervention between screening positive patients were neurological damage due to the trauma with 23 patients (37.1%) and prior psychiatric disorder with 18 (29%). Both variables were associated with substance consumption: negatives 9.9% vs positive 22.3% (P=.001) and negatives 3% vs positive 17.5% (P=.016) respectively. The number of candidates for motivational intervention was 41, 16.9% of all admitted patients. CONCLUSIONS: Almost 2 out of 10 patients were potential candidates. The factors that in a greater proportion precluded the intervention were the same as those associated with consumption. Mortality in ICU was associated with non-compliance with the screening protocol.