634 research outputs found

    Second generation of viral vectors based on pepino mosaic virus

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    [SPA] Nuestro grupo ya ha desarrollado un vector eficiente (pBPepGFP2a) basado en el Virus del mosaico de pepino dulce (PepMV; género Potexvirus) para la expresión de proteínas y el silenciamiento génico inducido por virus en Nicotiana benthamiana. El objetivo de este trabajo fue mejorar aún más pBPepGFP2a en términos de idoneidad y estabilidad en infecciones en tomate. Para evitar la recombinación homóloga debido a la duplicación de la secuencia, se diseñaron dos construcciones nuevas con diferente número de mutaciones, pBPepGFPm1 y pBPepGFPm2, utilizando pBPepGFP2a como molde. Los ensayos de estabilidad mostraron que PepGFPm2 era estable en plantas de N. benthamiana durante al menos 3 pases, pero no en plantas de tomate, en las que no se detectó expresión de GFP con ninguno de los vectores. Posteriormente, el gen gfp de pBPepGFPm2 fue reemplazado por los genes DsRed o mCherry, dando lugar a los vectores PepDsRed y PepmCherry. Ambos vectores fueron estables en N. benthamiana y PepDsRed fue también estable en tomate al menos durante dos pases. [ENG] Our group has already developed an efficient Pepino mosaic virus (PepMV; genus Potexvirus)-based vector (pBPepGFP2a) for protein expression and virus induced gene silencing in Nicotiana benthamiana. The aim of this work was to further improve pBPepGFP2a in terms of stability and suitability for reporting infections in tomato. To avoid homologous recombination due to sequence duplication, two new constructs with different number of mutations were designed, pBPepGFPm1 and pBPepGFPm2, using pBPepGFP2a as backbone. Stability assays showed that PepGFPm2 was stable in N. benthamiana plants for at least 3 passages, but not in tomato plants, in which no GFP expression was detected with any of the vectors. Then, the gfp gene from pBPepGFPm2 was replaced by the DsRed or mCherry genes, giving rise to the PepDsRed and PepmCherry vectors. Both vectors were stable in N. benthamiana and PepDsRed was also stable in tomato at least during two passages.Agradezco la financiación de la beca FPI a la Fundación Séneca de la Región de Murcia, y la financiación de los experimentos al proyecto de Plan Estatal AGL2015-65838-R (Ministerio de Economía y Competitividad)

    PepGPFm2, PepDsRed and PepBar, a new generation of viral vectors based on pepino mosaic virus

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    [SPA] Nuestro grupo ya ha desarrollado un vector eficiente (pBPepGFP2a) basado en el virus del mosaico de pepino dulce (PepMV; género Potexvirus) para la expresión de proteínas y el silenciamiento génico inducido por virus en Nicotiana benthamiana. Tras mejorar pJLPepGFP2a en términos de estabilidad e idoneidad para la monitorización de la infección tanto en N. benthamiana como en tomate, los vectores pJLPepGFPm2 y pJLPepDsRed han sido empleados para el estudio de localización subcelular de PepMV. A la vez hemos desarrollado un vector, PepBar, para su empleo en escrutinios masivos de susceptibilidad, en el que el gen gfp de PepGFPm2 fue sustituido por el gen bar, un gen de resistencia a herbicidas. Este vector fue estable durante al menos dos pasajes tanto en N. benthamiana como en plantas de tomate. [ENG] Our group has already developed an efficient Pepino mosaic virus (PepMV; genus Potexvirus)-based vector (pBPepGFP2a) for protein expression and virus-induced gene silencing in Nicotiana benthamiana. We have improved pJLPepGFP2a in terms of stability and suitability for reporting infections in N. benthamiana and tomato, producing pJLPepGFPm2 and pJLPepDsRed. These new vectors have been used for PepMV subcellular localization assays. Moreover, to be used in massive susceptibility screenings, we also developed PepBar, in which the gfp gene from PepGFPm2 was substituted by the bar gene, an herbicide resistance gene. This vector was stable for at less two passages both in N. benthamiana and tomato plants.Agradezco la financiación de la beca FPI a la Fundación Séneca de la Región de Murcia, y la financiación de los experimentos al proyecto de Plan Estatal AGL2015-65838-R (Ministerio de Economía y Competitividad)

    The Role of Pediatricians in Providing Greater-Quality Care for Children: An Ongoing Debate

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    Child healthcare frequently is regarded as a secondary objective in the development of public health policies and not as a central issue. Cost-containment policies instead of quality of healthcare frequently have inspired strategic decisions in public health investments for pediatric structures and workforce. The negative implications of this decisional approach have been shown by the substantial unpreparedness of virtually all European health systems during the coronavirus disease 2019 (COVID-19) pandemic,5 particularly in the area of primary care. The debate on the negative effects of cost-containment policies in the area of primary care in not new. In recent years, the role of pediatricians within the several European healthcare systems was variously reformed by governments, with an eye more to the budget than to the quality of care and its structural components, including staffing adequacy, training, and consistency as well as facility environment and size.1 As a result, the number of general pediatricians in Europe has decreased during the past 20 years.Data from the World Health Organization show that the average number of general pediatricians per 100 000 population in Europe declined 30.1% from 26.5% in 1998 to 18.5% in 2013.A further reduction in the pediatric workforce was reported in 2018 by the European Paediatric Association, the Union of National European Paediatric Societies and Associations (EPA-UNEPSA). During the past 20 years, due to different factors, including socioeconomic and political reasons, several European countries decided to reform their public health systems and the responsibility of pediatric healthcare delivery was moved from pediatricians to general and family practitioners. The aim of this commentary by the Spanish Primary Care Pediatrics Association (AEPap), in collaboration with the European Confederation of Primary Care Pediatricians and EPA-UNEPSA, is to discuss data supporting the essential role played in Europe by well-trained pediatricians vs other providers in delivering quality healthcare for children

    Pepino mosaic virus RNA-Dependent RNA Polymerase POL Domain Is a Hypersensitive Response-Like Elicitor Shared by Necrotic and Mild Isolate

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    [SPA] El virus del mosaico del pepino dulce (PepMV) es un patógeno emergente que representa una grave amenaza para la producción de tomate. Las enfermedades inducidas por PepMV se manifiestan con una amplia gama de síntomas, incluyendo la necrosis sistémica. Nuestros resultados muestran que la acumulación de PepMV depende tanto del aislado del virus, como de la variedad de tomate o las condiciones ambientales, asociado todo ello al desarrollo de la necrosis. La sustitución de una lisina por un ácido glutámico en la posición 67 del triple bloque de genes (TGB3), previamente descrita como un determinante de la necrosis, parece favorecer una mayor acumulación del virus pero no parece ser el elemento elicitor de la necrosis sistémica. La sobreexpresión tanto de TGB3 como del dominio polimerasa (POL) de la replicasa (RdRp) produjo necrosis, aunque sólo la expresión local de POL desencadenó síntomas caracteristos de HR. En conjunto, nuestros datos sugieren que el dominio RdRp-POL desempeña un papel importante en la inducción de necrosis de PepMV, dependiendo del nivel de acumulación del virus, que puede ser modulado por la naturaleza de TGB3, el genotipo del huesped y las condiciones ambientales. [ENG] Pepino mosaic virus (PepMV) is an emerging pathogen that represents a serious threat to tomato production worldwide. PepMV-induced diseases manifest with a wide range of symptoms, including systemic necrosis. Our results showed that PepMV accumulation depends on the virus isolate, tomato cultivar, and environmental conditions, and associates with the development of necrosis. Substitution of lysine for glutamic acid at position 67 in the triple gene block 3 (TGB3) protein, previously described as a necrosis determinant, led to increased virus accumulation and was necessary but not sufficient to induce systemic necrosis. Overexpression of both TGB3 and the polymerase domain (POL) of the RNA-dependent RNA polymerase (RdRp) resulted in necrosis, although only local expression of POL triggered HR-like symptoms. Altogether, our data suggest that the RdRp-POL domain plays an important role in PepMV necrosis induction, with necrosis development depending on the virus accumulation level, which can be modulated by the nature of TGB3, host genotype and environmental conditions.Agradecer la financiación del Ministerio de Economía y Competitividad (AGL2012-37390 ) y la fundación Séneca por la financiación de la Beca FPI

    Cambios en el perfil terpénico y en el crecimiento secundario en poblaciones en decaimiento de Pinus sylvestris L. bajo influencia mediterránea en respuesta a factores locales

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    The terpenoid profile could give information about the water status in Scots pine, especially for trees growing in the same geographical area but under contrasting local environmental conditions. Terpenes were analyzed by gas chromatography-mass spectrometry in needles, twigs and wood of ten affected and ten unaffected Scots pines in the southern “Sistema Ibérico” range (Teruel, Spain), where forest decline has been recently reported. Soil depth and secondary growth was also studied in both types of trees. Needles and twigs total resin acids were significantly higher in affected trees. The pimarane type resin acids were also higher in the twigs of affected trees. Secondary growth was lower in affected trees and it showed higher climate sensitivity. The use of the terpenoid profile may be used as an additional tool for the estimation of the water status, especially for situations inducing moderate but relatively prolonged stress conditions.El perfil terpénico podría dar información sobre el estado hídrico en el pino albar, especialmente cuando se comparen especímenes de una zona geográfica concreta afectados por factores ambientales locales. Los terpenos de acículas, brotes del año y madera fueron analizados en diez ejemplares afectados y otros tantos no afectados por el decaimiento mediante cromatografía de masas acoplada a espectrometría de masas. La serie de crecimiento secundario en ambos tipos de ejemplares fue también estudiada. La concentración total de ácidos resínicos aumentó de modo significativo en los árboles afectados tanto en brotes del año como en acículas. La cantidad de ácidos de tipo pimarano también aumentó en los brotes de los árboles afectados. La profundidad del suelo y el crecimiento secundario era menor en este tipo de ejemplares, que muestran una mayor sensibilidad en términos dendrocronológicos. El perfíl terpénico podría utilizarse como una herramienta adicional a la estimación del estado hídrico, especialmente cuando el árbol está afectado por condiciones de estrés moderadas pero prolongadas en el tiempo

    A novel targeted RNA-Seq panel identifies a subset of adult patients with acute lymphoblastic leukemia with BCR-ABL1-like characteristics

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    BCR-ABL1-like B-cell precursor acute lymphoblastic leukemia (BCP-ALL) remains poorly characterized in adults. We sought to establish the frequency and outcome of adolescent and adult BCR-ABL1-like ALL using a novel RNA-Seq signature in a series of patients with BCP-ALL. To this end, we developed and tested an RNA-Seq custom panel of 42 genes related to a BCR-ABL1-like signature in a cohort of 100 patients with BCP-ALL and treated with risk-adapted ALL trials. Mutations related to BCR-ABL1-like ALL were studied in a panel of 33 genes by next-generation sequencing (NGS). Also, CRLF2 overexpression and IKZF1/CDKN2A/B deletions were analyzed. Twenty out of 79 patients (12-84 years) were classified as BCR-ABL1-like (25%) based on heatmap clustering, with significant overexpression of ENAM, IGJ, and CRLF2 (P ≤ 0.001). The BCR-ABL1-like subgroup accounted for 29% of 15-60-year-old patients, with the following molecular characteristics: CRLF2 overexpression (75% of cases), IKZF1 deletions (64%), CDKN2A/B deletions (57%), and JAK2 mutations (57%). Among patients with postinduction negative minimal residual disease, those with the BCR-ABL1-like ALL signature had a higher rate of relapse and lower complete response duration than non-BCR-ABL1-like patients (P = 0.007). Thus, we have identified a new molecular signature of BCR-ABL1-like ALL that correlates with adverse prognosis in adult patients with ALL

    A novel targeted RNA-Seq panel identifies a subset of adult patients with acute lymphoblastic leukemia with BCR-ABL1-like characteristics

    Get PDF
    BCR-ABL1-like B-cell precursor acute lymphoblastic leukemia (BCP-ALL) remains poorly characterized in adults. We sought to establish the frequency and outcome of adolescent and adult BCR-ABL1-like ALL using a novel RNA-Seq signature in a series of patients with BCP-ALL. To this end, we developed and tested an RNA-Seq custom panel of 42 genes related to a BCR-ABL1-like signature in a cohort of 100 patients with BCP-ALL and treated with risk-adapted ALL trials. Mutations related to BCR-ABL1-like ALL were studied in a panel of 33 genes by next-generation sequencing (NGS). Also, CRLF2 overexpression and IKZF1/CDKN2A/B deletions were analyzed. Twenty out of 79 patients (12-84 years) were classified as BCR-ABL1-like (25%) based on heatmap clustering, with significant overexpression of ENAM, IGJ, and CRLF2 (P <= 0.001). The BCR-ABL1-like subgroup accounted for 29% of 15-60-year-old patients, with the following molecular characteristics: CRLF2 overexpression (75% of cases), IKZF1 deletions (64%), CDKN2A/B deletions (57%), and JAK2 mutations (57%). Among patients with postinduction negative minimal residual disease, those with the BCR-ABL1-like ALL signature had a higher rate of relapse and lower complete response duration than non-BCR-ABL1-like patients (P = 0.007). Thus, we have identified a new molecular signature of BCR-ABL1-like ALL that correlates with adverse prognosis in adult patients with ALL

    Cognitive clusters in first-episode psychosis

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    Impairments in a broad range of cognitive domains have been consistently reported in some individuals with first-episode psychosis (FEP). Cognitive deficits can be observed during the prodromal stage. However, the course of cognitive deficits is still unclear. The aim of this study was to identify cognitive subgroups over time and to compare their sociodemographic, clinical and functional profiles. A total of 114 patients with Schizophrenia Spectrum Disorders were included in the present study. We assessed subjects through psychiatric scales and eight neuropsychological tests at baseline and at two-year follow-up visit. We performed the Partition Around Medoids algorithm with all cognitive variables. Furthermore, we performed a logistic regression to identify the predictors related to the different cognitive clusters at follow-up. Two distinct subgroups were found: the first cluster characterized by cognitive impairment and a second cluster had relatively intact cognition in comparison with norms. Up to 54.7% of patients with cognitive deficits at baseline tended to improve during the first two years of treatment. Patients with intact cognition at follow-up had a higher socioeconomic status, later age of onset, lower negative symptoms and a higher cognitive reserve (CR) at baseline. CR and age of onset were the baseline variables that predicted cognitive impairment. This research allows us to obtain a better understanding of the heterogeneous profile of psychotic disorders. Identifying the characteristics of patients who will present a cognitive impairment could improve early detection and intervention. These results suggest that enhancing CR could contribute to improving the course of the illness. © 2021 Elsevier B.V

    Cognitive clusters in first-episode psychosis

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    Impairments in a broad range of cognitive domains have been consistently reported in some individuals with first-episode psychosis (FEP). Cognitive deficits can be observed during the prodromal stage. However, the course of cognitive deficits is still unclear. The aim of this study was to identify cognitive subgroups over time and to compare their sociodemographic, clinical and functional profiles. A total of 114 patients with Schizophrenia Spectrum Disorders were included in the present study. We assessed subjects through psychiatric scales and eight neuropsychological tests at baseline and at two-year follow-up visit. We performed the Partition Around Medoids algorithm with all cognitive variables. Furthermore, we performed a logistic regression to identify the predictors related to the different cognitive clusters at follow-up. Two distinct subgroups were found: the first cluster characterized by cognitive impairment and a second cluster had relatively intact cognition in comparison with norms. Up to 54.7% of patients with cognitive deficits at baseline tended to improve during the first two years of treatment. Patients with intact cognition at follow-up had a higher socioeconomic status, later age of onset, lower negative symptoms and a higher cognitive reserve (CR) at baseline. CR and age of onset were the baseline variables that predicted cognitive impairment. This research allows us to obtain a better understanding of the heterogeneous profile of psychotic disorders. Identifying the characteristics of patients who will present a cognitive impairment could improve early detection and intervention. These results suggest that enhancing CR could contribute to improving the course of the illness
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