Transformation and regeneration of the holoparasitic plant Phelipanche aegyptiaca

<p>Abstract</p> <p>Background</p> <p>Transformation and subsequent regeneration of holoparasitic plants has never been reported, in part due to challenges in developing transformation protocols, but also because regeneration of obligate parasites is difficult since their survival depends completely on successful haustorium penetration of a host and the formation of vascular connections. The recent completion of a massive transcriptome sequencing project (the Parasitic Plant Genome Project) will fuel the use of genomic tools for studies on parasitic plants. A reliable system for holoparasite transformation is needed to realize the full value of this resource for reverse genetics and functional genomics studies.</p> <p>Results</p> <p>Here we demonstrate that transformation of <it>Phelipanche aegyptiaca </it>is achieved by infection of 3 month-old <it>in vitro </it>grown <it>P. aegyptiaca </it>calli with <it>Agrobacterium rhizogenes </it>harboring the yellow fluorescent protein (YFP). Four months later, YFP-positive regenerated calli were inoculated onto tomato plants growing in a minirhizotron system. Eight days after inoculation, transgenic parasite tissue formed lateral haustoria that penetrated the host and could be visualized under UV illumination through intact host root tissue. YFP-positive shoot buds were observed one month after inoculation.</p> <p>Conclusions</p> <p>This work constitutes a breakthrough in holoparasitic plant research methods. The method described here is a robust system for transformation and regeneration of a holoparasitic plant and will facilitate research on unique parasitic plant capabilities such as host plant recognition, haustorial formation, penetration and vascular connection.</p

Differential expression proteomics to investigate responses and resistance to Orobanche crenata in Medicago truncatula

<p>Abstract</p> <p>Background</p> <p>Parasitic angiosperm <it>Orobanche crenata </it>infection represents a major constraint for the cultivation of legumes worldwide. The level of protection achieved to date is either incomplete or ephemeral. Hence, an efficient control of the parasite requires a better understanding of its interaction and associated resistance mechanisms at molecular levels.</p> <p>Results</p> <p>In order to study the plant response to this parasitic plant and the molecular basis of the resistance we have used a proteomic approach. The root proteome of two accessions of the model legume <it>Medicago truncatula </it>displaying differences in their resistance phenotype, in control as well as in inoculated plants, over two time points (21 and 25 days post infection), has been compared. We report quantitative as well as qualitative differences in the 2-DE maps between early- (SA 27774) and late-resistant (SA 4087) genotypes after Coomassie and silver-staining: 69 differential spots were observed between non-inoculated genotypes, and 42 and 25 spots for SA 4087 and SA 27774 non-inoculated and inoculated plants, respectively. In all, 49 differential spots were identified by peptide mass fingerprinting (PMF) following MALDI-TOF/TOF mass spectrometry. Many of the proteins showing significant differences between genotypes and after parasitic infection belong to the functional category of defense and stress-related proteins. A number of spots correspond to proteins with the same function, and might represent members of a multigenic family or post-transcriptional forms of the same protein.</p> <p>Conclusion</p> <p>The results obtained suggest the existence of a generic defense mechanism operating during the early stages of infection and differing in both genotypes. The faster response to the infection observed in the SA 27774 genotype might be due to the action of proteins targeted against key elements needed for the parasite's successful infection, such as protease inhibitors. Our data are discussed and compared with those previously obtained with pea <abbrgrp><abbr bid="B1">1</abbr></abbrgrp> and transcriptomic analysis of other plant-pathogen and plant-parasitic plant systems.</p

A Novel Electron Microscopic Characterization of Core/Shell Nanobiostimulator Against Parasitic Plants

Nanoencapsulation has proven to be an efficient route to increase significantly the solubility and bioavailability of organic compounds. This aspect of nanotechnology is illustrated for the case of phthalimide-lactone (PL), a recently synthesized strigolactone mimic whose very limited solubility in water, as a free chemical, precludes its practical use as an agrochemical in the fight against parasitic plants. Pluronic F-127 (P127) nanoparticles functionalized with PL have been synthesized and embedded in a polymeric matrix of poly(vinyl alcohol) (PVA). Low-voltage and medium voltage imaging and spectroscopic scanning electron microscopy (S(T)EM) techniques were combined to confirm the synthesis of multicore nanoparticles that were rich in nitrogen, a finding that is due to the successful encapsulation of PL. This PL@P127/PVA nanobiostimulator formulation has an impressive solubility in water, that is, 27 times higher than that of pure phthalimide-lactone. Also critical from the functional point of view, comparative bioassays clearly showed that the intrinsic stimulatory activity of this agrochemical is fully maintained in the nanoencapsulated formulation

Cobertura de la vacunación antigripal en los farmacéuticos comunitarios: razones aportadas y absentismo relacionado. Predisposición a vacunar a la población

Introducción: la vacuna antigripal está indicada entre el personal sanitario, aunque la cobertura antigripal en farmacéuticos comunitarios españoles actualmente se desconoce. Este trabajo pretende averiguar esta cobertura, las causas de los farmacéuticos para vacunarse o no de la gripe, el absentismo laboral que les ocasiona, si recomiendan la vacunación antigripal entre los grupos de riesgo y su disposición a administrarla en la farmacia. Métodos: estudio descriptivo transversal mediante un cuestionario online destinado a farmacéuticos comunitarios de toda España. Se realizó un análisis descriptivo de las variables estudiadas y de la asociación entre la vacunación y las variables cualitativas (test Chi-cuadrado y Fisher) y cuantitativas (ANOVA). Resultados: se obtuvo una tasa de respuesta del 9,4 % (n=1.436). Los farmacéuticos refirieron vacunarse en torno al 30 % en las tres temporadas estudiadas (2016-2019), siendo del 31-35,8 % (IC95 %) en la temporada 2018-2019. Ser titular/cotitular de la farmacia, la experiencia profesional, la edad, vacunarse para evitar la gripe y/o por responsabilidad, y vacunarse en temporadas anteriores son factores asociados a vacunarse (p<0,05). Ser socio de SEFAC también lo es. El absentismo por gripe en la temporada 2018-2019 fue del 9,5 %. Los farmacéuticos que más dijeron vacunarse recomiendan más la vacuna que los que no, y tendrían mayor predisposición a administrarla (p<0,05). Conclusiones: la cobertura vacunal antigripal entre los farmacéuticos comunitarios es baja a pesar del absentismo que les causa. Facilitar la vacunación a los farmacéuticos podría incrementar su cobertura. Los farmacéuticos recomiendan la vacunación contra la gripe a los pacientes de riesgo y estarían dispuestos a vacunar en farmacias autorizadas

The Assessment of Myocardial Longitudinal Strain in a Paediatric Spanish Population Using a New Software Analysis

Two-dimensional speckle-tracking echocardiography (2DSTE) has been present for years. However, it is underutilized due to the expertise and time requirements for its analysis. Our aims were to provide strain values in a paediatric Spanish population and to assess the feasibility and reproducibility of a new strain software analysis in our environment. Methods: A cross-sectional study of 156 healthy children aged 6 to 17 years. Longitudinal strain (LS) analysis of the left ventricle, right ventricle, and left atrium was performed. Feasibility and reproducibility were assessed. The associations of clinical and echocardiographic variables with strain values were investigated by multivariate analysis. Results: Mean age was 11 ± 3 years (50% female). Feasibility of LS measurement ranged from 94.2% for left ventricle global LS (LVGLS) to 98.1% for other chamber strain parameters. Strain values were 26.7 ± 2.3% for LVGLS; 30.5 ± 4.4% and 26.9 ± 4% for right ventricle free wall LS (RVFWLS) and four chambers view LS (RV4CLS) respectively; and 57.8 ± 10.5%, 44.9 ± 9.5%, and 12.9 ± 5.5% for left atrium LS reservoir phase (LALSr), conduct phase (LALScd) and contraction phase (LALSct), also respectively. Body surface area (BSA) and age presented a negative correlation with strain values. Higher values were found in females than in males, except for LALScd. Excellent intra- and inter-observer reproducibility were found for right and left ventricular strain measurement, with intraclass correlation coefficients (ICC) ranging from 0.88 to 0.98, respectively. In conclusion, we described strain values in a healthy Spanish paediatric population. LS assessment by this new strain analysis software by semi-automatic manner was highly feasible and reproducible

Factores de riesgo y diagnóstico citológico del cáncer cervico-uterino

Se analizaron los factores de riesgo y diagnóstico citológico del cáncer cervico-uterino en pacientes de una comunidad de Veracruz bajo un estudio analítico y transversal, en este se incluyó 132 pacientes mayores de 18 años que acudieron al Módulo de Atención a realizarse la citología cervical, las variables fueron factores de riesgo del cáncer cervico-uterino y diagnostico citológico. Se usó como instrumento el “formato de solicitud y reporte de resultados de citología cervical”, se utilizó la chi2 para la asociación de los factores de riesgo con el diagnostico citológico. Se halló que el 79.5% de las mujeres tenían el cuello aparentemente sano, el 9.8% erosión de cuello, 4.5% leucorrea, 2.3% de las mujeres no se observaba el cuello y tenían sangrado anormal, el .8% presento cuello anormal y cervicitis Conclusiones: La enfermera debe dar seguimiento a los casos, ya que es el primer contacto del paciente, así pueda brindar educación para la salud a todas las mujeres que acuden a realizarse esta prueba

Comorbidities in patients with Primary Sjögren's Syndrome and Systemic Lupus Erythematosus: A comparative registries-based study

OBJECTIVE: To compare the prevalence of the main comorbidities in 2 large cohorts of patients with primary Sjögren's syndrome (SS) and systemic lupus erythematosus (SLE), with a focus on cardiovascular (CV) diseases. METHODS: This was a cross-sectional multicenter study where the prevalence of more relevant comorbidities in 2 cohorts was compared. Patients under followup from SJOGRENSER (Spanish Rheumatology Society Registry of Primary SS) and RELESSER (Spanish Rheumatology Society Registry of SLE), and who fulfilled the 2002 American-European Consensus Group and 1997 American College of Rheumatology classification criteria, respectively, were included. A binomial logistic regression analysis was carried out to explore potential differences, making general adjustments for age, sex, and disease duration and specific adjustments for each variable, including CV risk factors and treatments, when appropriate. RESULTS: A total of 437 primary SS patients (95% female) and 2,926 SLE patients (89% female) were included. The mean age was 58.6 years (interquartile range [IQR] 50.0-69.9 years) for primary SS patients and 45.1 years (IQR 36.4-56.3 years) for SLE patients (P?<?0.001), and disease duration was 10.4 years (IQR 6.0-16.7 years) and 13.0 years (IQR 7.45-19.76 years), respectively (P?<?0.001). Smoking, dyslipidemia, and arterial hypertension were associated less frequently with primary SS (odds ratio [OR] 0.36 [95% confidence interval (95% CI) 0.28-0.48], 0.74 [95% CI 0.58-0.94], and 0.50 [95% CI 0.38-0.66], respectively) as were life-threatening CV events (i.e., stroke or myocardial infarction; OR 0.57 [95% CI 0.35-0.92]). Conversely, lymphoma was associated more frequently with primary SS (OR 4.41 [95% CI 1.35-14.43]). The prevalence of severe infection was lower in primary SS than in SLE (10.1% versus 16.9%; OR 0.54 [95% CI 0.39-0.76]; P?<?0.001). CONCLUSION: Primary SS patients have a consistently less serious CV comorbidity burden and a lower prevalence of severe infection than those with SLE. In contrast, their risk of lymphoma is greater

Tumor regression and resistance mechanisms upon CDK4 and RAF1 inactivation in KRAS/P53 mutant lung adenocarcinomas.

KRAS mutant lung adenocarcinomas remain intractable for targeted therapies. Genetic interrogation of KRAS downstream effectors, including the MAPK pathway and the interphase CDKs, identified CDK4 and RAF1 as the only targets whose genetic inactivation induces therapeutic responses without causing unacceptable toxicities. Concomitant CDK4 inactivation and RAF1 ablation prevented tumor progression and induced complete regression in 25% of KRAS/p53-driven advanced lung tumors, yet a significant percentage of those tumors that underwent partial regression retained a population of CDK4/RAF1-resistant cells. Characterization of these cells revealed two independent resistance mechanisms implicating hypermethylation of several tumor suppressors and increased PI3K activity. Importantly, these CDK4/RAF1-resistant cells can be pharmacologically controlled. These studies open the door to new therapeutic strategies to treat KRAS mutant lung cancer, including resistant tumors.We thank S. Ortega for the generation of the Cdk4FxKD mouse model; and M. San Roman, R. Villar, M. C. Gonzalez, A. Lopez, N. Cabrera, P. Villanueva, J. Condo, J. Klett, A. Cebria, A. Otero, O. Dominguez, G. Luengo, G. Garaulet, F. Mulero, and D. Megias for excellent technical support. This work was supported by European Research Council Grant ERC-2015-AdG/695566, THERACAN, Spanish Ministry of Science, Innovation, and Universities Grant RTC-2017-6576-1, and the Autonomous Community of Madrid Grant B2017/BMD-3884 iLUNG-CM (to M.B.); Spanish Ministry of Science, Innovation, and Universities Grant RTI2018-094664B-I00 (to M.B. and M.M.); and National Natural Science Foundation of China Grant 31771469 (to H.W.). M.B. is a recipient of an Endowed Chair from the AXA Research Fund. L.E.-B. is the recipient of an FPI fellowship from the Spanish Ministry of Economy and Competitiveness. F.F.-G., M.S., and P.N. were supported by an FPU fellowships from the Spanish Ministry of Education.S

Big Data en Educación III: recomendaciones personalizadas de actuación ante estudiantes en riesgo de abandono

En este proyecto se elaboran un conjunto de recomendaciones personalizadas para guiar la actuación del profesor con estudiantes que hayan sido detectados en riesgo de abandono de una asignatura o de la titulación en general. Con carácter previo, se desarrollan métodos estadísticos que permitan la detección del riesgo de abandono de un estudiante