Guía de seguimiento farmacoterapéutico sobre niño enfermo

Coordinadores de esta edición: Emilio García Jiménez y Martha Milena Silva CastroEsta guía tiene por objetivo facilitar la fase de estudio necesaria para realizar seguimiento farmacoterapéutico en el niño enfermo. En primer lugar se han tratado las características farmacocinéticas y farmacodinámicas especiales de los niños. Para, a continuación presentar las características de los problemas de salud más importantes en la población infantil, así como los tratamientos de tipo farmacológico o no, indicados para tratar estos problemas de salud

COP1 destabilizes DELLA proteins in Arabidopsis

DELLA transcriptional regulators are central components in the control of plant growth responses to the environment. This control is considered to be mediated by changes in the metabolism of the hormones gibberellins (GAs), which promote the degradation of DELLAs. However, here we show that warm temperature or shade reduced the stability of a GA-insensitive DELLA allele in Arabidopsis thaliana. Furthermore, the degradation of DELLA induced by the warmth preceded changes in GA levels and depended on the E3 ubiquitin ligase CONSTITUTIVELY PHOTOMORPHOGENIC1 (COP1). COP1 enhanced the degradation of normal and GA-insensitive DELLA alleles when coexpressed in Nicotiana benthamiana. DELLA proteins physically interacted with COP1 in yeast, mammalian, and plant cells. This interaction was enhanced by the COP1 complex partner SUPRESSOR OF phyA-105 1 (SPA1). The level of ubiquitination of DELLA was enhanced by COP1 and COP1 ubiquitinated DELLA proteins in vitro. We propose that DELLAs are destabilized not only by the canonical GA-dependent pathway but also by COP1 and that this control is relevant for growth responses to shade and warm temperature.This work was supported by the Spanish Ministry of Economy, Industry and Competitiveness and Agencia Española de Investigación/Fondo Europeo para el Desarrollo Regional/Unión Europea (grants BIO2016-79133-P to D.A. and BIO2013-46539-R and BIO2016-80551-R to V.R.); the European Union SIGNAT-Research and Innovation Staff Exchange (Grant H2020-MSCA-RISE-2014-644435 to M.A.B., D.A., and J.J.C.); the Argentinian Agencia Nacional de Promoción Científica y Tecnológica (Grant Proyectos de Investigación Científica y Tecnológica-2016-1459 to J.J.C.); Universidad de Buenos Aires (grant 20020170100505BA to J.J.C.); the National Institute of General Medical Sciences of the National Institutes of Health (awards R01GM067837 and R01GM056006 to S.A.K.); the German Research Foundation (DFG) under Germany’s Excellence Strategy/Initiative (Cluster of Excellence on Plant Sciences – Excellence Cluster EXC-2048/1, Project ID 390686111 to M.D.Z.); the International Max Planck Research School of the Max Planck Society; the Universities of Düsseldorf and of Cologne to T.B.; Nordrhein Westfalen Bioeconomy Science Center-FocusLabs CombiCom to N.H. and M.D.Z.; and Ministry of Education, Youth and Sports of the Czech Republic (Project LQ1601 Central European Institute of Technology 2020 to B.B. and M.C.). N.B.-T., E.I., and M.G.-L. were supported by Ministerio de Economía y Competitividad-Formación de Personal Investigador Program fellowships

COP1 destabilizes DELLA proteins in Arabidopsis

DELLA transcriptional regulators are central components in the control of plant growth responses to the environment. This control is considered to be mediated by changes in the metabolism of the hormones gibberellins (GAs), which promote the degradation of DELLAs. However, here we show that warm temperature or shade reduced the stability of a GA-insensitive DELLA allele in Arabidopsis thaliana. Furthermore, the degradation of DELLA induced by the warmth preceded changes in GA levels and depended on the E3 ubiquitin ligase CONSTITUTIVELY PHOTOMORPHOGENIC1 (COP1). COP1 enhanced the degradation of normal and GAinsensitive DELLA alleles when coexpressed in Nicotiana benthamiana. DELLA proteins physically interacted with COP1 in yeast, mammalian, and plant cells. This interaction was enhanced by the COP1 complex partner SUPRESSOR OF phyA-105 1 (SPA1). The level of ubiquitination of DELLA was enhanced by COP1 and COP1 ubiquitinated DELLA proteins in vitro. We propose that DELLAs are destabilized not only by the canonical GA-dependent pathway but also by COP1 and that this control is relevant for growth responses to shade and warm temperature.Fil: Blanco Touriñán, Noel. Universidad Politécnica de Valencia; EspañaFil: Legris, Martina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Minguet, Eugenio G.. Universidad Politécnica de Valencia; EspañaFil: Costigliolo Rojas, María Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Nohales, María A.. University of Southern California; Estados UnidosFil: Iniesto, Elisa. Consejo Superior de Investigaciones Científicas; EspañaFil: García León, Marta. Consejo Superior de Investigaciones Científicas; EspañaFil: Pacín, Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; ArgentinaFil: Heucken, Nicole. Universitat Dusseldorf; AlemaniaFil: Blomeier, Tim. Universitat Dusseldorf; AlemaniaFil: Locascio, Antonella. Universidad Politécnica de Valencia; EspañaFil: Cerný, Martin. Mendel University in Brno; República ChecaFil: Esteve Bruna, David. Universidad Politécnica de Valencia; EspañaFil: Díez Díaz, Mónica. Univerdiad Catolica de Valencia; EspañaFil: Brzobohatý, Bretislav. Mendel University in Brno; República ChecaFil: Frerigmann, Henning. Max Planck Institute for Plant Breeding Research; AlemaniaFil: Zurbriggen, Matías D.. Universitat Dusseldorf; AlemaniaFil: Kay, Steve A.. University of Southern California; Estados UnidosFil: Rubio, Vicente. Consejo Superior de Investigaciones Científicas; EspañaFil: Blázquez, Miguel A.. Universidad Politécnica de Valencia; EspañaFil: Casal, Jorge José. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; ArgentinaFil: Alabadí, David. Universidad Politécnica de Valencia; Españ

Akt regulation of glycolysis mediates bioenergetic stability in epithelial cells

Cells use multiple feedback controls to regulate metabolism in response to nutrient and signaling inputs. However, feedback creates the potential for unstable network responses. We examined how concentrations of key metabolites and signaling pathways interact to maintain homeostasis in proliferating human cells, using fluorescent reporters for AMPK activity, Akt activity, and cytosolic NADH/NAD+ redox. Across various conditions, including glycolytic or mitochondrial inhibition or cell proliferation, we observed distinct patterns of AMPK activity, including both stable adaptation and highly dynamic behaviors such as periodic oscillations and irregular fluctuations that indicate a failure to reach a steady state. Fluctuations in AMPK activity, Akt activity, and cytosolic NADH/NAD+ redox state were temporally linked in individual cells adapting to metabolic perturbations. By monitoring single-cell dynamics in each of these contexts, we identified PI3K/Akt regulation of glycolysis as a multifaceted modulator of single-cell metabolic dynamics that is required to maintain metabolic stability in proliferating cells

Paper dels productes avançats de la glicació (AGE) en l'envelliment del cor

L'envelliment disminueix la tolerància del cor a l'exercici i l'estrès i augmenta la incidència d'insuficiència cardíaca, una de les principals causes de mort i discapacitat a tot el món. Fins a un 80% dels pacients amb insuficiència cardíaca són ancians i aquesta condició empitjora significativament el pronòstic d'aquesta malaltia. Estudis preclínics indiquen que els cardiomiòcits envellits produeixen alguns canvis endògens, que contribueixen a una major susceptibilitat a desenvolupar disfuncions i errors, independentment d'altres factors com la taxa de comorbiditat present en l'envelliment. De fet, els cardiomiòcits envellits comparteixen alguns trets fenotípics comuns amb els cardiomiòcits malmesos, com ara el dany oxidatiu i el desajust energètic en augment de la demanda metabòlica. Estudis previs del nostre grup han demostrat que aquests trets fenotípics es poden explicar per una alteració en la comunicació anatòmica i funcional entre el reticle sarcoplasmàtic (RS) i els mitocondris, necessaris per regular la producció d'energia i la regeneració antioxidant. Tanmateix, no es coneixen els mecanismes de tal interrupció. Aquesta tesi investiga la contribució de la glicació intracel·lular avançada a la disfunció dels cardiomiòcits i a l'alteració mitocondrial durant l'envelliment.El envejecimiento disminuye la tolerancia del corazón al ejercicio y el estrés y aumenta la incidencia de insuficiencia cardiaca, una de las principales causas de muerte y discapacidad en todo el mundo. Hasta un 80% de los pacientes con insuficiencia cardíaca son ancianos y esta condición empeora significativamente el pronóstico de esta enfermedad. Estudios preclínicos indican que los cardiomiocitos envejecidos producen algunos cambios endógenos, que contribuyen a una mayor susceptibilidad a desarrollar disfunciones y errores, independientemente de otros factores como la tasa de comorbilidad presente en el envejecimiento. De hecho, los cardiomiocitos envejecidos comparten algunos rasgos fenotípicos comunes con los cardiomiocitos dañados, como el daño oxidativo y el desajuste energético en aumento de la demanda metabólica. Estudios previos de nuestro grupo han demostrado que estos rasgos fenotípicos pueden explicarse por una alteración en la comunicación anatómica y funcional entre el retículo sarcoplásmico (RS) y las mitocondrias, necesarios para regular la producción de energía y la regeneración antioxidante. Sin embargo, no se conocen los mecanismos de tal interrupción. Esta tesis investiga la contribución de la glicación intracelular avanzada a la disfunción de los cardiomiocitos y la alteración mitocondrial durante el envejecimiento. La glicación avanzada es una de las alteraciones químicas más relevantes y omnipresentes asociadas al envejecimiento biológico. Induce daños terminales a proteínas y otras macromoléculas, que en última instancia precipitan en forma de AGE (productos finales de glicación avanzada). La acumulación de AGE se ha documentado en varias enfermedades asociadas al envejecimiento, como el Alzheimer, el Parkinson, las cataratas y otros. Sin embargo, no se conoce su contribución a la fisiopatología del corazón envejecido.Aging decreases the tolerance of the heart to exercise and stress and increases the incidence of heart failure, a leading cause of death and disability worldwide. Up to 80% of heart failure patients are elderly and being old significantly worsens the prognosis of this condition. Preclinical studies indicate that aged cardiomyocytes develop some endogenous changes that contribute to their greater susceptibility to develop dysfunction and failure, independently of other factors, like higher comorbidity rate present in aging. Indeed, aged cardiomyocytes share some common phenotypic traits with failing cardiomyocytes, such as oxidative damage and energy mismatch under increased metabolic demand. Previous studies of our group have demonstrated that these phenotypic traits can be explained by an alteration in the anatomical and functional communication between the sarcoplasmic reticulum (SR) and the mitochondria, necessary to regulate energy production and antioxidant regeneration. However, the mechanisms of such disruption are not known. This thesis investigates the contribution of intracellular advanced glycation to cardiomyocyte dysfunction and mitochondrial alteration during aging

Receptor Level Mechanisms Are Required for Epidermal Growth Factor (EGF)-stimulated Extracellular Signal-regulated Kinase (ERK) Activity Pulses*

In both physiological and cell culture systems, EGF-stimulated ERK activity occurs in discrete pulses within individual cells. Many feedback loops are present in the EGF receptor (EGFR)-ERK network, but the mechanisms driving pulsatile ERK kinetics are unknown. Here, we find that in cells that respond to EGF with frequency-modulated pulsatile ERK activity, stimulation through a heterologous TrkA receptor system results in non-pulsatile, amplitude-modulated activation of ERK. We further dissect the kinetics of pulse activity using a combination of FRET- and translocation-based reporters and find that EGFR activity is required to maintain ERK activity throughout the 10-20-minute lifetime of pulses. Together, these data indicate that feedbacks operating within the core Ras-Raf-MEK-ERK cascade are insufficient to drive discrete pulses of ERK activity and instead implicate mechanisms acting at the level of EGFR

Cartas americanas

Alejandro de Humboldt, el último de los grandes hombres universales, nació en Berlín en 1769 y murió en la misma ciudad en 1859, después de haber realizado largos viajes de exploración científica. Su expedición a América del Sur duró cinco años, durante los cuales realizó un enorme acopio de información y datos con los que contribuyó a establecer las bases de la moderna geografía física y de la fitogeografía. En Europa, a su regreso de América, dedicó casi treinta y cinco años a escribir y publicar sus obras sobre este continente. De la enorme producción epistolar y científica de Humboldt, el presente volumen recoge lo más representativo de su correspondencia, escritos y proyectos vinculados con su viaje por las regiones equinocciales de América

Assessment of Cas12a‐mediated gene editing efficiency in plants

The CRISPR/Cas12a editing system opens new possibilities for plant genome engineering. To obtain a comparative assessment of RNA‐guided endonuclease (RGEN) types in plants, we adapted the CRISPR/Cas12a system to the GoldenBraid (GB) modular cloning platform and compared the efficiency of Acidaminococcus (As) and Lachnospiraceae (Lb) Cas12a variants with the previously described GB‐assembled Streptococcus pyogenes Cas9 (SpCas9) constructs in eight Nicotiana benthamiana loci using transient expression. All three nucleases showed drastic target‐dependent differences in efficiency, with LbCas12 producing higher mutagenesis rates in five of the eight loci assayed, as estimated with the T7E1 endonuclease assay. Attempts to engineer crRNA direct repeat (DR) had little effect improving on‐target efficiency for AsCas12a and resulted deleterious in the case of LbCas12a. To complete the assessment of Cas12a activity, we carried out genome editing experiments in three different model plants, namely N. benthamiana, Solanum lycopersicum and Arabidopsis thaliana. For the latter, we also resequenced Cas12a‐free segregating T2 lines to assess possible off‐target effects. Our results showed that the mutagenesis footprint of Cas12a is enriched in deletions of −10 to −2 nucleotides and included in some instances complex rearrangements in the surroundings of the target sites. We found no evidence of off‐target mutations neither in related sequences nor somewhere else in the genome. Collectively, this study shows that LbCas12a is a viable alternative to SpCas9 for plant genome engineering.ISSN:1467-7644ISSN:1467-765

Assessment of Cas12a‐mediated gene editing efficiency in plants

The CRISPR/Cas12a editing system opens new possibilities for plant genome engineering. To obtain a comparative assessment of RNA‐guided endonuclease (RGEN) types in plants, we adapted the CRISPR/Cas12a system to the GoldenBraid (GB) modular cloning platform and compared the efficiency of Acidaminococcus (As) and Lachnospiraceae (Lb) Cas12a variants with the previously described GB‐assembled Streptococcus pyogenes Cas9 (SpCas9) constructs in eight Nicotiana benthamiana loci using transient expression. All three nucleases showed drastic target‐dependent differences in efficiency, with LbCas12 producing higher mutagenesis rates in five of the eight loci assayed, as estimated with the T7E1 endonuclease assay. Attempts to engineer crRNA direct repeat (DR) had little effect improving on‐target efficiency for AsCas12a and resulted deleterious in the case of LbCas12a. To complete the assessment of Cas12a activity, we carried out genome editing experiments in three different model plants, namely N. benthamiana, Solanum lycopersicum and Arabidopsis thaliana. For the latter, we also resequenced Cas12a‐free segregating T2 lines to assess possible off‐target effects. Our results showed that the mutagenesis footprint of Cas12a is enriched in deletions of −10 to −2 nucleotides and included in some instances complex rearrangements in the surroundings of the target sites. We found no evidence of off‐target mutations neither in related sequences nor somewhere else in the genome. Collectively, this study shows that LbCas12a is a viable alternative to SpCas9 for plant genome engineering.ISSN:1467-7644ISSN:1467-765