Nitrogen sensing and regulatory networks: it's about time and space.

A plant's response to external and internal nitrogen signals/status relies on sensing and signaling mechanisms that operate across spatial and temporal dimensions. From a comprehensive systems biology perspective, this involves integrating nitrogen responses in different cell types and over long distances to ensure organ coordination in real time and yield practical applications. In this prospective review, we focus on novel aspects of nitrogen (N) sensing/signaling uncovered using temporal and spatial systems biology approaches, largely in the model Arabidopsis. The temporal aspects span: transcriptional responses to N-dose mediated by Michaelis-Menten kinetics, the role of the master NLP7 transcription factor as a nitrate sensor, its nitrate-dependent TF nuclear retention, its "hit-and-run" mode of target gene regulation, and temporal transcriptional cascade identified by "network walking." Spatial aspects of N-sensing/signaling have been uncovered in cell type-specific studies in roots and in root-to-shoot communication. We explore new approaches using single-cell sequencing data, trajectory inference, and pseudotime analysis as well as machine learning and artificial intelligence approaches. Finally, unveiling the mechanisms underlying the spatial dynamics of nitrogen sensing/signaling networks across species from model to crop could pave the way for translational studies to improve nitrogen-use efficiency in crops. Such outcomes could potentially reduce the detrimental effects of excessive fertilizer usage on groundwater pollution and greenhouse gas emissions.

Dynamic changes in mRNA nucleocytoplasmic localization in the nitrate response of Arabidopsis roots.

Nitrate is a nutrient and signal that regulates gene expression. The nitrate response has been extensively characterized at the organism, organ, and cell-type-specific levels, but intracellular mRNA dynamics remain unexplored. To characterize nuclear and cytoplasmic transcriptome dynamics in response to nitrate, we performed a time-course expression analysis after nitrate treatment in isolated nuclei, cytoplasm, and whole roots. We identified 402 differentially localized transcripts (DLTs) in response to nitrate treatment. Induced DLT genes showed rapid and transient recruitment of the RNA polymerase II, together with an increase in the mRNA turnover rates. DLTs code for genes involved in metabolic processes, localization, and response to stimulus indicating DLTs include genes with relevant functions for the nitrate response that have not been previously identified. Using single-molecule RNA FISH, we observed early nuclear accumulation of the NITRATE REDUCTASE 1 (NIA1) transcripts in their transcription sites. We found that transcription of NIA1, a gene showing delayed cytoplasmic accumulation, is rapidly and transiently activated; however, its transcripts become unstable when they reach the cytoplasm. Our study reveals the dynamic localization of mRNAs between the nucleus and cytoplasm as an emerging feature in the temporal control of gene expression in response to nitrate treatment in Arabidopsis roots.

Multiomics analyses reveal the central role of the nucleolus and its machinery during heat stress acclimation in Pinus radiata.

Global warming is causing rapid changes in mean annual temperature and more severe drought periods. These are major contributors of forest dieback, which is becoming more frequent and widespread. In this work, we investigated how the transcriptome of Pinus radiata changed during initial heat stress response and acclimation. To this end, we generated a high-density dataset employing Illumina technology. This approach allowed us to reconstruct a needle transcriptome, defining 12 164 and 13 590 transcripts as down- and up-regulated, respectively, during a time course stress acclimation experiment. Additionally, the combination of transcriptome data with other available omics layers allowed us to determine the complex inter-related processes involved in the heat stress response from the molecular to the physiological level. Nucleolus and nucleoid activities seem to be a central core in the acclimating process, producing specific RNA isoforms and other essential elements for anterograde-retrograde stress signaling such as NAC proteins (Pra_vml_051671_1 and Pra_vml_055001_5) or helicase RVB. These mechanisms are connected by elements already known in heat stress response (redox, heat-shock proteins, or abscisic acid-related) and with others whose involvement is not so well defined such as shikimate-related, brassinosteriods, or proline proteases together with their potential regulatory elements. This work provides a first in-depth overview about molecular mechanisms underlying the heat stress response and acclimation in P. radiata.

Fracaso multiorgánico secundario a infección por coronavirus no COVID-19 / Multiple-organ failure as a result of non-COVID-19 coronavirus infection

Las infecciones por coronavirus son habituales en los pacientes pediátricos. Por lo general, producen un cuadro clínico leve de infección del tracto respiratorio superior que no suele afectar a los pulmones, salvo en prematuros y niños con enfermedades crónicas de base. Excepcionalmente, afectan a otros órganos (corazón, cerebro, tracto gastrointestinal) e incrementan su gravedad.En relación con la coincidencia temporal con el inicio de la actual pandemia por el nuevo beta coronavirus (SARS-CoV-2), responsable de su enfermedad asociada (COVID-19), se presenta el caso clínico de un paciente de 5 años con fracaso multiorgánico y secuelas neurológicas por afectación bulbar y trombosis vascular ocasionados por un alfa coronavirus (CoV-NL63) debido a su gravedad y excepcionalidad

Coronavirus infections (CoV) are common in pediatric patients. In general, they produce a mild clinical presentation consisting of an upper respiratory tract infection that does not usually infect the lungs, with the exception of preterm infants and children with chronic diseases. These infections exceptionally affect other organs (heart, brain, gastrointestinal tract), thus increasing their severity.In relation to the temporal coincidence with the beginning of the current situation of pandemic by the new beta coronavirus SARS-CoV-2 responsible for its associated disease (COVID-19), this study presents a clinical case of a 5-year-old patient showing multiple-organ failure and neurological sequelae due to bulbar injury and vascular thrombosis caused by an alpha coronavirus (CoV-NL63) due to its severity and exceptionality

Competencias profesionales, gestión clínica y grupos relacionados de diagnósticos: el caso de hospitales públicos chilenos / Professional competences, clinical management and diagnostic related groups in Chilean public hospitals

RESUMEN Objetivo Definir un modelo competencias profesionales para el desarrollo de un sistema de información de apoyo a la Gestión Clínica basado en Grupos Relacionados de Diagnósticos-GRD en hospitales públicos chilenos. Método Mixta. Investigación cualitativa, descriptiva, basada en entrevistas focalizadas con un muestreo teórico o intencionado a cuatro líderes expertos en GRD en Chile, con análisis de contenido; Investigación cuantitativa, con uso de Método Delphi a 18 gestores encargados de la implementación de las unidades de GRD en Chile, con 3 rondas. El análisis de los datos cuantitativos se realizó por conglomerados. Resultados Luego de cinco iteraciones, se evaluaron 78 competencias de un total de 179 en nivel "alto", del tipo: Conocimientos del líder de los equipos, formación profesional preferentemente enfermeras, 15 actitudes y valores, 17 habilidades o destrezas y 12 competencias específicas relacionadas al sistema de codificación. Conclusión Existe tendencia en los profesionales, a requerir el máximo de competencias, se observó una conducta masificadora, con baja discriminación y priorización. Se propone analizar las causas que dificultan la toma de decisiones y priorizar las competencias requeridas; Determinar para cada competencia el nivel requerido, las brechas entre la oferta de competencias y su demanda, y finalmente diseñar un sistema de evaluación del impacto del modelo en el desarrollo de competencias de los equipos.(AU)

ABSTRACT Objective To define a professional skills model for the creation of an information system to support clinical management based on diagnosis related groups (DRG) in Chilean public hospitals. Methods Mixed methodology. Qualitative, descriptive research based on focused interviews, with a theoretical or intentional sample of four leading DRG experts from Chile, with content analysis. Quantitative research using the Delphi method on 18 managers in charge of the implementation of DRG units in Chile, with three rounds. The analysis of quantitative data was carried out by clusters. Results After five iterations, 78 skills were evaluated out of a total of 179 as "high", including knowledge of the team leader, professional training (preferably nurses), 15 attitudes and values, 17 skills and 12 specific skills related to the coding system. Conclusion There is a tendency among professionals to require the maximum skills; a massive behavior was observed, with low discrimination and prioritization. To analyze the causes that make decision-making difficult and to prioritize the required skills is proposed to determine the necessary level for each skill, the gaps between skill offer and demand, and to design a system for evaluating the impact of the model on the development of the skills among the teams.(AU)

Canalización vascular ecoguiada: Experiencia en el paciente pediátrico crítico / Ultrasound-guided vascular cannulation: Experience in critically-ill pediatric patients

Introducción. La canalización vascular central en niños presenta alta complejidad técnica y dificultades. La ecografía vascular puede facilitar este procedimiento. Objetivo. Describir las características de las canalizaciones vasculares ecoguiadas en el paciente pediátrico crítico. Población y métodos. Las variables de interés registradas prospectivamente fueron los vasos más comúnmente canalizados, su localización, la medición del diámetro/profundidad, la tasa de éxito y las complicaciones presentadas, entre otras. Resultados. En 86 pacientes pediátricos, se realizaron 124 punciones vasculares. Los accesos vasculares fueron la vena femoral (39,7%), seguida de la arteria femoral (27,2%) y la vena yugular interna (14,7%). Los vasos femorales se localizaron a una profundidad de 0,75 ± 0,25 mm con un diámetro medio de 0,31 ± 0,16 mm. La profundidad de los vasos venosos yugulares fue menor (0,64 ± 0,24 mm) y su diámetro global, mayor (0,44 ± 0,19 mm). El número medio de intentos en las canalizaciones vasculares ecoguiadas fue de 2,2 ± 1,3. La tasa de éxito fue del 79% asociada a un mayor diámetro del vaso (0,39 ± 0,20 mm vs. 0,28 ± 0,13 mm, p 0,01) y un menor número de intentos (1,90 ± 1,16 vs. 3,45 ± 1,77, p= 0,001). Las complicaciones, fueron la punción accidental de otro vaso (5,3%) y el desarrollo de un hematoma durante la punción (2,3%). Conclusiones. La canalización vascular ecoguiada en los pacientes pediátricos estudiados permite visualizar los vasos y medir su profundidad y diámetro; presenta una alta tasa de éxito y se asocia a una baja tasa de complicaciones.

Introduction. Central vascular cannulation in children is a highly complex technique and poses many difficulties. Vascular ultrasound can make this procedure easier. Objective. To describe the characteristics of ultrasound-guided vascular cannulation in critically-ill pediatric patients. Population and methods. Outcome measures prospectively recorded were vessels most frequently cannulated, their localization, the measurement of their diameter/depth, the success rate and complications developed, among others. Results. One hundred and twenty four vascular punctures were performed in 86 pediatric patients. Vascular accesses were the femoral vein (39.7%), followed by the femoral artery (27.2%) and the internal jugular vein (14.7%). Femoral vessels were localized at a depth of 0.75 ± 0.25 mm, with a mean diameter of 0.31 ± 0.16 mm. The depth of jugular vein vessels was smaller (0.64 ± 0.24 mm) and their overall diameter, larger (0.44 ± 0.19 mm). The mean number of attempts in ultrasound-guided cannulations was 2.2 ± 1.3. The success rate was 79% and was associated to a larger vessel diameter (0.39 ± 0.20 mm vs. 0.28 ± 0.13 mm, p= 0.01) and a lower number of attempts (1.90 ± 1.16 vs. 3.45 ± 1.77, p= 0.001). Complications were accidental puncture of another vessel (5.3%) and hematoma formation during puncture (2.3%). Conclusions. In the pediatric patients studied, ultrasound-guided vascular cannulation allowed vessel visualization and measurement of their depth and diameter; the success rate was high and it was associated to a low complication rate.

Evolution of subpatent parasitaemia in Trypanosoma cruzi chronically infected mice with the help of a cyclophosphamide amplification transfer assay

Avaliamos o potencial do ensaio classico de subinoculacao, modificado pelo tratamento com ciclofosfamida dos animais receptores, na deteccao de parasitemias ocultas em camundongos com infeccao cronica pelo Trypanosoma cruzi. O ensaio, alem de simples, mostrou ter uma alta sensibilidade; assim, utilizando-se parasitas da fase aguda, o tratamento com ciclofosfamida revelou parasitemias em 53,8 por cento dos animais infectados com um tripanosoma da cepa y, e em 20 por cento dos animais infectados com um tripanosoma da cepa CL. O tratamento com ciclofosfamida aumentou a sensibilidade do ensaio de subinoculacao nas infeccoes pela cepa CL, e resultou em igual sensibilidade quando utilizada a cepa Y. Nos camundongos de fase cronica, obtidos a partir de diversos esquemas de imunoprofilaxia (BCG, soro de camundongo imune) ou quimioterapia, o ensaio revelou parasitemias ocultas em 99 por cento dos animais. Auxiliados pelo metodo da subinoculacao-ciclofosfamida estudamos no espaco de um ano a evolucao das parasitemias ocultas em um grupo de camundongos infectados que soberviveram a fase aguda pelo tratamento com Benzonidazol. O ensaio revelou parasitemias ocultas em 100 por cento dos animais. Entretanto, padroes continuos e descontinuos de positividade puderam ser detectados.

Aspectos epidemiológicos sobre las vacunas incluidas en el esquema de vacunación de Venezuela / Epidemiological aspects on vaccines including in vaccination plan of Venezuela

Se hacen consideraciones epidemiológicas sobre las vacunas incluidas en el esquema rutinario de vacunación de Venezuela, suministrando algunos datos históricos así como también aspectos demográficos e información de los Indices Vitales en el país. Son estudiadas las inmunizaciones a través de los datos estadísticos, incluidos en los anuarios de Epidemiología y Estadistica Vital, publicados por el Ministerio de Sanidad y Asistencia Social, sintetizados en cuadros y expresados en gráficos que muestran los logros obtenidos. En ocasiones en forma exitosa, como en el caso de la vacuna antipoliomielítica y con altibajos, como en el caso de las vacunas DPT y antisarampionosa, que no han permitido un efectivo control de las enfermedades. Se hacen una serie de conclusiones y recomendaciones en donde se señalan entre otros temas, la importancia de la Educación Sanitaria y la participación comunal en el exito de los programas de vacunación