Methylation-Dependent Gene Silencing Induced by Interleukin 1β via Nitric Oxide Production

Interleukin (IL)-1β is a pleiotropic cytokine implicated in a variety of activities, including damage of insulin-producing cells, brain injury, or neuromodulatory responses. Many of these effects are mediated by nitric oxide (NO) produced by the induction of NO synthase (iNOS) expression. We report here that IL-1β provokes a marked repression of genes, such as fragile X mental retardation 1 (FMR1) and hypoxanthine phosphoribosyltransferase (HPRT), having a CpG island in their promoter region. This effect can be fully prevented by iNOS inhibitors and is dependent on DNA methylation. NO donors also cause FMR1 and HPRT gene silencing. NO-induced methylation of FMR1 CpG island can be reverted by demethylating agents which, in turn, produce the recovery of gene expression. The effects of IL-1β and NO appear to be exerted through activation of DNA methyltransferase (DNA MeTase). Although exposure of the cells to NO does not increase DNA MeTase gene expression, the activity of the enzyme selectively increases when NO is applied directly on a nuclear protein extract. These findings reveal a previously unknown effect of IL-1β and NO on gene expression, and demonstrate a novel pathway for gene silencing based on activation of DNA MeTase by NO and acute modification of CpG island methylation

Regulación de la Respuesta a Hipoxia y la Actividad Mitocondrial por el Óxido Nítrico en las Células Madre Embrionarias y Células de Pluripotencia Inducida

La expansión de las células pluripotentes (ESC e iPS) en condiciones que mantengan sus propiedades de autorenovación y pluripotencia es necesario en un programa de terapia celular. Previamente en nuestro laboratorio hemos observado que bajas concentraciones de óxido nítrico (NO) soportan la expansión de estos tipos celulares (Tejedo JR et al; 2010). Los mecanismos comprometidos en esta respuesta no son conocidos. En este trabajo presentamos evidencias de que cuando estas células crecen en condiciones de normoxia, el NO activa una respuesta similar a la hipoxia, consiguiendo así el mantenimiento de la pluripotencia y la autorrenovación (Catherine E Forristal et al ; 2010).Para comprobar que el NO activa la repuesta celular a hipoxia se ha estudiado la estabilidad y localización celular de la proteína HIFs (factor inductor de hipoxia) mediante western blotting e inmunocitoquímica en las condiciones experimentales de hipoxia, normoxia y normoxia en presencia de bajas concentraciones de NO. También se ha llevado a cabo el estudio de la estabilidad de la proteína von Hippel-Lindau tumor supresor (VHL) encargada de la  degradación de HIF. Por otro lado, dado la estrecha relación existente entre hipoxia y metabolismo celular ( Jin Zhang et al; 2012), hemos analizado la expresión de genes dianas de HIF mediante RT-PCR implicados en la vía glucolítica como son la hexoquinasa-2 (HK-2),  lactato deshidrogensa (LDH) y piruvato deshidrogenasa kinasa1 (PDK1), además del estudio de otros genes diana como, Nanog involucrado en el mantenimiento de la pluripotencia y VEGF cuya función está relacionada con la angiogénesis.Los resultados obtenidos hasta el momento muestran que las células mantenidas en cultivo bajo condiciones de normoxia con bajas concentraciones de DETA-NO (2 y 10 uM) adicionadas al medio para que liberen NO consiguen estabilizar la proteína HIF-1 alpha y tras el análisis de los genes diana de HIF-1 alpha implicados en la vía glucolítca como HK-2, LDH y PDK1 se ha podido comprobar que estos se encuentran sobreexpresados tanto en condiciones de hipoxia como en las condiciones de normoxia más DETA-NO.Tras estos estudios iniciales se puede concluir que bajas concentraciones de NO en condiciones de normoxia consiguen activar una respuesta celular biológica similar a la que ocurre en hipoxia, que se demuestra al observar niveles estables de la proteína que regula esa adaptación celular (HIF) y la sobreexpresión de sus genes diana

Technical and clinical evaluation of a closed loop TIVA system with SEDLineTM spectral density monitoring: Multicentric prospective cohort study

Introduction: Closed loop total intravenous anesthesia is a technique in which the patient’s hemodynamic and anesthetic depth variables are monitored, and based on this information, a computer controls the infusion rate of drugs to keep them within pre-established clinical parameters. Objective: To describe the technical and clinical performance of a closed loop system for total intravenous anesthesia with propofol and remifentanil, using the SEDLineTM monitor Design: Multicentric prospective cohort study Setting: Surgery room Patients: ASA I-II undergoing elective surgery Measurements: The authors designed a closed loop system that implements a control algorithm based on anesthetic depth monitoring and the Patient State Index (PSITM) of the SEDLine monitor for propofol, and on hemodynamic variables for remifentanil. The measurement of clinical performance was made based on the percentage of PSITM maintenance time in the range 20–50. Precision analysis was evaluated by measuring median performance error (MDPE) can be defined as the median difference between actual and desired values, which refers to the degree of precision in which the controller is able to maintain the control variable within the objective set by the anesthesiologist; it represents the direction (over-prediction or underprediction) of performance error (PE) rather than size of errors, which is represented by MDAPE, median absolute percentage error, Wobble index, which is used for measuring the intrasubject variability in performance error. Results: Data were obtained from 93 patients in three healthcare centers. The percentage of PSITM maintenance time in the 20–50 range was 92% (80.7–97.0). MDPE was 10.7 (− 11.0–18.0), MDAPE 21.0 (14.2– 26.8) and wobble 10.7 (7.0–16.9). No adverse surgical or anesthetic events were found

Cluster Analysis of Physical Activity Patterns, and Relationship with Sedentary Behavior and Healthy Lifestyles in Prepubertal Children: Genobox Cohort

Sedentary habits during childhood are associated with adverse health outcomes. The aim of this work was to cluster lifestyle behaviors and metabolic biomarkers to establish different patterns in children. Their physical and sedentary activities were evaluated by accelerometry, and questionnaires that included lifestyle behaviors, such as adherence to a Mediterranean diet, anthropometry and blood biochemical markers. Cluster analysis was performed to establish different groups based on physical activity levels. A total of 489 children were finally selected. Cluster 1 included children with a mostly sedentary state, whereas Cluster 3 included the most active children and Cluster 2 included children that did not fit into either the sedentary or the highly active groups. In Cluster 3, 56% of children were in a sports club, and a lower percentage used electronic devices in their rooms compared to the other groups. Cluster 1 children exhibited higher insulin, HOMA-IR and triacylglycerides with respect to the other groups. No differences were found regarding adherence to a Mediterranean diet. The choice to practice an extracurricular sport could be an influencing factor to increase exercise and ensure an active lifestyle in children. Reducing or limiting screen time mainly in children''s rooms could contribute to an active lifestyle

Nitric Oxide And Hypoxia Response In Pluripotent Stem Cells

The expansion of pluripotent cells (ESCs and iPSCs) under conditions that maintain their pluripotency is necessary to implement a cell therapy program. Previously, we have described that low nitric oxide (NO) donor diethylenetriamine/nitric oxide adduct (DETA-NO) added to the culture medium, promote the expansion of these cell types. The molecular mechanisms are not yet known. We present evidences that ESC and iPSCs in normoxia in presence of low NO triggers a similar response to hypoxia, thus maintaining the pluripotency. We have studied the stability of HIF-1α (Hypoxia Inducible Factor) in presence of low NO. Because of the close relationship between hypoxia, metabolism, mitochondrial function and pluripotency we have analyzed by q RT-PCR the expression of genes involved in the glucose metabolism such as: HK2, LDHA and PDK1; besides other HIF-1α target gene. We further analyzed the expression of genes involved in mitochondrial biogenesis such as PGC1α, TFAM and NRF1 and we have observed that low NO maintains the same pattern of expression that in hypoxia. The study of the mitochondrial membrane potential using Mito-Tracker dye showed that NO decrease the mitochondrial function. We will analyze other metabolic parameters, to determinate if low NO regulates mitochondrial function and mimics Hypoxia Response. The knowledge of the role of NO in the Hypoxia Response and the mechanism that helps to maintain self-renewal in pluripotent cells in normoxia, can help to the design of culture media where NO could be optimal for stem cell expansion in the performance of future cell therapies

Evaluación de la actividad alelopática del extracto en acetato de etilo de Miconia caudata (bonpl.) dc.

El objetivo de este trabajo fue evaluar el potencial alelopático del extracto en acetato de etilo (AcOEt) y sus fracciones obtenidas de las hojas de Miconia caudata (melastomataceae), a través del bioensayo de germinación y crecimiento en semillas de Lactuca sativa L. Inicialmente, el extracto en AcOEt se separó en columna cromatográfica sobre DIAION HP-20, utilizando un sistema de elución agua-isopropanol, obteniendo 20 fracciones las cuales fueron monitoreadas mediante la medición de la absorbancia a 280 nm y por cromatografía en capa delgada en fase normal y reversa. Adicionalmente, se realizaron pruebas químicas para la identificación de diferentes núcleos fitoquímicos. Los resultados obtenidos mostraron que la fracción F1M presentó un efecto estimulador de crecimiento en el hipocótilo con un máximo valor de 54.39%, dicho efecto se puede atribuir a la presencia de taninos condensados. Las fracciones F1L y F1N tuvieron un comportamiento de actividad alelopática contrario, inhibiendo el crecimiento del hipocótilo de L. sativa. La fracción F1R presentó un mayor efecto inhibidor sobre las plántulas, con valores máximos de 86.67% y 93.78%. A la fracción F1R se le realizaron pruebas químicas que indicaron la presencia de lactonas sesquiterpénicas, las cuales pueden estar involucradas en la actividad alelopática presentada

Development of a Fixed-Wing Drone System for Aerial Insect Sampling

Most current insect research techniques are ground-based and provide scarce information about flying insects in the planetary boundary layer (PBL), which remains a poorly studied ecological niche. To address this gap, we developed a new insect-sampling method consisting of a fixed-wing drone platform with net traps attached to the fuselage, a mobile design that has optimal aerodynamic characteristics for insect capture in the PBL. We tested the proposed device on 16 flights in Doñana National Park (Spain) with two different trap designs fitted on the fuselage nose and wing. We collected 34 insect specimens belonging to four orders with a representation of twelve families at mean altitudes below 23 m above ground level and sampling altitudes between 9 and 365 m. This drone insect-sampling design constitutes a low-cost and low-impact method for insect monitoring in the PBL, especially in combination with other remote sensing technologies that directly quantify aerial insect abundance but do not provide taxonomic information, opening interesting possibilities for ecology and entomological research, with the possibility of transfer to economically important sectors, such as agriculture and health

Regulación de la Expresión del Factor de Transcripción Pdx1 por Altas Concentraciones de Óxido Nítrico en Células Madre Embrionarias de Ratón

Estudios previos en nuestro laboratorio muestran que el óxido nítrico a altas concentraciones promueve la diferenciación de células madre embrionarias de ratón (mESC), e induce la expresión de genes de endodermo, entre ellos el factor de transcripción Pdx1, “Pancreas/duodenum homeobox protein 1”. Pdx1, tiene un rol importante en el desarrollo embrionario del páncreas y en la células beta madura. Con el fin de conocer los mecanismos por los que el NO incrementa la expresión de Pdx1, hemos estudiado la región promotora de este gen, encontrando que la expresión de Pdx1 se asocia con la liberación del factor de transcripción Egr1. Además, análisis epigenéticos del promotor muestran que la expresión de Pdx1 va asociada a un cambio en el patrón de metilación de su promotor y a la ocupación de marcas de histonas activadoras (H3Ac y H3K4me3) y a la desocupación de modificadores represores de histonas (HDAC) en el promotor de Pdx1

Extreme, wintertime Saharan dust intrusion in the Iberian Peninsula: Lidar monitoring and evaluation of dust forecast models during the February 2017 event

The research leading to these results has received funding from the H2020 program from the European Union (grant agreement no. 654109, 778349) and also from the Spanish Ministry of Industry, Economy and Competitiviness (MINECO, ref. CGL2013-45410-R, CGL2016-81092-R, CGL2017-85344-R, TEC2015-63832-P), the Spanish Ministry of Science, Innovation and Universities (ref. CGL2017-90884-REDT); the CommSensLab "Maria de Maeztu" Unity of Excellence (ref. MDM-2016-0600) financed by the Spanish Agencia Estatal de Investigación. Co-funding was also provided by the European Union through the European Regional Development Fund (ref. POCI-01-0145-FEDER-007690, ALT20-03-0145-FEDER-000004, ALT20-03-0145-FEDER-000011); by the Andalusia Regional Government (ref. P12-RNM-2409); by the Madrid Regional Government (projects TIGAS-CM, ref. Y2018/EMT-5177 and AIRTEC-CM, ref. P2018/EMT4329); by the University of Granada through “Plan Propio. Programa 9 Convocatoria 2013” and by the Portuguese Foundation for Science and Technology and national funding (ref. SFRH/BSAB/143164/2019). The BSC-DREAM8b and NNMB/BSC-Dust (now NMMB-MONARCH) model simulations were performed by the Mare Nostrum supercomputer hosted by the Barcelona Supercomputer Center (BSC). S. Basart acknowledges the AXA Research Fund for supporting aerosol research at the BSC through the AXA Chair on Sand and Dust Storms Fund, as well as the InDust project (COST Action CA16202). The authors gratefully acknowledge the NOAA Air Resources Laboratory (ARL) for the provision of the HYSPLIT transport and dispersion model and/or READY website (http://www.ready.noaa.gov) used in this publication.An unprecedented extreme Saharan dust event was registered in winter time from 20 to 23 February 2017 over the Iberian Peninsula (IP). We report on aerosol optical properties observed under this extreme dust intrusion through passive and active remote sensing techniques. For that, AERONET (AErosol RObotic NETwork) and EARLINET (European Aerosol Research LIdar NETwork) databases are used. The sites considered are: Barcelona (41.38°N, 2.17°E), Burjassot (39.51°N, 0.42°W), Cabo da Roca (38.78°N, 9.50°W), Évora (38.57°N, 7.91°W), Granada (37.16°N, 3.61°W) and Madrid (40.45°N, 3.72°W). Large aerosol optical depths (AOD) and low Ångström exponents (AE) are observed. An AOD of 2.0 at 675 nm is reached in several stations. A maximum peak of 2.5 is registered in Évora. During and around the peak of AOD, AEs close to 0 and even slightly negative are measured. With regard to vertically-resolved aerosol optical properties, particle backscatter coefficients as high as 15 Mm−1 sr−1 at 355 nm are recorded at the lidar stations. Layer-mean lidar ratios are found in the range 40–55 sr at 355 nm and 34–61 sr at 532 nm during the event. The particle depolarization ratios are found to be constant inside the dust layer, and consistent from one site to another. Layer-mean values vary in the range 0.19–0.31. Another remarkable aspect of the event is the limited vertical distribution of the dust plume which never exceeds 5 km. The extreme aspect of the event also presented a nice case for testing the ability of two dust forecast models, BSC-DREAM8b and NMMB/BSC-Dust, to reproduce the arrival, the vertical distribution and the intensity of the dust plume over a long-range transport region. In the particular case of the February 2017 dust event, we found a large underestimation in the forecast of the extinction coefficient provided by BSC-DREAM8b at all heights independently of the site. In contrast NMMB/BSC-Dust forecasts presented a better agreement with the observations, especially in southwestern part of the IP. With regard to the forecast skill as a function of lead time, no clear degradation of the prognostic is appreciated at 24, 48 and 72 h for Évora and Granada stations (South). However the prognostic does degrade (bias increases and/or correlation decreases) for Barcelona (North), which is attributed to the fact that Barcelona is at a greater distance from the source region and to the singularity of the event.Funding from the H2020 program from the European Union (grant agreement no. 654109, 778349)Spanish Ministry of Industry, Economy and Competitiviness (MINECO, ref. CGL2013-45410-R, CGL2016-81092-R, CGL2017-85344-R, TEC2015-63832-P)Spanish Ministry of Science, Innovation and Universities (ref. CGL2017-90884-REDT)CommSensLab "Maria de Maeztu" Unity of Excellence (ref. MDM-2016-0600) financed by the Spanish Agencia Estatal de InvestigaciónCo-funding was also provided by the European Union through the European Regional Development Fund (ref. POCI-01-0145-FEDER-007690, ALT20-03-0145-FEDER-000004, ALT20-03-0145-FEDER-000011)Andalusia Regional Government (ref. P12-RNM-2409); by the Madrid Regional Government (projects TIGAS-CM, ref. Y2018/EMT-5177 and AIRTEC-CM, ref. P2018/EMT4329)Portuguese Foundation for Science and Technology and national funding (ref. SFRH/BSAB/143164/2019

Pdx1 is transcriptionally regulated by EGR-1 during nitric oxide-induced endoderm differentiation of mouse embryonic stem cells

The transcription factor, early growth response-1 (EGR-1), is involved in the regulation of cell differentiation, proliferation, and apoptosis in response to different stimuli. EGR-1 is described to be involved in pancreatic endoderm differentiation, but the regulatory mechanisms controlling its action are not fully elucidated. Our previous investigation reported that exposure of mouse embryonic stem cells (mESCs) to the chemical nitric oxide (NO) donor diethylenetriamine nitric oxide adduct (DETA-NO) induces the expression of early differentiation genes such as pancreatic and duodenal homeobox 1 (Pdx1). We have also evidenced that Pdx1 expression is associated with the release of polycomb repressive complex 2 (PRC2) and P300 from the Pdx1 promoter; these events were accompanied by epigenetic changes to histones and site-specific changes in the DNA methylation. Here, we investigate the role of EGR-1 on Pdx1 regulation in mESCs. This study reveals that EGR-1 plays a negative role in Pdx1 expression and shows that the binding capacity of EGR-1 to the Pdx1 promoter depends on the methylation level of its DNA binding site and its acetylation state. These results suggest that targeting EGR-1 at early differentiation stages might be relevant for directing pluripotent cells into Pdx1-dependent cell lineages.C.S.-A. was a doctoral PFIS fellow of ISCIII (FI11/00301). This study was supported by grants from Consejería de Igualdad, Salud y Políticas Sociales, Junta de Andalucía (PI105/2010, TCMR06/009, and PI-0022) to F.J.B. and F.M.; from Consejería de Economía, Innovación, Ciencia y Empleo, Junta de Andalucía (CTS-7127/2011) to F.J.B.; from ISCIII cofunded by Fondos FEDER (RED-TERCEL RD 16-011-0034, along with PICI PICI21/00016 and GVA-COVID19/2021/047 to B.S.); from Servicio Andaluz de Salud (SAS 11245), and from Ministerio de Economía y Competitividad- Secretaría de Estado de Investigación Desarrollo e Innovación (IPT-2011-1615-900000). Along with this, there was support to PAIDI group CTS576, and by the European Regional Development Fund (FEDER) and the Consejería de Economía, Conocimiento, Empresas y Universidades de la Junta de Andalucía, within the framework of the operational program FEDER Andalucía 2014–2020. Specific Objective 1.2.3 “Promotion and generation of frontier knowledge and knowledge oriented to the challenges of society, development of emerging technologies” led the reference research project (UPO-1381598) of J.R.T.Peer reviewe