Análisis del perfil de expresión de microRNAs en tejido embrionario sano y ectópico

La placentación es un proceso fundamental en el éxito de la reproducción, sometido a una estrecha regulación donde el trofoblasto juega una función clave. Este proceso se realiza adecuadamente gracias a la relación estrictamente regulada entre los tejidos materno-fetales, donde intervienen multitud de moléculas y receptores; por tanto es necesario un correcto desarrollo de dicho proceso para asegurar una gestación a término saludable, y la invasión anómala del trofoblasto conducirá a un gran abanico de complicaciones, entre ellas el embarazo ectópico. El proceso de placentación está alterado en las gestaciones ectópicas, produciéndose la implantación fuera de la cavidad endometrial. Las alteraciones en este proceso pueden ser origen o debidas a la aparición de diferentes marcadores tisulares con distintos patrones de expresión en las gestaciones ectópicas, a diferencia de las gestaciones evolutivas, que podrían modificar las vías fundamentales que intervienen en la regulación del proceso y la correcta placentación e implantación. Por otro lado, los microRNAs son moléculas con gran capacidad reguladora de múltiples dianas y multitud de procesos biológicos; además, algunos estudios preliminares de perfiles de expresión de microRNAs en tejido humano mediante microarrays identifican microRNAs selectivamente expresados en tejido placentario. Nuestra hipótesis por tanto plantearía que, dado que existen distintos mecanismos que jugarían un papel en el proceso de regulación de la placentación de los embarazos ectópicos respecto de gestaciones normales evolutivas, existiría un patrón de expresión de microRNAs diferencial entre ambas gestaciones, con lo que el análisis de tal perfil de expresión en ambos tipos de embarazo en las mismas etapas de la gestación podría proporcionarnos nuevos conocimientos y ayudarnos a conocer los mecanismos implicados en la fisiopatología de la gestación ectópica en el ser humano. Nuestro objetivo fue investigar el perfil global de expresión de microRNAs en tejido embrionario procedente de embarazos ectópicos y de interrupciones voluntarias del embarazo. 23 pacientes con embarazo ectópico tubárico y 29 pacientes con interrupción voluntaria del embarazo fueron reclutados. Las muestras de tejido embrionario fueron analizadas por microarrays de microRNA y los resultados obtenidos fueron validados por PCR en tiempo real. Los estudios de microarrays mostraron que cuatro microRNAs mostraron unos niveles de expresión diferencialmente disminuidos (hsa-mir-196b, hsa-mir-30a, hsa-mir-873 y hsa-mir-337-3p) y tres microRNAs mostraron unos niveles de expresión aumentados (hsa-mir-1288, hsa-mir -451, y hsa-mir-223) en muestras procedentes de tejido ectópico comparado con muestras procedentes de tejido control. Hsa-miR-196, hsa-miR-223 y hsa-miR-451 fueron posteriormente validados por PCR en tiempo real en una población más amplia de pacientes compuesta por 15 muestras procedentes de embarazos ectópicos y 21 muestras control. También se realizó un análisis computacional para identificar los genes diana y las vías de señalización que podrían ser moduladas por estos microRNAs diferencialmente expresados. Las vías más significativas encontradas fueron la biosíntesis de O-glicano de tipo mucina y las vías de interacción del receptor de la matriz extracelular. También se comprobó que la desregulación de estos tres microRNAs fue capaz de alterar la expresión de los genes dianas en los tejidos embrionarios incluidos en estas vías (como los genes GALNT13 e ITGA2). En conclusión, el análisis de del perfil de expresión de microRNAs en tejido embrionario procedente de embarazos ectópicos y eutópicos mostró diferentes patrones de expresión que podrían modificar vías de señalización y genes diana que son críticos para la correcta implantación implantación del blastocisto, ayudando a comprender y proporcionando nuevos conocimientos sobre la implantación ectópica en los seres humanos.Placentation is a fundamental process in the success of reproduction, with a strict regulation where the trophoblast has an important role. This process is properly performed because of the strictly regulated relationship between maternal and fetal tissues, where a multitude of molecules and receptors intervene; therefore, a proper development of this process is necessary to ensure a healthy pregnancy at term, and the anomalous invasion of the trophoblast will lead to a large range of complications, including ectopic pregnancy. The placentation process is altered in ectopic gestations, with an implantation outside the endometrial cavity. Alterations in this process may be cause or consequence of different tissue markers with different expression patterns in ectopic gestations, unlike the evolutionary gestations, which could modify the fundamental pathways involved in the regulation of the process and the correct placentation and implantation. On the other hand, microRNAs are molecules with great regulatory capacity of multiple dianas and multitude of biological processes; in addition, some preliminary studies of microRNA expression profiles in human tissue using microarrays identify microRNAs selectively expressed in placental tissue. Our hypothesis is, since there are different mechanisms that would play a role in the process of regulating the placentation of ectopic pregnancies in relation to normal pregnancies, there would be a pattern of expression of differential microRNAs between both gestations, and the analysis of such expression profile in both types of pregnancy in the same stages of gestation could provide us new knowledge and help us to know the mechanisms involved in the pathophysiology of ectopic gestation in humans. Our objective was to investigate the microRNA profile of embryonic tissues in ectopic pregnancies and controlled abortions (voluntary termination of pregnancy). Twenty-three patients suffering from tubal ectopic pregnancies and twenty-nine patients with a normal ongoing pregnancy scheduled for a voluntary termination of pregnancy were recruited. Embryonic tissue samples were analyzed by microRNAs microarray and further validated by real time PCR. Microarray studies showed that four microRNAs were differentially downregulated (hsa-mir-196b, hsa-mir-30a, hsa-mir-873, and hsa-mir-337-3p) and three upregulated (hsa-mir-1288, hsa-mir-451, and hsa-mir-223) in ectopic pregnancies compared to control tissue samples. Hsa-miR-196, hsa-miR-223, and hsa-miR-451 were further validated by real time PCR in a wider population of ectopic pregnancies and control samples. We also performed a computational analysis to identify the gene targets and pathways which might be modulated by these three differentially expressed microRNAs. The most significant pathways found were the mucin type O-glycan biosynthesis and the ECM-receptor-interaction pathways. We also checked that the dysregulation of these three microRNAs was able to alter the expression of the gene targets in the embryonic tissues included in these pathways such as GALNT13 and ITGA2 genes. In conclusion, analysis of microRNAs in ectopic and eutopic embryonic tissues shows different expression patterns that could modify pathways which are critical for correct implantation, providing new insights into the understanding of ectopic implantation in humans

Fenoles totales y capacidad antioxidante estimada con los ensayos DPPH/ABTS en rosas en soluciones preservantes

Tallos de rosa ¿Freedom¿ se evaluaron en dos soluciones preservantes: 8-citrato de hidroxiquinoleina (HQC) y Chrysal CLEAR Professional 2® T-bag (CHRYSAL) en un pulso de 24 h a temperatura ambiente (24 ± 2 °C, 75 % HR) y agua como testigo. Se utilizó un diseño completamente al azar con cuatro repeticiones, la unidad experimental fue un tallo floral. Se evaluó la vida de florero, el contenido de fenoles totales con el método colorimétrico de Folin-Ciocalteauy la capacidad antioxidante total (caT) con los ensayos DPPH y ABTS tanto en hoja como en pétalo. Se realizó el ANOVA y correlaciones simples entre la caT y el contenido de fenoles, y entre ambos ensayos. Los resultados muestran que las soluciones preservantes promueven un aumento en la caT y contenido de fenoles totales en hoja, pero no en pétalo. Las hojas de los tallos florales tratados con CHRYSAL presentaron los mayores contenidos de fenoles totales y caT, mientras que el testigo los menores. La vida media de florero fue de: 13, 11 y 9 días para los tratados con CHRYSAL, HQC y el testigo, respectivamente. El contenido de fenoles totales presentó una estrecha relación positiva (a= 0.01) con la caT: 0.87 y 0.85 medida con el ABTS y 0.92 y 0.85 con el DPPH en hoja y pétalo respectivamente y también ambos métodos se correlacionaron entre sí positiva y significativamente (r= 0.91) en hoja y (r= 0.93) en pétalo.Rose stems �Freedom� were evaluated in two preservative solutions: 8-hydroxyquinoline citrate (HQC) and Chrysal CLEAR® Professional 2 T-bag (CHRYSAL) in a pulse of 24 h at room temperature (24 ± 2 °C, 75% HR) and water as control. A completely randomized design with four replications was used; the experimental unit was a flower stem. Vase life, total phenolic content with the colorimetric method Folin-Ciocalteauy, total antioxidant capacity (caT) with DPPH and ABTS assays in both leaf and petal was evaluated. ANOVA and simple correlations between caT and content of phenols and between both assays was made. The results show that preservative solutions promote an increase in caT and total phenolic content in leaf, but not in petal. The leaves of the flowering stems treated with CHRYSAL had the highest contents of total phenols and caT, while control the lowest. The average vase life was 13, 11 and 9 days for those treated with CHRYSAL, HQC and control, respectively. The total phenolic content showed a strong positive relation (a =0.01) with caT: 0.87 and 0.85 measured with ABTS and 0.92 and 0.85 with DPPH in leaf and petal respectively and also both methods were correlated positively and significantly with each other (r= 0.91) in leaf and (r= 0.93) in petal

The Lin28/Let-7 System in Early Human Embryonic Tissue and Ectopic Pregnancy

Our objective was to determine the expression of the elements of the Lin28/Let-7 system, and related microRNAs (miRNAs), in early stages of human placentation and ectopic pregnancy, as a means to assess the potential role of this molecular hub in the pathogenesis of ectopic gestation. Seventeen patients suffering from tubal ectopic pregnancy (cases) and forty-three women with normal on-going gestation that desired voluntary termination of pregnancy (VTOP; controls) were recruited for the study. Embryonic tissues were subjected to RNA extraction and quantitative PCR analyses for LIN28B, Let-7a, miR-132, miR-145 and mir-323-3p were performed. Our results demonstrate that the expression of LIN28B mRNA was barely detectable in embryonic tissue from early stages of gestation and sharply increased thereafter to plateau between gestational weeks 7–9. In contrast, expression levels of Let-7, mir-132 and mir-145 were high in embryonic tissue from early gestations (≤6-weeks) and abruptly declined thereafter, especially for Let-7. Opposite trends were detected for mir-323-3p. Embryonic expression of LIN28B mRNA was higher in early stages (≤6-weeks) of ectopic pregnancy than in normal gestation. In contrast, Let-7a expression was significantly lower in early ectopic pregnancies, while miR-132 and miR-145 levels were not altered. Expression of mir-323-3p was also suppressed in ectopic embryonic tissue. We are the first to document reciprocal changes in the expression profiles of the gene encoding the RNA-binding protein, LIN28B, and the related miRNAs, Let-7a, mir-132 and mir-145, in early stages of human placentation. This finding suggests the potential involvement of LIN28B/Let-7 (de)regulated pathways in the pathophysiology of ectopic pregnancy in humans

Análisis comparativo de la participación por género en la olimpiada estatal de química Chihuahua, México

The disciplines in science, technology, engineering and mathematics (STEM), present a direct contribution in the fulfillment of the 17 objectives presented by the 2030 Agenda adopted by the General Assembly of the United Nations (UN) in September 2015, for which It is essential to promote the scientific vocation among girls, boys and young people. In addition, the Sustainable Development Goals (SDG) 4 and 5 are aimed at quality in education and gender equality. Taking into account the aforementioned, the main purpose of this research is to carry out a comparative analysis to determine the degree of participation by gender of the students in the State Chemistry Olympiad Chihuahua Mexico, as well as the trend that exists in the same indicator by gender. campus and by city, in addition to the influence of the event in promoting scientific and/or technological vocationsLas disciplinas en ciencias, tecnología, ingeniería y matemáticas, STEM acrónimo de Science, Technology, Engineering and Mathematics presentan una contribución directamente en el cumplimiento a los 17 objetivos presentados por la Agenda 2030 adoptada por la Asamblea General de las Naciones Unidas (ONU) en septiembre de 2015, por lo que es imprescindible fomentar la vocación científica entre niñas, niños y jóvenes. Además, los Objetivos de Desarrollo Sostenible (ODS) 4 y 5 están orientados a la calidad en la educación e igualdad de género. Teniendo en cuenta lo antes mencionado, la finalidad principal de la presente investigación es realizar un análisis comparativo para determinar el grado de participación por género de los estudiantes en la Olimpiada Estatal de Química Chihuahua México, así como la tendencia que existe en el mismo indicador por plantel y por ciudad, además de la influencia del certamen en fomentar las vocaciones científicas y/o tecnológicas

Deregulation of miR-324/KISS1/kisspeptin in early ectopic pregnancy: mechanistic findings with clinical and diagnostic implications

[Abstract] BACKGROUND: Ectopic pregnancy is a life-threatening condition for which novel screening tools that would enable early accurate diagnosis would improve clinical outcomes. Kisspeptins, encoded by KISS1, play an essential role in human reproduction, at least partially by regulating placental function and possibly embryo implantation. Kisspeptin levels are elevated massively in normal pregnancy and reportedly altered in various gestational pathologic diseases. Yet, the pathophysiologic role of KISS1/kisspeptin in ectopic pregnancy has not been investigated previously. OBJECTIVE: The purpose of this study was to evaluate changes of KISS1/kisspeptin levels in ectopic pregnancy and their underlaying molecular mechanisms and to ascertain the diagnostic implications of these changes. STUDY DESIGN: A total of 122 women with normal pregnancy who underwent voluntary termination of pregnancy and 84 patients who experienced tubal ectopic pregnancy were recruited. Measurements of plasma kisspeptins and KISS1 expression analyses in human embryonic/placental tissue were conducted in ectopic pregnancy and voluntary termination of pregnancy control subjects during the early gestational window (<12 weeks). Putative microRNA regulators of KISS1 were predicted in silico, followed by expression analyses of selected microRNAs and validation of repressive interactions in vitro. Circulating levels of these microRNAs were also assayed in ectopic pregnancy vs voluntary termination of pregnancy. RESULTS: Circulating kisspeptins gradually increased during the first trimester of normal pregnancy but were reduced markedly in ectopic pregnancy. This profile correlated with the expression levels of KISS1 in human embryonic/placental tissue, which increased in voluntary termination of pregnancy but remained suppressed in ectopic pregnancy. Bioinformatic predictions and expression analyses identified miR-27b-3p and miR-324-3p as putative repressors of KISS1 in human embryonic/placental tissue at <12 weeks gestation, when expression of microRNAs was low in voluntary termination of pregnancy control subjects but significantly increased in ectopic pregnancy. Yet, a significant repressive interaction was documented only for miR-324-3p, occurring at the predicted 3'-UTR of KISS1. Interestingly, circulating levels of miR-324-3p, but not of miR-27b-3p, were suppressed distinctly in ectopic pregnancy, despite elevated tissue expression of the pre-microRNA. A decision-tree model that used kisspeptin and miR-324-3p levels was successful in discriminating ectopic pregnancy vs voluntary termination of pregnancy, with a receiver-operating characteristic area under the curve of 0.95±0.02 (95% confidence interval). CONCLUSION: Our results document a significant down-regulation of KISS1/kisspeptins in early stages of ectopic pregnancy via, at least partially, a repressive interaction with miR-324-3p. Our data identify circulating kisspeptins and miR-324-3p as putative biomarkers for accurate screening of ectopic pregnancy at early gestational ages.Ministerio de E$conomía y Competitividad (España); BFU2014-57581-PMinisterio de Economía y Competitividad ; BFU2017-83934-PInstituto de Salud Carlos III; PIE-00005Junta de Andalucía; P08-CVI-03788Junta de Andalucía; P12-FQM-0194

Utilización de las Redes sociales en el aprendizaje colaborativo y transversal de la Farmacovigilancia Veterinaria

Sección Deptal. de Farmacología y Toxicología (Veterinaria)Fac. de VeterinariaFALSEsubmitte

Identification of clusters of asthma control: A preliminary analysis of the inspirers studies

This work was funded by ERDF (European Regional Development Fund) through the operations: POCI- -01-0145-FEDER-029130 (“mINSPIRERS—mHealth to measure and improve adherence to medication in chronic obstructive respiratory diseases - generalisation and evaluation of gamification, peer support and advanced image processing technologies”) co-funded by the COMPETE2020 (Programa Operacional Competitividade e Internacionalização), Portugal 2020 and by Portuguese Funds through FCT (Fundação para a Ciência e a Tecnologia).© 2020, Sociedade Portuguesa de Alergologia e Imunologia Clinica. All rights reserved. Aims: To identify distinct asthma control clusters based on Control of Allergic Rhinitis and Asthma Test (CARAT) and to compare patients’ characteristics among these clusters. Methods: Adults and adolescents (≥13 years) with persistent asthma were recruited at 29 Portuguese hospital outpatient clinics, in the context of two observational studies of the INSPIRERS project. Demographic and clinical characteristics, adherence to inhaled medication, beliefs about inhaled medication, anxiety and depression, quality of life, and asthma control (CARAT, >24 good control) were collected. Hierarchical cluster analysis was performed using CARAT total score (CARAT-T). Results: 410 patients (68% adults), with a median (percentile 25–percentile 75) age of 28 (16-46) years, were analysed. Three clusters were identified [mean CARAT-T (min-max)]: cluster 1 [27(24-30)], cluster 2 [19(14-23)] and cluster 3 [10(2-13)]. Patients in cluster 1 (34%) were characterised by better asthma control, better quality of life, higher inhaler adherence and use of a single inhaler. Patients in clusters 2 (50%) and 3 (16%) had uncontrolled asthma, lower inhaler adherence, more symptoms of anxiety and depression and more than half had at least one exacerbation in the previous year. Further-more, patients in cluster 3 were predominantly female, had more unscheduled medical visits and more anxiety symp-toms, perceived a higher necessity of their prescribed inhalers but also higher levels of concern about taking these inhalers. There were no differences in age, body mass index, lung function, smoking status, hospital admissions or specialist physician follow-up time among the three clusters. Conclusion: An unsupervised method based on CARAT--T, identified 3 clusters of patients with distinct, clinically meaningful characteristics. The cluster with better asthma control had a cut-off similar to the established in the validation study of CARAT and an additional cut-off seems to distinguish more severe disease. Further research is necessary to validate the asthma control clusters identified.publishersversionpublishe

EDUCACIÓN AMBIENTAL Y SOCIEDAD. SABERES LOCALES PARA EL DESARROLLO Y LA SUSTENTABILIDAD

Este texto contribuye al análisis científico de varias áreas del conocimiento como la filosofía social, la patología, la educación para el cuidado del medio ambiente y la sustentabilidad que inciden en diversas unidades de aprendizaje de la Licenciatura en Educación para la Salud y de la Maestría en Sociología de la SaludLas comunidades indígenas de la sierra norte de Oaxaca México, habitan un territorio extenso de biodiversidad. Sin que sea una área protegida y sustentable, la propia naturaleza de la región ofrece a sus visitantes la riqueza de la vegetación caracterizada por sus especies endémicas que componen un paisaje de suma belleza