Invasive and non-invasive methods to study inflammatory parameters in premature newborns with nrcrotizing enterocolitis

Introducción: La inflamación que acontece en el parto, mayor en el caso de la prematuridad, puede perdurar tras el nacimiento. Esto, unido a otros factores como la inmadurez del sistema inmunológico de la barrera intestinal puede hacer que estos recién nacidos sean susceptibles de presentar diversas enfermedades como la enterocolitis necrotizante (ECN), con una morbimortalidad muy elevada. El objetivo de este proyecto es profundizar en el conocimiento de la relación de la función de la barrera intestinal y la ECN a través del estudio de marcadores inflamatorios intestinales y sistémicos, realizando también una aproximación al conocimiento de las vías metabólicas implicadas en el desarrollo y regulación intestinal del recién nacido a través del estudio de miRNAs en muestras de leche materna de distintos grupos de recién nacidos. Metodología: Es un estudio observacional, descriptivo, analítico de casos y controles. Se establecieron tres grupos: un grupo de neonatos nacidos ≥37 semanas de edad gestacional (EG) con peso al nacer ≥ 2,500 g (grupo T), un grupo de neonatos prematuros nacidos antes de las 37 semanas de EG (grupo PT) y un tercer grupo que desarrolló ECN siguiendo los criterios de Bell. Se les realizó una evaluación clínica completa y se recogieron muestras de meconio en las primeras 48 h de vida realizando también una extracción sanguínea para estudio de diversos marcadores bioquímicos como citoquinas. En el grupo ECN se recogieron muestras una vez iniciada la clínica. En otro subgrupo de niños PT y a T se recogieron muestras de leche materna en el día 0 y día 15 para estudio de miRNAs. Resultados: En el grupo PT, al compararlos con los T se observaron valores más elevados de calprotectina, MPO y PMN-E en heces y menores en la actividad de la fosfatasa alcalina. Y al comparar grupo PT con los niños del grupo ECN, se observaron valores más elevados en este último grupo en los siguientes parámetros: calprotectina, PMN-E, TNF-α, IL-17, MIP, IL-1 β, IL-A y E-selectina, y de forma similar, a excepción de la fosfatasa alcalina también al compararlos con el grupo T. En sangre, se detectaron valores más elevados de IL-1 β, HGF, IL-8 y fosfatasa alcalina en el grupo PT con respecto al grupo T, y el TNF-α y la fosfatasa alcalina fueron significativamente más elevados en pacientes con ECN al compararlos con el grupo PT, y notable aumento en la concentración de IL-1β, IL-8, TNF-α, IL-6, fosfatasa alcalina y NGF cuando comparamos con los T. En la leche materna de niños PT y T se determinaron 379 miRNAs que no se modifican en abundancia en función de la edad gestacional ni de la madurez de la leche participando fundamentalmente en la regulación del desarrollo del sistema inmunológico. La función de los 5 miRNAs más abundantes y más aumentados en la leche inmadura parece estar relacionada con el desarrollo del tubo intestinal, del endotelio y de la función neurológica. Conclusiones: Los neonatos con ECN muestran elevación de parámetros proinflamatorios en heces confirmando la existencia de inflamación en el intestino y la validez de este tipo de muestras para el estudio de diversos biomarcadores. Parece que TNF-α, MIP e IL-1β son parámetros de posible utilidad para detectar la ECN temprana, mientras que IL-17, IL-1α y E-selectina son los marcadores más específicos. En sangre, sólo los niveles circulantes de TNF-α y la actividad de la fosfatasa alcalina parece que podrían ser útiles en la caracterización en la ECN, aunque serían necesarios más estudios para definir un perfil diagnóstico, así como conocer la fisiopatología de la barrera intestinal y su estado en la prematuridad y condiciones de enfermedad.Introduction: The inflammation that occurs during childbirth, greater in prematurity, can persist after birth. This, together with other factors such as the immaturity of the intestinal barrier immune system, can make these newborns susceptible to various diseases such as necrotizing enterocolitis (NEC), with very high morbidity and mortality. The objective of this project is to deepen the knowledge of the relationship between the function of the intestinal barrier and NEC through the study of intestinal and systemic inflammatory markers, also making an approximation to the knowledge of the metabolic pathways involved in the development and intestinal regulation of the newborn through the study of miRNAs in breast milk samples from different groups of newborns. Methodology: It is an observational, descriptive, analytical study of cases and controls. Three groups were established: a group of neonates born ≥37 weeks gestational age (GA) with birth weight ≥ 2,500 g (T group), a group of preterm neonates born before 37 weeks GA (PT group) and a third group that developed NEC following the Bell criteria. A complete clinical evaluation was carried out and meconium samples were collected in the first 48 h of life, also performing a blood extraction to study various biochemical markers such as cytokines. In the ECN group, samples were collected once the symptoms began. In another subgroup of PT and T children, breast milk samples were collected on day 0 and day 15 for the study of miRNAs. Results: In the PT group, when compared with the T group, higher values of calprotectin, MPO and PMN-E were observed in faeces and lower values of alkaline phosphatase activity. And when comparing the PT group with the ECN group, higher values were observed in the latter group in the following parameters: calprotectin, PMN-E, TNF-α, IL-17, MIP, IL-1β, IL-A and E-selectin, and similarly, with the exception of alkaline phosphatase also when compared with the T group. In blood, higher values of IL-1β, HGF, IL-8 and alkaline phosphatase were detected in the PT group with respect to the T group, and TNF-α and alkaline phosphatase were significantly higher in patients with NEC when compared with the PT group, and a notable increase in the concentration of IL-1β, IL-8, TNF-α, IL- 6, alkaline phosphatase and NGF when compared with the T. In the breast milk of PT and T children, 379 miRNAs were determined that do not change in abundance depending on the gestational age or the maturity of the milk, participating in the regulation of development of the immune system. The function of the 5 most abundant and increased miRNAs in immature milk seems to be related to the development of the intestinal tract, the endothelium, and neurological function. Conclusions: Neonates with NEC show elevated proinflammatory parameters in faeces, confirming the existence of inflammation in the intestine and the validity of this type of sample for the study of various biomarkers. It seems that TNF-α, MIP, and IL-1β are potentially useful parameters for detecting early NEC, while IL-17, IL-1α, and E-selectin are the most specific markers. In blood, only the circulating levels of TNF-α and the activity of alkaline phosphatase seem to be useful in the characterization of NEC, although more studies would be necessary to define a diagnostic profile, as well as to know the pathophysiology of the intestinal barrier and their status in prematurity and disease conditions

Sustainability of traditional ecological knowledge: importance, distribution, endemicity and conservation of Spanish medicinal plants

Trabajo presentado en la 58th Annual Meeting of the Society for Economic Botany (Living in a global world: local knowledge ans sustainability), celebrada en Braganza (Portugal) del 4 al 9 de junio de 2017.-- IECTB authors: L Aceituno, R Acosta, A Alvarez, E Barroso, J Blanco, MA Bonet, L Calvet, E Carrio, R Cavero, U DAmbrosio , L Delgado, J Fajardo, I Fernandez-Ordonez, J Garcia, T Garnatje, JA Gonzalez, R Gonzalez-Tejero, A Gras, E Hernandez-Bermejo, E Laguna, JA Latorre, C. Lopez, MJ Macia, E Marcos, V Martinez, G Menendez, M Molina, R Morales, LM Munoz, C Obon, R Ontillera, M Parada, A Perdomo, I Perez, MP Puchades, V Reyes-Garcia, M Rigat, S Rios, D Rivera, R Rodriguez, O Rodriguez, R Roldan, L San Joaquin, FJ Tardio, JR Vallejo, J Valles, H Velasco and A Verde.More than 17,000 of the plant species of the world have been used as medicines. The Mediterranean basin, and specifically Spain, has a great floristic and ethnobotanical richness, comprising its useful flora around 3,000 plant species. This paper studies medicinal plants traditionally used in Spain in order to analyze the sustainability of their exploitation. Given that sustainability is related to the amount of the resource and its gathering pressure, its availability and cultural importance were analysed based on: the number of papers cited from a selection of over 180 papers, the number of 10x10 km UTM grid cells in which the plants were represented, the number of phytosociological inventories in which the presence of the plant has been registered, and searched on their current conservation status in European, national and regional legislations. The total number of wild or naturalized medicinal species in Spain reaches 1,393, 15% of them being endemic. A positive correlation was found among cultural importance and abundance (ρ=0.48) and among cultural importance and distribution (ρ=0.502), showing that abundant widely distributed species are those more commonly used. Most of the medicinal plants (72%) do not appear on the consulted regulations and do not have any legal protection or known threat and only 11 species are registered in any of the annexes of the European Habitats directive. While this study confirms that people tend to select as medicinal abundant and widely distributed species, many other criteria are used for selecting them.Peer reviewe