MAL regulates clathrin-mediated endocytosis at the apical surface of Madin–Darby canine kidney cells

MAL is an integral protein component of the machinery for apical transport in epithelial Madin–Darby canine kidney (MDCK) cells. To maintain its distribution, MAL cycles continuously between the plasma membrane and the Golgi complex. The clathrin-mediated route for apical internalization is known to differ from that at the basolateral surface. Herein, we report that MAL depends on the clathrin pathway for apical internalization. Apically internalized polymeric Ig receptor (pIgR), which uses clathrin for endocytosis, colocalized with internalized MAL in the same apical vesicles. Time-lapse confocal microscopic analysis revealed cotransport of pIgR and MAL in the same endocytic structures. Immunoelectron microscopic analysis evidenced colabeling of MAL with apically labeled pIgR in pits and clathrin-coated vesicles. Apical internalization of pIgR was abrogated in cells with reduced levels of MAL, whereas this did not occur either with its basolateral entry or the apical internalization of glycosylphosphatidylinositol-anchored proteins, which does not involve clathrin. Therefore, MAL is critical for efficient clathrin-mediated endocytosis at the apical surface in MDCK cells

Social Farming in Catalonia: diagnosis of an emerging phenomenon

Social Farming (SF) engages groups at risk of social exclusion in agricultural activities with the aim of including them in society, providing them with job opportunities, and empowering them. This social economic field, according to key agents, helps to sustain small farms, which contributes to local development and territorial cohesion from the viewpoint of multifunctional agriculture. SF, in its various types and models, has already existed in different countries of Europe since more than 30 years. In Catalonia, scope of our study, it is a phenomenon that has begun its development in late twentieth century. In this paper, we show some theoretical and conceptual elements about SF, describing the types of entities that exist in Catalonia, their activities, and the main stakeholders that shape the SF sector from the economic, social and environmental perspectives. Finally, we make a diagnosis about this social and economic sector and sketch out some future challenges. In order to conduct a thorough diagnosis of the phenomenon, we combined quantitative and qualitative methodology

Pulsational frequencies of the eclipsing delta-Scuti star HD 172189

The eclipsing delta-Scuti star HD 172189 is a probable member of the open cluster IC 4756 and a promising candidate target for the CoRoT mission. The detection of pulsation modes is the first step in the asteroseismological study of the star. Further, the calculation of the orbital parameters of the binary system allows us to make a dynamical determination of the mass of the star, which works as an important constraint to test and calibrate the asteroseismological models. From a detailed frequency analysis of 210 hours of photometric data of HD 172189 obtained from the STEPHI XIII campaign we have identified six pulsation frequencies with a confidence level of 99% and a seventh with a 65% confidence level in the range between 100-300 uHz. In addiction, three eclipses were observed during the campaign, allowing us to improve the determination of the orbital period of the system.Comment: 6 pages, 7 figure

MDCK Cystogenesis Driven by Cell Stabilization within Computational Analogues

The study of epithelial morphogenesis is fundamental to increasing our understanding of organ function and disease. Great progress has been made through study of culture systems such as Madin-Darby canine kidney (MDCK) cells, but many aspects of even simple morphogenesis remain unclear. For example, are specific cell actions tightly coupled to the characteristics of the cell's environment or are they more often cell state dependent? How does the single lumen, single cell layer cyst consistently emerge from a variety of cell actions? To improve insight, we instantiated in silico analogues that used hypothesized cell behavior mechanisms to mimic MDCK cystogenesis. We tested them through in vitro experimentation and quantitative validation. We observed novel growth patterns, including a cell behavior shift that began around day five of growth. We created agent-oriented analogues that used the cellular Potts model along with an Iterative Refinement protocol. Following several refinements, we achieved a degree of validation for two separate mechanisms. Both survived falsification and achieved prespecified measures of similarity to cell culture properties. In silico components and mechanisms mapped to in vitro counterparts. In silico, the axis of cell division significantly affects lumen number without changing cell number or cyst size. Reducing the amount of in silico luminal cell death had limited effect on cystogenesis. Simulations provide an observable theory for cystogenesis based on hypothesized, cell-level operating principles

A Computational Approach to Understand In Vitro Alveolar Morphogenesis

Primary human alveolar type II (AT II) epithelial cells maintained in Matrigel cultures form alveolar-like cysts (ALCs) using a cytogenesis mechanism that is different from that of other studied epithelial cell types: neither proliferation nor death is involved. During ALC formation, AT II cells engage simultaneously in fundamentally different, but not fully characterized activities. Mechanisms enabling these activities and the roles they play during different process stages are virtually unknown. Identifying, characterizing, and understanding the activities and mechanisms are essential to achieving deeper insight into this fundamental feature of morphogenesis. That deeper insight is needed to answer important questions. When and how does an AT cell choose to switch from one activity to another? Why does it choose one action rather than another? We report obtaining plausible answers using a rigorous, multi-attribute modeling and simulation approach that leveraged earlier efforts by using new, agent and object-oriented capabilities. We discovered a set of cell-level operating principles that enabled in silico cells to self-organize and generate systemic cystogenesis phenomena that are quantitatively indistinguishable from those observed in vitro. Success required that the cell components be quasi-autonomous. As simulation time advances, each in silico cell autonomously updates its environment information to reclassify its condition. It then uses the axiomatic operating principles to execute just one action for each possible condition. The quasi-autonomous actions of individual in silico cells were sufficient for developing stable cyst-like structures. The results strengthen in silico to in vitro mappings at three levels: mechanisms, behaviors, and operating principles, thereby achieving a degree of validation and enabling answering the questions posed. We suggest that the in silico operating principles presented may have a biological counterpart and that a semiquantitative mapping exists between in silico causal events and in vitro causal events

A meta-analysis of pre-pregnancy maternal body mass index and placental DNA methylation identifies 27 CpG sites with implications for mother-child health

Higher maternal pre-pregnancy body mass index (ppBMI) is associated with increased neonatal morbidity, as well as with pregnancy complications and metabolic outcomes in offspring later in life. The placenta is a key organ in fetal development and has been proposed to act as a mediator between the mother and different health outcomes in children. The overall aim of the present work is to investigate the association of ppBMI with epigenome-wide placental DNA methylation (DNAm) in 10 studies from the PACE consortium, amounting to 2631 mother-child pairs. We identify 27 CpG sites at which we observe placental DNAm variations of up to 2.0% per 10 ppBMI-unit. The CpGs that are differentially methylated in placenta do not overlap with CpGs identified in previous studies in cord blood DNAm related to ppBMI. Many of the identified CpGs are located in open sea regions, are often close to obesity-related genes such as GPX1 and LGR4 and altogether, are enriched in cancer and oxidative stress pathways. Our findings suggest that placental DNAm could be one of the mechanisms by which maternal obesity is associated with metabolic health outcomes in newborns and children, although further studies will be needed in order to corroborate these findings.We would like to thank the Pregnancy and Childhood Epigenetics (PACE) consortium, as well as all the families that participated in these studies for their generous contribution. This work was partially funded by GVSAN2018111086 from the Basque Department of Health and PI18/01142 from ISCIII - Spanish Ministry of Science and Innovation - cofounded by the ERDF “A way to make Europe” to JRB and LSM, respectively; and by the Joint Programming Initiative – A Healthy Diet for a Healthy Life (JPI HDHL) (NutriPROGRAM). ACP was supported by grant GVSAN2019111085 from the Basque Department of Health to NFJ. Detailed acknowledgements and funding for each participating cohort are described in Supplementary Note 1

Basolateral Sorting of Syntaxin 4 Is Dependent on Its N-terminal Domain and the AP1B Clathrin Adaptor, and Required for the Epithelial Cell Polarity

Generation of epithelial cell polarity requires mechanisms to sort plasma membrane proteins to the apical and basolateral domains. Sorting involves incorporation into specific vesicular carriers and subsequent fusion to the correct target membranes mediated by specific SNARE proteins. In polarized epithelial cells, the SNARE protein syntaxin 4 localizes exclusively to the basolateral plasma membrane and plays an important role in basolateral trafficking pathways. However, the mechanism of basolateral targeting of syntaxin 4 itself has remained poorly understood. Here we show that newly synthesized syntaxin 4 is directly targeted to the basolateral plasma membrane in polarized Madin-Darby canine kidney (MDCK) cells. Basolateral targeting depends on a signal that is centered around residues 24–29 in the N-terminal domain of syntaxin 4. Furthermore, basolateral targeting of syntaxin 4 is dependent on the epithelial cell-specific clathrin adaptor AP1B. Disruption of the basolateral targeting signal of syntaxin 4 leads to non-polarized delivery to both the apical and basolateral surface, as well as partial intercellular retention in the trans-Golgi network. Importantly, disruption of the basolateral targeting signal of syntaxin 4 leads to the inability of MDCK cells to establish a polarized morphology which suggests that restriction of syntaxin 4 to the basolateral domain is required for epithelial cell polarity

A blood microRNA classifier for the prediction of ICU mortality in COVID-19 patients: a multicenter validation study

Background: The identification of critically ill COVID-19 patients at risk of fatal outcomes remains a challenge. Here, we first validated candidate microRNAs (miRNAs) as biomarkers for clinical decision-making in critically ill patients. Second, we constructed a blood miRNA classifier for the early prediction of adverse outcomes in the ICU. Methods: This was a multicenter, observational and retrospective/prospective study including 503 critically ill patients admitted to the ICU from 19 hospitals. qPCR assays were performed in plasma samples collected within the first 48 h upon admission. A 16-miRNA panel was designed based on recently published data from our group. Results: Nine miRNAs were validated as biomarkers of all-cause in-ICU mortality in the independent cohort of critically ill patients (FDR < 0.05). Cox regression analysis revealed that low expression levels of eight miRNAs were associated with a higher risk of death (HR from 1.56 to 2.61). LASSO regression for variable selection was used to construct a miRNA classifier. A 4-blood miRNA signature composed of miR-16-5p, miR-192-5p, miR-323a-3p and miR-451a predicts the risk of all-cause in-ICU mortality (HR 2.5). Kaplan‒Meier analysis confirmed these findings. The miRNA signature provides a significant increase in the prognostic capacity of conventional scores, APACHE-II (C-index 0.71, DeLong test p-value 0.055) and SOFA (C-index 0.67, DeLong test p-value 0.001), and a risk model based on clinical predictors (C-index 0.74, DeLong test-p-value 0.035). For 28-day and 90-day mortality, the classifier also improved the prognostic value of APACHE-II, SOFA and the clinical model. The association between the classifier and mortality persisted even after multivariable adjustment. The functional analysis reported biological pathways involved in SARS-CoV infection and inflammatory, fibrotic and transcriptional pathways. Conclusions: A blood miRNA classifier improves the early prediction of fatal outcomes in critically ill COVID-19 patients.11 página

Slavery and the african cultural legacy in the Caribbean

Con autorización de la editorial para este libro.[EN] The purpose of this book is to raise awareness among a wide audience of one of the most significant and shameful phenomena for humanity, as was the enslavement of over twelve and a half million Africans who were brought to America and forced to work and live as slaves. Many countries participated in the slave trade at different times and withvaried intensity (Great Britain, Portugal, France, Spain, Denmark, Netherlands, Germany, United States...).[ES] El propósito de esta obra es dar a conocer a un público amplio uno de los fenómenos de mayor trascendencia y vergüenza para la humanidad como fue la esclavización de más de doce millones y medio de africanos que fueron trasladados a América, obligados a trabajar y vivir como esclavos. Muchos países participaron en la trata de esclavos en distintos momentos y con diferente intensidad (Gran Bretaña, Portugal, Francia, España, Dinamarca, Países Bajos, Alemania, Estados Unidos…).Connected Worlds: The Caribbean, Origin of Modern World. This project has received funding from the European Union´s Horizon 2020 research and innovation programme under the Marie Sklodowska Curie grant agreement Nº 823846. This project is directed by professor Consuelo Naranjo Orovio, Institute of History-CSIC.Peer reviewe

Anales de Edafología y Agrobiología Tomo 47 Número 3-4

Suelos. Física. Resistencia del suelo y susceptibilidad a la compactación en terrenos a monte sometidos a pastoreo. Por R. Pérez Moreira y F. Diaz-Fierros Viqueira.-- La reserva de agua útil de los suelos de Galicia. l. Relación con la textura y el contenido de materia orgánica. Por A. M. Martinez Cortizas.-- Química Empleo de aminas alifáticas en el estudio de asociaciones haloisita-esmectita en suelos. Por F. J. Aragoneses, J. Casas, y J. L. Martin de Vidales.-- Quelación por EDDHA de micronutrientes en suelos calizos. Ecuación de límite máximo. Por M. Juárez, J. Sánchez-Andréu, L. Pla y J. Jorda.-- Quelación por EDDHA de micronutrientes en suelos calizos. Ecuación de orden "n ".Por J. Sánchez-Andréu, M. Juarez, L. Pla y J. Jordá.-- Génesis, Clasificación y Cartografía Caracterización de un podsol ferro - húmico en el Puerto de la Quesera (Sierra del Ayllón). Por R. Espejo Serrano, F. Guerrero López y A. Saa Requejo.-- Natrixerales en el Baix Segre (Lleida). Por J. Bech i Borrás, J. Garrigo i Reixach y J. R. Torrento i Marselles.-- Aspectos micromorfológicos del horizonte superior en suelos artificiales (Sorribas) de las Islas Canarias. Por A. Rodríguez Rodríguez y J. M. Ontañón Sánchez.-- Fertilidad Influencia de diferentes factores del suelo sobre su contenido en microelementos asimilables: Mn, Fe, Cu y Zn. Por B. C. Ortega, Ma C. Ortega y J. G. de las Heras.--Incidencia de la salinidad del agua de riego en la mineralización del nitrógeno orgánico en suelos calizos del sureste español. Por J. García-Serna, J. Sánchez Andréu, M. Juárez y J. Mataix.-- Biología Vegetal-Fisiología Efectos de la toxicidad del flúor sobre el ciclo biológico en especies herbáceas dicotiledoneas. Por M. Ibarra, F. López Belmonte y Ma A. Diez.-- Efectos de la toxicidad del flúor sobre el ciclo biológico de especies de monocotiledoneas. Por M. Ibarra, F. López-Belmonte y Mª A . Diez.-- Proteasas ácidas en uvas Vitis vinifera (variedad Macabeo). l. Actividad proteásica durante su maduración. Por J. Marín Expósito, C. Miguel Gordillo, J. l. Maynar Mariño y J. L. Mesias Iglesias.-- Agro biología Efectos de enmiendas calcáreas en suelos fijadores de fósforo. Por S. G. Ramos Hernández y N. Aguilera Herrera.-- III. Trabajo Recapitulativo. Una deriva hacia hemiparasftismo de los líquenes epifitos: Análisis fisiológico de las relaciones con sus fitoforos. Por C. VicentePeer reviewed2019-08.- CopyBook.- Libnova.- Biblioteca IC