Iron and Sphingolipids as Common Players of (Mal)Adaptation to Hypoxia in Pulmonary Diseases

Hypoxia, or lack of oxygen, can occur in both physiological (high altitude) and pathological conditions (respiratory diseases). In this narrative review, we introduce high altitude pulmonary edema (HAPE), acute respiratory distress syndrome (ARDS), Chronic Obstructive Pulmonary Disease (COPD), and Cystic Fibrosis (CF) as examples of maladaptation to hypoxia, and highlight some of the potential mechanisms influencing the prognosis of the affected patients. Among the specific pathways modulated in response to hypoxia, iron metabolism has been widely explored in recent years. Recent evidence emphasizes hepcidin as highly involved in the compensatory response to hypoxia in healthy subjects. A less investigated field in the adaptation to hypoxia is the sphingolipid (SPL) metabolism, especially through Ceramide and sphingosine 1 phosphate. Both individually and in concert, iron and SPL are active players of the (mal)adaptation to physiological hypoxia, which can result in the pathological HAPE. Our aim is to identify some pathways and/or markers involved in the physiological adaptation to low atmospheric pressures (high altitudes) that could be involved in pathological adaptation to hypoxia as it occurs in pulmonary inflammatory diseases. Hepcidin, Cer, S1P, and their interplay in hypoxia are raising growing interest both as prognostic factors and therapeutical targets

El valor de los paradigmas de la historia del arte en la práctica del diseño gráfico

La historia del arte, en su camino para erigirse como una ciencia, ha ido adoptando y adaptando distintas metodologías de trabajo dependientes de los conocimientos y la cultura de cada época. En otras palabras, ha ido añadiendo prismas bajo los que observar su objeto de estudio, -la obra de arte-, que han dotado a la teoría del arte de una riqueza de interpretaciones y formas de acercarse a las imágenes de las que la teoría del diseño carece o bien aplica de forma más bien intuitiva y sin fundamentación teórica. Sin embargo, resulta algo evidente que las imágenes, sus formas, sus significados, así como lo que comunican son una parte consustancial del diseño gráfico. En este trabajo tratamos de descubrir si el conocimiento de la historia del arte resulta útil a los diseñadores. Creemos interesante saber si la investigación que proponemos armoniza con las necesidades profesionales del diseño y para observar si los métodos de análisis de la obra de arte son aplicables al diseño gráfico pretendemos conocer la opinión de los diseñadores sobre la utilidad de los diferentes paradigmas metodológicos del estudio del arte en relación a su actividad profesional. Por tanto, realizamos un cuestionario online a 274 diseñadores gráficos repartidos principalmente en Catalunya, Madrid, País Vasco, Valencia y Andalucía que nos ha permitido descubrir que los diseñadores gráficos se interesan especialmente por aquellos métodos que contribuyen a aportar conocimientos críticos sobre la realización formal de las piezas -ya sean pinturas o infografías- y sobre sus contenidos y significados.Art history, on its way to establish itself as a science, has been adopting and adapting different methods of work dependent knowledge and culture of each era. In other words, it has been adding prisms under which observe its subject, -the work of art, which have endowed art theory of a wealth of interpretations and ways of approaching the images that theory no design or applied rather intuitively and without theoretical foundation. However, it is evident that the images, forms, their meanings and what they communicate is an essential part of graphic design.In this paper we try to discover if knowledge of art history is useful to designers. We believe interesting to know whether the research we propose harmonizes with professional design needs and to see if the methods of analysis of the artwork are applicable to graphic design pretend to know the opinion of the designers on the usefulness of the different methodological paradigms of the study art in relation to their professional activity. Therefore, we conducted an online questionnaire to 274 graphic designers spread mainly in Catalonia, Madrid, Basque Country, Valecia and Andalusia that has allowed us to discover that graphic designers are especially interested in methods that help provide critical knowledge of the formal aspects of the pieces are paintings or-and infografías- and their content and meaning

Lights and Shadows in the Use of Mesenchymal Stem Cells in Lung Inflammation, a Poorly Investigated Topic in Cystic Fibrosis

Mesenchymal stem cells (MSCs) are multipotent non-hematopoietic stem cells residing in many tissues, including the lung. MSCs have long been regarded as a promising tool for cell-based therapy because of their ability to replace damaged tissue by differentiating into the resident cell and repopulating the injured area. Their ability to release soluble factors and extracellular vesicles has emerged as crucial in the resolution of inflammation and injury. There is a growing literature on the use of MSCs and MSC secretome to hamper inflammation in different lung pathologies, including: asthma, pneumonia, acute lung injury (ALI), pulmonary hypertension, and chronic obstructive pulmonary disease (COPD). However, their potential therapeutic role in the context of Cystic Fibrosis (CF) lung inflammation is still not fully characterized. CF morbidity and mortality are mainly due to progressive lung dysfunction. Lung inflammation is a chronic and unresolved condition that triggers progressive tissue damage. Thus, it becomes even more important to develop innovative immunomodulatory therapies aside from classic anti-inflammatory agents. Here, we address the main features of CF and the implications in lung inflammation. We then review how MSCs and MSC secretome participate in attenuating inflammation in pulmonary pathologies, emphasizing the significant potential of MSCs as new therapeutic approach in CF

Valoración de los profesores y padres de alumnos con retraso mental de las aulas de aprendizaje de tareas de Álava

El objetivo de este trabajo es: a) presentar un cuestionario elaborado para valorar aspectos sobre la satisfacción, la intervención, y las creencias y expectativas acerca de la integración de los padres (n=68) y profesores (n=56) de los jóvenes con retraso mental que cursan el itinerario educativo de las Aulas de Aprendizaje de Tareas de Álava; y b) exponer los resultados más significativos obtenidos. Asimismo, se discuten las implicaciones teóricas y prácticas de dichos resultados

Transcriptional control of the B3GALT5 gene by a retroviral promoter and methylation of distant regulatory elements

We focused on transcription factors and epigenetic marks that regulate the B3GALT5 gene through its retroviral long terminal repeat (LTR) promoter. We compared the expression levels of the B3GALT5 LTR transcript, quantitated by competitive RT-PCR, with those of the candidate transcription factors HNF1\u3b1/\u3b2 and Cdx1/2, determined by Western blot analysis, in colon cancer biopsies, various cell lines, and cell models serving as controls. We found that HNF1\u3b1/\u3b2 were easily detected, irrespective of the amount of LTR transcript expressed by the source, whereas Cdx1/2 were undetectable, and no sample lacking HNF1\u3b1/\u3b2 expressed the LTR transcript. On transfection in proper host cells, both HNF1\u3b1 and HNF1\u3b2 provided detectable LTR transcript, whereas shRNA-mediated silencing of HNF1\u3b2 impaired transcription. Treating cells with 5\u2032-aza-2\u2032-deoxycytidine (5AZA) strongly reduced expression, without affecting HNF1\u3b1/\u3b2, despite the lack of CpG islands in the LTR and proximal sequences. By electrophoresis mobility shift and luciferase reporter assays, the LTR promoter binding and activity did not correlate with the amounts of LTR transcript expressed in the cells and depended on the levels of the transcription factors. We conclude that HNF1\u3b1/\u3b2 are necessary but insufficient to activate and regulate B3GALT5 LTR transcription, which depends on unknown regulatory elements that are active when methylated and located outside of and far from the LTR promoter

TRANSCRIPTIONAL REGULATION OF THE B3GALT5 GENE

RIASSUNTO Introduzione. La \u3b21,3 galattosiltransferasi (B3GALT5) \ue8 responsabile della sintesi della catena oligosaccaridica di tipo 1, tra cui gli antigeni Lewis come il sialil-Lewis a, epitope del marcatore tumorale CA19.9 e ligando della E-selectina, potenzialmente coinvolto nella malignit\ue0 tumorale. La sua trascrizione \ue8 regolata da molteplici promotori. In alcuni epiteli essa \ue8 sotto il controllo di un promotore debole, chiamato nativo, modulato epigeneticamente e tramite il fattore nucleare NF-Y. In alcuni organi e cellule di origine gastrointestinale \ue8 attivo inoltre un altro promotore, pi\uf9 forte e chiamato LTR per la sua origine retrovirale, che secondo la letteratura dovrebbe essere regolato attraverso il fattore nucleare epatocitario HNF1\u3b1/\u3b2 e quello analogo al Caudale di drosofila Cdx1/2. Tuttavia la B3GALT5 \ue8 repressa nel cancro del colon, il trascritto LTR non \ue8 rilevante nell\u2019intestino tenue, e Cdx1/2 risultano assenti in una linea cellulare che esprime grandi quantit\ue0 di tale trascritto. Scopi. Scoprire i meccanismi che controllano la trascrizione di B3GALT5 attraverso il suo promotore LTR, con l\u2019obiettivo di spiegare la specificit\ue0 tissutale e la repressione negli adenocarcinomi del colon, nonch\ue9 di capire il processo di stabilizzazione evolutiva del trasposone in alcuni primati. Metodi. A questo scopo abbiamo quantificato l\u2019espressione di HNF1\u3b1/\u3b2 e Cdx1/2, tramite Western Blot, e quella del trascritto B3GALT5 LTR, tramite RT-PCR competitiva, in tessuti tumorali e linee cellulari. Inoltre, abbiamo silenziato HNF1\u3b1 o \u3b2 in diverse cellule, tramite shRNA, e li abbiamo espressi in un\u2019altra, tramite trasfezione del cDNA. Abbiamo quindi trattato delle cellule con l'agente demetilante il DNA 5'-AZA-2'-desossicitidina, misurandone poi i livelli di trascritto LTR. Abbiamo infine valutato il promotore LTR in vitro, mediante saggi di EMSA e di luciferasi. Risultati. Cdx1/2 risultano immisurabili in cellule e tessuti che pur esprimono alti livelli di trascritto LTR, mentre HNF1\u3b1/\u3b2 sono presenti anche in cellule e tumori che esprimono una quantit\ue0 bassa o nulla del trascritto. Nelle cellule che non esprimono HNF1\u3b1/\u3b2 per\uf2 non c\u2019\ue8 espressione alcuna del trascritto LTR. Tra queste, le MDA-MB-231, dopo transfezione con HNF1\u3b1 o \u3b2, esprimono il trascritto LTR, ma a bassi livelli, paragonabili a quelli dei tumori del colon. Il silenziamento di HNF1\u3b1 in una linea cellulare che esprime entrambi i fattori HNF1\u3b1 e \u3b2 non ha effetti nell\u2019espressione del trascritto LTR, mentre quello di HNF1\u3b2 in un\u2019altra linea che esprime solo HNF1\u3b2 produce forte riduzione del trascritto. Pure il trattamento con 5'-AZA-2'-desossicitidina riduce il trascritto in cellule che ne esprimono alti livelli, portandolo ai livelli dei tumori del colon, e senza effetto sulla quantit\ue0 di HNF1. Usando i saggi delle luciferasi, abbiamo visto che la luciferasi sotto il controllo del promotore LTR \ue8 pi\uf9 attiva in cellule e cloni che esprimono alte quantit\ue0 di HNF1, anche se non esprimono o esprimono poco trascritto LTR, che in quelle cellule che esprimono poco HNF1 ma magari una grande quantit\ue0 di trascritto. Usando la sequenza del promotore LTR in saggi di EMSA, abbiamo visto che forma dei complessi specifici con estratti di proteine nucleari di tutte le linee cellulari che esprimono HNF1, indipendentemente dei livelli di espressione del trascritto B3GALT5 LTR. Conclusione. I nostri risultati suggeriscono che HNF1\u3b1 e -\u3b2 sono necessari ma non sufficienti a regolare l'espressione del promotore LTR, mentre Cdx1/2 non sono coinvolti. HNF1\u3b1/\u3b2 svolgono un ruolo intercambiabile e non cumulativo, e non sono immediatamente responsabili della regolazione negativa che avviene nel cancro, che invece dipende da elementi regolatori distanti attivi solo se metilati. Il successo della inserzione e l'attivazione del promotore B3GALT5 LTR durante l'evoluzione dipendono quindi non solo dal suo sito di legame a HNF1, ma anche da questi elementi distanti attualmente sconosciuti.ABSTRACT Background. \u3b21,3 galactosyltransferase (B3GALT5) is responsible for the synthesis of type 1 chain oligosaccharides, including Lewis antigens as sialyl-Lewis a, the epitope of tumor marker CA19.9 and an E-selectin ligand potentially involved in cancer malignancy. Transcription occurs through multiple promoters. In some epithelia it is driven by a weak promoter, known as the native promoter that is epigenetically modulated and sensitive to nuclear factor NF-Y. In some organs of the gastrointestinal tract (as the colon, stomach, pancreas and related cell lines) another stronger promoter is active and named the LTR promoter after its retroviral origin. It was supposed to be regulated through a set of homeoproteins: hepatocyte nuclear factor HNF1\u3b1/\u3b2 and caudal-related homeobox Cdx1/2. Surprisingly, B3GALT5 is strongly down regulated in colon cancer, the LTR transcript is not relevant in the small intestine, and Cdx1/2 were reported absent from a cell line expressing large amount of such transcript. Aims. To elucidate the mechanisms controlling transcription of B3GALT5 through its retroviral LTR promoter, in order to explain the tissue specificity and down-regulation in colon adenocarcinomas, and to understand the evolutionary stabilization of the transposon in some primates. Methods. To this aim, we determined the expression levels of putative transcription factors by western blot and the amounts of B3GALT5 LTR transcript by competitive RT-PCR in cancer tissues and cell lines. Moreover, we silenced HNF1\u3b1 or \u3b2 in different cell lines, through an shRNA approach, expressed them in another by permanent cDNA transfection, and treated cells with the DNA demethylating agent 5\u2019-AZA-2\u2019-deoxycitydine and in all cases, we measure the effects on LTR transcript levels. We also evaluated the behavior of the LTR promoter in vitro, through electrophoresis mobility shift and reporter luciferase assays. Results. We found that Cdx1/2 are not detectable in cells and tissues expressing high amount of B3GALT5 LTR transcript, while HNF1\u3b1/\u3b2 are well detectable, but even in cells and cancers expressing very low or undetectable levels of the transcript, which is absent in all cells lacking HNF1\u3b1/\u3b2. Among them, the cell line MDA-MB-231, upon transfection with HNF1\u3b1 or \u3b2, became able to express B3GALT5 LTR transcript, but a very low levels, similar to those found in colon cancers. Transient silencing of HNF1\u3b1 in cells expressing both HNF1\u3b1 and \u3b2, has no effect on LTR transcript, while similar silencing of HNF1\u3b2 in cells expressing HNF1\u3b2 only, determines strong reduction of the transcript. Cell lines expressing high levels of B3GALT5 LTR transcript are affected by the demethylating agent 5AZA that determines strong down regulation of the transcript, falling down to the amounts found in colon cancers, while HNF1 levels remain unaffected. In vitro, luciferase placed under the control of LTR promoter is more active in cells or clones expressing high HNF1 and low or no LTR transcript than in those expressing low HNF1 and high transcript. The same promoter, when used as a probe in EMSA, forms specific complex with nuclear protein extracted from all cells expressing HNF1, irrespectively of the levels of B3GALT5 LTR transcript. Conclusion. Our results suggest that HNF1\u3b1 and HNF1\u3b2 are necessary but not sufficient to drive expression of LTR promoter, while Cdx1/2 are not involved. HNF1\u3b1/\u3b2 play an interchangeable and not cumulative role and are not immediately responsible for cancer down-regulation, which depends on a distal regulatory element(s) active when methylated. The successful insertion and activation of B3GALT5 LTR promoter during evolution depended not only on its HNF1 binding site, but even on such distal element(s) unknown at present

An Innovative Lipidomic Workflow to Investigate the Lipid Profile in a Cystic Fibrosis Cell Line

Altered lipid metabolism has been associated to cystic fibrosis disease, which is characterized by chronic lung inflammation and various organs dysfunction. Here, we present the validation of an untargeted lipidomics approach based on high-resolution mass spectrometry aimed at identifying those lipid species that unequivocally sign CF pathophysiology. Of n.13375 mass spectra recorded on cystic fibrosis bronchial epithelial airways epithelial cells IB3, n.7787 presented the MS/MS data, and, after software and manual validation, the final number of annotated lipids was restricted to n.1159. On these lipids, univariate and multivariate statistical approaches were employed in order to select relevant lipids for cellular phenotype discrimination between cystic fibrosis and HBE healthy cells. In cystic fibrosis IB3 cells, a pervasive alteration in the lipid metabolism revealed changes in the classes of ether-linked phospholipids, cholesterol esters, and glycosylated sphingolipids. Through functions association, it was evidenced that lipids variation involves the moiety implicated in membrane composition, endoplasmic reticulum, mitochondria compartments, and chemical and biophysical lipids properties. This study provides a new perspective in understanding the pathogenesis of cystic fibrosis and strengthens the need to use a validated mass spectrometry-based lipidomics approach for the discovery of potential biomarkers and perturbed metabolism

Spatial Mapping and Modeling of Reported Dengue Incidences in Luzon

Dengue, the most rapidly spreading mosquito-borne viral infection, has significantly spread worldwide in recent decades - flourishing both in affluent and impoverished locations of tropical and subtropical countries. In 2012, the Philippines ranked fourth out of the ten Association of the Southeast Asian Nations (ASEAN) countries in having the highest number of dengue cases. The following study intends to analyze the spatial distribution of dengue incidences across all Luzon provinces in 2018. It aims to determine significant correlates that affect dengue incidences, map the incidence rate of dengue cases, and explore the clustering of recorded dengue cases. Poisson and Negative Binomial (NB) regression analyses and Multiple Linear Regression Models (MLRM) were applied to determine the significant correlations affecting dengue incidence rates. Simultaneously, spatial mapping was utilized to visualize and detect clustering in the provinces through dengue count, incidence ratios, and standard incidence ratios (SIR). MLRM and NB showed that rainfall and poverty incidence are significant correlates of dengue counts and incidence, and Nueva Ecija and Tarlac were observed to be provinces with distinct dengue count and SIR greater than 1, as well as provinces found in clusters. With the provided results, health organizations can provide health programs and allocate more funds in areas with SIR greater than 1 to prevent dengue spreading

Use of Gene Therapy in a Subcutaneous Murine Model of Lung Cancer

OBJECTIVE: To assess the effectiveness of in vivo gene therapy to treat subcutaneous tumors generated from murine lung cancer cells. MATERIAL AND METHODS: C57BL/6 mice received subcutaneus injections of 5×105 cells from the murine Lewis lung cancer cell line. By 10 days, subcutaneous tumors of approximately 5 mm diameter were formed. At that point, treatment was provided by intratumor injection of a replication-defective recombinant adenovirus carrying the gene for thymidine kinase (AdCMV-Tk) or interleukin (IL) 12 (AdCMV-IL12), or by injection of syngeneic dendritic cells previously transduced with adenovirus containing the IL-12 gene (DC-IL12). Control groups were treated with saline or adenovirus containing the gene for β-galactosidase (AdCMV-LacZ), which functions as a reporter gene and does not have a therapeutic effect. The number of animals in each group ranged from 14 to 25 in experiments using adenovirus and from 10 to 12 in experiments using dendritic cells. Tumor size was followed for 3 weeks in the case of treatment with adenovirus and 4 weeks for treatment with dendritic cells. RESULTS: A significant reduction in subcutaneous tumor growth was observed in the groups treated with AdCMVTk, AdCMV-IL12, and DC-IL12 compared with control groups treated with saline or AdCMV-LacZ. The difference was statistically significant from day 7 of treatment in the AdCMV-Tk group, from day 9 in the AdCMV-IL12 group, and from day 10 in the DC-IL12 group, and in all cases it was maintained until the end of the follow-up period. CONCLUSIONS: Gene therapy with AdCMV-Tk, AdCMVIL12, or DC-IL12 is effective in our model of subcutaneous tumors arising from cells of the Lewis lung cancer cell line. The treatment leads to a significant reduction in tumor growth compared with control groups