Physiological Plasticity of Neural-Crest-Derived Stem Cells in the Adult Mammalian Carotid Body

Adult stem cell plasticity, or the ability of somatic stem cells to cross boundaries and differentiate into unrelated cell types, has been amatter of debate in the last decade. Neural-crest-derived stem cells (NCSCs) display a remarkable plasticity during development. Whether adult populations of NCSCs retain this plasticity is largely unknown. Herein, we describe that neural-crest-derived adult carotid body stem cells (CBSCs) are able to undergo endothelial differentiation in addition to their reported role in neurogenesis, contributing to both neurogenic and angiogenic processes taking place in the organ during acclimatization to hypoxia. Moreover, CBSC conversion into vascular cell types is hypoxia inducible factor (HIF) dependent and sensitive to hypoxiareleased vascular cytokines such as erythropoietin. Our data highlight a remarkable physiological plasticity in an adult population of tissue-specific stem cells and could have impact on the use of these cells for cell therapy.ERC Starting Grant: CBSCsMinisterio de Economía y Competitividad SAF2013-48535-P and SAF2016-80412-

Plasticidad fisiológica de las células madre del cuerpo carotídeo adulto: caracterización del linaje mesectodérmico

Falta palabras claveEl cuerpo carotídeo (CB), un órgano quimiosensor derivado de la cresta neural, permite la adaptación a la hipoxia crónica mediante neurogénesis y angiogénesis. Recientemente se ha descrito en nuestro grupo que ambos procesos son resultado de la actividad de una población de células madre neurales que residen en este nicho germinal. Éstas dan lugar a neuroesferas (NS) in vitro. Mediante análisis génico por microarrays de NS de CB enriquecidas en células indiferenciadas versus diferenciadas hemos hallado a la proteína CD10 como un marcador de progenitores de CB. CD10 es una metaloendopeptidasa que inactiva péptidos señalizadores a través de su dominio catalítico exterior, y paralelamente, interviene en proliferación, diferenciación endotelial y migración gracias a su dominio citoplasmático. La expresión de CD10 en CB co-localiza parcialmente con Nestina y/o GFAP, y los análisis del trazado del linaje celular han confirmado que las células CD10+ provienen del linaje de la célula madre del CB (CBSC) GFAP+. El aislamiento de células CD10+ mediante citometría de flujo incrementa la eficiencia de formación de NS. De manera relevante, los progenitores CD10+ se diferencian hacia células de músculo liso (SMA+) y células endoteliales (GSA I+), pero no se diferencian hacia células neuronales (TH+), indicando que los progenitores CD10+ están restringidos hacia el linaje mesectodérmico del CB. Dado que CD10 es un regulador angiogénico en otros tejidos, y la importante angiogénesis que tiene lugar en el CB en hipoxia mantenida, realizamos experimentos funcionales de esta proteína. Estos han revelado que la expresión de CD10 está regulada por la hipoxia en CB, y que CD10 controla la diferenciación endotelial de los progenitores CD10+, al menos a través de su actividad catalítica. Uno de los principales sustratos de CD10 es la endotelina-1 (ET-1), que es liberada en el CB en hipoxia. Es una citoquina pro-proliferativa y pro-angiogénica que incrementa el número de células CD10+ y los niveles de expresión de CD10 in vitro, además mediante análisis génico de mciroarrays hemos visto que ET-1 induce la diferenciación mesectodérmica de las CBSCs. En definitiva, los progenitores CD10+, restringidos al linaje mesectodérmico, tras el estímulo hipóxico, ven reducidos los niveles de expresión de CD10, reproduciéndose la activación de dichos progenitores y su consecuente diferenciación hacia células vasculares, que contribuyen a la angiogénesis que tiene lugar en el órgano en hipoxia crónica.We have described a novel neurogenic niche in the adult carotid body (CB), an oxygen-sensing organ derived from the neural crest that is able to adapt to chronic hypoxemia by increasing the number of chemosensory neuronal cells and vascular cells. These neurogenesis and angiogenesis depend on the activity of a resident population of neural progenitors, which are also able to form neurospheres (NS) in vitro. Gene expression analysis comparing NS cultures enriched in undifferentiated versus differentiated cells displayed CD10 as a candidate marker for CB progenitor cells. CD10 is a metalloendopeptidase that inactivates signalling peptides, modifying the extracellular environment. In parallel, CD10 protein has an intracellular domain that mediates proliferation, endothelial differentiation and migration processes, by direct protein-protein associations. In the CB, CD10 expression partially co-localizes with Nestin and/or GFAP, and a cell-fate mapping analysis has confirmed that CD10+ cells lie within the CB progenitor lineage. Sorting for CD10+ cells increases NS-forming efficiency. Interestingly, CD10+ progenitors can differentiate into smooth muscle (SMA+) or endothelial (GSA I+) cells, but are not able to convert into neuronal (TH+) cells, indicating a mesectodermal commitment in these cells. Since a profound angiogenesis occurs in the CB in response to hypoxia and it is described that CD10 is a key regulator of angiogenesis, we have tested whether CD10 is having a role in regulating this process. Our functional study revealed that CD10 protein controls endothelial differentiation of the CD10+ progenitor subpopulation, likely through the catalytic activity of CD10. One of the typical molecules cleaved by CD10 is the pro-angiogenic peptide endothelin-1 (ET-1). ET-1 significantly increases CD10 expression and the amount of CD10+ cells in vitro, and gene expression analysis shows that ET-1 induces an increase in mesectodermal differentiation of CBSCs. Taking together, our results suggest that GFAP+ stem cells give rise to mesectoderm-committed CD10+ progenitors, and that CD10 could have a functional role cleaving peptides such as ET-1, regulating the angiogenesis that accompany the neurogenesis suffered in hypoxemia. A deeper knowledge of the molecular mechanisms governing CB progenitor cell fate decisions will improve our understanding of the physiology of this important chemoreceptor organ

Rol de las enfermeras en las escuelas saludables: revisión bibliográfica

The School Nurse is a professional who performs her role in the educational community, contributing to its full development and physical, mental and social well-being; A bibliographic review was carried out in national and international journals by consulting Science Direct, Scopus, ProQuest central, SciELO, and Dialnet, with full text availability limiting the search to the last 10 years; Articles published in the language ESP, ING, POR were included. A total of 210 articles were counted, after reading and considering the inclusion criteria and their relevance to answer the research question, the final sample was 29 articles. After analyzing the results, these were grouped into three different topics: progress of school nursing after the passage of time, school nursing in European countries and school nursing in Latin America. The lack of commitment of the State to execute programs and personnel that help to improve the stay in educational establishments was evidenced by promoting, maintaining health and preventing disease.La Enfermera Escolar es un profesional que realiza su rol en el ámbito de la comunidad educativa contribuyendo al pleno desarrollo y el bienestar físico, mental y social de la misma; se realizó una revisión bibliográfica en revistas nacionales e internacionales a través de la consulta en Science Direct, Scopus, ProQuest central, SciELO, y Dialnet, con disponibilidad de texto completo limitando la búsqueda a los últimos 10 años; Se incluyeron artículos publicados en idioma ESP, ING, POR. Se contabilizaron un total de 210 artículos, posterior a la lectura y considerando los criterios de inclusión y su relevancia para responder a la pregunta de investigación, la muestra final fueron 29 artículos, Tras el análisis de los resultados, estos fueron agrupados en tres temas diferentes: avance de la enfermería escolar tras el paso del tiempo, enfermería escolar en países europeos y enfermería escolar en Latinoamérica. Se evidenció la falta de compromiso del Estado para ejecutar programas y personal que ayude a mejorar la estancia en los planteles educativos haciendo promoción, mantenimiento de la salud y prevención de la enfermedad

Phenotypic Characterization of Macrophages from Rat Kidney by Flow Cytometry

There is increasing evidence suggesting the important role of inflammation and, subsequently, macrophages in the development and progression of renal disease. Macrophages are heterogeneous cells that have been implicated in kidney injury. Macrophages may be classified into two different phenotypes: classically activated macrophages (M1 macrophages), that release pro-inflammatory cytokines and promote fibrosis; and alternatively activated macrophages (M2 macrophages) that are associated with immunoregulatory and tissue-remodeling functions. These macrophage phenotypes need to be discriminated and analyzed to determine their contribution to renal injury. However, there are scarce studies reporting consistent phenotypic and functional information about macrophage subtypes in inflammatory renal disease models, especially in rats. This fact may be related to the limited macrophage markers used in rats, contrary to mice. Therefore, novel strategies are necessary to quantify and characterize the renal content of these infiltrating cells in a reliable way. This manuscript details a protocol for kidney digestion and further phenotypic and quantitative analysis of macrophages from rat kidneys by flow cytometry. Briefly, kidneys were incubated with collagenase and total macrophages were identified according to the dual presence of CD45 (leukocytes common antigen) and CD68 (PAN macrophage marker) in live cells.This was followed by surface staining of CD86 (M1 marker) and CD163 (M2 marker). Rat peritoneal macrophages were used as positive control for macrophage marker detection by flow cytometry. Our protocol resulted in low cellular mortality and allowed characterization of different intracellular and surface protein markers, thus limiting the loss of cellular integrity observed in other protocols. Moreover, this procedure allows the use of macrophages for further techniques, including cell sorting and mRNA or protein expression studies, among others.This work was supported by grants from FIS/FEDER (Programa Miguel Servet: CP10/00479, PI13/00802 and PI14/00883), Spanish Society of Atherosclerosis, Spanish Society of Nephrology and Fundaciòn Renal Iñigo Alvarez de Toledo (FRIAT) to Juan Antonio Moreno. FIS/FEDER funds PI14/00386 and Instituto Reina Sofìa de Investigaciòn Nefrològica to Jesús Egido. Fundaciòn Conchita Rabago to Melania Guerrero Hue. Fundaciòn Renal Iñigo Alvarez de Toledo (FRIAT) to Alfonso Rubio Navarro.S

Therapeutic Management and Long-Term Outcome of Hy-Perthyroidism in Patients with Antithyroid-Induced Agranu-Locytosis: A Retrospective, Multicenter Study

Background: Antithyroid drug-induced agranulocytosis (AIA) (neutrophils <500/mu L) is a rare but serious complication in the treatment of hyperthyroidism. Methodology: Adult patients with AIA who were followed up at 12 hospitals in Spain were retrospectively studied. A total of 29 patients were studied. The etiology of hyperthyroidism was distributed as follows: Graves' disease (n = 21), amiodarone-induced thyrotoxicosis (n = 7), and hyperfunctioning multinodular goiter (n = 1). Twenty-one patients were treated with methimazole, as well as six patients with carbimazole and two patients with propylthiouracil. Results: The median (IQR) time to development of agranulocytosis was 6.0 (4.0-11.5) weeks. The most common presenting sign was fever accompanied by odynophagia. All of the patients required admission, reverse isolation, and broad-spectrum antibiotics; moreover, G-CSF was administered to 26 patients (89.7%). Twenty-one patients received definitive treatment, thirteen patients received surgery, nine patients received radioiodine, and one of the patients required both treatments. Spontaneous normalization of thyroid hormone values occurred in six patients (four patients with amiodarone-induced thyrotoxicosis and two patients with Graves' disease), and two patients died of septic shock secondary to AIA. Conclusions: AIA is a potentially lethal complication that usually appears around 6 weeks after the initiation of antithyroid therapy. Multiple drugs are required to control hyperthyroidism before definitive treatment; additionally, in a significant percentage of patients (mainly in those treated with amiodarone), hyperthyroidism resolved spontaneously

Single nucleotide variations in ZBTB46 are associated with post-thrombolytic parenchymal haematoma

Haemorrhagic transformation is a complication of recombinant tissue-plasminogen activator treatment. The most severe form, parenchymal haematoma, can result in neurological deterioration, disability, and death. Our objective was to identify single nucleotide variations associated with a risk of parenchymal haematoma following thrombolytic therapy in patients with acute ischaemic stroke. A fixed-effect genome-wide meta-analysis was performed combining two-stage genome-wide association studies (n = 1904). The discovery stage (three cohorts) comprised 1324 ischaemic stroke individuals, 5.4% of whom had a parenchymal haematoma. Genetic variants yielding a P-value < 0.05 1 x 10(-5) were analysed in the validation stage (six cohorts), formed by 580 ischaemic stroke patients with 12.1% haemorrhagic events. All participants received recombinant tissue-plasminogen activator; cases were parenchymal haematoma type 1 or 2 as defined by the European Cooperative Acute Stroke Study (ECASS) criteria. Genome-wide significant findings (P < 5 x 10(-8)) were characterized by in silica functional annotation, gene expression, and DNA regulatory elements. We analysed 7 989 272 single nucleotide polymorphisms and identified a genome-wide association locus on chromosome 20 in the discovery cohort; functional annotation indicated that the ZBTB46 gene was driving the association for chromosome 20. The top single nucleotide polymorphism was rs76484331 in the ZBTB46 gene [P = 2.49 x 10(-8); odds ratio (OR): 11.21; 95% confidence interval (CI): 4.82-26.55]. In the replication cohort (n = 580), the rs76484331 polymorphism was associated with parenchymal haematoma (P = 0.01), and the overall association after meta-analysis increased (P = 1.61 x 10(-8), OR: 5.84; 95% CI: 3.16-10.76). ZBTB46 codes the zinc finger and BTB domain-containing protein 46 that acts as a transcription factor. In silica studies indicated that ZBTB46 is expressed in brain tissue by neurons and endothelial cells. Moreover, rs76484331 interacts with the promoter sites located at 20q13. In conclusion, we identified single nucleotide variants in the ZBTB46 gene associated with a higher risk of parenchymal haematoma following recombinant tissue-plasminogen activator treatment.Peer reviewe

From magma source to volcanic sink under Tagoro Volcano (El hierro, Canary Islands): Petrologic, Geochemical and Physiographic Evolution of the 2011-2012 Submarine Eruption

Active volcanoes are key laboratories to carry out detailed research -and monitoring- about the history of magmas before, during and after eruptions. Tagoro, the submarine active vol- cano at El Hierro Island (Canary archipelago), is a highly favorable case to assess and monitor its daily ongoing behaviour, as well as to study the links between the processes of magma genesis occurring at depth and their derived eruptive events at the surface. In this interdisciplinary research we combine new results of classical petrology (petrography, geochemistry, and thermodynamics) on the volcanic products expelled by Tagoro during the 2011–2012 eruption, with a high- resolution (5 m grid) bathymetry model car- ried out during 2017, and recent data from magnetometry, to refine the current knowl- edge of this eruption. Our results mainly reveal (i) slight magma differentiation and mixing processes at c. 12 km depth during a continuous eruptive pulse; (ii) a similar mag- matic evolution and residence times at depth between previous and 2011–2012 eruptions on the island; (iii) an insignificant interaction of external fluids with the magma at depth or within the ascent conduit; (iv) a present-day magnetometric anomaly under the Tagoro’s area; (v) a minimum volume estimate for the magma withdrawn from the plumbing system at depth.MINECO and FEDER: VULCANO I (CTM2012-36317). Instituto Español de Oceanografía.VULCANA (Vulcana IEO-2015-2017). Instituto Español de OceanografíaMINECO AND MEC. EXPLORA-CIENCIA (CGL2014—61775-EXP)MINECO AND MEC. EXPLORA-CIENCIA (CAS14-00189; MEC)Programa Propio mod. 1B— 2019 (USAL)EC Grant EVE (DG ECHO H2020 826292)PhD grant “Programa Propio III Universidad de Salamanca, cofounded by Banco de Santander”The PTI VOLCAN research initiatives.Peer reviewe

Table_3_A Polygenic Risk Score Based on a Cardioembolic Stroke Multitrait Analysis Improves a Clinical Prediction Model for This Stroke Subtype.DOCX

[Background] Occult atrial fibrillation (AF) is one of the major causes of embolic stroke of undetermined source (ESUS). Knowing the underlying etiology of an ESUS will reduce stroke recurrence and/or unnecessary use of anticoagulants. Understanding cardioembolic strokes (CES), whose main cause is AF, will provide tools to select patients who would benefit from anticoagulants among those with ESUS or AF. We aimed to discover novel loci associated with CES and create a polygenetic risk score (PRS) for a more efficient CES risk stratification.[Methods] Multitrait analysis of GWAS (MTAG) was performed with MEGASTROKE-CES cohort (n = 362,661) and AF cohort (n = 1,030,836). We considered significant variants and replicated those variants with MTAG p-value < 5 × 10−8 influencing both traits (GWAS-pairwise) with a p-value < 0.05 in the original GWAS and in an independent cohort (n = 9,105). The PRS was created with PRSice-2 and evaluated in the independent cohort.[Results] We found and replicated eleven loci associated with CES. Eight were novel loci. Seven of them had been previously associated with AF, namely, CAV1, ESR2, GORAB, IGF1R, NEURL1, WIPF1, and ZEB2. KIAA1755 locus had never been associated with CES/AF, leading its index variant to a missense change (R1045W). The PRS generated has been significantly associated with CES improving discrimination and patient reclassification of a model with age, sex, and hypertension.[Conclusion] The loci found significantly associated with CES in the MTAG, together with the creation of a PRS that improves the predictive clinical models of CES, might help guide future clinical trials of anticoagulant therapy in patients with ESUS or AF.Peer reviewe

Natural History of MYH7-Related Dilated Cardiomyopathy

BACKGROUND Variants in myosin heavy chain 7 (MYH7) are responsible for disease in 1% to 5% of patients with dilated cardiomyopathy (DCM); however, the clinical characteristics and natural history of MYH7-related DCM are poorly described. OBJECTIVES We sought to determine the phenotype and prognosis of MYH7-related DCM. We also evaluated the influence of variant location on phenotypic expression. METHODS We studied clinical data from 147 individuals with DCM-causing MYH7 variants (47.6% female; 35.6 +/- 19.2 years) recruited from 29 international centers. RESULTS At initial evaluation, 106 (72.1%) patients had DCM (left ventricular ejection fraction: 34.5% +/- 11.7%). Median follow-up was 4.5 years (IQR: 1.7-8.0 years), and 23.7% of carriers who were initially phenotype-negative developed DCM. Phenotypic expression by 40 and 60 years was 46% and 88%, respectively, with 18 patients (16%) first diagnosed at <18 years of age. Thirty-six percent of patients with DCM met imaging criteria for LV noncompaction. During follow-up, 28% showed left ventricular reverse remodeling. Incidence of adverse cardiac events among patients with DCM at 5 years was 11.6%, with 5 (4.6%) deaths caused by end-stage heart failure (ESHF) and 5 patients (4.6%) requiring heart transplantation. The major ventricular arrhythmia rate was low (1.0% and 2.1% at 5 years in patients with DCM and in those with LVEF of <= 35%, respectively). ESHF and major ventricular arrhythmia were significantly lower compared with LMNA-related DCM and similar to DCM caused by TTN truncating variants. CONCLUSIONS MYH7-related DCM is characterized by early age of onset, high phenotypic expression, low left ventricular reverse remodeling, and frequent progression to ESHF. Heart failure complications predominate over ventricular arrhythmias, which are rare. (C) 2022 The Authors. Published by Elsevier on behalf of the American College of Cardiology Foundation

Treatment with tocilizumab or corticosteroids for COVID-19 patients with hyperinflammatory state: a multicentre cohort study (SAM-COVID-19)

Objectives: The objective of this study was to estimate the association between tocilizumab or corticosteroids and the risk of intubation or death in patients with coronavirus disease 19 (COVID-19) with a hyperinflammatory state according to clinical and laboratory parameters. Methods: A cohort study was performed in 60 Spanish hospitals including 778 patients with COVID-19 and clinical and laboratory data indicative of a hyperinflammatory state. Treatment was mainly with tocilizumab, an intermediate-high dose of corticosteroids (IHDC), a pulse dose of corticosteroids (PDC), combination therapy, or no treatment. Primary outcome was intubation or death; follow-up was 21 days. Propensity score-adjusted estimations using Cox regression (logistic regression if needed) were calculated. Propensity scores were used as confounders, matching variables and for the inverse probability of treatment weights (IPTWs). Results: In all, 88, 117, 78 and 151 patients treated with tocilizumab, IHDC, PDC, and combination therapy, respectively, were compared with 344 untreated patients. The primary endpoint occurred in 10 (11.4%), 27 (23.1%), 12 (15.4%), 40 (25.6%) and 69 (21.1%), respectively. The IPTW-based hazard ratios (odds ratio for combination therapy) for the primary endpoint were 0.32 (95%CI 0.22-0.47; p < 0.001) for tocilizumab, 0.82 (0.71-1.30; p 0.82) for IHDC, 0.61 (0.43-0.86; p 0.006) for PDC, and 1.17 (0.86-1.58; p 0.30) for combination therapy. Other applications of the propensity score provided similar results, but were not significant for PDC. Tocilizumab was also associated with lower hazard of death alone in IPTW analysis (0.07; 0.02-0.17; p < 0.001). Conclusions: Tocilizumab might be useful in COVID-19 patients with a hyperinflammatory state and should be prioritized for randomized trials in this situatio