Extracellular vesicles from airway secretions: New insights in lung diseases

Lung diseases (LD) are one of the most common causes of death worldwide. Although it is known that chronic airway inflammation and excessive tissue repair are processes associated with LD such as asthma, chronic obstructive pulmonary disease (COPD) or idiopathic pulmonary fibro-sis (IPF), their specific pathways remain unclear. Extracellular vesicles (EVs) are heterogeneous na-noscale membrane vesicles with an important role in cell-to-cell communication. EVs are present in general biofluids as plasma or urine but also in secretions of the airway as bronchoalveolar lavage fluid (BALF), induced sputum (IS), nasal lavage (NL) or pharyngeal lavage. Alterations of airway EV cargo could be crucial for understanding LD. Airway EVs have shown a role in the pathogenesis of some LD such as eosinophil increase in asthma, the promotion of lung cancer in vitro models in COPD and as biomarkers to distinguishing IPF in patients with diffuse lung diseases. In addition, they also have a promising future as therapeutics for LD. In this review, we focus on the importance of airway secretions in LD, the pivotal role of EVs from those secretions on their pathophysiology and their potential for biomarker discovery

Circulating Exosomal Mir21 And Mir320 In Obstructive Sleep Apnea

Rational. Epidemiological studies indicate that there may be an association between obstructive sleep apnea (OSA) and cardiovascular and metabolic diseases. Some pro-inflammatory miRs (miR-21, miR320) critical for the immune response or hypoxia are often overexpressed in cancers and atherosclerosis. Aim. To examine the expression of miR-21& miR320 in circulating exosomes from patients with OSA. Methods: From a Sleep Unit and in the frame of a long-term longitudinal cohort study we selected 65 non-smokers OSA patients (apnea-hypopnea index -AHI- 30 events/ti) and 26 age, gender and BMI-matched controls (AHI 0.85 mm. Plasma-derived exosomes were isolated by precipitation using miRCURY, , , Exosome Isolation Kit. Exosomes were characterized by transmission electron microscopy, dynamic light scattering assay and Western Blot analysis using CD63 and HSP70. Exosome total RNA was obtained using miRCURY"* RNA isolation kit. miR-21 -5p and miR-320-3p were analysed by real time quantitative PCR (RT-qPCR) using miRCURY LNA~ technology..

Increased circulating microRNAs miR-342-3p and miR-21-5p in natural sheep prion disease

Scrapie is a transmissible spongiform encephalopathy (TSE), or prion disease, of sheep and goats. As no simple diagnostic tests are yet available to detect TSEs in vivo, easily accessible biomarkers could facilitate the eradication of scrapie agents from the food chain. To this end, we analysed by quantitative reverse transcription PCR a selected set of candidate microRNAs (miRNAs) from circulating blood plasma of naturally infected, classical scrapie sheep that demonstrated clear scrapie symptoms and pathology. Significant scrapie-associated increase was repeatedly found for miR-342-3p and miR-21-5p. This is the first demonstration, to our knowledge, of circulating miRNA alterations in any animal suffering from TSE. Genome-wide expression studies are warranted to investigate the true depth of miRNA alterations in naturally occurring TSEs, especially in presymptomatic animals, as the presented study demonstrates the potential feasibility of miRNAs as circulating TSE biomarkers

Heterogeneity of melanoma cell responses to sleep apnea-derived plasma exosomes and to intermittent hypoxia

Obstructive sleep apnea (OSA) is associated with increased cutaneous melanoma incidence and adverse outcomes. Exosomes are secreted by most cells, and play a role in OSA-associated tumor progression and metastasis. We aimed to study the effects of plasma exosomes from OSA patients before and after adherent treatment with continuous positive airway pressure (CPAP) on melanoma cells lines, and also to identify exosomal miRNAs from melanoma cells exposed to intermittent hypoxia (IH) or normoxia. Plasma-derived exosomes were isolated from moderate-to-severe OSA patients before (V1) and after (V2) adherent CPAP treatment for one year. Exosomes were co-incubated with three3 different melanoma cell lines (CRL 1424; CRL 1619; CRL 1675) that are characterized by genotypes involving different mutations in BRAF, STK11, CDKN2A, and PTEN genes to assess the effect of exosomes on cell proliferation and migration, as well as on pAMK activity in the presence or absence of a chemical activator. Subsequently, CRL-1424 and CRL-1675 cells were exposed to intermittent hypoxia (IH) and normoxia, and exosomal miRNAs were identified followed by GO and KEG pathways and gene networks. The exosomes from these IH-exposed melanoma cells were also administered to THP1 macrophages to examine changes in M1 and M2 polarity markers. Plasma exosomes from V1 increased CRL-1424 melanoma cell proliferation and migration compared to V2, but not the other two cell lines. Exposure to CRL-1424 exosomes reduced pAMPK/tAMPK in V1 compared to V2, and treatment with AMPK activator reversed the effects. Unique exosomal miRNAs profiles were identified for CRL-1424 and CRL-1675 in IH compared to normoxia, with six miRNAs being regulated and several KEGG pathways were identified. Two M1 markers (CXCL10 and IL6) were significantly increased in monocytes when treated with exosomes from IH-exposed CRL-1424 and CRL-1625 cells. Our findings suggest that exosomes from untreated OSA patients increase CRL-1424 melanoma malignant properties, an effect that is not observed in two other melanoma cell lines. Exosomal cargo from CRL-1424 cells showed a unique miRNA signature compared to CRL-1675 cells after IH exposures, suggesting that melanoma cells are differentially susceptible to IH, even if they retain similar effects on immune cell polarity. It is postulated that mutations in STK-11 gene encoding for the serine/threonine kinase family that acts as a tumor suppressor may underlie susceptibility to IH-induced metabolic dysfunction, as illustrated by CRL-1424 cells. © 2021 by the authors. Licensee MDPI, Basel, Switzerland

Soluble RAGE in COPD, with or without coexisting obstructive sleep apnoea

Background: Hypoxia can reduce the levels of soluble receptor for advanced glycation end-products (sRAGE), a new anti-inflammatory biomarker of COPD. We assessed sRAGE in patients with hypoxia-related diseases such as COPD, OSA and OSA-COPD overlap. Methods: Plasma levels of sRAGE were measured in 317 subjects at baseline (57 heathy nonsmokers HNS], 84 healthy smokers HS], 79 OSA, 62 COPD and 35 OSA-COPD overlap patients) and in 294 subjects after one year of follow-up (50 HNS, 74 HS, 77 OSA, 60 COPD and 33 overlap). Results: After adjusting for age, sex, smoking status and body mass index, sRAGE levels showed a reduction in OSA (- 12.5%, p = 0.005), COPD (- 14.8%, p < 0.001) and OSA-COPD overlap (- 12.3%, p = 0.02) compared with HNS. There were no differences when comparing sRAGE plasma levels between overlap patients and those with OSA or COPD alone. At follow-up, sRAGE levels did not change significantly in healthy subjects, COPD and OSA or OSA-COPD overlap nontreated with continuous positive airway pressure (CPAP). Moreover, in patients with OSA and OSA-COPD overlap who were treated with CPAP, sRAGE increased significantly. Conclusions: The levels of sRAGE are reduced in COPD and OSA. Treatment with CPAP appears to improve sRAGE levels in patients with OSA who also had COPD

L-Arginine Ameliorates Defective Autophagy in GM2 Gangliosidoses by mTOR Modulation.

AIMS: Tay-Sachs and Sandhoff diseases (GM2 gangliosidosis) are autosomal recessive disorders of lysosomal function that cause progressive neurodegeneration in infants and young children. Impaired hydrolysis catalysed by β-hexosaminidase A (HexA) leads to the accumulation of GM2 ganglioside in neuronal lysosomes. Despite the storage phenotype, the role of autophagy and its regulation by mTOR has yet to be explored in the neuropathogenesis. Accordingly, we investigated the effects on autophagy and lysosomal integrity using skin fibroblasts obtained from patients with Tay-Sachs and Sandhoff diseases. RESULTS: Pathological autophagosomes with impaired autophagic flux, an abnormality confirmed by electron microscopy and biochemical studies revealing the accelerated release of mature cathepsins and HexA into the cytosol, indicating increased lysosomal permeability. GM2 fibroblasts showed diminished mTOR signalling with reduced basal mTOR activity. Accordingly, provision of a positive nutrient signal by L-arginine supplementation partially restored mTOR activity and ameliorated the cytopathological abnormalities. INNOVATION: Our data provide a novel molecular mechanism underlying GM2 gangliosidosis. Impaired autophagy caused by insufficient lysosomal function might represent a new therapeutic target for these diseases. CONCLUSIONS: We contend that the expression of autophagy/lysosome/mTOR-associated molecules may prove useful peripheral biomarkers for facile monitoring of treatment of GM2 gangliosidosis and neurodegenerative disorders that affect the lysosomal function and disrupt autophagy

El cabalgamiento Finiserravallense del Norte de Sierra Arana (Cordillera Bética). Observaciones sobre la caracterización bioestratigráfica del Serravallense

Al N. de Sierra Arana se reconocen una serie de ventanas tectónicas en las que afloran margas Serravallenses y calcarenitas del Langhense superior-Serravallense inferior, bajo unas dolomías que presentan, en la base, intercalaciones de rocas subvolcánicas básicas. Los materiales cabalgantes parecen pertenecer a la unidad de Sierra Arana, o a alguna otra directamente relacionada con ella, aún cuando están separados del conjunto por un importante accidente de desgarre que discurre al pie de la ladera N. de dicha sierra. El cabalgamiento, de una magnitud mínima estimada de 2'5 Km., se produjo entre el Serravallense superior y el Tortonense basal.In the northern part of Sierra Arana several tectonic windows have been recognized. Serravallian marls and upper Langhian - lower Serravallian calcarenites are overthrusted by dolomites with basic subvolcanic rocks interbedded in their lower part. The allochtonous rocks must belong to the Sierra Arana unit, or to some other unit' directly connected with it, even if no stratigrafic continuity between them can be ascertained because of an important strike-slip fault bordering the northern foot of the Sierra Arana massif. Tectonic superposition has an amplitude of 2'5 Km. or more and took place during the upper Serravallian or the lowermost Tortonian

Evolution of the incidence of colonized and infected patients by VIM carbapenemase-producing bacteria in a pediatric hospital in Spain

OBJECTIVE: The aim of this study is to describe the evolution of the incidence of infected and colonized patients with carbapenemase VIM-producing bacteria (CPB-VIM) at a national referral pediatric center in Madrid, Spain, between 2012 and 2015. METHODS: Descriptive epidemiological surveillance study. The surveillance system included case detection (screening for BPC colonization in all admitted patients, with periodicity according to the ward) and control measures (contact precautions, identification of previously colonized patients at admission, environmental cleaning, education, supervision of contact precautions, and patient cohort). All hospitalized patients with first positive microbiological sample for CPB-VIM in 2012-2015 were included. Colonized patients were followed through clinical history to evaluate later infection. RESULTS: We found 239 colonized and 51 infected patients with CPB-VIM (49.3% women, 47.6% were 5 months old or younger, 52.1% admitted at Intensive Care Unit). Infection and colonization incidence were, respectively, 2.6 and 6.7 cases per one thousand hospitalized patients in 2012, 1.8 and 10.0 in 2014 and 0.3 and 5.0 in 2015. Within these patients, 84.4% shared ward with other patient with previous positive sample. 13.0% (31/239) of colonized patients had a subsequent infection. CONCLUSIONS: We have shown data of pediatric patients affected by BPC-VIM, collected from an epidemiological surveillance system that included systematic screening at a national referral center. After an epidemic period, the incidence of cases went down. The surveillance and infection control measures intensification, as well as coordination with involved departments, were key in the handling of the situation.S

Manejo antibiótico en neumonía adquirida en la comunidad en la hospitalización a domicilio: ¿Hay margen de mejora?

Introducción: La neumonía adquirida en la comunidad (NAC) es una enfermedad frecuente que puede serabordada en hospitalización a domicilio (HAD). En el presente trabajo evaluamos el margen de mejoracon el uso de antibióticos en la NAC en HAD. Métodos: Se reclutaron retrospectivamente todos los pacientes con NAC en dos hospitales españolesdesde el 01 de enero de 2018 al 30 de octubre de 2019. Se registraron variables demográficas, clínicas y sobre calidad de prescripción antibiótica. Posteriormente se construyó una variable que recogía seis indi-cadores de calidad asistencial, categorizando y comparando a los pacientes en dos grupos: buena calidadasistencial (cuatro o más indicadores realizados) o mala calidad asistencial (tres o menos indicadoresrealizados). Resultados: Obtuvimos una muestra de 260 pacientes. La solicitud de pruebas diagnósticas y la adecuacióna las guías de práctica clínica fue del 85, 4 y 85, 8%, respectivamente. Los porcentajes de realización dedesescalada (53, 7%) y terapia secuencial (57, 7%) cuando estaban indicadas fueron bajos. La duraciónmedia del tratamiento fue de 7, 3 días para el intravenoso y 9, 5 días para el total. La calidad de prescripciónfue buena en 134 (63, 2%) pacientes, siendo más frecuente en aquellos que ingresaron directamente enHAD desde urgencias. También se asoció a menor reingreso a 30 días. Conclusión: Existe un amplio margen de mejora en algunos aspectos con el uso de antimicrobianos enHAD, que podría motivar la implementación de programas de optimización del uso de antibióticos. Introduction: Community Acquired Pneumonia (CAP) is common disease that can be treated in Hospital At Home (HAH). In this paper we evaluate the room of improvement in the use of antibiotics in CAP in HH. Methods: Patients with CAP were retrospectively recruited in two Spanish hospitals from 1/1/18 to 10/30/19. Demographic, clinical and quality of antibiotic prescription variables were recorded. Subsequently, we created a new variable that collected six quality of care indicator, categorizing and comparing patients into two groups: good quality of care (4 or more indicators performed) or poor quality of care (3 or less indicators performed). Results: We recruited 260 patients. The request for diagnostic tests and the adequacy to Clinical Practice Guidelines were 85.4% and 85.8% respectively. Percentages of de-escalation (53.7%) and sequential therapy (57.7%) when indicated were low. The average length of treatment was 7.3 days for intravenous and 9.5 days for total. Quality of prescription was good in 134 (63.2%) patients, being more frequent in those who were admitted directly to HAD from the emergency room. It was also associated with less readmission at 30 days. Conclusion: There is a wide room for improvement in some fields of antimicrobials use in HAH that could stimulate the implementation of Antimicrobial Stewardship Programs

El Neógeno del Valle de los Guájares (Cordillera Bética, Granada)

El estudio llevado a cabo sobre las características litoestratigráficas, el contenido faunístico y la edad de los materiales neógenos del Valle de los Guájares, pone de manifiesto la presencia de materiales del Serravallense y del Tortonense, además de los correspondientes al Plioceno (?) - Cuaternario. Las deformaciones de tales materiales se deben esencialmente a fracturas con saltos tanto horizontales como verticales, que controlaron en buena medida el relieve y la sedimentación. A partir de estos datos y de otros de carácter regional se aborda la evolución geológica del sector, posterior a la estructuración esencial de la Cordillera Bética durante el Mioceno inferior. A este respecto se pueden destacar dos fases deformacionales: una fase finiserravallense, responsable de algunas fallas inversas que afectan a las margas pelágicas serravallenses y a los materiales alpujárrides y, probablemente, también responsable del nacimiento de sistemas de fallas, las cuales podrían ya controlar la sedimentación; segunda fase intratortonense que produce discordancia angular entre los materiales tortonenses así como una nueva actividad, principalmente con desplazamientos verticales, de los sistemas de fallas previamente formados. Los materiales del Plioceno (?) - Cuaternario muestran igualmente evidencias de movimientos tectónicos recientes en los que prevalecen los desplazamientos verticales sobre los horizontales.The geological study, lead upon the lithostratigraphic characteristics, the faunistic content and the age of the neogene materials from the Guájares valley reveals the presence of terranes belonging to the Serravallian and the Tortonian, besides the ones corresponding to the Pliocene (?) - Quaternary. The deformation features of such materials are essentially due to faults showing both horizontal and vertical slips, which strongly controlled the relief and the deposition. From these data and others of regional character we envisage the geological evolution of the area, once the main structuration of the Betic Cordilleras during the lower Miocene was already accomplished. In this regard two deformational phases stand out: a finiserravallian phase, responsible for sorne reverse faults concerning serravallian pelagic marls and alpujarride materials and, probably also and for the birth of important fault systems which could already control the sedimentation; which a second one, the intratortonian phase, which yields an angular unconformity within the tortonian materials as well as a new activity, mainly with vertical displacements, of the previous fault systems. The Pliocene (?)- Quaternary deposits provide also evidence of recent tectonic movements, the vertical slips being prevalent over the horizontal ones.Trabajo realizado dentro del Proyecto «El Borde Mediterráneo español: Evolución del Orógeno bético y geodinámica de las depresiones neógenas», realizado por el Departamento de Investigaciones Geológicas, C.S.I.C. y financiado por C.A.I.C.YT. y C.S.I.C