Caracterización de un modelo in vitro para el estudio de daño cardíaco

La insuficiencia cardiaca es un síndrome clínico heterogéneo que supone un problema de salud a nivel global. En esta patología se han reconocido mecanismos de inflamación, estrés oxidativo y apoptosis, por lo que nos proponemos caracterizar la línea celular H9c2 de cardiomioblastos y diferentes estímulos para generar un modelo in vitro que nos permita emular estas condiciones en el laboratorio. Estudiamos además si se producen cambios en el citoesqueleto como consecuencia de dichas alteraciones metabólicas. Tras los estudios comprobamos que los estímulos para estudiar el perfil inflamatorio, LPS y TNFαα, no son capaces de poner en marcha las rutas de señalización esperadas, mientras que el estímulo mimético de hipoxia, CoCl2, sí que es capaz de activar la línea de cardiomiocitos. Sería necesaria la realización de más experimentos para una completa caracterización y uso del modelo propuesto para el estudio de la insuficiencia cardiaca in vitro.Departamento de Bioquímica y Biología Molecular y FisiologíaMáster en Investigación Biomédica y Terapias Avanzada

Immune signaling kinases in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD)

Amyotrophic lateral sclerosis (ALS) is the most common neurodegenerative disorder of motor neurons in adults, with a median survival of 3–5 years after appearance of symptoms, and with no curative treatment currently available. Frontotemporal dementia (FTD) is also an adult-onset neurodegenerative disease, displaying not only clinical overlap with ALS, but also significant similarities at genetic and pathologic levels. Apart from the progressive loss of neurons and the accumulation of protein inclusions in certain cells and tissues, both disorders are characterized by chronic inflammation mediated by activated microglia and astrocytes, with an early and critical impact of neurodegeneration along the disease course. Despite the progress made in the last two decades in our knowledge around these disorders, the underlying molecular mechanisms of such non-cell autonomous neuronal loss still need to be clarified. In particular, immune signaling kinases are currently thought to have a key role in determining the neuroprotective or neurodegenerative nature of the central and peripheral immune states in health and disease. This review provides a comprehensive and updated view of the proposed mechanisms, therapeutic potential, and ongoing clinical trials of immune-related kinases that have been linked to ALS and/or FTD, by covering the more established TBK1, RIPK1/3, RACK I, and EPHA4 kinases, as well as other emerging players in ALS and FTD immune signaling.Ministerio de Economía y Competitividad RTI2018- 098432-B-I00, RYC-2017-23127Junta de Andalucía US-1265227, PY20_0109

Carbon nanodot–based electrogenerated chemiluminescence biosensor for miRNA-21 detection

A simple carbon nanodot–based electrogenerated chemiluminescence biosensor is described for sensitive and selective detection of microRNA-21 (miRNA-21), a biomarker of several pathologies including cardiovascular diseases (CVDs). The photoluminescent carbon nanodots (CNDs) were obtained using a new synthesis method, simply by treating tiger nut milk in a microwave reactor. The synthesis is environmentally friendly, simple, and efficient. The optical properties and morphological characteristics of the CNDs were exhaustively investigated, confirming that they have oxygen and nitrogen functional groups on their surfaces and exhibit excitation-dependent fluorescence emission, as well as photostability. They act as co-reactant agents in the anodic electrochemiluminescence (ECL) of [Ru(bpy)3]2+, producing different signals for the probe (single-stranded DNA) and the hybridized target (double-stranded DNA). These results paved the way for the development of a sensitive ECL biosensor for the detection of miRNA-21. This was developed by immobilization of a thiolated oligonucleotide, fully complementary to the miRNA-21 sequence, on the disposable gold electrode. The target miRNA-21 was hybridized with the probe on the electrode surface, and the hybridization was detected by the enhancement of the [Ru(bpy)3]2+/DNA ECL signal using CNDs. The biosensor shows a linear response to miRNA-21 concentration up to 100.0 pM with a detection limit of 0.721 fM. The method does not require complex labeling steps, and has a rapid response. It was successfully used to detect miRNA-21 directly in serum samples from heart failure patients without previous RNA extraction neither amplification processThis study is funded by the Comunidad Autónoma de Madrid (Spain) projects (TRANSNANOAVANSENS, S2018/NMT-4349, CAM/B2017/BMD-3686) and Ministerio de Economía, Industria y Competitividad (Spanish Government) projects: CTQ2015-71955-REDT (ELECTROBIONET), CTQ2014-53334-C2-1-R and PID2020-116728RB-I0

Clinical, Molecular and Genetic Characteristics of Early Onset Gastric Cancer: Analysis of a Large Multicenter Study

Gastric adenocarcinoma (GC) is a common tumor with high morbidity and mortality. Only 7% of patients with GC are diagnosed before age 50 (early onset gastric cancer (EOGC)), and their characteristics have been poorly described. We aimed to describe clinical, molecular, and genetic characteristics of EOGC. A total of 309 patients with EOGC were retrospectively studied in four Spanish centers. Personal information, family history, and tumor information were registered. Germinal genetic analysis was performed in patients who met current criteria of a hereditary syndrome at the time of diagnosis. The median age at diagnosis was 44 years. The majority (73.3%) of tumors were diffuse, and 78.3% were diagnosed in an advanced stage. Familial aggregation of GC was present in 18/117 (15.4%) cases, and 5/117 (4.3%) met criteria for familial GC. MMR-IHC was performed in 126/309 (40.7%) tumors: 4/126 (3.1%) had loss of expression in MLH1/PMS2, without an associated germline mutation. Sixteen germline genetic analyses were performed, detecting a pathogenic variant in four (25%) cases: one in BRCA2, one in TP53, and two in CDH1. Most EOGC are diffuse and diagnosed in an advanced stage. In these patients, DNA MMR system deficiency is uncommon. Although familial aggregation was observed in only 15% of cases, a germline mutation was found in 25% of patients tested with clinical criteria. This demonstrates that EOGC has a marked genetic heterogeneity, reinforcing the importance of an accurate genetic counseling and enhancing the emerging use of multigene panels

Pathogen sensing device based on 2D MoS2/graphene heterostructure

In this work we propose a new methodology for selective and sensitive pathogen detection based on a 2D layered heterostructured biosensing platform. As a proof of concept, we have chosen SARS-CoV-2 virus because the availability of new methods to detect this virus is still a great deal of interest. The prepared platform is based on the covalent immobilization of molybdenum disulphide functionalized with a diazonium salt (f-MoS2) onto graphene screen-printed electrodes (GPH SPE) by electrografting of the diazonium salt. This chemistry-based method generates an improved heterostructured biosensing platform for aptamer immobilization and aptasensor development. Electrochemical impedance spectroscopy (EIS) is used to obtain the signal response of the device, proving the ability of the sensor platform to detect the virus. SARS-CoV-2 spike RBD recombinant protein (SARS-CoV-2 S1 protein) has been detected and quantified with a low detection limit of 2.10 fg/mL. The selectivity of the developed biosensor has been confirmed after detecting the S1 protein even in presence of other interfering proteins. Moreover, the ability of the device to detect SARS-CoV-2 S1 protein has been also tested in nasopharyngeal swab samplesThis work has been financially supported by the Spanish Ministry of Economy and Competitiveness (PID2020-116728RB-I00, PID2020- 116661RB-I00, CTQ2015-71955-REDT (ELECTROBIONET)) and Community of Madrid (TRANSNANOAVANSENS, S2018/NMT-4349, and PhotoArt P2018/NMT-4367). E. Enebral thank the financial support of “Nanotecnología para detección del SARS-CoV-2 y sus variantes. NANOCOV” project. IMDEA Nanociencia receives support from the “Severo Ochoa” Programme for Centres of Excellence in R&D (MINECO, Grant CEX2020-001039-S). We also thank the Spanish Ministry of Universities for supporting Laura Gutiérrez-Galvez with the Formación del Profesorado Universitario (FPU) grant (FPU19/06309

Índices de shock prehospitalario y hospitalario como predictores de transfusión masiva en la atención inicial del paciente politraumático

Objectives. To explore a possible association between the shock index and a need for massive blood transfusion, duration of hospital stay in the critical care unit, and mortality. Methods. Observational study of data for all patients over the age of 18 years with multiple high-energy injuries included in the TraumCat Registry who were treated in Hospital Universitario de Bellvitge between 2012 and 2016.We calculated shock index values before hospital emergency department arrival, on arrival at the hospital, and on admission to the critical care unit for resuscitation. The amount of blood transfused in the first 24 hours was also obtained from the registry. Results. Of 184 polytrauma patients, 75 (41%) received blood transfusions. Median (interquartile range) shock indices were as follows: prehospital, 0.77 (0.61-1.01); on hospital arrival, 0.78 (0.64-1); and on critical care admission, 0.92 (0.76-1.13). Forty-six patients (25%) died. A prehospital shock index of 0.9 was significant, differentiating the amount of blood transfused. The specificity and sensitivity of the cut off were 73% and 66%,respectively, at the prehospital recording and 74% and 80% on hospital arrival. The areas under the receiver operating characteristic curve and 95% CIs were as follows for prehospital and on-arrival shock indices: 68% (61%-75%) and 72% (65%-79%). Mortality and hospital stay were not significantly associated with shock indices. Conclusions: The shock index is a useful, easy-to-obtain predictor to identify polytrauma patients who need early blood transfusion for optimal treatment. Hospital stay and mortality might be better predicted by other indicators

Asthma outcomes improve with continuous positive airway pressure for obstructive sleep apnea

Continuous positive airway pressure () in asthma patients with concomitant obstructive sleep apnea syndrome () seems to have a favorable impact on asthma, but data are inconsistent due to methodological limitations of previous studies. Prospective, multicenter study. We examined asthma outcomes after 6 months of in 99 adult asthma patients (mean age 57 years) with (respiratory disturbance index ≥20). Asthma control and quality of life were assessed with the Asthma Control Questionnaire () and the Mini Asthma Quality of Life Questionnaire (Mini), respectively. Data were analyzed by intention-to-treat basis. The mean ± score of the decreased from 1.39 ± 0.91 at baseline to 1.0 ± 0.78 at 6 months (P = 0.003), the percentage of patients with uncontrolled asthma from 41.4% to 17.2% (P = 0.006), and the percentage of patients with asthma attacks in the 6 months before and after treatment from 35.4% to 17.2% (P = 0.015). The score of the increased from 5.12 ± 1.38 to 5.63 ± 1.17 (P = 0.009). There were also significant improvements in symptoms of gastroesophageal reflux and rhinitis, bronchial reversibility, and exhaled nitric oxide values (all P < 0.05). No significant changes were observed in drug therapy for asthma or their comorbidities nor in the patients' weight. Asthma control (both actual and future risk), quality of life, and lung function improved after starting continuous positive airway pressure in asthmatics with moderate to severe obstructive sleep apnea syndrome

Asthma outcomes improve with continuous positive airway pressure for obstructive sleep apnea

Continuous positive airway pressure () in asthma patients with concomitant obstructive sleep apnea syndrome () seems to have a favorable impact on asthma, but data are inconsistent due to methodological limitations of previous studies. Prospective, multicenter study. We examined asthma outcomes after 6 months of in 99 adult asthma patients (mean age 57 years) with (respiratory disturbance index ≥20). Asthma control and quality of life were assessed with the Asthma Control Questionnaire () and the Mini Asthma Quality of Life Questionnaire (Mini), respectively. Data were analyzed by intention-to-treat basis. The mean ± score of the decreased from 1.39 ± 0.91 at baseline to 1.0 ± 0.78 at 6 months (P = 0.003), the percentage of patients with uncontrolled asthma from 41.4% to 17.2% (P = 0.006), and the percentage of patients with asthma attacks in the 6 months before and after treatment from 35.4% to 17.2% (P = 0.015). The score of the increased from 5.12 ± 1.38 to 5.63 ± 1.17 (P = 0.009). There were also significant improvements in symptoms of gastroesophageal reflux and rhinitis, bronchial reversibility, and exhaled nitric oxide values (all P < 0.05). No significant changes were observed in drug therapy for asthma or their comorbidities nor in the patients' weight. Asthma control (both actual and future risk), quality of life, and lung function improved after starting continuous positive airway pressure in asthmatics with moderate to severe obstructive sleep apnea syndrome

The therapeutic relationship from the perspective of patients and nurses in the first days of admission: A cross‐sectional study in acute mental health units

The therapeutic relationship (TR) is essential in mental health nursing care and plays a fundamental role in the understanding and treatment of the patient's health status. Despite being a bidirectional construct, limited evidence is available to shed light on this issue in mental health units and even less so in the first days of admission. This study aimed to examine the association and differences between nurses' and patients' perspectives on the establishment of the therapeutic relationship in acute mental health units during the first days of hospitalization. A cross-sectional study was carried out in 12 Spanish mental health units. Data were collected from patients and nurses using the Working Alliance Inventory-Short (WAI-S) questionnaire. A total of 234 cases were analysed, including 234 patients and 58 nurses. The results showed a positive association between nurses' and patients' perspectives on the therapeutic relationship, but also revealed significant differences on each WAI-S dimension. Nurses assigned higher scores compared to patients on the perception of the quality of the therapeutic relationship. The dimensions with the greatest weight from the patients' perspective regarding the quality of the therapeutic relationship were the perception of greater agreement on goals and tasks among nurses. This study demonstrates the importance of establishing shared goals and tasks with nurses from the first days of hospitalization to improve the quality of the therapeutic relationship as perceived by patients. These findings underline the need to consider the different perspectives of both parties to promote a high-quality therapeutic relationship

Germline Mutations in FAF1 Are Associated With Hereditary Colorectal Cancer

Background & aims: A significant proportion of colorectal cancer (CRC) cases have familial aggregation but little is known about the genetic factors that contribute to these cases. We performed an exhaustive functional characterization of genetic variants associated with familial CRC. Methods: We performed whole-exome sequencing analyses of 75 patients from 40 families with a history of CRC (including early-onset cases) of an unknown germline basis (discovery cohort). We also sequenced specific genes in DNA from an external replication cohort of 473 families, including 488 patients with colorectal tumors that had normal expression of mismatch repair proteins (validation cohort). We disrupted the Fas-associated factor 1 gene (FAF1) in DLD-1 CRC cells using CRISPR/Cas9 gene editing; some cells were transfected with plasmids that express FAF1 missense variants. Cells were analyzed by immunoblots, quantitative real-time polymerase chain reaction, and functional assays monitoring apoptosis, proliferation, and assays for Wnt signaling or nuclear factor (NF)-kappa-B activity. Results: We identified predicted pathogenic variant in the FAF1 gene (c.1111G>A; p.Asp371Asn) in the discovery cohort; it was present in 4 patients of the same family. We identified a second variant in FAF1 in the validation cohort (c.254G>C; p.Arg85Pro). Both variants encoded unstable FAF1 proteins. Expression of these variants in CRC cells caused them to become resistant to apoptosis, accumulate beta-catenin in the cytoplasm, and translocate NF-kappa-B to the nucleus. Conclusions: In whole-exome sequencing analyses of patients from families with a history of CRC, we identified variants in FAF1 that associate with development of CRC. These variants encode unstable forms of FAF1 that increase resistance of CRC cells to apoptosis and increase activity of beta-catenin and NF-kappa-B