Targeted Next-Generation Sequencing in a Large Cohort of Genetically Undiagnosed Patients with Neuromuscular Disorders in Spain

This article belongs to the Special Issue Genetic Advances in Neuromuscular Disorders: From Gene Identification to Gene Therapy.The term neuromuscular disorder (NMD) includes many genetic and acquired diseases and differential diagnosis can be challenging. Next-generation sequencing (NGS) is especially useful in this setting given the large number of possible candidate genes, the clinical, pathological, and genetic heterogeneity, the absence of an established genotype-phenotype correlation, and the exceptionally large size of some causative genes such as TTN, NEB and RYR1. We evaluated the diagnostic value of a custom targeted next-generation sequencing gene panel to study the mutational spectrum of a subset of NMD patients in Spain. In an NMD cohort of 207 patients with congenital myopathies, distal myopathies, congenital and adult-onset muscular dystrophies, and congenital myasthenic syndromes, we detected causative mutations in 102 patients (49.3%), involving 42 NMD-related genes. The most common causative genes, TTN and RYR1, accounted for almost 30% of cases. Thirty-two of the 207 patients (15.4%) carried variants of uncertain significance or had an unidentified second mutation to explain the genetic cause of the disease. In the remaining 73 patients (35.3%), no candidate variant was identified. In combination with patients’ clinical and myopathological data, the custom gene panel designed in our lab proved to be a powerful tool to diagnose patients with myopathies, muscular dystrophies and congenital myasthenic syndromes. Targeted NGS approaches enable a rapid and cost-effective analysis of NMD- related genes, offering reliable results in a short time and relegating invasive techniques to a second tier.This study was granted by FIS PI15/01898, funded by ISCIII and FEDER, ‘Una manera de hacer Europa’ and by Fundación Mutua Madrileña in the “Convocatoria de ayudas a la Investigación en Salud 2015”. It was also funded by an ACCI grant from CIBERER. Daniel Natera-de Benito is the recipient of a grant from the Instituto de Salud Carlos III (Contrato Rio Hortega, CM17/00044)

Synergetic monitoring of Saharan dust plumes and potential impact on surface: a case study of dust transport from Canary Islands to Iberian Peninsula

The synergetic use of meteorological information, remote sensing both ground-based active (lidar) and passive (sun-photometry) techniques together with backtrajectory analysis and in-situ measurements is devoted to the characterization of dust intrusions. A case study of air masses advected from the Saharan region to the Canary Islands and the Iberian Peninsula, located relatively close and far away from the dust sources, respectively, was considered for this purpose. The observations were performed over three Spanish geographically strategic stations within the dust-influenced area along a common dust plume pathway monitored from 11 to 19 of March 2008. A 4-day long dust event (13–16 March) over the Santa Cruz de Tenerife Observatory (SCO), and a linked short 1-day dust episode (14 March) in the Southern Iberian Peninsula over the Atmospheric Sounding Station "El Arenosillo" (ARN) and the Granada station (GRA) were detected.This work has been supported by the Spanish Ministry for Science and Innovation (MICINN) under the Complementary Actions CGL2008-01330-E (2009) in the frame of the Spanish and Portuguese Aerosol LIdar NETwork (SPALINET), and CGL2010-10012-E (Evaluation of lidar observations in the frame of CIEMAT/EARLINET intercomparison by airborne in-situ measurements of trace gases and aerosols, MISPA-LIDAR, 2010); projects CGL2008-05939-C03-03/CLI, CGL2007-66477-C02-01, CSD2007-00067 and CGL2010-18782 of the Spanish Ministry of Education; projects P08-RNM-3568 and P10-RNM-6299 of the Autonomous Government of Andalusia; and the EARLINET-ASOS project (EU Coordination Action, contract no. 025991 (RICA)). JLG-R thanks the Spanish Ministry of Education and the Portuguese Fundaçao para a Ciencia e a Tecnologia for supporting under grants EX2009-0700 and SFRH/BPD/63090/2009, respectively

MicroRNA-223 is a novel negative regulator of HSP90B1 in CLL

This is an Open Access article distributed under the terms of the Creative Commons Attribution License.-- et al.[Background]: MicroRNAs are known to inhibit gene expression by binding to the 3'UTR of the target transcript. Downregulation of miR-223 has been recently reported to have prognostic significance in CLL. However, there is no evidence of the pathogenetic mechanism of this miRNA in CLL patients. [Methods]: By applying next-generation sequencing techniques we have detected a common polymorphism (rs2307842), in 24% of CLL patients, which disrupts the binding site for miR-223 in HSP90B1 3'UTR. We investigated whether miR-223 directly targets HSP90B1 through luciferase assays and ectopic expression of miR-223. Quantitative real-time polymerase chain reaction and western blot were used to determine HSP90B1 expression in CLL patients. The relationship between rs2307842 status, HSP90B1 expression and clinico-biological data were assessed. [Results]: HSP90B1 is a direct target for miR-223 by interaction with the putative miR-223 binding site. The analysis in paired samples (CD19+ fraction cell and non-CD19+ fraction cell) showed that the presence of rs2307842 and IGHV unmutated genes determined HSP90B1 overexpression in B lymphocytes from CLL patients. These results were confirmed at the protein level by western blot. Of note, HSP90B1 overexpression was independently predictive of shorter time to the first therapy in CLL patients. By contrast, the presence of rs2307842 was not related to the outcome. [Conclusions]: HSP90B1 is a direct target gene of miR-223. Our results provide a plausible explanation of why CLL patients harboring miR-223 downregulation are associated with a poor outcome, pointing out HSP90B1 as a new pathogenic mechanism in CLL and a promising therapeutic target.This work was partially supported by grants from the Spanish Fondo de Investigaciones Sanitarias FIS 09/01543 and PI12/00281, Proyectos de Investigación del SACYL 355/A/09, COST Action EuGESMA (BM0801), Fundación Manuel Solórzano, Obra Social Banca Cívica (Caja Burgos), Fundación Española de Hematología y Hemoterapia (FEHH) and by a grant (RD12/0036/0069) from the Red Temática de Investigación Cooperativa en Cáncer (RTICC), Instituto de Salud Carlos III (ISCIII), Spanish Ministry of Economy and Competitiveness & European Regional Development Fund (ERDF) “Una manera de hacer Europa” (Innocampus). The research leading to these results has received funding from the European Union Seventh Framework Programme [FP7/2007-2013] under Grant Agreement n°306242-NGS-PTL. MHS is fully supported by an Ayuda predoctoral de la Junta de Castilla y Leon by the Fondo Social Europeo. ME Sarasquete is supported by Contrato Miguel Servet (CP13/00080).Peer Reviewe

Impact of COVID-19 on the degree of compliance with hand hygiene: a repeated cross-sectional study

Hand hygiene (HH) is the paramount measure used to prevent healthcare associated infections. A repeated cross-sectional study was undertaken with direct observation of the degree of compliance on HH of healthcare personnel during the SARS-CoV-2 pandemic. Between, 2018-2019, 9,083 HH opportunities were considered, and 5,821 in 2020-2022. Chi squared tests were used to identify associations. The crude and adjusted odds ratios were used along with a logistic regression model for statistical analyses. Compliance on HH increased significantly (p < 0.001) from 54.5% (95% CI: 53.5, 55.5) to 70.1% (95% CI: 68.9, 71.2) during the COVID-19 pandemic. This increase was observed in four of the five key moments of HH established by the World Health Organization (WHO) (p<0.05), except at moment 4. The factors that were significantly and independently associated with compliance were the time period considered, type of healthcare-personnel, attendance at training sessions, knowledge of HH and WHO guidelines, and availability of hand disinfectant alcoholic solution in pocket format. Highest HH compliance occurred during the COVID-19 pandemic, reflecting a positive change in healthcare-personnel’s behavior regarding HH recommendations.We received funding through the Alicante Institute for Health and Biomedical Research (ISABIAL) plan for scientific and technical research and innovation project number 2021-0392

Distinct patterns of APP processing in the CNS in autosomal-dominant and sporadic Alzheimer disease

The online version of this article (doi:10.1007/s00401-012-1062-9) contains supplementary material, which is available to authorized users

Type II InAs/GaAsSb quantum dots: Highly tunable exciton geometry and topology

External control over the electron and hole wavefunctions geometry and topology is investigated in a p-i-n diode embedding a dot-in-a-well InAs/GaAsSb quantum structure with type II band alignment. We find highly tunable exciton dipole moments and largely decoupled exciton recombination and ionization dynamics. We also predicted a bias regime where the hole wavefunction topology changes continuously from quantum dot-like to quantum ring-like as a function of the external bias. All these properties have great potential in advanced electro-optical applications and in the investigation of fundamental spin-orbit phenomena

miniBELEN: a modular neutron counter for (a, n) reactions

miniBELEN is a modular and transportable neutron moderated counter with a nearly flat neutron detection efficiency up to 10 MeV. Modularity implies that the moderator can be reassembled in different ways in order to obtain different types of response. The detector has been developed in the context of the Measurement of Alpha Neutron Yields (MANY) collaboration, which is a scientific effort aiming to carry out measurements of (a, n) production yields, reaction cross-sections and neutron energy spectra. In this work we present and discuss several configurations of the miniBELEN detector. The experimental validation of the efficiency calculations using 252Cf sources and the measurement of the 27Al(a, n) 30P reaction is also presented.This work has been supported by the Spanish Ministerio de Economía y Competitividad under grants FPA2017-83946- C2-1 & C2-2 and PID2019-104714GB-C21 & C22, the Generalitat Valenciana Grant PROMETEO/2019/007, both cofounded by FEDER (EU), and the SANDA project funded under H2020-EURATOM-1.1 Grant No. 847552. The authors acknowledge the support from Centro de Microanálisis de Materiales (CMAM) - Universidad Autónoma de Madrid, for the beam time proposal (Comissioning of neutron detector systems for (a,݊n) reaction measurements) with code P01156, and its technical staff for their contribution to the operation of the accelerator.Article signat per 41 autors/es: N. Mont-Geli, A. Tarifeño-Saldivia, L.M. Fraile, S. Viñals, A. Perea, M. Pallàs, G. Cortés, E. Nácher, J.L. Tain, V. Alcayne, A. Algora, J. Balibrea-Correa, J. Benito, M.J.G. Borge, J.A. Briz, F. Calviño, D. Cano-Ott, A. De Blas, C. Domingo-Pardo, B. Fernández, R. Garcia, G. García, J. Gómez-Camacho, E.M. González-Romero, C. Guerrero, J. Lerendegui-Marco, M. Llanos, T. Martínez, E. Mendoza, J.R. Murias, S.E.A. Orrigo, A. Pérez de Rada, V. Pesudo, J. Plaza, J.M. Quesada, A. Sánchez, V. Sánchez-Tembleque, R. Santorelli, O. Tengblad, J.M. Udías and D. Villamarín.Postprint (published version

An Objective Scatter Index Based on Double-Pass Retinal Images of a Point Source to Classify Cataracts

PURPOSE: To propose a new objective scatter index (OSI) based in the analysis of double-pass images of a point source to rank and classify cataract patients. This classification scheme is compared with a current subjective system. METHODS: We selected a population including a group of normal young eyes as control and patients diagnosed with cataract (grades NO2, NO3 and NO4) according to the Lens Opacities Classification System (LOCS III). For each eye, we recorded double-pass retinal images of a point source. In each patient, we determined an objective scatter index (OSI) as the ratio of the intensity at an eccentric location in the image and the central part. This index provides information on the relevant forward scatter affecting vision. Since the double-pass retinal images are affected by both ocular aberrations and intraocular scattering, an analysis was performed to show the ranges of contributions of aberrations to the OSI. RESULTS: We used the OSI values to classify each eye according to the degree of scatter. The young normal eyes of the control group had OSI values below 1, while the OSI for subjects in LOCS grade II were around 1 to 2. The use of the objective index showed some of the weakness of subjective classification schemes. In particular, several subjects initially classified independently as grade NO2 or NO3 had similar OSI values, and in some cases even higher than subjects classified as grade NO4. A new classification scheme based in OSI is proposed. CONCLUSIONS: We introduced an objective index based in the analysis of double-pass retinal images to classify cataract patients. The method is robust and fully based in objective measurements; i.e., not depending on subjective decisions. This procedure could be used in combination with standard current methods to improve cataract patient surgery scheduling

Trifaceted Mickey Mouse Amphiphiles for Programmable Self-Assembly, DNA Complexation and Organ-Selective Gene Delivery

Instilling segregated cationic and lipophilic domains with an angular disposition in a trehalose-based trifaceted macrocyclic scaffold allows engineering patchy molecular nanoparticles leveraging directional interactions that emulate those controlling self-assembling processes in viral capsids. The resulting trilobular amphiphilic derivatives, featuring a Mickey Mouse architecture, can electrostatically interact with plasmid DNA (pDNA) and further engage in hydrophobic contacts to promote condensation into transfectious nanocomplexes. Notably, the topology and internal structure of the cyclooligosaccharide/pDNA co-assemblies can be molded by fine-tuning the valency and characteristics of the cationic and lipophilic patches, which strongly impacts the transfection efficacy in vitro and in vivo. Outstanding organ selectivities can then be programmed with no need of incorporating a biorecognizable motif in the formulation. The results provide a versatile strategy for the construction of fully synthetic and perfectly monodisperse nonviral gene delivery systems uniquely suited for optimization schemes by making cyclooligosaccharide patchiness the focus.Peer reviewe