Incremento del diagnóstico mas allá del análisis estándar del exoma en una cohorte de pacientes con LGMD

[ES] La distrofia muscular es un grupo heterogéneo de trastornos neuromusculares. Muchos pacientes presentan debilidad progresiva y desgaste muscular, y pueden tener una esperanza de vida reducida debido a comorbilidades como la insuficiencia respiratoria y cardíaca. Dentro de los diferentes tipos de distrofias se encuentra la distrofia de cinturas (LGMD, Limb-girdle muscular dystrophy) cuya prevalencia se estima en alrededor de 20-40/1.000.000, frecuencia probablemente subestimada por la falta de diagnóstico debido a la heterogeneidad genética y clínica. En el año 2014 se propuso realizar un proyecto colaborativo europeo, basado en utilizar la secuenciación del exoma completo en una cohorte de 1466 pacientes con distrofia de cinturas sin diagnóstico, para ayudar a comprender mejor la etiología de la enfermedad y como iniciativa de emplear esta técnica en el diagnóstico rutinario, proyecto denominado MYO-SEQ. La tasa de casos diagnosticados después de cuatro años ha sido del 54%, estando dentro de los valores esperados y observados en otros estudios. El objetivo de nuestro proyecto se ha centrado en profundizar en el análisis de datos y comprender mejor la falta de diagnóstico en el resto de pacientes, enunciando diferentes hipótesis explicativas: herencia digénica, presencia de variantes intrónicas, papel modificador de variantes patogénicas y estudio funcional de nuevos genes candidatos utilizando el sistema modelo Danio rerio. Para ello hemos analizado los datos de la secuenciación del exoma desde la plataforma Seqr, integrando al mismo tiempo la información clínica de los médicos de cada paciente obtenida del software PhenoTips, permitiendo reevaluar los hallazgos clínicos, histopatológicos y de resonancia magnética. Los resultados obtenidos indican la necesidad de profundizar el análisis en los pacientes todavía no diagnosticados utilizando otras técnicas de secuenciación para dilucidar los posibles mecanismos involucrados en la LGMD, para asegurar en un futuro un diagnóstico más preciso y, por lo tanto, un asesoramiento genético y tratamiento que permita mejorar la calidad de vida de estos pacientes.[EN] Muscular dystrophy is a heterogeneous group of neuromuscular disorders. Many patients present with proximal muscle weakness and wasting, and may have a reduced life expectancy due to respiratory and cardiac failure. Among the different types of dystrophies is the limbgirdle muscular disease (LGMD), whose prevalence is estimated at 20-40/1.000.000, a frequency probably underestimated because of the lack of diagnosis due to genetic and clinical heterogeneity. In 2014, it was proposed to carry out a European collaborative project, based on applying whole exome sequencing to a cohort of 1466 patients with unexplained limb‐girdle weakness, to help better understand disease aetiology and as an initiative to use this technique in routine diagnosis; a project called MYO-SEQ. The diagnostic rate has been 54%, being within the values observed in other studies. The aim of our project has focused on deepening data analysis and better understanding the lack of diagnosis in the rest of the patient cohort, establishing different explanatory hypotheses: digenic inheritance, presence of intronic variants, genetic modifiers and functional work of new candidate genes using Danio rerio as a model organism. For this, we have analyzed the exome sequencing data from the Seqr platform, while integrating the clinical information of the reference physicians of each patient obtained from the PhenoTips software, which allows to reevaluate the clinical, histopathological and magnetic resonance imaging reports. Our findings suggest the need to deepen the analysis in undiagnosed patients using other sequencing techniques to elucidate the possible mechanisms involved in LGMD, to ensure in the future a more accurate diagnosis and, therefore, a genetic counseling and treatment that improves the quality of life of these patients.Casasús Tomás, AB. (2019). Incremento del diagnóstico mas allá del análisis estándar del exoma en una cohorte de pacientes con LGMD. http://hdl.handle.net/10251/129808TFG

Unravelling opportunities, synergies, and barriers for enhancing silvopastoralism in the Mediterranean

Silvopastoral systems combine wood perennials with forage and livestock. These multipurpose wood-pasture habitats represent an important part of European bio-cultural and ecological heritage. However, their gradual disappearance due to processes of farm abandonment and intensification as well as forestry abandonment threatens biodiversity conservation and bio-cultural heritages. The behaviours of forest owners and livestock farmers determine the success of silvopastoral systems since a productive coherence between forest management and livestock grazing is required for their optimal functioning. In this study, we investigate the livestock farmers’ and forest owners’ attitudes and opinions towards wood pasture grazing and their relationship with structural factors and farming objectives in two Spanish regions. We used data collected through surveys to identify opportunities, synergies, and barriers in the integration of these actors in joint silvopastoralism. The results reveal a relationship between production objectives and positive attitudes towards silvopastoralism and the environmental functions provided by this activity, in both farmers and forest owners. Cattle farmers express a greater economic interest in wood pastures, as compared to sheep farmers who perceive more difficulties in using them. Acknowledgement of the role of grazing in landscape maintenance is positively correlated with a wide spectrum of objectives of forest owners, from the economic-productivist to the more altruistic profiles. The most synergies are found between cattle farmers and small forest owners for joint silvopastoral management. However, the incorporation of sheep grazing is advisable for the sustainable management of these systems, and hence additional efforts may be required to integrate such farming systems into silvopastoral management. As it currently stands, the EU Common Agricultural Policy appears to be inadequate for maintaining silvopastoral systems.info:eu-repo/semantics/publishedVersio

Unravelling opportunities, synergies, and barriers for enhancing silvopastoralism in the Mediterranean

Silvopastoral systems combine wood perennials with forage and livestock. These multipurpose wood-pasture habitats represent an important part of European bio-cultural and ecological heritage. However, their gradual disappearance due to processes of farm abandonment and intensification as well as forestry abandonment threatens biodiversity conservation and bio-cultural heritages. The behaviours of forest owners and livestock farmers determine the success of silvopastoral systems since a productive coherence between forest management and livestock grazing is required for their optimal functioning. In this study, we investigate the livestock farmers’ and forest owners’ attitudes and opinions towards wood pasture grazing and their relationship with structural factors and farming objectives in two Spanish regions. We used data collected through surveys to identify opportunities, synergies, and barriers in the integration of these actors in joint silvopastoralism. The results reveal a relationship between production objectives and positive attitudes towards silvopastoralism and the environmental functions provided by this activity, in both farmers and forest owners. Cattle farmers express a greater economic interest in wood pastures, as compared to sheep farmers who perceive more difficulties in using them. Acknowledgement of the role of grazing in landscape maintenance is positively correlated with a wide spectrum of objectives of forest owners, from the economic-productivist to the more altruistic profiles. The most synergies are found between cattle farmers and small forest owners for joint silvopastoral management. However, the incorporation of sheep grazing is advisable for the sustainable management of these systems, and hence additional efforts may be required to integrate such farming systems into silvopastoral management. As it currently stands, the EU Common Agricultural Policy appears to be inadequate for maintaining silvopastoral systems.The research leading to these results received funding from the Instituto Nacional de Investigacion y Tecnología Agraria y Alimentaria (INIA) (Spain) under Grant Agreement No RTA-2017-00036-C01-0

Opciones para la producción de corderos ligeros

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Efecto de las prácticas forestales y ganaderas en la provisión de bienes y servicios de los sistemas silvopastorales de la media montaña mediterránea, una evaluación aplicando el método Delphi

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Drivers of change in mountain agriculture: A thirty-year analysis of trajectories of evolution of cattle farming systems in the Spanish Pyrenees

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Amidase-responsive controlled release of antitumoral drug into intracellular media using gluconamide-capped mesoporous silica nanoparticles

MCM-41 silica nanoparticles were used as inorganic scaffolding to prepare a nanoscopic-capped hybrid material S1, which was able to release an entrapped cargo in the presence of certain enzymes, whereas in the absence of enzymes, a zero release system was obtained. S1 was prepared by loading nanoparticles with Safranine O dye and was then capped with a gluconamide derivative. In the absence of enzymes, the release of the dye from the aqueous suspensions of S1 was inhibited as a result of the steric hindrance imposed by the bulky gluconamide derivative, the polymerized gluconamide layer and the formation of a dense hydrogen-bonded network around the pore outlets. Upon the addition of amidase and pronase enzymes, delivery of Safranine O dye was observed due to the enzymatic hydrolysis of the amide bond in the anchored gluconamide derivative. S1 nanoparticles were not toxic for cells, as demonstrated by cell viability assays using HeLa and MCF-7 cell lines, and were associated with lysosomes, as shown by confocal microscopy. Finally, the S1¿CPT material loaded with the cytotoxic drug camptothecin and capped with the gluconamide derivative was prepared. The HeLa cells treated with S1¿CPT underwent cell death as a result of material internalization, and of the subsequent cellular enzyme-mediated hydrolysis and aperture of the molecular gate, which induced the release of the camptothecin cargo.We thank the Spanish Government (Project MAT2009-14564-C04 and SAF2010-15512) and the Generalitat Valenciana (Project PROMETEO/2009/016and/2010/005) for support. I. C. thanks the Universitat Politecnica de Valencia for her fellowship. L. M. thanks the Generalitat Valenciana for her post-doctoral VALi+d contract. E. A. and C. T. also thank the CIBER-BBN for contracts. We thank Eva Maria Lafuente Villarreal and Alberto Hernandez Cano from the Confocal Microscopy service of CIPF and the Electronic Microscopy service of UPV for their technical support.Candel Busquets, I.; Aznar Gimeno, E.; Mondragón Martínez, L.; De La Torre Paredes, C.; Martínez Mañez, R.; Sancenón Galarza, F.; Marcos Martínez, MD.... (2012). Amidase-responsive controlled release of antitumoral drug into intracellular media using gluconamide-capped mesoporous silica nanoparticles. Nanoscale. 4(22):7237-7245. https://doi.org/10.1039/c2nr32062bS7237724542

Dystrophinopathy Phenotypes and Modifying Factors in Exon 45-55 Deletion

Duchenne muscular dystrophy (DMD) exon 45-55 deletion (del45-55) has been postulated as a model that could treat up to 60% of DMD patients, but the associated clinical variability and complications require clarification. We aimed to understand the phenotypes and potential modifying factors of this dystrophinopathy subset. This cross-sectional, multicenter cohort study applied clinical and functional evaluation. Next generation sequencing was employed to identify intronic breakpoints and their impact on the Dp140 promotor, intronic long noncoding RNA, and regulatory splicing sequences. DMD modifiers (SPP1, LTBP4, ACTN3) and concomitant mutations were also assessed. Haplotypes were built using DMD single nucleotide polymorphisms. Dystrophin expression was evaluated via immunostaining, Western blotting, reverse transcription polymerase chain reaction (PCR), and droplet digital PCR in 9 muscle biopsies. The series comprised 57 subjects (23 index) expressing Becker phenotype (28%), isolated cardiopathy (19%), and asymptomatic features (53%). Cognitive impairment occurred in 90% of children. Patients were classified according to 10 distinct index-case breakpoints; 4 of them were recurrent due to founder events. A specific breakpoint (D5) was associated with severity, but no significant effect was appreciated due to the changes in intronic sequences. All biopsies showed dystrophin expression of >67% and traces of alternative del45-57 transcript that were not deemed pathogenically relevant. Only the LTBP4 haplotype appeared associated the presence of cardiopathy among the explored extragenic factors. We confirmed that del45-55 segregates a high proportion of benign phenotypes, severe cases, and isolated cardiac and cognitive presentations. Although some influence of the intronic breakpoint position and the LTBP4 modifier may exist, the pathomechanisms responsible for the phenotypic variability remain largely unresolved. ANN NEUROL 2022;92:793-80

Modulation of folic acid bioaccessibility by encapsulation in pH-responsive gated mesoporous silica particles

[EN] A study on the controlled release of folic acid (FA) from pH-responsive gated mesoporous silica particles (MSP) is reported. The MCM-41 support was synthesized using tetraethyl orthosilicate (TEOS) as hydrolytic inorganic precursor and the surfactant hexadecyltrimethylammonium bromide (CTAB) as porogen species. Calcination of the mesostructured phase resulted in the starting solid. This solid was loaded with FA to obtain the initial support S0. Moreover, this FA-loaded material was further functionalized with 3-[2-(2-aminoethylamino)ethylamino]propyltrimethoxysilane (N3) in order to obtain the gated polyamine-functionalised material S1. Solids S0 and S1 were characterized using standard solid state procedures. It was found that the functionalization process and the inclusion of FA on the pores did not modify the mesoporous structure of the starting material. FA delivery studies in water with solids S0 and S1 were carried out in water at pH 2 and 7.5. S0 was not able to completely inhibit FA delivery at acidic pH yet a rapid FA release at neutral pH was observed in few minutes. In contrast, S1 was tightly capped at pH 2 and displayed a sustained delivery of FA when the pH was switched to 7.5. In the second part of the study, FA loading and functionalization of S1-like supports was optimized. In particular, solids loaded with FA in phosphate buffered saline (PBS) and capped with N3 in acetate buffer at pH 2 exhibited a delivery capacity up to 95 μg FA/mg solid. Finally, FA release from the selected optimized supports was studied following an in vitro digestion procedure. The results showed that amine-capped MSP were not only able to hinder the release of the vitamin in gastric fluids (pH 2), but were also capable of deliver progressively the FA in presence of a simulated intestinal juice (pH 7.5) offering a suitable mechanism to control the bioaccessibility of the vitaminAuthors gratefully acknowledge the financial support from the Ministerio de Economia y Competitividad (Projects AGL201239597-C02 and MAT2012-38429-004-01) and the Generalitat Valenciana (project PROMETEO/2009/016). E.P. is grateful to the Ministerio de Ciencia e Innovacion for his Grant (AP2008-00620). C.C. thanks the Generalitat Valenciana for her post-doctoral contract VALi+D.Pérez-Esteve, É.; Fuentes López, A.; Coll Merino, MC.; Acosta, C.; Bernardos Bau, A.; Amoros Del Toro, PJ.; Marcos Martínez, MD.... (2015). Modulation of folic acid bioaccessibility by encapsulation in pH-responsive gated mesoporous silica particles. Microporous and Mesoporous Materials. 202:124-132. https://doi.org/10.1016/j.micromeso.2014.09.049S12413220

Enrichment of stirred yogurts with folic acid encapsulated in pH-responsive mesoporous silica particles: Bioaccessibility modulation and physico-chemical characterization

[EN] In this work, we have studied the ability of a mesoporous silica support loaded with folic acid and functionalized with amines (S1) to modulate the bioaccessibility of the vitamin after its incorporation in stirred yoghurts with different fat contents. Due to the novelty of using mesoporous silica supports in food matrixes, the influence of S1 addition on the physicochemical, rheological and lactic acid bacteria viability of these yoghurts during 21 days of refrigerated storage at 4 °C was also evaluated. The in vitro digestion procedure showed that S1 was capable of inhibiting the release of folic acid in acidic solution at pH 2 (stomach) and controllably release their contents in neutral pH (intestine), thereby modulating the bioaccessibility. Moreover, the physicochemical and microbiological assays revealed that enrichment generally does not alter the physicochemical properties (pH, colour, syneresis and rheology) of either type of yoghurt and does not cause any effect on lactic acid bacteria survival.Authors gratefully acknowledge the financial support from the Ministerio de Economia y Competitividad (Projects AGL2012-39597-C02-01, AGL2012-39597-C02-02 and MAT2012-38429-C04-01) and the Generalitat Valenciana (project PROMETEO/2009/016). E.P.E. and M.R.R. are grateful to the Ministerio de Ciencia e Innovacion for their grants (AP2008-00620, AP2010-4369). Electron Microscopy Service of the UPV is also acknowledged.Pérez-Esteve, É.; Ruiz Rico, M.; Fuentes López, A.; Marcos Martínez, MD.; Sancenón Galarza, F.; Martínez-Máñez, R.; Barat Baviera, JM. (2016). Enrichment of stirred yogurts with folic acid encapsulated in pH-responsive mesoporous silica particles: Bioaccessibility modulation and physico-chemical characterization. Food Science and Technology. (72):351-360. https://doi.org/10.1016/j.lwt.2016.04.061S3513607