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Objective The surgical management of nasal septal defects due to perforations, malformations, congenital cartilage absence, traumatic defects, or tumors would benefit from availability of optimally matured septal cartilage substitutes. Here, we aimed to improve in vitro maturation of 3-dimensional (3D)-printed, cell-laden polycaprolactone (PCL)-based scaffolds and test their in vivo performance in a rabbit auricular cartilage model. Design Rabbit auricular chondrocytes were isolated, cultured, and seeded on 3D-printed PCL scaffolds. The scaffolds were cultured for 21 days in vitro under standard culture media and normoxia or in prochondrogenic and hypoxia conditions, respectively. Cell-laden scaffolds (as well as acellular controls) were implanted into perichondrium pockets of New Zealand white rabbit ears ( N = 5 per group) and followed up for 12 weeks. At study end point, the tissue-engineered scaffolds were extracted and tested by histological, immunohistochemical, mechanical, and biochemical assays. Results Scaffolds previously matured in vitro under prochondrogenic hypoxic conditions showed superior mechanical properties as well as improved patterns of cartilage matrix deposition, chondrogenic gene expression ( COL1A1, COL2A1, ACAN, SOX9, COL10A1), and proteoglycan production in vivo, compared with scaffolds cultured in standard conditions. Conclusions In vitro maturation of engineered cartilage scaffolds under prochondrogenic conditions that better mimic the in vivo environment may be beneficial to improve functional properties of the engineered grafts. The proposed maturation strategy may also be of use for other tissue-engineered constructs and may ultimately impact survival and integration of the grafts in the damaged tissue microenvironment.

Innovative methodologies and new emerging technologies have significant potential for education. Therefore, their use is becoming more and more common in the educational society. This research analyses the integration of gamification through ICT in natural sciences in primary education to improve environmental education. Two didactic units of renewable energies are implemented through a control group which uses a traditional learning method and an experimental group which uses the gamification method and ICT as an educational resource. By means of a mixed methodology, data from 36 students of 6th grade of primary school are analysed, applying the Mann Whitney U-test, a questionnaire and systematic observation. It is highlighted that the work and integration of the gamification methodology, through ICT, does not significantly improve academic performance. However, statistically significant improvements are stressed in satisfaction and motivation in learning processes. The results underline the need for teachers to receive technological and pedagogical training to improve the use of technology in their classrooms.

En la presente investigación se analiza la integración del método de aprendizaje basado en proyectos, a través de las TIC como método emergente con impacto significativo en el aprendizaje. Se implementan dos unidades didácticas de la higiene de las manos a través de un método tradicional y un método de aprendizaje basado en proyectos y TIC como recurso educativo. Mediante un diseño cuasi experimental se analizan los datos de 39 alumnos de 2º de primaria, aplicando el test U de Mann Whitney, un cuestionario y la observación participante. Se resalta que el trabajo e integración de la metodología de aprendizaje basado en proyectos, a través de las TIC no mejora significativamente en el rendimiento académico. Sin embargo, se subrayan mejoras estadísticamente significativas en el reconocimiento de los contenidos, la participación activa, la colaboración, satisfacción y la motivación en los procesos de aprendizaje. A través de este método de aprendizaje y la utilización de las TIC, los estudiantes investigan de primera mano y reflexionan sobre el tema, realizando un aprendizaje significativo.

Autophagy is a mechanism responsible for the degradation of cellular components to maintain their homeostasis. However, autophagy is commonly altered and compromised in several diseases, including neurodegenerative disorders. Parkinson’s disease (PD) can be considered a multifactorial disease because environmental factors, genetic factors, and aging are involved. Several genes are involved in PD pathology, among which the LRRK2 gene and its mutations, inherited in an autosomal dominant manner, are responsible for most genetic PD cases. The R1441G LRRK2 mutation is, after G2019S, the most important in PD pathogenesis. Our results demonstrate a relationship between the R1441G LRRK2 mutation and a mechanistic dysregulation of autophagy that compromises cell viability. This altered autophagy mechanism is associated with organellar stress including mitochondrial (which induces mitophagy) and endoplasmic reticulum (ER) stress, consistent with the fact that patients with this mutation are more vulnerable to toxins related to PD, such as MPP+.

The effect of vanadia loading on the acidic and redox properties of VOx/TiO2 catalyst and their role in the simultaneous reduction of NO with NH3 and oxidation of o-DCB is studied. Catalyst samples with different proportions of VOx species, ranging from monomerics to V2O5 octahedral crystals have been prepared. A relationship between VOx species and their acidic and redox properties was found. At low temperature, o-DCB is adsorbed mainly on Lewis sites, associated to monomeric species, but at high temperature, o-DCB and NH3 compete for Bronsted sites associated to polymeric species.

Resumen basado en el de la publicación Resumen en inglés Se presenta un estudio que tiene como objetivo analizar la autopercepción de la competencia digital del alumnado en el grado en Educación Social de la Universidad de Deusto. Para ello, se utiliza el test Ikanos. En el estudio participaron 266 estudiantes de tres cursos. Los resultados indican una autopercepción baja de la competencia digital. Los ítems con menor puntuación se corresponden con las competencias de innovación, seguridad de los datos personales y privacidad, y programación. Sin embargo, las competencias de almacenamiento y recuperación de la información, e interactuar son las puntuaciones medias más altas ESP

The research work conducted by Orue et al. [1] aims to analyze the impact of protective capacity on the application of the third step of the drum-buffer-rope methodology of the theory of constraints (subordinate everything to the bottleneck) in make-to-order environments. To analyze the impact of protective capacity, a discrete event simulation is performed. Experimental results suggest that, at different levels of variability, different levels of protective capacity are needed to keep the system stable. The higher the variability, the higher the level of protection needed. It can be concluded that the definition of the level of protective capacity required is a strategic decision for the organization. For managers to know and control how much protection is necessary and how much is overcapacity or waste, monitoring and managing the use of protective capacity is paramount. This concept of protection monitoring and control is relatively new in the theory of constraints methodology, but it is applied in the demand-driven adaptive enterprise model [2]. Therefore, it is advisable to manage capacity buffers as is done in the methodology. Keywords: theory of constraints, drum-buffer-rope, protective capacity, make-to-order, capacity buffer, demand-driven adaptive enterprise

This study uses simulation to analyse the impact of protective capacity in the implementation of the step 3 of the theory of constraints (TOC) methodology in make-to-order (MTO) environments. The results suggest that at different levels of variability, different levels of protective capacity are needed to keep a system stable and meet the bottleneck (BN) schedule. The higher the variability is, the higher the level of protection needed. The definition of the level of protective capacity required is a strategic decision for the organisation, as it must be emphasised that with high levels of variability in a system, the protective capacity must be very high, which contradicts the efficient use of resources. However, for managers to know and control how much protection is needed without needing additional capacity, it is advisable to monitor the use of protective capacity. This concept of protection monitoring and control is relatively new in the TOC–drum-buffer-rope (TOC-DBR) methodology, but it is applied in the demand-driven adaptive business model, and therefore, should be integrated into the TOC-DBR methodology.

19 páginas, 4 figuras, 2 tablas; material suplementario con 3 figuras. 2 tablas de material suplementario en: http://dx.doi.org/10.18632/aging.103992, The research of new biomarkers for Parkinson's disease is essential for accurate and precocious diagnosis, as well as for the discovery of new potential disease mechanisms and drug targets. The main objective of this work was to identify metabolic changes that might serve as biomarkers for the diagnosis of this neurodegenerative disorder. For this, we profiled the plasma metabolome from mice with neurotoxin-induced Parkinson's disease as well as from patients with familial or sporadic Parkinson's disease. By using mass spectrometry technology, we analyzed the complete metabolome from healthy volunteers compared to patients with idiopathic or familial (carrying the G2019S or R1441G mutations in the LRRK2 gene) Parkinson's disease, as well as, from mice treated with 6-hydroxydopamine to induce Parkinson disease. Both human and murine Parkinson was accompanied by an increase in plasma levels of unconjugated bile acids (cholic acid, deoxycholic acid and lithocholic acid) and purine base intermediary metabolites, in particular hypoxanthine. The comprehensive metabolomic analysis of plasma from Parkinsonian patients underscores the importance of bile acids and purine metabolism in the pathophysiology of this disease. Therefore, plasma measurements of certain metabolites related to these pathways might contribute to the diagnosis of Parkinson's Disease., This research was supported by “Instituto de Salud Carlos III”, “Fondo de Investigaciones Sanitarias” (PI15/0034), “CIBERNED” (CB06/05/0041 and 2015/03), “MINECO” (SAF2014-52940-R and SAF2017-85199-P) and partially supported by “European Regional Development Fund (ERDF)” from the European Union. SMSY-D was supported by “Isabel Gemio Foundation”. JMB-SP and MN-S were funded by “Ramon y Cajal Program” (RYC-2018-025099-I) and (RYC-2016-20883), respectively. GM-C was supported by “ONCE Foundation”. EU-C is supported by an FPU predoctoral fellowship (FPU16/00684) from “Ministerio de Educación, Cultura y Deporte”. JMF received research support from the “Instituto de Salud Carlos III”; “Fondo de Investigaciones Sanitarias” (PI15/0034) and CIBERNED (CB06/05/0041 and 2015/03). AP-C was supported by MINECO (SAF2014-52940-R and SAF2017-85199- P). JR-M and MZ were supported by Institute de Salud Carlos III, Fondo de Investigaciones Sanitarias (PI19/01653), CIBERNED and PPMI Initiative. JP-T received funding from CIBERNED (CB06/05/1123 and 2015/03). This work was also partially supported by “Fondo Europeo de Desarrollo Regional” (FEDER) from the European Union. GK is supported by the Ligue contre le Cancer (équipe labellisée); Agence National de la Recherche (ANR) – Projets blancs; ANR under the frame of E-Rare-2, the ERA-Net for Research on Rare Diseases; AMMICa US23/CNRS UMS3655; Association pour la recherche sur le cancer (ARC); Association “Le Cancer du Sein, Parlons-en!”; Cancéropôle Ile-de-France; Chancelerie des universités de Paris (Legs Poix), Fondation pour la Recherche Médicale (FRM); a donation by Elior; European Research Area Network on Cardiovascular Diseases (ERA-CVD, MINOTAUR); Gustave Roussy Odyssea, the European Union Horizon 2020 Project Oncobiome; Fondation Carrefour; High-end Foreign Expert Program in China (GDW20171100085), Institut National du Cancer (INCa); Inserm (HTE); Institut Universitaire de France; LeDucq Foundation; the LabEx Immuno-Oncology (ANR-18-IDEX-0001); the RHU Torino Lumière; the Seerave Foundation; the SIRIC Stratified Oncology Cell DNA Repair and Tumor Immune Elimination (SOCRATE); and the SIRIC Cancer Research and Personalized Medicine (CARPEM).

Dichloromethane (CH2Cl2) hydrodechlorination to chloromethane (CH3Cl) is a pivotal step toward the successful implementation of chlorine-mediated methane-to-liquid technologies. This study investig...

Uno de los principales elementos de la teoria de las limitaciones (TOC) es su metodo de gestion de la produccion Drum-Buffer-Rope (DBR), que controla el lanzamiento del trabajo al sistema. La secuencia en la que se lanzan las ordenes de produccion esta determinada por la programacion del Cuello de Botella (Drum). Si bien la literatura reconoce la validez del metodo, la mayoria de los trabajos se centra en estudios analiticos que no tienen en cuenta casos reales de produccion bajo pedido en plantas de fabricacion versatiles. En este estudio exponemos un proceso sistematico para la implementacion de los dos primeros pasos de TOC en sistemas productivos. Lo hacemos desde una perspectiva estrategica, sosteniendo que el recurso que controla el lanzamiento y los ritmos de produccion debe ser el resultado de una decision gerencial.

Limb-girdle muscular dystrophy recessive 1 (LGMDR1) represents one of the most common types of LGMD in the population, where patients develop a progressive muscle degeneration. The disease is caused by mutations in calpain 3 gene, with over 500 mutations reported to date. However, the molecular events that lead to muscle wasting are not clear, nor the reasons for the great clinical variability among patients, and this has so far hindered the development of effective therapies. Here we generate human induced pluripotent stem cells (iPSCs) from skin fibroblasts of 2 healthy controls and 4 LGMDR1 patients with different mutations. The generated lines were able to differentiate into myogenic progenitors and myotubes in vitro and in vivo, upon a transient PAX7 overexpressing protocol. Thus, we have generated myogenic cellular models of LGMDR1 that harbor different CAPN3 mutations within a human genetic background, and which do not derive from muscular biopsies. These models will allow us to investigate disease mechanisms and test therapies. Despite the variability found among iPSC lines that was unrelated to CAPN3 mutations, we found that patient-derived myogenic progenitors and myotubes express lower levels of DMD, which codes a key protein in satellite cell regulation and myotube maturation. This work has been funded by grants from Ilundain Foundation, Isabel Gemio Foundation, Fundació La Caixa, Basque Government (2015111038), Catalan Government (2017-SGR-899 and CERCA Programme), Provincial Council of Gipuzkoa (A.LdM 114/17), and Instituto de Salud Carlos III (PI14/00436, PS09/00660 and RD16/0011/0024). A.M.-A and N.N.-G. received a studentship from the Department of Education, University and Research of the Basque Government (BFI-2012-19, PRE2013-1-1168)

BACKGROUND: In recent years, three-dimensional (3D)-printing of tissue-engineered cartilaginous scaffolds is intended to close the surgical gap and provide bio-printed tissue designed to fit the specific geometric and functional requirements of each cartilage defect, avoiding donor site morbidity and offering a personalizing therapy. METHODS: To investigate the role of 3D—bioprinting scaffolding for nasal cartilage defects repair a systematic review of the electronic databases for 3D-Bioprinting articles pertaining to nasal cartilage bio-modelling was performed. The primary focus was to investigate cellular source, type of scaffold utilization, biochemical evaluation, histological analysis, in-vitro study, in-vivo study, animal model used, length of research, and placement of experimental construct and translational investigation. RESULTS: From 1011 publications, 16 studies were kept for analysis. About cellular sources described, most studies used primary chondrocyte cultures. The cartilage used for cell isolation was mostly nasal septum. The most common biomaterial used for scaffold creation was polycaprolactone alone or in combination. About mechanical evaluation, we found a high heterogeneity, making it difficult to extract any solid conclusion. Regarding biological and histological characteristics of each scaffold, we found that the expression of collagen type I, collagen Type II and other ECM components were the most common patterns evaluated through immunohistochemistry on in-vitro and in-vivo studies. Only two studies made an orthotopic placement of the scaffolds. However, in none of the studies analyzed, the scaffold was placed in a subperichondrial pocket to rigorously simulate the cartilage environment. In contrast, scaffolds were implanted in a subcutaneous plane in almost all of the studies included. CONCLUSION: The role of 3D—bioprinting scaffolding for nasal cartilage defects repair is growing field. Despite the amount of information collected in the last years and the first surgical applications described recently in humans. Further investigations are needed due to the heterogeneity on mechanical evaluation parameters, the high level of heterotopic scaffold implantation and the need for quantitative histological data.

Several studies and publications support the idea that it is critical for companies to manage projects successfully, thus gaining a competitive edge. The effort made in the last decades to develop project management has led to a dramatic progress of this discipline. As a result, the theories, know-how, means and capabilities available today are superior by far to those available a few years ago. However, it has also been concluded that projects continue to struggle to achieve their objectives. Furthermore, some authors consider that the respective approach in itself is one of the main causes for this situation. This study is a real-world based comparison of two approaches, applied to the same project. In particular, the regular approach of the company, based on a traditional project management perspective, has been confronted with a holistic perspective. The results suggest that aspects which are generally considered to be adequate can be counterproductive.

In certain project-based contexts, the most effective managerial approach may be different from a “pure” Project Management approach. Thus, under certain circumstances, a Production Management approach may be more suitable. This research offers some important insights into the problem of addressing contexts in which both, Project and Production Management characteristics, coexist. This paper presents a real-world case study in an industrial company devoted to designing and manufacturing special cutting tools. Depending on the items in the orders, different lead times, resources, and batch sizes occur, leading to different levels of complexity. Production Management approaches are preferred for standard items, whereas complex items are better managed by applying a Project Management approach. In an attempt to maximize effectiveness, the management followed the suggestion to apply the DBR production approach in order to harmonize the capacity of the plant and the requirements of the orders. Thus, this research deals with the implementation of this methodology in the abovementioned environment. The consequences, limitations, and conclusions are discussed, offering insights for further research.

Change and high uncertainty levels are the main characteristics of current project contexts. Years ago, the traditional project management faced problems when operating in these environments. Thus, at the end of the 20th century, new project management approaches were conceived to provide a more effective answer to such contexts. These methods propose a different approach, aimed at promoting the project flow by focusing on the short term. However, their adoption involves certain adjustments from a managerial perspective. The influence on the way resources are used is of special interest, as it may cause unexpected behaviors and reactions. The literature vastly analyzes the features and benefits of these methods. Nevertheless, there is a lack of empirical evidence about both the practical implications of the transition process toward them and their superior performance. Thus, this research aims to contribute to filling this gap by providing real-world based evidence related to the change process from a traditional project management approach to a flow-driven one. With this objective in mind, we analyzed the transition processes experienced by two design departments from companies of different industries. The results of the study confirm that the adoption of this type of approach can improve an organization’s performance and simplify its project management system. Therefore, we consider that our findings are useful for anybody interested in these methodologies. From the academic perspective, the evidence obtained in this study contributes to supporting the research works suggested by the literature. Furthermore, it can be helpful to guide further research and extend current knowledge. Additionally, they can assist companies in the improvement of their current project management approaches.

Drum-buffer-rope (DBR) is a key element of Goldratt’s theory of constraints (TOC) in which the launch of production orders into the system is controlled according to the schedule of the system’s bottleneck. The existing literature has discussed the validity of TOC-DBR as a production planning and control system in make-to-order (MTO) environments, but most studies have taken an analytical approach; real-world empirical cases are scarce. In fact, the literature has recognised that any implementation of production planning and control systems in MTO environments is complex to manage, and this could also be the main cause of the lack of studies on this subject. Additionally, a literature review showed a lack of understanding regarding selecting and exploiting the bottleneck (a crucial part of DBR) in these MTO manufacturing contexts. Therefore, the researcher aimed to provide the following novel guidelines for MTO environments: 1) How to select the bottleneck: The research provided a set of four criteria inspired by the resource-based view and Barney’s VRIO concept. 2) How to exploit the bottleneck: Three operative sub-steps to exploit the bottleneck were included; these were partly inspired by a strategic perspective based on the practice-based view. The selection and exploitation of the bottleneck was included in a systematic decision-making process that enabled the operationalisation of the first two steps of Goldratt’s DBR. This systematic process was developed and tested on four real-world cases using action research (as well as some aspects of constructive research). Given the practical origin of this research, action research was an appropriate methodology since it enabled contributions to academic research while solving real-world problems. The research provided several theoretical contributions (e.g., how to select and exploit the bottleneck in MTO environments) as well as practical contributions (i.e., a systematic process for operationalising the first two steps of TOC-DBR and determining how the company performance could be impacted). Additional (non-expected) contributions also emerged from the research. Among others, a key contribution was related to the different management required by the two types of MTO environments, as follows: • Repeat-business customizers (medium product variety and medium volume production): The research showed that exploiting the bottleneck could be a source of better performance; these findings aligned with the practice-based view. • Versatile manufacturing companies (high product variety and low volume production): The research showed that approaches based on the resource-based view and practice-based view could improve firms’ performance. The practice-based view applied to the exploitation of the bottleneck, while the selection of the bottleneck could be determined using the resource-based view’s VRIO concept (especially concerning resources that were rare and inimitable). Drum-buffer-rope (DBR) Goldratten murrizketen teoriaren (Theory of Constraints-TOC) funtsezko elementu bat da. DBR-ren arabera, botila-lepoaren programak kontrolatzen du ekoizpen-aginduak abiaraztea. Egungo literaturan jadanik aztertuta dago TOC-DBRren baliotasuna eskaeraren araberako testuinguruetan (Make to Order-MTO), hau da, ekoizpen ingurunea planifikatzeko eta kontrolatzeko sistema gisa balio ote duen. Egindako azterketa gehienek, ordea, ikuspegi analitikoa hartu dute, mundu errealeko kasu enpiriko gutxi daudelarik. Are gehiago, literaturak onartu du MTO inguruneetan ekoizpena planifikatzeko eta kontrolatzeko sistemak ezartzea zaila dela, eta hori izan liteke, hain zuzen, azterketa enpirikorik ez egotearen arrazoi nagusia. Bestalde, MTO inguruneen gainean egindako literaturaren berrikuspenak erakutsi du botila-lepoaren hautaketa eta ustiapenari buruz nahikoa ikerketarik ez dagoela, hori DBR kudeaketaren funtsezko zati bat izanik. Beraz, ikertzaileak MTO inguruneetarako jarraibide berri hauek ematea izan du helburu: 1) Botila-lepoa nola aukeratu: ikerketak lau irizpide eman ditu botila-lepoa aukeratzeko. Irizpide horiek baliabideetan oinarritutako ikuspegiarekin (Resource Based View-RBV) bat datoz, eta bereziki Barneyren VRIO kontzeptuarekin. 2) Botila-lepoa nola ustiatu: botila-lepoa ustiatzeko hiru urrats operatibo garatu dira, praktikan oinarritutako ikuspegi estrategikoan finkaturik (Practice Based View-PBV). Lanaren emaitza botila-lepoaren aukeraketa eta ustiapena gauzatzeko prozesu sistematiko bat da. Goldratten DBR metodoaren lehen bi urratsak operazionalizatzea ahalbidetzen du prozesu horrek, eta lau kasu errealen bitartez garatu eta probatu da, ekintza bidezko ikerketa (Action Research-AR) eta ikerketa konstruktiboaren alderdi batzuk baliatuz. AR metodologiak mundu errealeko arazoak konpontzen dituen bitartean ikerketa akademikoari ekarpenak egiteko aukera ematen du. Ondorioz, izaera praktikoa duten ikerketetarako egokia da. Ikerketa horrek zenbait ekarpen teoriko eman ditu (adibidez, nola hautatu eta ustiatu botila-lepoa MTO inguruneetan), baita ekarpen praktiko batzuk ere (hau da, TOC-DBR-ren lehenengo bi urratsak aplikatzeko prozesu sistematikoa, eta enpresaren emaitzetan zer nolako eragina duen erakutsi). Azkenik, espero ez ziren zenbait ekarpen gehigarri ere erdietsi dira. Horietako batek berebiziko garrantzia dauka, eta bi MTO ingurune mota desberdinentzat identifikatutako kudeaketa bereiztua da, zehazki:  Produktu errepikakorren enpresa (Repeat-business customizers; produktu aniztasun ertaina eta bolumen ertainen ekoizpena): ikerketaren arabera, botila-lepoa ustiatzea errendimendu hobeko iturria izan daiteke; emaitza horiek bat etorri dira praktikan oinarritutako ikuspegiarekin (PBV).  Fabrikazio-enpresa aldakorrak (Versatile manufacturing companies; produktu-aukera handia eta bolumen txikiko ekoizpena): ikerketak erakutsi duenez, baliabideetan oinarritutako ikuspegiak (RBV) eta praktikan oinarritutako ikuspegiak (PBV) enpresen errendimendua hobetu dezakete. Botila-lepoa hautatzeko baliabideetan oinarritutako ikuspegiaren VRIO kontzeptua erabil daiteke (bereziki, baliabide arraroak eta imitaezinak kontzeptuak). Praktikan oinarritutako ikuspegiak (PBV), berriz, botila-lepoaren ustiapenari eragiten dio. Uno de los principales elementos de la teoría de las limitaciones (TOC) de Goldratt es su método de gestión de la producción Drum-Buffer-Rope (DBR), que controla el lanzamiento del trabajo al sistema. La secuencia en la que se lanzan las órdenes de producción está determinada por la programación del Cuello de Botella (Drum). La literatura existente ha discutido la validez de TOC-DBR como un sistema de planificación y control de la producción en un entorno de fabricación bajo pedido (MTO), pero la mayor parte de los trabajos tienen un enfoque analítico y los casos empíricos del mundo real son escasos. De hecho, la literatura ha reconocido que cualquier intento de poner en práctica sistemas de planificación y control de la producción en entornos MTO es complejo de gestionar y, por lo tanto, esta podría ser también la causa principal de la falta de estudios en la literatura. Además, la revisión de la literatura ha mostrado una falta de entendimiento respecto a cómo seleccionar y explotar el cuello de botella (una parte esencial de DBR) en contextos de fabricación MTO. Por lo tanto, el investigador ha desarrollado su trabajo para proporcionar los siguientes aspectos novedosos aplicados a los entornos MTO: 1) Cómo seleccionar el cuello de botella: nuestra investigación proporciona cuatro criterios para determinar la ubicación del cuello de botella del sistema. Estos criterios se inspiran en la perspectiva basada en los recursos (Resource Based View-RBV) y en el marco VRIO de Barney. 2) Cómo explotar el cuello de botella: hemos incluido tres sub-pasos como guía operativa para la explotación el cuello de botella, inspirados en parte en la perspectiva estratégica de la visión basada en la práctica (Practice Based View-PBV). La selección y explotación del cuello de botella se han incluido en un proceso sistemático de toma de decisiones que facilita la puesta en marcha de los dos primeros pasos del DBR de Goldratt. Este proceso sistemático ha sido desarrollado y probado en cuatro casos reales mediante la Investigación en Acción, combinada con ciertos aspectos de la Investigación Constructiva. Dada la naturaleza práctica de esta investigación, la Investigación en Acción es una metodología apropiada, ya que permite integrar la investigación académica y la resolución problemas en contextos reales. Como resultado de nuestra investigación se han realizado diversas contribuciones, tanto teóricas (por ejemplo, cómo seleccionar y explotar el cuello de botella en los entornos MTO) como prácticas (proceso sistemático para hacer operativa la aplicación de los dos primeros pasos del TOC-DBR o cómo impactan en los resultados operativos de la empresa). Como resultado de esta investigación también se han producido ciertas contribuciones inesperadas. Una de estas contribuciones, de especial interés, es la identificación de una gestión diferenciada para los dos tipos de escenarios MTO analizados: • Producción customizada repetitiva (Repeat-business customizers (RBC): variedad de productos medio y producción de volumen medio): La investigación demostró que la explotación del cuello de botella podría ser una fuente de mejor rendimiento; estos hallazgos coinciden con la PBV. • Empresas manufactureras versátiles (Versatile manufacturing companies (VMC): gran variedad de productos y producción de bajo volumen): La investigación ha demostrado que los enfoques RBV y PVB pueden mejorar el rendimiento de las empresas. Así, mientras la selección del cuello de botella puede determinarse utilizando el concepto VRIO de RBV (especialmente en lo que respecta al concepto de recursos que son “raros” e “inimitables”), la PBV se aplica a la explotación del cuello de botella.

The research of new biomarkers for Parkinson's disease is essential for accurate and precocious diagnosis, as well as for the discovery of new potential disease mechanisms and drug targets. The main objective of this work was to identify metabolic changes that might serve as biomarkers for the diagnosis of this neurodegenerative disorder. For this, we profiled the plasma metabolome from mice with neurotoxin-induced Parkinson's disease as well as from patients with familial or sporadic Parkinson's disease. By using mass spectrometry technology, we analyzed the complete metabolome from healthy volunteers compared to patients with idiopathic or familial (carrying the G2019S or R1441G mutations in the LRRK2 gene) Parkinson's disease, as well as, from mice treated with 6-hydroxydopamine to induce Parkinson disease. Both human and murine Parkinson was accompanied by an increase in plasma levels of unconjugated bile acids (cholic acid, deoxycholic acid and lithocholic acid) and purine base intermediary metabolites, in particular hypoxanthine. The comprehensive metabolomic analysis of plasma from Parkinsonian patients underscores the importance of bile acids and purine metabolism in the pathophysiology of this disease. Therefore, plasma measurements of certain metabolites related to these pathways might contribute to the diagnosis of Parkinson's Disease.

Uncertainty and change are two features of modern project management. They strongly influence the project management needs to operate in such contexts. This is the case when a complete and accurate definition of the scope of a project is not available. Those situations require a project management approach capable of dealing with the special conditions that characterise said contexts. This study focuses on the application of the progressive elaboration approach and the Critical Chain method. We analysed the implementation process of the new procedure in a company that produces capital goods for the automotive industry. The work’s main focus is on the e ect of this change from the multi-project perspective. We found that the change had a larger impact than was expected by the company. Firstly, we found that the new approach provided an opportunity to improve the performance of the company. Besides, the new approach uncovered significant problems that previously were ignored, as well as problems and obstacles to the change. Based on the results and findings of this work, we conclude that shifting to this kind of approach requires a global managerial perspective, and strong support from the management.

International Conference on Industrial Engineering and Industrial Management (13th. 2019. Gijón)

Parkinson's disease (PD) is a chronic and progressive neurodegenerative disorder. While most PD cases are idiopathic, the known genetic causes of PD are useful to understand common disease mechanisms. Recent data suggests that autophagy is regulated by protein acetylation mediated by histone acetyltransferase (HAT) and histone deacetylase (HDAC) activities. The changes in histone acetylation reported to be involved in PD pathogenesis have prompted this investigation of protein acetylation and HAT and HDAC activities in both idiopathic PD and G2019S leucine-rich repeat kinase 2 (LRRK2) cell cultures. Fibroblasts from PD patients (with or without the G2019S LRRK2 mutation) and control subjects were used to assess the different phenotypes between idiopathic and genetic PD. G2019S LRRK2 mutation displays increased mitophagy due to the activation of class III HDACs whereas idiopathic PD exhibits downregulation of clearance of defective mitochondria. This reduction of mitophagy is accompanied by more reactive oxygen species (ROS). In parallel, the acetylation protein levels of idiopathic and genetic individuals are different due to an upregulation in class I and II HDACs. Despite this upregulation, the total HDAC activity is decreased in idiopathic PD and the total HAT activity does not significantly vary. Mitophagy upregulation is beneficial for reducing the ROS-induced harm in genetic PD. The defective mitophagy in idiopathic PD is inherent to the decrease in class III HDACs. Thus, there is an imbalance between total HATs and HDACs activities in idiopathic PD, which increases cell death. The inhibition of HATs in idiopathic PD cells displays a cytoprotective effect.

Differential interactions of wildtype and phospho-mimetic Rab8a mutants with GDI1/2 and Rabin8, effects on centrosome splitting and subcellular localization. (DOCX 824 kb)

Distinct pathogenic LRRK2 mutants cause deficits in centrosome cohesion in transfected HEK293T cells. (DOCX 1160 kb)

Golgi dispersal/disruption has no effect on LRRK2-mediated pericentrosomal/centrosomal accumulation of Rab8a. (DOCX 1670 kb)

Background Mutations in LRRK2 are a common genetic cause of Parkinson’s disease (PD). LRRK2 interacts with and phosphorylates a subset of Rab proteins including Rab8a, a protein which has been implicated in various centrosome-related events. However, the cellular consequences of such phosphorylation remain elusive. Methods Human neuroblastoma SH-SY5Y cells stably expressing wildtype or pathogenic LRRK2 were used to test for polarity defects in the context of centrosomal positioning. Centrosomal cohesion deficits were analyzed from transiently transfected HEK293T cells, as well as from two distinct peripheral cell types derived from LRRK2-PD patients. Kinase assays, coimmunoprecipitation and GTP binding/retention assays were used to address Rab8a phosphorylation by LRRK2 and its effects in vitro. Transient transfections and siRNA experiments were performed to probe for the implication of Rab8a and its phosphorylated form in the centrosomal deficits caused by pathogenic LRRK2. Results Here, we show that pathogenic LRRK2 causes deficits in centrosomal positioning with effects on neurite outgrowth, cell polarization and directed migration. Pathogenic LRRK2 also causes deficits in centrosome cohesion which can be detected in peripheral cells derived from LRRK2-PD patients as compared to healthy controls, and which are reversed upon LRRK2 kinase inhibition. The centrosomal cohesion and polarity deficits can be mimicked when co-expressing wildtype LRRK2 with wildtype but not phospho-deficient Rab8a. The centrosomal defects induced by pathogenic LRRK2 are associated with a kinase activity-dependent increase in the centrosomal localization of phosphorylated Rab8a, and are prominently reduced upon RNAi of Rab8a. Conclusions Our findings reveal a new function of LRRK2 mediated by Rab8a phosphorylation and related to various centrosomal defects., S.H. was supported by the Michael J. Fox Foundation, the BBVA Foundation, FEDER, and the Spanish Ministry of Economy and Competitiveness (SAF2014–58653-R). E.G. was funded by the Michael J. Fox Foundation. J-M.T. and M.-C. C.-H. were funded by the Michael J. Fox Foundation, and M.-C.C.-H. was supported by Inserm, CHU de Lille, Lille University and the Ministère de la Recherche et de la Santé (PHRC Convergence). We gratefully acknowledge funding from the European Union’s Horizon 2020 research and innovation programme (Marie Sklodowska-Curie Action Individual Fellowship to J.-M.T.). A.C.N. was supported by the Dept. of the Army (USAMRAA W23RYX-9049-N610) and NIH (DA10044). This research was supported in part by the Intramural Research Program of the NIH, National Institute on Aging (to M.R.C.).

Golgi dispersal/disruption has no effect on LRRK2-mediated pericentrosomal/centrosomal accumulation of Rab8a. (DOCX 1670 kb)

Differential interactions of wildtype and phospho-mimetic Rab8a mutants with GDI1/2 and Rabin8, effects on centrosome splitting and subcellular localization. (DOCX 824 kb)

Pathogenic LRRK2 disturbs centrosome cohesion in a kinase-dependent manner. (DOCX 1155 kb)

Limb-girdle muscular dystrophy type 2A (LGMD2A) is characterised by muscle wasting and progressive degeneration of proximal muscles because of mutations in the CAPN3 gene. However, the underlying pathophysiological mechanisms of muscle degeneration are still not well understood. The objective of this study was to assess the relevance of genes with differential expression in the muscle of LGMD2A patients. For this purpose, we analysed their in vitro expression in primary cultures of human myoblasts and myotubes. Abnormal fusion was observed in the myotubes of these patients, which may be explained by the lack of physiological replacement of integrin β1D. Owing to this observation, we focused on deregulated genes coding proteins that directly interact with integrin, ITGB1BP2 and CD9, as well as FRZB gene, because of its in vitro upregulation in myotubes. Silencing studies established that these genes are closely regulated, CD9 and FRZB being positive regulators of the expression of ITGB1BP2, and in turn, this gene being a negative regulator of the expression of FRZB. Interestingly, we observed that FRZB regulates integrin β1D expression, its silencing increasing integrin β1D expression to levels similar to those in controls. Finally, the administration of LiCl, an enhancer of the Wnt-signalling pathway showed similar experimentally beneficial effects, suggesting FRZB silencing or LiCl administration as potential therapeutic targets, though further studies are required.

Parkinson's disease (PD) is a neurodegenerative disorder of unknown etiology. It is considered as a multifactorial disease dependent on environmental and genetic factors. Deregulation in cell degradation has been related with a significant increase in cell damage, becoming a target for studies on the PD etiology. In the present study, we have characterized the parkinsonian toxin 1-methyl-4-phenylpyridinium ion (MPP(+))-induced damage in fibroblasts from Parkinson's patients with the mutation G2019S in leucine-rich repeat kinase 2 protein (LRRK2) and control individuals without this mutation. The results reveal that MPP(+) induces mTOR-dependent autophagy in fibroblasts. Moreover, the effects of caspase-dependent cell death to MPP(+) were higher in cells with the G2019S LRRK2 mutation, which showed basal levels of autophagy due to the G2019S LRRK2 mutation (mTOR-independent). The inhibition of autophagy by 3-methyladenine (3-MA) treatment reduces these sensitivity differences between both cell types, however, the inhibition of autophagosome-lysosome fusion by bafilomycin A1 (Baf A1) increases these differences. This data confirm the importance of the combination of genetic and environmental factors in the PD etiology. Thereby, the sensitivity to the same damage may be different in function of a genetic predisposition, reason why individuals with certain mutations can develop some early-onset diseases, such as individuals with G2019S LRRK2 mutation and PD.

Inflammatory mechanisms are activated in aging and late-onset neurodegenerative diseases, such as Parkinson's disease (PD). Mutations in leucine-rich repeat kinase 2 (LRRK2) contribute to both idiopathic and familial forms of PD. Here, we investigated the involvement of LRRK2 in inflammatory pathways using primary dermal fibroblasts from patients with 2 common mutations in LRRK2 (G2019S and R1441G), idiopathic PD and age-matched healthy individuals. Basal cyclooxygenase (COX)-2 RNA levels were very high in the fibroblasts of all patients. Remarkably, LRRK2 silencing experiments significantly reduced basal COX-2 levels and COX-2 induction after a pro-inflammatory stimulus. Additionally, in samples from patients with the R1441G mutation and with idiopathic PD, we found a prominent cytoplasmic re-distribution of human antigen R, a protein that, among others, stabilizes COX-2 RNA. Furthermore, the response to lipopolysaccharide was defective in these 2 groups, which showed weak induction of pro-inflammatory cytokines and reduced NFκB transcriptional activation. In summary, we describe multiple defects in inflammatory pathways in which LRRK2 appears to be critically involved. Further studies are required to establish the therapeutic implications of inflammatory dysregulation in the pathophysiology of Parkinson's disease.

Scope The potential effects of yerba mate (YM) on mitochondrial biogenesis and thermogenesis are evaluated. Methods and results The in vitro effects of YM on mitochondrial respiration are assessed in C2C12 cells. The expression of genes related to mitochondrial biogenesis and thermogenesis are analyzed by quantitative PCR. The in vivo experiments are performed on mice fed a high-fat diet (HFD) and treated with YM extract. Indirect calorimetry was performed, and the expression of genes and proteins related to mitochondrial biogenesis, thermogenesis, and de novo lipogenesis is determined by quantitative PCR and western blot. Our in vitro data indicate that YM increases mtDNA copy number as well as mitochondrial spare respiratory capacity and coupling efficiency. The gene expression profile reinforces this evidence, indicating a modulation of genes downstream of Ampk. In vivo, it is found that YM partially prevents diet-induced obesity by increasing energy expenditure and enhancing mitochondrial biogenesis via the AMPK/SIRT1/PGC1α pathway. Conclusions YM stimulates mitochondriogenesis and Ucp expression, leading to an increase in the spare respiratory capacity and energy dissipation. These effects may help to better understand the potential use of YM for obesity treatment.

The management of municipal solid waste (MSW), particularly the role of incineration, is currently a subject of public debate. Incineration shows to be a good alternative of reducing the volume of waste and eliminating certain infectious components. Moreover, Municipal Waste Incinerators (MWI), are reported to be highly hygienic and apart from that MWIs are immediately effective in terms of transport (incinerators can be built close to the waste sources) and incineration's nature. Nevertheless, the emissions of many hazardous substances make the Municipal Waste Incineration (MWI) plants to be unpopular. Metals (especially lead, manganese, cadmium, chromium and mercury) are concentrated in fly and bottom ashes. Furthermore, incomplete combustion produces a wide variety of potentially hazardous organic compounds, such as aldehydes, polycyclic aromatic hydrocarbons (PAH), chlorinated hydrocarbons including polychlorinated dibenzodioxins (PCDD) and dibenzofurans (PCDF), and even acid gases, including NOx. Many of these hazardous substances are carcinogenic and some have direct systemic toxicity.

Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene cause late-onset autosomal dominant Parkinson's disease (PD), and sequence variations at the LRRK2 locus are associated with increased risk for sporadic PD. LRRK2 contains both GTPase and kinase domains flanked by protein interaction motifs, and mutations associated with familial PD have been described for both catalytic domains. LRRK2 has been implicated in diverse cellular processes, and recent evidence pinpoints to an important role for LRRK2 in modulating a variety of intracellular membrane trafficking pathways. However, the underlying mechanisms are poorly understood. Here, by studying the classical, well-understood, degradative trafficking pathway of the epidermal growth factor receptor (EGFR), we show that LRRK2 regulates endocytic membrane trafficking in an Rab7-dependent manner. Mutant LRRK2 expression causes a slight delay in early-to-late endosomal trafficking, and a pronounced delay in trafficking out of late endosomes, which become aberrantly elongated into tubules. This is accompanied by a delay in EGFR degradation. The LRRK2-mediated deficits in EGFR trafficking and degradation can be reverted upon coexpression of active Rab7 and of a series of proteins involved in bridging the EGFR to Rab7 on late endosomes. Effector pulldown assays indicate that pathogenic LRRK2 decreases Rab7 activity both in cells overexpressing LRRK2, as well as in fibroblasts from pathogenic mutant LRRK2 PD patients when compared with healthy controls. Together, these findings provide novel insights into a previously unknown regulation of Rab7 activity by mutant LRRK2 which impairs membrane trafficking at very late stages of the endocytic pathway.

Limb-girdle muscular dystrophy type 2A (LGMD2A) is the most frequent autosomal recessive muscular dystrophy. It is caused by mutations in the calpain-3 (CAPN3) gene. The majority of the mutations described to date are located in the coding sequence of the gene. However, it is estimated that 25% of the mutations are present at exon-intron boundaries and modify the pre-mRNA splicing of the CAPN3 transcript. We have previously described the first deep intronic mutation in the CAPN3 gene: c.1782+1072G>C mutation. This mutation causes the pseudoexonization of an intronic sequence of the CAPN3 gene in the mature mRNA. In the present work, we show that the point mutation generates the inclusion of the pseudoexon in the mRNA using a minigene assay. In search of a treatment that restores normal splicing, splicing modulation was induced by RNA-based strategies, which included antisense oligonucleotides and modified small-nuclear RNAs. The best effect was observed with antisense sequences, which induced pseudoexon skipping in both HeLa cells cotransfected with mutant minigene and in fibroblasts from patients. Finally, transfection of antisense sequences and siRNA downregulation of serine/arginine-rich splicing factor 1 (SRSF1) indicate that binding of this factor to splicing enhancer sequences is involved in pseudoexon activation.

Myotonic dystrophy type 1 (DM1) is caused by the expansion of CTG repeats in the 3' untranslated region of the DMPK gene. Several missplicing events and transcriptional alterations have been described in DM1 patients. A large number of these defects have been reproduced in animal models expressing CTG repeats alone. Recent studies have also reported miRNA dysregulation in DM1 patients. In this work, a Drosophila model was used to investigate miRNA transcriptome alterations in the muscle, specifically triggered by CTG expansions. Twenty miRNAs were differentially expressed in CTG-expressing flies. Of these, 19 were down-regulated, whereas 1 was up-regulated. This trend was confirmed for those miRNAs conserved between Drosophila and humans (miR-1, miR-7 and miR-10) in muscle biopsies from DM1 patients. Consistently, at least seven target transcripts of these miRNAs were up-regulated in DM1 skeletal muscles. The mechanisms involved in dysregulation of miR-7 included a reduction of its primary precursor both in CTG-expressing flies and in DM1 patients. Additionally, a regulatory role for Muscleblind (Mbl) was also suggested for miR-1 and miR-7, as these miRNAs were down-regulated in flies where Mbl had been silenced. Finally, the physiological relevance of miRNA dysregulation was demonstrated for miR-10, since over-expression of this miRNA in Drosophila extended the lifespan of CTG-expressing flies. Taken together, our results contribute to our understanding of the origin and the role of miRNA alterations in DM1. © The Author 2012. Published by Oxford University Press. All rights reserved., Fundacion Ramon Areces; Generalitat Valenciana (Prometeo/2010/081); Ministerio de Ciencia e Innovacion (SAF2006-09121) in collaboration with the biotechnology company Sistemas Genomicos S.L.; FIS (FIS09-00660) ; Isabel Gemio Foundation; Accion Especial de Enfermedades Raras ‘Cetegen’ by Genoma Espana Foundation; Generalitat Valenciana; Fundacion Ramon Areces; Banca Civica; Basque Government (AE-BFI-08.164); ISCIII; Ministerio de Economia y Competitividad

Skeletal muscle can be engineered by converting dermal precursors into muscle progenitors and differentiated myocytes. However, the efficiency of muscle development remains relatively low and it is currently unclear if this is due to poor characterization of the myogenic precursors, the protocols used for cell differentiation, or a combination of both. In this study, we characterized myogenic precursors present in murine dermospheres, and evaluated mature myotubes grown in a novel three-dimensional culture system. After 5-7 days of differentiation, we observed isolated, twitching myotubes followed by spontaneous contractions of the entire tissue-engineered muscle construct on an extracellular matrix (ECM). In vitro engineered myofibers expressed canonical muscle markers and exhibited a skeletal (not cardiac) muscle ultrastructure, with numerous striations and the presence of aligned, enlarged mitochondria, intertwined with sarcoplasmic reticula (SR). Engineered myofibers exhibited Na(+)- and Ca(2+)-dependent inward currents upon acetylcholine (ACh) stimulation and tetrodotoxin-sensitive spontaneous action potentials. Moreover, ACh, nicotine, and caffeine elicited cytosolic Ca(2+) transients; fiber contractions coupled to these Ca(2+) transients suggest that Ca(2+) entry is activating calcium-induced calcium release from the SR. Blockade by d-tubocurarine of ACh-elicited inward currents and Ca(2+) transients suggests nicotinic receptor involvement. Interestingly, after 1 month, engineered muscle constructs showed progressive degradation of the myofibers concomitant with fatty infiltration, paralleling the natural course of muscular degeneration. We conclude that mature myofibers may be differentiated on the ECM from myogenic precursor cells present in murine dermospheres, in an in vitro system that mimics some characteristics found in aging and muscular degeneration.

Myotonic dystrophy type 1 (DM1) is caused by the expansion of CTG repeats in the 3' untranslated region of the DMPK gene. Several missplicing events and transcriptional alterations have been described in DM1 patients. A large number of these defects have been reproduced in animal models expressing CTG repeats alone. Recent studies have also reported miRNA dysregulation in DM1 patients. In this work, a Drosophila model was used to investigate miRNA transcriptome alterations in the muscle, specifically triggered by CTG expansions. Twenty miRNAs were differentially expressed in CTG-expressing flies. Of these, 19 were down-regulated, whereas 1 was up-regulated. This trend was confirmed for those miRNAs conserved between Drosophila and humans (miR-1, miR-7 and miR-10) in muscle biopsies from DM1 patients. Consistently, at least seven target transcripts of these miRNAs were up-regulated in DM1 skeletal muscles. The mechanisms involved in dysregulation of miR-7 included a reduction of its primary precursor both in CTG-expressing flies and in DM1 patients. Additionally, a regulatory role for Muscleblind (Mbl) was also suggested for miR-1 and miR-7, as these miRNAs were down-regulated in flies where Mbl had been silenced. Finally, the physiological relevance of miRNA dysregulation was demonstrated for miR-10, since over-expression of this miRNA in Drosophila extended the lifespan of CTG-expressing flies. Taken together, our results contribute to our understanding of the origin and the role of miRNA alterations in DM1.

The link between the deregulation of autophagy and cell death processes can be essential in the development of several neurodegenerative diseases, such as Parkinson disease (PD). However, the molecular mechanism of deregulation of this degradative process in PD patients is unknown. The leucine-rich repeat kinase 2 (LRRK2) gene is related to PD and its implication in autophagy regulation has been described. Our recent work shows that the presence of the G2019S LRRK2 mutation, one of the most prevalent in LRRK2, is accompanied by a deregulation of autophagy basal levels dependent on the MAPK1/3 (ERK2/1) pathway.

Fibroblasts are ubiquitous cells essential to tissue homeostasis. Despite their nonphagocytic nature, fibroblasts restrain replication of intracellular bacterial pathogens such as Salmonella enterica serovar Typhimurium. The extent to which the entry route of the pathogen determines this intracellular response is unknown. Here, we analyzed S. Typhimurium invasion in fibroblasts obtained from diverse origins, including primary cultures and stable nontransformed cell lines derived from normal tissues. Features distinct to the invasion of epithelial cells were found in all fibroblasts tested. In some fibroblasts, bacteria lacking the type III secretion system encoded in the Salmonella pathogenicity island 1 displayed significant invasion rates and induced the formation of lamellipodia and filopodia at the fibroblast-bacteria contact site. Other bacterial invasion traits observed in fibroblasts were the requirement of phosphatidylinositol 3-kinase, mitogen-activated protein kinase MEK1, and both actin filaments and microtubules. RNA interference studies showed that different Rho family GTPases are targeted by S. Typhimurium to enter into distinct fibroblasts. Rac1 and Cdc42 knockdown affected invasion of normal rat kidney fibroblasts, whereas none of the GTPases tested (Rac1, Cdc42, RhoA, or RhoG) was essential for invasion of immortalized human foreskin fibroblasts. Collectively, these data reveal a marked diversity in the modes used by S. Typhimurium to enter into fibroblasts.

Our group has recently derived skeletal muscle from dermis-derived cells, by using an extracellular matrix that recreates the myogenic niche. After one week of differentiation, we observed isolated, twitching myotubes followed by spontaneous contractions of the entire tissue-engineered muscle construct. In vitro engineered myofibers expressed canonical markers, ultrastructure and electrophysiological characteristics of skeletal muscle. Interestingly, after one-month engineered muscle constructs showed progressive degradation of the myofibers concomitant with fatty infiltration, paralleling the natural course of muscular degeneration. However, we do not yet know how dermis-resident precursors are related with our isolated-myogenic precursors, a critical point for extrapolating our results to the human system. Knowing that dermal cells and muscle cells share a common embryonic origin from somite-derived dermomyotome and taking into account that there are different types of cells within the skin that have myogenic potential, our main objective is to identify and characterize the origin of murine dermal subpopulation with a myogenic potential, hypothesizing that may be (i) satellite cells from the Panniculus carnosus, (ii) dermomyotome-derived adult stem cells, (iii) perivascular cells and/or (iv) neural crest-derived precursor cells. For this end, lineage tracing experiments [(i) Pax3-GFP, (ii) Pax7CE, (iii) Myf5-Cre, (iv) CSPG4-Cre, and (v) Sox10-Cre] combined with FACS strategies and cellular differentiation assays have been developed. Our results show a high contribution of Myf5+ cells, a low contribution of Cspg4+ cells, and no contribution of Sox10+ cells to dermis-derived myogenic precursor cell subset. In principle, that means that dermomyotome-derived but not neural crest-derived cells are involved in subsequent engineered muscle differentiation.

Detection of phospho-Rab8a in pathogenic LRRK2-expressing cells as well as in cells co-transfected with wildtype LRRK2 and wildtype Rab8a, but not phospho-deficient Rab8a. (DOCX 958 kb)

Rab8a protein levels and pericentrosomal/centrosomal accumulation of phosphorylated Rab8a in lymphoblasts from control and G2019S mutant LRRK2 PD patients. (DOCX 636 kb)

Rab8a protein levels and pericentrosomal/centrosomal accumulation of phosphorylated Rab8a in lymphoblasts from control and G2019S mutant LRRK2 PD patients. (DOCX 636 kb)

Detection of phospho-Rab8a in pathogenic LRRK2-expressing cells as well as in cells co-transfected with wildtype LRRK2 and wildtype Rab8a, but not phospho-deficient Rab8a. (DOCX 958 kb)

100 p.