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Graphene derivatives are expected to have a great impact in a wide range of applications, among them as food packaging materials. This is one of the sources of potential human oral exposure to them. However, studies devoted to investigating their putative toxic effects at the intestinal level are underrepresented in the scientific literature. Thus, this study aimed to investigate the in vitro toxicity of reduced graphene oxide (rGO) and gra- phene oxide (GO) in the human intestinal Caco-2 cell line. rGO and GO were firstly characterized and later, cell viability was assessed after exposure to 0–250 μg/mL rGO/GO for 24 and 48 h. Internalization was evidenced for both materials using transmission electron microscopy. A mean effective concentration (24 h) of 176.3 ± 7.6 μg/ mL for cytotoxicity was obtained for rGO, whereas GO did not induce any change at the concentration range evaluated. However, both of them altered oxidative stress biomarkers, causing increased reactive oxygen species (ROS) and depletion of the glutathione content (GSH) after exposures up to 24 h. Further studies, particularly with rGO, are required to elucidate their toxicity profile in experimental models relevant for oral exposures. Fondo Europeo de Desarrollo Regional (FEDER) y la Consejería de Economía, Conocimiento, Empresas y Universidad de la Junta de Andalucía - FEDER 2014–2020/US-1259106 Consejería de Transformación Económica, Industria, Conocimiento y Universidades, Junta de Andalucía (PAIDI-2020/FEDER)-P18-RT- 1993 Plan VI Propio de Investigación de la Universidad de Sevilla

Recent advances in molecular biology have led to the CRISPR revolution, but the lack of an efficient and safe delivery system into cells and tissues continues to hinder clinical translation of CRISPR approaches. Polymeric vectors offer an attractive alternative to viruses as delivery vectors due to their large packaging capacity and safety profile. In this paper, we have demonstrated the potential use of a highly branched poly(β-amino ester) polymer, HPAE-EB, to enable genomic editing via CRISPRCas9-targeted genomic excision of exon 80 in the COL7A1 gene, through a dual-guide RNA sequence system. The biophysical properties of HPAE-EB were screened in a human embryonic 293 cell line (HEK293), to elucidate optimal conditions for efficient and cytocompatible delivery of a DNA construct encoding Cas9 along with two RNA guides, obtaining 15–20% target genomic excision. When translated to human recessive dystrophic epidermolysis bullosa (RDEB) keratinocytes, transfection efficiency and targeted genomic excision dropped. However, upon delivery of CRISPR–Cas9 as a ribonucleoprotein complex, targeted genomic deletion of exon 80 was increased to over 40%. Our study provides renewed perspective for the further development of polymer delivery systems for application in the gene editing field in general, and specifically for the treatment of RDEB.

Current efforts to find specific genodermatoses treatments and define precise pathogenesis mechanisms require appropriate surrogate models with human cells. Although transgenic and gene knockout mouse models for several of these disorders exist, they often fail to faithfully replicate the clinical and histopathological features of the human skin condition. We have established a highly efficient method for precise deletion of critical gene sequences in primary human keratinocytes, based on CRISPR-Cas9-mediated gene editing. Using this methodology, in the present study we generated a model of Netherton syndrome by disruption of SPINK5. Gene-edited cells showed absence of LEKTI expression and were able to recapitulate a hyperkeratotic phenotype with most of the molecular hallmarks of Netherton syndrome, after grafting to immunodeficient mice and in organotypic cultures. To validate the model as a platform for therapeutic intervention, we tested an ex vivo gene therapy approach using a lentiviral vector expressing SPINK5. Re-expression of SPINK5 in an immortalized clone of SPINK5-knockout keratinocytes was capable of reverting from Netherton syndrome to a normal skin phenotype in vivo and in vitro. Our results demonstrate the feasibility of modeling genodermatoses, such as Netherton syndrome, by efficiently disrupting the causative gene to better understand its pathogenesis and to develop novel therapeutic approaches., Graphical Abstract, Exploiting the advantages of their highly efficient CRISPR-Cas9 gene editing strategy, in this study, Gálvez et al. abrogate SPINK5, the Netherton syndrome-causing gene, in primary human keratinocytes. SPINK5-knockout cells show disease hallmarks in vitro and in vivo, enabling the development of trustworthy models suitable for the evaluation of advanced therapies for genodermatoses.

Recessive dystrophic epidermolysis bullosa, a devastating skin fragility disease characterized by recurrent skin blistering, scarring, and a high risk of developing squamous cell carcinoma is caused by mutations in

Summary Background Recessive dystrophic epidermolysis bullosa (RDEB), Kindler syndrome (KS) and xeroderma pigmentosum complementation group C (XPC) are three cancer‐prone genodermatoses whose causal genetic mutations cannot fully explain, on their own, the array of associated phenotypic manifestations. Recent evidence highlights the role of the stromal microenvironment in the pathology of these disorders. Objectives To investigate, by means of comparative gene expression analysis, the role played by dermal fibroblasts in the pathogenesis of RDEB, KS and XPC. Methods We conducted RNA‐Seq analysis, which included a thorough examination of the differentially expressed genes, a functional enrichment analysis and a description of affected signalling circuits. Transcriptomic data were validated at the protein level in cell cultures, serum samples and skin biopsies. Results Interdisease comparisons against control fibroblasts revealed a unifying signature of 186 differentially expressed genes and four signalling pathways in the three genodermatoses. Remarkably, some of the uncovered expression changes suggest a synthetic fibroblast phenotype characterized by the aberrant expression of extracellular matrix (ECM) proteins. Western blot and immunofluorescence in situ analyses validated the RNA‐Seq data. In addition, enzyme‐linked immunosorbent assay revealed increased circulating levels of periostin in patients with RDEB. Conclusions Our results suggest that the different causal genetic defects converge into common changes in gene expression, possibly due to injury‐sensitive events. These, in turn, trigger a cascade of reactions involving abnormal ECM deposition and underexpression of antioxidant enzymes. The elucidated expression signature provides new potential biomarkers and common therapeutic targets in RDEB, XPC and KS. What's already known about this topic? Recessive dystrophic epidermolysis bullosa (RDEB), Kindler syndrome (KS) and xeroderma pigmentosum complementation group C (XPC) are three genodermatoses with high predisposition to cancer development.Although their causal genetic mutations mainly affect epithelia, the dermal microenvironment likely contributes to the physiopathology of these disorders. What does this study add? We disclose a large overlapping transcription profile between XPC, KS and RDEB fibroblasts that points towards an activated phenotype with high matrix‐synthetic capacity.This common signature seems to be independent of the primary causal deficiency, but reflects an underlying derangement of the extracellular matrix via transforming growth factor‐β signalling activation and oxidative state imbalance. What is the translational message? This study broadens the current knowledge about the pathology of these diseases and highlights new targets and biomarkers for effective therapeutic intervention.It is suggested that high levels of circulating periostin could represent a potential biomarker in RDEB., Linked Comment: https://doi.org/10.1111/bjd.18104. https://doi.org/10.1111/bjd.18271 available online

Hotel firms can enhance their performance by accessing external resources through their inter-personal and inter-organizational ties. However, neither has a repertory been compiled of relevant external resources in the sector, nor are appropriate diagnostic and analytical tools available to improve the way these resources may be used. The hotel resource generator is an instrument adapted from the field of sociology that is used here to measure access to those network resources. It is presented in this study as a tool to measure the external resources that condition the performance of hotel firms and is tested on a sample of hotels from Andalusia (Spain). JEL classification: L83, M10, Keywords: Hotel resources, Network resources, Resource generator, Social capital

Alliances in the airline industry operate as a basic strategy to stimulate competition and both the number and the types of alliances have increased over time. Each airline maintains alliances simultaneously with a variable number of partners. The set of airline's alliance partners constitutes its alliance portfolio. A central theme of these portfolios is the way in which partner selection improve performance. Is the alliance portfolio configuration based on either similar or different partners? We examine how the differences can arise from differences between the partners themselves, from resource complementarity, and from the partners structural positions in the network. The relations between these types of differences and whether they affect firm performance are studied. Codeshare alliances established between airline companies at a global level are analyzed to establish their effects on performance. In particular, the study comprises 135 alliance portfolios all of which from major airlines. The results suggest that structural homogeneity and complementarity improve performance and that alliance portfolio diversity favors network resource complementarity.

Firms establish a presence in a wide range of alliances to access the critical resources that they may need at any one time. Although inter-organizational relationships constitute the channels through which social capital flows, firms should consider which mechanisms or characteristics of the relations facilitate their actual mobilization. Accordingly, the final purpose of alliances cannot simply be potential access to partner resources, it must also be their real use. A definition of alliance types yielded the parameters for an ordinary least squares regression of a sample from top international airline databases. The results showed that certain kind of alliances favored the actual mobilization of social capital. Accordingly, cooperation as a structural organizational form, and in particular certain types of alliances such as global or multilateral, makes it easier to cope with increasingly volatile, uncertain, complex and ambiguous environments -VUCA conditions.

Genome-editing technologies that enable the introduction of precise changes in DNA sequences have the potential to lead to a new class of treatments for genetic diseases. Epidermolysis bullosa (EB) is a group of rare genetic disorders characterized by extreme skin fragility. The recessive dystrophic subtype of EB (RDEB), which has one of the most severe phenotypes, is caused by mutations in COL7A1. In this study, we report a gene-editing approach for ex vivo homology-directed repair (HDR)-based gene correction that uses the CRISPR-Cas9 system delivered as a ribonucleoprotein (RNP) complex in combination with donor DNA templates delivered by adeno-associated viral vectors (AAVs). We demonstrate sufficient mutation correction frequencies to achieve therapeutic benefit in primary RDEB keratinocytes containing different COL7A1 mutations as well as efficient HDR-mediated COL7A1 modification in healthy cord blood-derived CD34+ cells and mesenchymal stem cells (MSCs). These results are a proof of concept for HDR-mediated gene correction in different cell types with therapeutic potential for RDEB. This work was supported by Spanish grants PI17/01747, PI20/00615, AC17/00054 (MutaEB-E-rare), and CIBERER ER18TRL714 from the Instituto de Salud Carlos III and grant SAF2017-86810-R from the Ministry of Economy and Competitiveness , all co-funded with European Regional Development Funds , and Avancell-CM grant ( S2017/BMD-3692 ). Authors are indebted to Almudena Holguín and Nuria Illera for grafting experiments, and to Jesus Martínez and Edilia De Almeida for animal maintenance and care.

The analysis of relations of dependency is widespread in tourism research. However, there are a series of questions related to endogeneity, such as dependence on the past and reverse causality, clo...

This paper analyses the virtualization of the airline industry as a strategic response to competitive pressures. Increased competition has driven the associative behavior of airlines, fostered the development of global alliances and the use of codeshare agreements, substantially modifying the structure of the industry. These changes have resulted in greater efficiency and division of labor in the industry, and have caused airlines to significantly modify their operational strategies. The analysis of the evolution of operational and associative strategies in the airline industry during the past decade (2005–2015) shows the growing virtualization of the sector, derived from the use of codeshare agreements and other alliances between companies. The results show that the associative behavior allows airlines to specialize their operational strategy and to focus their resources and capabilities on a certain type of routes or origin-destination (OD) markets, with a wide variety of strategic behaviors in the industry.

Los riesgos emergentes son aquellos derivados de un nuevo peligro para el que puede producirse una exposición significativa, o derivado de una exposición y/o susceptibilidad nueva o incrementada significativamente a un peligro conocido. La identificación de estos riesgos puede realizarse utilizando distintas fuentes de información y herramientas, que pueden ir desde el análisis de bibliografía científica publicada, e incluso de la llamada literatura gris (documentos que no son editados o que se distribuyen a través de canales poco convencionales como tesis doctorales, actas de congresos, informes de investigación, etc.), hasta el estudio de las alertas alimentarias. La evaluación de los riesgos emergentes es realizada por los especialistas en cada área de conocimiento, pero en su identificación pueden utilizarse herramientas comunes a todos ellos de manera que se detecten y caractericen de forma eficaz antes de su evaluación. Por ello, dado que La Ley 17/2011, de Seguridad Alimentaria y Nutrición, establece un mandato para el Comité Científico en relación a la identificación de los riesgos emergentes alimentarios, se solicita a la Sección de Seguridad Alimentaria y Nutrición que elabore un informe sobre las estrategias aplicables para la identificación de riesgos emergentes alimentarios. Teniendo como base la definición de la Autoridad Europea de Seguridad Alimentaria (EFSA) en relación a riesgo emergente es importante identificar si hay un nuevo peligro al que estamos expuestos, o si ha surgido una nueva exposición o se incrementado la exposición a uno conocido. Al mismo tiempo que valorar el incremento de la susceptibilidad a un peligro conocido. Una vez revisadas las distintas estrategias posibles, el Comité Científico considera las siguientes como de utilidad: • Análisis de la bibliografía científica. Para ello es de utilidad el desarrollo o implementación de sistemas potentes de gestión de datos y plataformas de colaboración digital para gestionar la gran cantidad de datos e información disponibles. • Análisis de la literatura gris. Al igual que en el caso anterior se necesitarían herramientas informáticas específicas que permitan extraer información de interés. • Análisis de las alertas y de los resultados de los programas de control oficial. Estas herramientas controlan aquellos riesgos conocidos y sobre los que existe legislación de control. Podrían ser útiles para detectar aumentos de exposición por incrementos en la presencia de algún contaminante a lo largo del tiempo. • Análisis de los sistemas de vigilancia epidemiológica que podrían servir para detectar incrementos de susceptibilidad o exposición. • Análisis de tendencias de consumo. Serviría para detectar incrementos de exposición, siempre que la metodología de recogida de datos sea comparable entre años. • Análisis de la información recabada de estudios de percepción del riesgo por parte de la población. • Análisis de la información procedente de expertos e investigadores. Sería deseable mejorar la colaboración de la comunidad científica con las Instituciones y organizaciones nacionales e internacionales. Emerging risks are those derived from a new hazard for which a significant exposure could be produced, or derived from an exposure and/or new or significantly increased susceptibility to a known hazard. Identifying these risks may be done using different information sources and tools, which may range from the analysis of the published scientific bibliography, even including what is known as grey literature (documents that are unedited or that are distributed by means of unconventional channels such as doctoral theses, conference proceedings, research reports, etc.), to the study of food alerts. The assessment of emerging risks is done by specialists in each area of expertise, but in identifying them they may use common tools in such a way that they may detect and characterise them more efficiently prior to their assessment. Therefore, given that Law 17/2011, on Food Safety and Nutrition, establishes a mandate for the Scientific Committee, regarding the identification of emerging food risks, the Section of Food Safety and Nutrition is asked to prepare a report about the applicable strategies for the identification of emerging food risks. Having as a base the definition of the European Food Safety Authority (EFSA) regarding emerging risks it is important to identify if there is a new hazard to which we are exposed, or if a new exposure has come about, or if there has been an increase in the exposure of a known one. At the same time evaluating the increase in susceptibility to a known hazard. After reviewing the different possible strategies, the Scientific Committee considers the following to be useful: • Analysis of the scientific bibliography. For this the development or implementation of powerful data management systems and digital collaboration platforms is of use in order to manage the large quantity of available data and information. • Analysis of grey literature. Just as in the previous case, specific IT tools would be needed that allow for information of interest to be extracted. • Analysis of the alerts and the results of the official control programmes. These tools control those known risks and about which there is control legislation. They could be useful for detecting incremental exposure increases in the presence of some contaminant over time. • Analysis of the epidemiological monitoring systems, which would help detect increases in susceptibility or exposure. • Analysis of consumption trends. It would help detect exposure increases, provided that the methodology for collecting data is comparable between years. • Analysis of the information gathered from studies on the risk perception by the population. • Analysis of information provided by experts and researchers. It would be desirable to improve the collaboration of the scientific community with the national and international institutions and organisations.

Gene editing constitutes a novel approach for precisely correcting disease-causing gene mutations. Frameshift mutations in COL7A1 causing recessive dystrophic epidermolysis bullosa are amenable to open reading frame restoration by non-homologous end joining repair-based approaches. Efficient targeted deletion of faulty COL7A1 exons in polyclonal patient keratinocytes would enable the translation of this therapeutic strategy to the clinic. In this study, using a dual single-guide RNA (sgRNA)-guided Cas9 nuclease delivered as a ribonucleoprotein complex through electroporation, we have achieved very efficient targeted deletion of COL7A1 exon 80 in recessive dystrophic epidermolysis bullosa (RDEB) patient keratinocytes carrying a highly prevalent frameshift mutation. This ex vivo non-viral approach rendered a large proportion of corrected cells producing a functional collagen VII variant. The effective targeting of the epidermal stem cell population enabled long-term regeneration of a properly adhesive skin upon grafting onto immunodeficient mice. A safety assessment by next-generation sequencing (NGS) analysis of potential off-target sites did not reveal any unintended nuclease activity. Our strategy could potentially be extended to a large number of COL7A1 mutation-bearing exons within the long collagenous domain of this gene, opening the way to precision medicine for RDEB., Highly efficient and safe approaches for precise correction of pathogenic mutations are required to realize the full potential of gene-editing protocols for clinical practice. Here we describe a single-step NHEJ-based CRISPR/Cas9 strategy of COL7A1 mutation-bearing exon deletion with unprecedented efficacy for ex vivo correction of recessive dystrophic epidermolysis bullosa.

The programme aimed at training the fellow in the risk assessment guidelines proposed by the EFSA in the field of contaminants, food contact materials, technological ingredients and nutritional risks. It had a modular ‘learning by doing’ approach and a balanced learning/case studies and theory. Module 1 offered an insight into chemical risk assessment and conferred transferable skills for a proper application of the framework. The hands-on activities consisted of three case studies that went from a simple exercise on an official opinion, to working in a team with experts to produce a new opinion, to an individual work to obtain a publishable review manuscript. Module 2 was a training in experimental toxicology designed to create a toxicological basis and to enable the fellow to perform toxicological studies for risk assessment purposes. She joined the team working on cyanotoxins, gained experience with both EFSA and Organization of Economic Cooperation and Development (OECD) guidelines on genotoxicity and an insight into the developing of analytical methods suitable for risk assessment purposes. During module 3, the fellow was trained in nutritional risk assessment and involved in experimental work in chemical characterisation, biomarkers and mechanisms of action of bioactive compounds. This developed the critical perspective when assessing nutritional and health claims related the design of experiments, methods used, interpretation of results and human relevance. Module 4 provided a ‘hand-on experience’ in scientific risk communication as the fellow was encouraged and supported in the participation at local, national and international workshops and congresses presenting the outcomes of the three modules. Thus, the fellow was successfully integrated in the day-by-day workflow of the department, gaining first-hand practical experience in risk assessment in a multicultural and interdisciplinary context. This enabled a productive exchange of good practices and contributed to building a European risk assessment community. European Food Safety Authority (EEFSA)-EU-FORA

The important links that are formed between tourist destinations and the airline industry are evident in the previous literature. These links are active in both directions. The characteristics and the policies of tourist destinations can affect such relevant aspects as the organization of airline route networks, the inauguration of new routes and their expansion. In this study, it is proposed that the value of a tourist destination and the inclusion of airports close to the tourist destination on the routes of an airline are also related to whether that airline decides to plan its route networks with its own resources or to complement those resources with assistance from other partner airlines through cooperative alliances, specifically code-share agreements. Taking into account the assumptions of the Resource-Based and the Relational views, the value of a tourist destination can be a relevant factor in the cooperative behaviour of airlines. Moreover, the routes within which the most valuable tourist destinations are embedded can be of different types. It is therefore of interest to know whether the type of route plays a relevant role in the preference for cooperative behaviour among the routes that include tourist destinations. Around 50,000 routes and the most important global tourist destinations were considered to analyse this behaviour. The results showed that the airlines under study preferred to operate the routes that included valuable tourist destinations with their own resources, although this behaviour was more variable when differentiating between types of routes.

espanolLas alianzas son una herramienta estrategica que permite colaborar y atraer recursos no disponibles de otra forma. Gracias a la complementariedad de los recursos de red y a la experiencia previa en alianzas, se puede maximizar el rendimiento que obtiene la organizacion y ello repercute de forma directa en el resultado empresarial. Asimismo, la complementariedad viene dada por la variedad de recursos que aportan socios heterogeneos. Tambien, consideramos que la experiencia en gestion de alianzas es un factor importante que influye en la obtencion de un mayor rendimiento. De este modo, se pueden crear diversas ventajas competitivas para adaptarse a las circunstancias del mercado. EnglishStrategic alliances are a tool that permits collaborate and attract resources that they are not available otherwise. Due to network resources complementary and previous alliance experience, it can maximize the alliance performance and it has direct repercussions on firm results. Furthermore, resource complementarity is given by heterogeneous partners. Moreover, we think alliance management experience is an important factor to obtain a better firm performance. In this way, they can create diverse competitive advantages to adapt itself to the market.

La almorta (Lathyrus sativus) es una legumbre utilizada como tal o en forma de harina en diferentes países tanto para consumo humano, como para alimentación animal. Pese al arraigo del consumo de su harina en algunas zonas para la preparación del plato tradicional de las gachas, su utilización para consumo humano está prohibida actualmente en España. El Comité Científico ha realizado una revisión del estado de esta cuestión desde el punto de vista de la seguridad del consumo de la harina de almortas, especialmente en lo referido a su ingesta en forma de gachas. Los estudios toxicológicos en animales aportan una información limitada respecto a los niveles de ingesta y no permiten determinar valores de ingesta tolerable del aminoácido β-N-oxalil-L-α, β-diamino-propiónico (beta-ODAP) presente en las almortas. Hay diferencias en la metabolización del beta-ODAP entre los animales y los humanos que dificultan la estimación de los niveles de consumo de harina de almortas que resultan seguros. Los casos de neurolatirismo se han producido en el marco de un consumo elevado y prolongado de almortas, en el que representaban la base de la alimentación. En ausencia de esas condiciones, no se desarrolla esta enfermedad. El Comité Científico de la Agencia Española de Consumo, Seguridad Alimentaria y Nutrición (AECOSAN) concluye que, de acuerdo con la información disponible en este momento, el riesgo para la salud de la población general del consumo de harina de almortas puede considerarse despreciable (excluidas las personas con dificultades metabólicas para destoxicar el beta-ODAP) en las condiciones habituales de consumo de esta harina en forma de gachas por la población española. Se considerará una ingesta esporádica de raciones de harina, cuando ésta no supere el límite de 25 g de harina de almortas/ración diaria, con contenido del aminoácido beta-ODAP reducido, no superior a un 1 % de beta-ODAP, y en el marco de una dieta variada que incluya aminoácidos azufrados presentes en alimentos de origen animal como la carne, el pescado, los huevos y los productos lácteos, y de origen vegetal como los cereales integrales (especialmente avena) y frutos secos. The grass pea (Lathyrus sativus) is a legume that is used in different countries in its natural state or in the form of flour, for human consumption as well as for animal feed. Despite the popularity of its consumption in the form of flour in some areas for the preparation of the traditional dish, gachas (porridge), its use for human consumption is currently forbidden in Spain. The Scientific Committee has carried out a review of this issue from the point of view of the safety of the consumption of grass pea flour, especially as regards its intake in the form of gachas. Toxicological studies on animals provide limited information regarding intake levels and they do not allow us to determine tolerable intake values of the β-N-oxalyl-α,β-diaminopropionic (betaODAP) amino acid present in grass peas. There are differences in the metabolisation of beta-ODAP between animals and humans, which hinders the estimation of safe levels of grass pea flour consumption. The cases of neurolathyrism occurred in the context of a high and prolonged grass peas intake, in which they were the staple food of the diet. In the absence of these conditions, this disease does not occur. The Scientific Committee of the Spanish Agency for Consumer Affairs, Food Safety and Nutrition (AECOSAN) has concluded that, in accordance with the information available at this point, the risk of the consumption of grass pea flour for the health of the general population may be considered negligible (excluding those with metabolic difficulties in detoxifying beta-ODAP) under the usual conditions of this flour in the form of gachas by the Spanish population. Intake of portions of flour will be considered occasional when the limit of 25 g of grass pea flour/daily portion is not exceeded, with a low content of beta-ODAP amino acid, not higher than 1 % beta-ODAP, and in the context of a balanced diet that includes sulphur amino acids present in foods of animal origin such as meat, fish, eggs and dairy products and of plant origin such as wholegrain cereals (especially oats) and nuts.

Los alcaloides de la pirrolizidina son toxinas naturales, producto del metabolismo secundario de las plantas, que se producen como mecanismo de defensa frente a herbívoros. Su estructura química está basada en un anillo de pirrolizidina, que consiste en dos anillos fusionados con un átomo de nitrógeno como puente. Se conocen cerca de 600 de estos alcaloides, el 95 % se encuentran en cinco familias de plantas: Asteraceae, Boraginaceae, Fabaceae, Orchidaceae y Apocynaceae. Los alcaloides de la pirrolizidina tienen un perfil común de toxicidad, los principales son diversos grados de daño hepático (necrosis hepatocelular centrolobular) y enfermedad venooclusiva. Además, la Agencia Internacional para la Investigación del Cáncer (IARC) los ha clasificado como “posiblemente carcinógenos para el ser humano” (grupo 2B). La Autoridad Europea de Seguridad Alimentaria (EFSA) ha elaborado dictámenes sobre distintos aspectos de estos alcaloides en 2011, 2016 y 2017 y la Comisión Europea ha seleccionado provisionalmente 17 de ellos como relevantes en alimentos. A falta de un límite legal que permita a las autoridades competentes en inspección tomar decisiones basadas en dicho límite ante la detección de estos alcaloides en el polen, se solicita a la Sección de Seguridad Alimentaria y Nutrición del Comité Científico de la Agencia Española de Seguridad Alimentaria y Nutrición (AESAN) que realice una evaluación del riesgo asociado a la presencia de alcaloides de la pirrolizidina en polen destinado al consumo humano que sirva de base a dicha toma de decisiones. Conforme a los datos disponibles actualmente y la estimación realizada teniendo en cuenta todas las incertidumbres identificadas en el proceso de evaluación de riesgos, la ingesta de alcaloides de la pirrolizidina a través del consumo de polen puede dar lugar a riesgos crónicos en la población española. No se considera probable, sin embargo, la aparición de riesgos agudos. Para llevar a revista del comité científico nº 28 128 cabo una evaluación del riesgo más realista se recomienda realizar estudios de consumo de polen en la población española. Igualmente, se estima la conveniencia de establecer límites legales en cuanto al contenido de alcaloides de la pirrolizidina con el fin de limitar la exposición. Pyrrolizidine alkaloids are natural toxins, the product of the secondary metabolism of plants which are produced as a defence mechanism against herbivores. Their chemical structure is based on a pyrrolizidine ring, consisting of two rings fused together by a bridge nitrogen atom. There are approximately 600 known alkaloids, 95 % of which are found in five plant families: Asteraceae, Boraginaceae, Fabaceae, Orchidaceae and Apocynaceae. The pyrrolizidine alkaloids have a common toxicity profile, the main ones being various degrees of liver damage (centrolobular hepatocellular necrosis) and veno occlusive disease. In addition, the International Agency for Research on Cancer (IARC) has classified them as “possibly carcinogenic to humans” (group 2B). The European Food Safety Authority (EFSA) issued opinions on different aspects of these alkaloids in 2011, 2016 and 2017, and the European Commission has provisionally selected 17 of these as relevant in food. In the absence of a legal limit which enables the competent inspection authorities to make decisions based on this limit in the event of the detection of these alkaloids in pollen, the Section of Food Safety and Nutrition of the Scientific Committee of the Spanish Agency for Food Safety and Nutrition (AESAN) is asked to conduct an assessment of the risk associated with the presence of pyrrolizidine alkaloids in pollen intended for human consumption to serve as a basis for taking such decisions. According to currently available data and the estimations made, and considering all the uncertainties identified in the risk assessment process, the intake of pyrrolizidine alkaloids through the consumption of pollen may lead to chronic risk for the Spanish population. However, the appearance of acute risks is not considered likely. In order to conduct a more realistic risk assessment, studies of pollen consumption among the Spanish population are recommended. Similarly, the establishment of legal limits for pyrrolizidine alkaloid content is considered to be appropriate in order to limit the risk of exposure.

A lo largo de la cadena alimentaria pueden estar presentes, incorporarse o producirse distintos peligros de tipo químico que podrían suponer un riesgo para el consumidor.El Comité Científico ha realizado una revisión de los peligros químicos de mayor interés para la seguridad alimentaria en España que no cuentan con una regulación específica, identificándolos y señalando aquellos alimentos o condiciones que, a priori, podrían implicar un mayor riesgo para el consumidor, con el fin de realizar, eventualmente, estudios prospectivos.En el informe se han contemplado los siguientes peligros químicos y matrices: Cilindrospermopsina (toxina de cianobacterias) en agua de bebida (no envasada), Cloropropanoles y Glicidol en alimentos infantiles, Furano y derivados en alimentos procesados en general y alimentos infantiles en particular, Hidrocarburos de aceites minerales, Micotoxinas producidas por hongos del género Claviceps en cereales y alimentos derivados, Toxinas de Alternaria en frutas, hortalizas, cereales y tomate, Micotoxinas de Fusarium (Eniatinas, Nivalenol) en cereales y Alcaloides de la pirrolizidina en alimentos infantiles, complementos alimenticios, miel, polen, té, infusiones y cereales.De cada uno de ellos se describe su identificación y caracterización, la evaluación de la exposición, y se incluyen así mismo recomendaciones y consideraciones futuras.Por otro lado, la identificación de nuevos peligros para los que puede producirse una exposición significativa, o la evaluación del riesgo derivado de una exposición o susceptibilidad nuevas o incrementadas significativamente a un peligro conocido es importante, no solo a efectos de un eventual control de estos peligros emergentes, sino también de promover la investigación y mejorar su conocimiento por parte de los consumidores y de la comunidad científica. Along the food chain different chemical hazards may be present, incorporated or produced that could pose a risk to the consumer.The Scientific Committee has reviewed the chemical hazards of most concern for food safety in Spain that are not specifically regulated, identifying them and drawing attention to those foods or conditions which, a priori, may involve a greater risk to consumers, with the purpose of eventually carrying out prospective studies.The following chemical hazards and matrices have been considered in the report: Cylinderper-mopsin (cyanobacteria toxin) in drinking water (not bottled), Chloropropanols and Glycidol in baby food, Furan and derivatives in processed foods in general, and particularly in baby food, Hydrocar-bons of mineral oils, Mycotoxins produced by fungi of the Claviceps genus in cereals and derived foods, Alternaria toxins in fruits, vegetables, cereals and tomatoes, Fusarium mycotoxins (Enniatins, Nivalenol) in cereals and Pyrrolizidine alkaloids in baby food, food supplements, honey, pollen, tea, infusions and cereals. The identification, characterisation, and exposure assessment of each are described, and re-commendations and future considerations are also included.Furthermore, the identification of new hazards which may have a significant exposure, or the risk assessment derived from a new or substantially increased exposure or susceptibility to a known hazard is important in order to not only eventually control these emerging hazards, but to also promote research and improve the knowledge of both consumers and the scientific community.

Purpose– This paper aims to advance a model that will explain how hotel firms access and mobilize external resources. Hotel operators and firms need to complement their internal resources with external resources, which they can access through their personal and organizational ties, so as to compete and to achieve success.Design/methodology/approach– A framework is proposed, on the basis of the resource-based view and network theory, to explain the process of access and mobilization of available external resources thanks to the professional and social ties of the managers of hotel firms.Findings– This framework distinguishes between access to network resources and their mobilization. This paper introduces network resource mobilization capability as an adaptive capability of managers and employees that can improve hotel firm performance. Previous experience and contextual factors such as the type of property and the management style all influence the nature of this capability.Practical implications– This work proposes a repertory of relevant resources in hotels and the preparation of an instrument to measure access to those resources and their mobilization through social capital. It also proposes the need to develop a new dynamic capability: the capability to mobilize network resources in hotel firms through their managers. Finally, it proposes that social capital is a valuable resource for both hotel firms and their managers.Originality/value– This theoretical approach makes a key distinction between access to and mobilization of network resources, which leads to a better understanding of the potential of the individual social capital of hotel managers. Network resource mobilization capability is introduced as an adaptive capability of managers.

Dystrophic epidermolysis bullosa is a rare blistering condition caused by mutations in the COL7A1 gene. Different clinical variants have been described, with dominant and recessive inheritance, but no consistent findings have been elucidated to establish a genotype-phenotype correlation. We present three unrelated patients with two identical pathogenic compound heterozygous mutations in the COL7A1 gene that developed different clinical forms of dystrophic epidermolysis bullosa-epidermolysis bullosa pruriginosa and mild recessive non-Hallopeau-Siemens-raising the possibility of other genetic or environmental modifying factors responsible for the phenotype of the disease.

La empresa SmartWash Solutions, LLC ha solicitado una evaluación de la seguridad del uso como coadyuvante tecnológico de una solución acuosa de ácido fosfórico (18-25 %) y propilenglicol (5-10 %) en el procesado de vegetales frescos cortados y hortalizas de hoja de IV gama. Ambas sustancias se encuentran autorizadas en la Unión Europea como aditivos alimentarios en diversas categorías de alimentos. El uso propuesto para el coadyuvante tecnológico es como solución estabilizadora del pH para favorecer la actividad antimicrobiana del cloro utilizado para la desinfección del agua de lavado utilizada en el procesado de vegetales frescos cortados y hortalizas de hoja de IV gama. Con esta solución se pretende reducir el pH de las aguas de lavado a niveles de entre 5,5 a 6,5. La concentración de uso varía dependiendo de los requerimientos del producto y la composición química de las aguas de lavado, siendo la concentración máxima solicitada 0,1 %. El solicitante realizó análisis de residuos en muestras de lechugas y espinacas, no detectándose residuos de propilenglicol en ninguna de las muestras analizadas. En el caso de los fosfatos, las concentraciones detectadas en las muestras lavadas con el coadyuvante tecnológico no fueron significativamente distintas de las detectadas en las mismas muestras lavadas sin el coadyuvante. El Comité Científico concluye que, basándose en la información facilitada por el solicitante y teniendo en cuenta la composición y condiciones de uso propuestas, el uso del coadyuvante tecnológico no implica riesgo para la salud del consumidor. The company SmartWash Solutions, LLC has requested a safety assessment of the use of an aqueous solution of phosphoric acid (18-25 %) and propylene glycol (5-10 %) as a processing aid in the processing of fresh cut vegetables and ready-to-eat leafy vegetables. Both substances are authorised in the European Union as food additives in different food categories. The proposed use for the processing aid is as a pH stabilising solution in order to enhance the antimicrobial activity of the chlorine used for the washing water disinfection required in the processing of fresh cut vegetables and ready-to-eat leafy vegetables. The desired effect of this solution is to reduce the pH levels of the washing waters to levels between 5.5-6.5. The use concentration varies, depending on the requirements of the product and the chemical composition of the washing waters, with 0.1 % as the maximum concentration requested. The applicant analysed the residues on lettuce and spinach samples, and did not detect propylene glycol residues in any of the samples tested. In the case of phosphates, the detected concentrations in the samples washed with the processing aid were not significantly different from those detected in the same washed samples without the processing aid. The Scientific Committee concludes that, based on the information provided by the applicant and taking into account the proposed composition and conditions of use, the use of this processing aid does not involve a health risk for the consumer.

La Agencia Española de Consumo, Seguridad Alimentaria y Nutrición (AECOSAN) y las comuni dades autónomas han elaborado un Documento de priorización y distribución de muestras para control oficial de peligros químicos con el propósito de mejorar la calidad y homogeneidad de los controles oficiales en el marco del Plan Nacional de Control Oficial de la Cadena Alimentaria (PN COCA). Este Documento de priorización pretende distribuir la toma de muestras para control oficial en toda España, previa evaluación de los riesgos y de la capacidad analítica de los laboratorios. Ade más servirá de apoyo y orientación a las comunidades autónomas para que ejecuten sus progra mas de control oficial. El Documento de priorización define un modelo semicuantitativo, en el que se establecen las variables a las que se les asigna un valor numérico relativo para obtener una calificación final. La metodología de jerarquización se basa en aproximarse a la medida del riesgo dependiente de la presencia de un contaminante en un alimento, siendo el riesgo la suma de dos variables, el impacto en la salud de la población y la prevalencia en los productos de consumo. El Comité Científico concluye que el Documento de priorización y distribución de muestras de control oficial dirigidas a la determinación de peligros químicos es adecuado, en el momento actual, para el fin previsto. Este Documento deberá actualizarse periódicamente a la luz de la experiencia de su aplicación, el avance en los conocimientos científicos, los cambios en la legislación y las directrices y herramientas sobre priorización y muestreo que puedan desarrollarse a nivel nacional o de la Unión Europea. The Spanish Agency for Consumer Affairs, Food Safety and Nutrition (AECOSAN) and the autonomous communities have prepared a Document for the prioritisation and distribution of samples for the official control of chemical hazards with the aim of improving the quality and uniformity of official controls in the framework of the National Food Chain Official Control Plan (PNCOCA). The prioritisation Document aims to distribute the sampling for official controls throughout Spain, following prior assessment of the risks and the analytical capacity of the laboratories. It will also serve to provide support and guidance to the autonomous communities in the implementation of their official control programmes. The prioritisation Document defines a semi-quantitative model which sets out the variables to which a relative numerical value is allocated in order to obtain a final qualification. The hierarchical methodology is based on estimating the extent of the risk depending on the presence of a contaminant in a food, where the risk is the sum of two variables, the impact on the health of the population and the prevalence in consumer products. The Scientific Committee concludes that the Document for the prioritisation and distribution of official control samples aimed at determining chemical hazards is adequate, at present, for the intended purpose. This Document should be regularly updated in light of the experience obtained from its application, progress in scientific knowledge, changes in the legislation and the directives and tools on prioritisation and sampling which may be developed at national or European Union level.

La empresa Herba Ricemills S.L.U. ha solicitado la autorización de la comercialización en la Unión Europea de semillas de chía (Salvia hispanica) como ingrediente de platos preparados esterilizados elaborados a base de granos de cereales, pseudocereales y/o legumbres. Se trataría de una extensión de los usos autorizados para este nuevo alimento en 2009, 2013 y 2015. El Comité Científico considera que de la información aportada no se deduce que el consumo de las semillas de chía (Salvia hispanica) en platos preparados esterilizados basados en granos de cereales, pseudocereales y/o legumbres, en las condiciones propuestas por el solicitante, pueda producir efectos negativos para la salud, concluyendo que el nuevo alimento cumple los criterios de aceptación establecidos por el Reglamento (CE) Nº 258/97 sobre nuevos alimentos y nuevos ingredientes alimentarios (UE, 1997a). The company Herba Ricemills S.L.U. requested authorization to market chia (Salvia hispanica) seeds in sterilized ready to serve meals based on cereal, pseudocereals and/or pulse grains in the European Union. This would be an extension of use of the novel food authorized in 2009, 2013 and 2015. The AECOSAN Scientific Committee takes the view that, according to the information provided, there is no indication that consumption of chia (Salvia hispanica) seeds in ready to serve meals based on cereal, pseudocereals and/or pulse grains, under the conditions proposed by the applicant, can produce adverse effects on health. The Committee concludes that the novel food presented for assessment meets the criteria for acceptance laid down by Regulation (EC) No 258/97 concerning novel foods and novel food ingredients (UE, 1997a).

La empresa Sanchis Mira S.A. ha solicitado la autorización de la comercialización en la Unión Europea de semillas de chía (Salvia hispanica) en chocolate en tabletas. Se trataría de una extensión de los usos autorizados para este nuevo alimento en 2009, 2013 y 2015. El Comité Científico considera que de la información aportada no se deduce que el consumo de las semillas de chía (Salvia hispanica) en chocolate en tabletas, en las condiciones propuestas por el solicitante, pueda producir efectos negativos para la salud, concluyendo que el nuevo alimento cumple los criterios de aceptación establecidos por el Reglamento (CE) Nº 258/97 sobre nuevos alimentos y nuevos ingredientes alimentarios (UE, 1997a). The company Sanchis Mira S.A requested authorization to market chia (Salvia hispanica) seeds in chocolate bars in the European Union. This would be an extension of use of the novel food authorized in 2009, 2013 and 2015. The AECOSAN Scientific Committee takes the view that, according to the information provided, there is no indication that consumption of chia (Salvia hispanica) in chocolate bars, under the conditions proposed by the applicant, can produce adverse effects on health. The Committee concludes that the novel food presented for assessment meets the criteria for acceptance laid down by Regulation (EC) No 258/97 concerning novel foods and novel food ingredients (UE, 1997a).

Elements such as tourists, service-oriented organizations, tourism resources, destinations and public institutions all have a role in tourism activity. If we are to understand the world of tourism, we need to examine the ties between these elements. Social network analysis (SNA) provides tourism research with a set of methods and tools that allow us to comprehend the patterns and the structures of these ties. The objective of this work is to establish how SNA is being applied to tourism, to describe its principal elements and to inquire into its potential in the development of tourism research. To do so, the articles that apply SNA research methods, published in tourism and hospitality journals, and the network of citations between their authors are all analysed. It was found that the application of SNA in tourism-related contexts is rare and very recent, although a cohesive and relevant group of authors is currently applying it. An exciting opportunity for tourism research becomes apparent with the use o...

Airline alliances represent examples of resource utilization across a network. This paper examines the need to distinguish between access to and mobilization of the resources held by allies. An ordinary least squares regression was applied to a sample from the Top International Airlines database. Our findings show that mobilization of the destinations of the partner companies through codeshare alliances has a positive and significant influence on the performance of airlines. Moreover, they suggest that the development of network resources mobilizing capability in increasingly dynamic and global environments will only generate an important source of competitive advantage when acted upon.

The behaviour of early exporters not only challenges the perspective of the sequential process of internationalization, but it also questions general concepts in the field of management, such as the liability of newness and the liability of foreignness. This study analyses how firms initiate their export activities and proposes differences between early exporters and other firms. The results of our empirical investigation demonstrate that early exporters begin exporting to a greater number of countries than late exporters. However, in the case of early exporters, those countries are institutionally closer to the country of origin of the firm. Finally, we analyze the role played by financial resources in the choice of markets for those initial exports.

The airline sector is a mature industry in which a wide range of competitive practices reflect highly intense levels of rivalry. This is evident from the alliance portfolios of airline companies that are composed of mutually advantageous agreements between firms in the aviation sector. In this context, we examine the influence that access to valuable network resources may have on company performance (in operational and financial terms). Moreover, as network resources come under the resource dimension of a firm's social capital, we also study the way the resource dimension is affected by other dimensions of social capital (network position, network structure formed by the alliance portfolio and the quality of the relations). We study the alliances agreed between 214 mainstream airlines, up until 2007, during which time 351 airlines formed various types of partnerships. The results suggest, on the one hand, that access to partner resources is influenced by structural and relational factors. However, on the other hand, the structure of each airline's alliance portfolio has more influence than its position in the global network of alliances. These results point to the need for satisfactory governance of the composition and management of an airline's alliance portfolio.

Clonal gene therapy protocols based on the precise manipulation of epidermal stem cells require highly efficient gene-editing molecular tools. We have combined adeno-associated virus (AAV)-mediated delivery of donor template DNA with transcription activator-like nucleases (TALE) expressed by adenoviral vectors to address the correction of the c.6527insC mutation in the COL7A1 gene, causing recessive dystrophic epidermolysis bullosa in a high percentage of Spanish patients. After transduction with these viral vectors, high frequencies of homology-directed repair were found in clones of keratinocytes derived from a recessive dystrophic epidermolysis bullosa (RDEB) patient homozygous for the c.6527insC mutation. Gene-edited clones recovered the expression of the COL7A1 transcript and collagen VII protein at physiological levels. In addition, treatment of patient keratinocytes with TALE nucleases in the absence of a donor template DNA resulted in nonhomologous end joining (NHEJ)-mediated indel generation in the vicinity of the c.6527insC mutation site in a large proportion of keratinocyte clones. A subset of these indels restored the reading frame of COL7A1 and resulted in abundant, supraphysiological expression levels of mutant or truncated collagen VII protein. Keratinocyte clones corrected both by homology-directed repair (HDR) or NHEJ were used to regenerate skin displaying collagen VII in the dermo-epidermal junction.

T cells recognize antigens via their cell surface TCR and are classified as either αβ or γδ depending on the variable chains in their TCR, α and β or γ and δ, respectively. Both αβ and γδ TCRs also contain several invariant chains, including CD3δ, which support surface TCR expression and transduce the TCR signal. Mutations in variable chains would be expected to affect a single T cell lineage, while mutations in the invariant chains would affect all T cells. Consistent with this, all CD3δ-deficient patients described to date showed a complete block in T cell development. However, CD3δ-KO mice have an αβ T cell-specific defect. Here, we report 2 unrelated cases of SCID with a selective block in αβ but not in γδ T cell development, associated with a new splicing mutation in the CD3D gene. The patients' T cells showed reduced CD3D transcripts, CD3δ proteins, surface TCR, and early TCR signaling. Their lymph nodes showed severe T cell depletion, recent thymus emigrants in peripheral blood were strongly decreased, and the scant αβ T cells were oligoclonal. T cell-dependent B cell functions were also impaired, despite the presence of normal B cell numbers. Strikingly, despite the specific loss of αβ T cells, surface TCR expression was more reduced in γδ than in αβ T cells. Analysis of individuals with this CD3D mutation thus demonstrates the contrasting CD3δ requirements for αβ versus γδ T cell development and TCR expression in humans and highlights the diagnostic and clinical relevance of studying both TCR isotypes when a T cell defect is suspected.

Many studies have examined the relations between individuals and organizations and their influence on innovativeness. Some have looked at networks that improve innovative activity at an organizational, departmental and individual level, using in the latter case the individual's egonet. This study explores the way in which an individual's social capital and each of its three dimensions affect innovativeness. Having assessed the entire network of a university department and calculated the social capital of its members, their innovativeness was compared on the basis of their scientific production. The results show that the positions of researchers in the network structure and network quality are less important than the resources that they are able to access through their relations.

Here we report the functional assessment of two novel deafness-associated gamma-actin mutants, K118N and E241K, in a spectrum of different situations with increasing biological complexity by combining biochemical and cell biological analysis in yeast and mammalian cells. Our in vivo experiments showed that while the K118N had a very mild effect on yeast behaviour, the phenotype caused by the E241K mutation was very severe and characterized by a highly compromised ability to grow on glycerol as a carbon source, an aberrant multi-vacuolar pattern and the deposition of thick F-actin bundles randomly in the cell. The latter feature is consistent with the highly unusual spontaneous tendency of the E241K mutant to form bundles in vitro, although this propensity to bundle was neutralized by tropomyosin and the E241K filament bundles were hypersensitive to severing in the presence of cofilin. In transiently transfected NIH3T3 cells both mutant actins were normally incorporated into cytoskeleton structures, although cytoplasmic aggregates were also observed indicating an element of abnormality caused by the mutations in vivo. Interestingly, gene-gun mediated expression of these mutants in cochlear hair cells results in no gross alteration in cytoskeletal structures or the morphology of stereocilia. Our results provide a more complete picture of the biological consequences of deafness-associated gamma-actin mutants and support the hypothesis that the post-lingual and progressive nature of the DFNA20/26 hearing loss is the result of a progressive deterioration of the hair cell cytoskeleton over time.

Miguel Moreno-Pelayo and colleagues report mutations in the seed region of human miR-96 segregating with progressive hearing loss in two families. In an accompanying paper, Karen Steel and colleagues show that the mouse diminuendo mutant, which also shows progressive hearing loss, carries a similar mutation in the seed region of mouse miR-96. MicroRNAs (miRNAs) bind to complementary sites in their target mRNAs to mediate post-transcriptional repression1,2, with the specificity of target recognition being crucially dependent on the miRNA seed region3. Impaired miRNA target binding resulting from SNPs within mRNA target sites has been shown to lead to pathologies associated with dysregulated gene expression4,5,6,7. However, no pathogenic mutations within the mature sequence of a miRNA have been reported so far. Here we show that point mutations in the seed region of miR-96, a miRNA expressed in hair cells of the inner ear8, result in autosomal dominant, progressive hearing loss. This is the first study implicating a miRNA in a mendelian disorder. The identified mutations have a strong impact on miR-96 biogenesis and result in a significant reduction of mRNA targeting. We propose that these mutations alter the regulatory role of miR-96 in maintaining gene expression profiles in hair cells required for their normal function.

The TECTA gene encodes alpha-tectorin (TECTA), a major noncollagenous component of the tectorial membrane (TM). In humans, mutations in TECTA lead to either dominant (DFNA8/A12) or recessive (DFNB21) forms of nonsyndromic hearing loss. All missense mutations in TECTA that have been reported thus far are associated with the dominant subtype, whereas those leading to recessive deafness are all inactivating mutations. In this paper, we characterize a spontaneous missense mutation (c.1046C > A, p.A349D) arising in the mouse Tecta gene that is, unlike all previously reported missense mutations in TECTA, recessive. The morphological phenotype of the TectaA349D/A349D mouse resembles but is not identical to that previously described for the \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$Tecta^{{{\Delta {\text{ENT}}} \mathord{\left/ {\vphantom {{\Delta {\text{ENT}}} {\Delta {\text{ENT}}}}} \right. \kern-\nulldelimiterspace} {\Delta {\text{ENT}}}}} $$\end{document} mouse. As in the \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$Tecta^{{{\Delta {\text{ENT}}} \mathord{\left/ {\vphantom {{\Delta {\text{ENT}}} {\Delta {\text{ENT}}}}} \right. \kern-\nulldelimiterspace} {\Delta {\text{ENT}}}}} $$\end{document} mouse, the TM is completely detached from the surface of the organ of Corti and spiral limbus, lacks a striated-sheet matrix, and is deficient in both beta-tectorin (Tectb) and otogelin. A significant amount of Tecta is, however, detected in the TM of the TectaA349D/A349D mouse, and numerous, electron-dense matrix granules are seen interspersed among the disorganized collagen fibrils. Mutated TectaA349D is therefore incorporated into the TM but presumably unable to interact with either Tectb or otogelin. The TectaA349D/A349D mouse reveals that missense mutations in Tecta can be recessive and lead to TM detachment and suggests that should similar mutations arise in the human population, they would likely cause deafness.

Mutations in the potassium channel gene KCNQ4 underlie DFNA2, a subtype of autosomal dominant progressive, high-frequency hearing loss. Based on a phenotype-guided mutational screening we have identified a novel mutation c.886G>A, leading to the p.G296S substitution in the pore region of KCNQ4 channel. The possible impact of this mutation on total KCNQ4 protein expression, relative surface expression and channel function was investigated. When the G296S mutant was expressed in Xenopus oocytes, electrophysiological recordings did not show voltage-activated K+ currents. The p.G296S mutation impaired KCNQ4 channel activity in two manners. It greatly reduced surface expression and, secondarily, abolished channel function. The deficient expression at the cell surface membrane was further confirmed in non-permeabilized NIH-3T3 cells transfected with the mutant KCNQ4 tagged with the hemagglutinin epitope in the extracellular S1-S2 linker. Co-expression of mutant and wild type KCNQ4 in oocytes was performed to mimic the heterozygous condition of the p.G296S mutation in the patients. The results showed that the G296S mutant exerts a strong dominant-negative effect on potassium currents by reducing the wild type KCNQ4 channel expression at the cell surface. This is the first study to identify a trafficking-dependent dominant mechanism for the loss of KCNQ4 channel function in DFNA2. © Springer-Verlag 2007.

The biological role in vivo of the homologous CD3 gamma and delta invariant chains within the human TCR/CD3 complex is a matter of debate, as murine models do not recapitulate human immunodeficiencies. We have characterized, in a Turkish family, two new patients with complete CD3 gamma deficiency and SCID symptoms and compared them with three CD3 gamma-deficient individuals belonging to two families from Turkey and Spain. All tested patients shared similar immunological features such as a partial TCR/CD3 expression defect, mild alpha beta and gamma delta T lymphocytopenia, poor in vitro proliferative responses to Ags and mitogens at diagnosis, and very low TCR rearrangement excision circles and CD45RA(+) alpha beta T cells. However, intrafamilial and interfamilial clinical variability was observed in patients carrying the same CD3G mutations. Two reached the second or third decade in healthy conditions, whereas the other three showed lethal SCID features with enteropathy early in life. In contrast, all reported human complete CD3 delta (or CD3 epsilon) deficiencies are in infants with life-threatening SCID and very severe alpha beta and gamma delta T lymphocytopenia. Thus, the peripheral T lymphocyte pool was comparatively well preserved in human CD3 gamma deficiencies despite poor thymus output or clinical outcome. We propose. a CD3 gamma >> CD3 gamma hierarchy for the relative impact of their absence on the signaling for T cell production in humans.

Epidermolysis bullosa with pyloric atresia (EB-PA) is a rare autosomal recessive hereditary disease with a variable prognosis from lethal to very mild. EB-PA is classified into Simplex form (EBS-PA: OMIM #612138) and Junctional form (JEB-PA: OMIM #226730), and it is caused by mutations in ITGA6, ITGB4 and PLEC genes. We report the analysis of six patients with EB-PA, including two dizygotic twins. Skin immunofluorescence epitope mapping was performed followed by PCR and direct sequencing of the ITGB4 gene. Two of the patients presented with non-lethal EB-PA associated with missense ITGB4 gene mutations. For the other four, early postnatal demise was associated with complete lack of β4 integrin due to a variety of ITGB4 novel mutations (2 large deletions, 1 splice-site mutation and 3 missense mutations). One of the deletions spanned 278 bp, being one of the largest reported to date for this gene. Remarkably, we also found for the first time a founder effect for one novel mutation in the ITGB4 gene. We have identified 6 novel mutations in the ITGB4 gene to be added to the mutation database. Our results reveal genotype-phenotype correlations that contribute to the molecular understanding of this heterogeneous disease, a pivotal issue for prognosis and for the development of novel evidence-based therapeutic options for EB management.

Los complementos alimenticios son alimentos cuyo fin es complementar la dieta normal y que consisten en fuentes concentradas de nutrientes (vitaminas y minerales) o de otras sustancias que tienen un efecto nutricional o fisiológico, en forma simple o combinada. Los complementos se comercializan en forma dosificada, se entregan al consumidor final únicamente preenvasados. En ningún caso, deben sustituir al uso de medicamentos sin una supervisión médica adecuada. Sólo deben utilizarse para complementar la dieta y, de forma general, su uso no es necesario si se sigue una dieta variada y equilibrada, a la que no pueden reemplazar. En España los complementos alimenticios están regulados por el Real Decreto 1487/2009 que traspuso a la legislación española la Directiva 2002/46/CE relativa a la aproximación de las legislaciones de los estados miembros en materia de complementos alimenticios. Sin embargo, actualmente sólo está regulado el uso de vitaminas y minerales, por lo que se ha solicitado al Comité Científico que realice una valoración de la propuesta de autorización de la utilización de determinadas sustancias distintas de vitaminas y minerales en la fabricación de complementos alimenticios. Las sustancias propuestas por la Agencia Española de Consumo, Seguridad Alimentaria y Nutrición (AECOSAN) son: ácido L-aspártico, L-citrulina, glicina, L-prolina, L-serina, L-arginina-L-aspartato, L-lisina-L-aspartato, L-lisina-L-glutamato, N-acetil-L-cisteína, N-acetil-L-metionina, hidroximetilbu- tirato, ácido lipoico, Monascus purpureus, carbón activo y lactulosa. El Comité Científico ha valorado cada propuesta, analizando las características y fuentes de cada sustancia, así como la nutrición, metabolismo y seguridad y ha concluido, en cada caso, si la presentada por la AECOSAN era aceptable desde el punto de vista de su seguridad en su uso como complemento alimenticio. En ningún caso, la evaluación realizada supone un aval de la eficacia biológica de las sustancias y dosis valoradas. El Comité Científico indica que, en todo caso es necesario que las personas que estén sometidas a tratamientos con medicamentos consulten con su médico la oportunidad o conveniencia de consumir complementos alimenticios dada la posibilidad de que existan interferencias en algunos casos. Food supplements are foods, the purpose of which is to supplement the normal diet and which consist of concentrated nutrient sources (vitamins and minerals) or other substances with a nutritional or physiological effect, alone or in combination. The supplements are marketed in dosage form and are only supplied to the end consumer prepacked. In no event should they replace the use of medicines without suitable medical supervision. They should only be used to supplement the diet and, on the whole, their usage is not required if the individual has a varied and balanced diet, which cannot be replaced. In Spain, food supplements are regulated by Royal Decree 1487/2009, which transposed Directive 2002/46/EC on the approximation of the laws of the Member States relating to food supplements into Spanish law. However, only the use of vitamins and minerals is currently regulated. Therefore the Scientific Committee has been asked to make an assessment of the proposal to authorise certain substances other than vitamins and minerals in the manufacture of food supplements. The substances proposed by the Spanish Agency for Consumer Affairs, Food Safety and Nutrition (AECOSAN) are food supplements, L-aspartic acid, L-citrulline, glycine, L-proline, L-serine, L-arginine-L-aspar- tate, L-lysine L-aspartate, L-lysine-L-glutamate, N-acetyl-L-cysteine, N-acetyl-L-methionine, hidro- xymethylbutyrate, lipoic acid, Monascus purpureus, activated carbon and lactulose. The Scientific Committee has assessed each proposal, analysing the characteristics and sources of each substance, and the nutrition, metabolism and safety and has concluded, in each case, whether that submitted by the AECOSAN is acceptable from a safety viewpoint for use as a food supplement. In no event is the assessment intended as a guarantee of the biological efficiency of the substances and doses assessed. The Scientific Committee states that, in any case individuals undergoing medical treatment must seek medical advice as to the suitability of taking food supplements, given the possibility of interactions in certain cases.

Tribulus terrestris L. es una planta de la familia Zygophyllaceae cuyo uso en complementos alimenticios está autorizado en varios países de la Unión Europea y que contiene de manera natural varias sustancias activas, entre las que destacan saponósidos esteroídicos, alcaloides b-carbolínicos, favonoides y lignanamidas. La toxicidad de Tribulus terrestris en animales ha sido ampliamente documentada a lo largo de su historia como planta medicinal y se describen efectos negativos a nivel neuronal, muscular, hepático y renal. Algunos países consideran que utilizar partes de la planta de Tribulus terrestris en la elaboración de complementos alimenticios no es seguro, por lo que impiden su comercialización y lo comunican al sistema de alertas europeo RASFF (Rapid Alert System for Food and Feed). El Comité Científico ha realizado una evaluación del riesgo a fin de determinar si el consumo de frutos, partes aéreas de la planta y sus extractos de Tribulus terrestris en complementos alimenticios es seguro y ha concluido que no hay disponibles datos toxicológicos suficientes que permitan evaluar la seguridad del uso de partes de la planta de Tribulus terrestris en complementos alimenticios. En cualquier caso, se considera que las cantidades máximas diarias de Tribulus terrestris en complementos alimenticios no deberían superar las dosis de uso farmacológico y que en los complementos debería constar la parte de la planta empleada, y si se trata de un extracto u otra forma de preparación, así como el contenido en saponósidos. Tribulus terrestris L. is a plant from the Zygophyllaceae family whose use in food supplements is authorised in various European Union countries. In its natural form, it contains various active substances, the most notable of which are steroidal saponins, b-Carboline alkaloids, flavonoids and lignanamides. Tribulus terrestris’ toxicity to animals has been widely documented throughout its history as a medicinal plant and it has been reported to have negative neuronal, muscular, hepatic and renal effects. Some countries consider using parts of the Tribulus terrestris plant to produce food supplements to be unsafe so they do not allow such supplements to be sold. This is reported through the European Rapid Alert System for Food and Feed (RASFF). The Scientific Committee has conducted a risk assessment with the aim of determining whether consuming fruit, plant shoots and their extracts from Tribulus terrestris in food supplements is safe, concluding that there is insufficient toxicological data to assess how safe using parts of the Tribulus terrestris plant in food supplements is. In any case, the maximum daily quantity of Tribulus terrestris in food supplements should not exceed the dose used for pharmacological purposes and the part of the plant used, whether it was extracted or prepared in some other way and its saponin content should all be made clear on the supplement itself.

Alliance Portfolio Configuration (APC) conditions access to network resources, however, not all access to partner resources is finally mobilized by the firm. Our paper contributes to the understanding of alliance portfolio performance by examining how an acceptable configuration of the alliance portfolio will be conditioned by the level of resource mobilization that the firm really achieves. A variance-based structural equation modelling (Partial Least Squares) has been applied to a sample from the Top International Airlines database. Results from the data analysis show that the Level of Network Resource Mobilization (LNRM) (a) fully mediate the effect of alliance portfolio configuration on the operating performance; and (b) partially mediate this effect on financial performance of airlines.

T lymphocytes play a crucial role in the pathogenesis of inflammatory bowel disease. Achieving stable T-cell lines, rather than continuous bleeding of patients, is desirable in order to dissect their implication in the disease.Long-lasting T-cell lines from patients with Crohn disease and ulcerative colitis and from healthy volunteers have been obtained by transformation of T lymphocytes using the lymphotropic Herpesvirus saimiri. Lines were subjected to phenotypic and functional analyses, and the results compared with freshly isolated peripheral blood mononuclear cells.Fresh cells revealed only minor differences between patients and controls, with regard to phenotype and proliferative capacity. In contrast, the use of T-cell lines showed that cells from Crohn disease patients, but not ulcerative colitis patients, over-responded to several membrane or cytoplasmic stimuli when compared to control T-cell lines. Thus, higher responses were found when stimulated with alphaCD3 and IL2, alphaCD3 and alphaCD28, IL2 alone, phorbol esters (PMA) and alphaCD3 and, finally, PMA and alphaCD2 (P0.05 in all instances). Further, lines from patients with Crohn disease responded more vigorously to alphaCD3 and alphaCD28 or alphaCD3 and PMA when compared to ulcerative colitis (P0.05 in both instances).The data obtained with these lines suggest that T cells from patients with Crohn disease differ in vivo in their proliferative capacity, as compared with those from ulcerative colitis patients, a finding that may reflect the clear Th-1 phenotype found in the former and absent in the latter.

Hereditary non-syndromic sensorineural hearing loss (NSSHL) is a genetically highly heterogeneous group of disorders. Autosomal dominant forms account for up to 20% of cases. To date, 39 loci have been identified by linkage analysis of affected families that segregate NSSHL forms in the autosomal dominant mode (DFNA). Investigation of a large Spanish pedigree with autosomal dominant inheritance of bilateral and progressive NSSHL of postlingual onset excluded linkage to known DFNA loci and, in a subsequent genome-wide scan, the disorder locus was mapped to 3q28-29. A maximum two-point LOD score of 4.36 at theta=0 was obtained for marker D3S1601. Haplotype analysis placed the novel locus, DFNA44, within a 3-cM genetic interval defined by markers D3S1314 and D3S2418. Heteroduplex analysis and DNA sequencing of coding regions and exon/intron boundaries of two genes (CLDN16 and FGF12) in this interval did not reveal disease-causing mutations.

To dissect the phenotypic and functional features of mucosal T lymphocytes in patients with gastric adenocarcinoma, we have used the Herpesvirus saimiri transformation procedure to achieve T-cell lines from gastric origin. Once achieved, cell function was assessed by in vitro stimulation with mitogens. CD2-specific monoclonal antibodies (alpha-CD2), alone or in combination with interleukin (IL)-2, rendered fewer counts in patients (34 408+/-3965 and 52 157+/-6473 c.p.m., respectively) than in controls (67 471+/-11 755 c.p.m., P

et al., Recessive dystrophic epidermolysis bullosa (RDEB) is caused by deficiency of type VII collagen due to COL7A1 mutations such as c.6527insC, recurrently found in the Spanish RDEB population. Assessment of clonal correction-based therapeutic approaches for RDEB requires large expansions of cells, exceeding the replication capacity of human primary keratinocytes. Thus, immortalized RDEB cells with enhanced proliferative abilities would be valuable. Using either the SV40 large T antigen or papillomavirus HPV16-derived E6-E7 proteins, we immortalized and cloned RDEB keratinocytes carrying the c.6527insC mutation. Clones exhibited high proliferative and colony-forming features. Cytogenetic analysis revealed important differences between T antigen-driven and E6-E7-driven immortalization. Immortalized cells responded to differentiation stimuli and were competent for epidermal regeneration and recapitulation of the blistering RDEB phenotype in vivo. These features make these cell lines useful to test novel therapeutic approaches including those aimed at editing mutant COL7A1., FL was supported by grants from Instituto de Salud Carlos III (PI11/01225) and Comunidad de Madrid (S2010/BMD- 2359; SKINMODEL). MDR was supported by grants from the Science and Innovation Ministry of Spain (SAF2010-16976) and Comunidad de Madrid (S2010/BMD-2420; CELLCAM). MG is supported in part by ERA-NET grant: E-Rare-2 (SpliceEB).