This dissertation studies the formation of the 1959 Obscene Publications Act with specific focus on the role of the publisher in its development. The 1950s contained a spate of obscenity trials alongside a five year struggle by the Society of Authors and MP Roy Jenkins to reform the Act and pass it through Parliament. A central question of this dissertation is with what power were publishers able to make an impact on the Act? Utilising the theories of Bourdieu and Foucault, this study aims to show that cultural and political capital were absolutely necessary to be able to be qualified to make significant societal contributions, particularly in the law. With Bourdieu’s theory of cultural capital and symbolic power, it is clear that publishers in the 1950s were able to use their positions as cultural agents to influence the public’s and the press’ attitudes to obscenity and censorship. Publishers studied in this dissertation are Alexander Stuart Frere, Fredric Warburg, Stanley Unwin, George Weidenfeld, and Nigel Nicolson. Using archival material like letters, court transcripts, articles and memoranda, this dissertation shows how they used their power and relationships with other important figures to both influence the outcome of their obscenity trials and change opinions on obscenity. Through exploring the material and analysing the role of the publisher—as well as the politician—this dissertation places these figures within the context of the 1950s and the development of the 1959 Obscene Publications Act, and looks forward to how their work against censorship and for freedom of expression manifested in its final form and influenced the 1960 trial of Lady Chatterley’s Lover.
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Reportatge en memòria del filòsof Josep Ferrater Mora. Recull la inauguració de la Càtedra Ferrater Mora de Pensament Contemporani, lliçò a càrrec del professor W.V.Quine, i les cròniques de la mort del filòsof a la televisió autonòmica, TV3, i a l'estatal, TV2. Finalitza amb una entrevista al director de la càtedra, el professor Josep Mª Terricabras que parla del llegat de Ferrater 7.mp4 7.mp3
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Polymer networks featuring dynamic bonds offer a promising solution to address key challenges in polymer materials, especially in terms of sustainability. The dynamic covalent chemistries enhance processability, enable recycling of the reversibly cross- linked thermosets and elastomers, and offer repairability. Over the last 10 years, the maturity of these networks has exponentially increased. However, they still haven’t made it into the mass consumer market. To stand a chance in the competitive polymer industry, dynamic polymer networks need to further exploit their advantageous properties and enhanced performance in combination with a focus on sustainability. This overarching objective has been divided in three stages, where the results of each stage were sequentially added to the next one. The first main focus is sustainability in all its aspects. Dynamic covalent polymer networks with self-healing properties were developed using the 12 principles of green chemistry as the designing driving force. The sustainability was tackled from a holistic point of view, taking into account the whole life cycle of the material (circular economy). The developed self-healing elastomers account for the sustainability of (1) the raw materials and (2) solvents, (3) the reusability, (4) the (re)processability, and (5) the end of life of the material. They are synthesized by a simple one-pot, solventless synthesis from commercially available reagents, they can be reprocessed and recycled, they autonomously heal at room temperature, and they can be hydrolytically degraded at the end of their service life. The objective of the second stage was to enhance the performance of dynamic covalent networks. Starting from the materials and the synthesis processes developed in the first stage, the trade-off between self-healing/reprocessability and creep resistance was minimized. Solving this trade-off has been a priority in the field, and it is currently the main performance challenge of dynamic polymer networks. This was achieved by combining a dissociative and an associative dynamic covalent chemistry in the same polymer network. This combination enables the fabrication of materials where the timescale at which they relax can be designed independently of its mechanical properties and tailored for the application and lifetime of the material. Finally, these materials were used in several applications where the dynamics bonds bring a performance advantage over traditional polymers. The materials were used in soft robotics, smart textiles, electronic skin, and 3D printing, proving its applicability in key technologies for the future without giving up on sustainability.
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handle: 10481/86920
This article proposes a hierarchy of translator and reviser competences in prototypical scenarios in legal translation with a view to determining the most appropriate revision foci to ensure translation quality. Built on a prior characterisation of the most common professional translator profiles in legal translation, the proposal for a hierarchy of competences derives from two premises: (1) The professional profile of those who translate and revise legal documents is very diverse in terms of competence and qualifications (training and experience), and (2) translation competence and specialist knowledge in legal fields (i.e., domain competence) are fundamental when revising to guarantee the quality of legal translations. The proposal is framed by quality assurance in legal translation through a revision process based on (1) the coherent management of the work of the translators and revisers involved in the translation project, and (2) the appropriate methodology for revision applied to legal translation by adapting the revision mode’s focus to ensure its effectiveness. Six common scenarios are identified in light of the translators’ profiles, for which revisers’ profiles are then proposed in order to detect any legal translation competence deficiencies among translators, and thus ensure quality. Firmé un contrato de cesión de derechos de autor a la editorial John Benjamins Publishing Company. No obstante, el artículo está disponible utilizando la VPN. Espero poder obtener el handle de los metadatos del artículo o de la copia off-print que he subido porque lo necesito para indicarlo en la solicitud de un sexenio de investigación. El archivo con la copia off-print del artículo que he subido indica lo siguiente: This electronic file is a contribution from Target 33:2 and may not be altered in any way. The author of this article is permitted to use this PDF file to generate printed copies to be used by way of offprints, for their personal use only. Permission is granted by the publishers to post this file on a closed server which is accessible onlty to members (students and faculty) of the author's institute. It is not permitted to post this PDF on teh internet, or to share it on sites such as Medeley, ResearchGate, Academia.edu. For any other use of this material prior written permission should be obtained from the publishers (John Benjamins Publishing Company) or through the Copyright Clearance Center. Gracias por su colaboración.
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Several studies have described the potential role of microRNA (miRNAs) in migraine but studies are scarcely reproducible primarily due to the heterogeneous variability of participants. Hence, the main objective of this project is to identify and validate differentially expressed miRNAs (DE miRNAs) in peripheral blood mononuclear cells (PBMCs) of patients with migraine compared to healthy controls (HC) in a well-controlled homogeneous cohort of participants using a three-step methodology: (a) exploratory study, (b) main study and (c) validation study. During the exploratory study, effect size was estimated in order to calculate the required sample size for the main and validation studies Patients diagnosed with migraine according to the International Classification of Headache Disorders and HC without family history of headache disorders were recruited. All participants completed a very thorough questionnaire and structured-interview in order to control environmental factors. RNA was extracted from PBMC and a microarray system was used to determine differential miRNA profiles between study groups in both exploratory and main studies. Principal components analysis and hierarchical clustering analysis were performed to study samples' distribution and to identify potential Batch effect. DE miRNAs were identified adjusting linear model with empirical Bayes moderated t-statistic and a classification task was performed for predictors selection using a supervised machine learning method widely used in the microarray analysis (RF, random forest models), with a bootstrapping algorithm in order to evaluate the stability of classification's results. During the validation study, a quantitative RT-PCR (qRT-PCR) procedure was performed of the DE miARNs identified during the main study and their relative quantification (RQ) was computed in terms of the difference between DE miRNAs and the housekeeping transcript (hsa-miR-93-5p) between study groups. Then, multivariate generalized mixed-effect regression models were estimated in order to study whether there were differences in the RQ of each DE miRNA between study groups, adjusted by covariates. Finally, a functional enrichment analysis of selected targets was computed through protein-protein interaction networks. A total of 313 participants (108 HC and 205 patients) were included during all the project and 23 DE miRNA (5 over-expressed and 18 under-expressed) were identified after the classification task as predictors which differentiate study groups (HC vs. patients with migraine). The RF model showed an accuracy [95% CI] of 87.8% [73.8% - 95.9%] with a 100.0% sensitivity, 84.4% specificity and kappa coefficient of 0.703. The area under ROC curve obtained was 0.821. Then, the top three miARNs (miR-DOWN006, miR-DOWN002 and miR-DOWN017) from this signature were validated through qRT-PCR in an external cohort of participants. Only miR-DOWN006 and miR-DOWN002 were under-expressed in the 97.2% and 87.3% of participants, respectively. Afterwards, multivariate mixed-effect regression models were fitted for each miRNA and we found that the group variable resulted statistically significant for both transcripts. Enrichment functional analysis showed that these 2 miARNs were closely associated with known migraine pathophysiology: hormone (estrogens, prolactin, oxytocin and cortisol) signaling pathways, dopaminergic and glutamatergic synapses, inflammatory signaling pathways (TGF-, chemokines, apelin, renin and Toll-like receptors), energy homeostasis (AMPK and mTOR signaling pathways), neuronal development (axon guidance and Notch signaling pathway) and environmental adaptation mechanisms (circadian rhythms). Hence, the results of this project will help us to develop the first kit for migraine diagnosis relying on quantitative indicators. This way we will be able (i) to reduce the stigma associated with the disease, (ii) to better phenotype patients and (iii) to treat them earlier. Moreover, the data gathered in this thesis will allow us not only to improve our understanding of migraine pathophysiology but also to identify new targets in order to develop future therapeutic approaches. Varios estudios piloto han estudiado el rol de los microARNs (miARNs) en la migraña. Sin embargo, los estudios son poco comparables y reproducibles debido al alto grado de heterogeneidad que existe entre los grupos. Así pues, el objetivo de la presente tesis es identificar y validar un patrón de expresión diferencial de miARNs en células mononucleares de la sangre periférica (PBMC) en una cohorte muy bien controlada de individuos con migraña y controles sanos (HC). Para llevar a cabo este objetivo, se ha planteado un diseño experimental basado en tres estudios secuenciales: (a) una prueba exploratoria para estimar el efecto del análisis y ajustar el tamaño de la muestra en (b) el estudio principal y, finalmente, (c) replicar los hallazgos en una tercera etapa, en un estudio de validación. Se reclutó individuos diagnosticados de migraña y HC que negaban tener antecedentes familiares de primer o segundo grado con cefalea recurrente. Todos los participantes completaron también un cuestionario muy exhaustivo para controlar los factores ambientales. Se realizó la extracción del ARN y se estudió el perfil de expresión diferencial de los miARNs mediante microchips específicos durante los estudios exploratorio y principal. Realizamos un análisis de las componentes principales y un análisis de agrupamiento jerárquico para estudiar la distribución de las muestras y detectar un posible efecto Batch. El análisis diferencial se realizó mediante el ajuste de modelos lineales basados en un enfoque Bayesiano de la prueba t de Student moderada. La selección de los mejores transcritos se llevó a cabo mediante una tarea de clasificación basada en árboles de decisión que recibe el nombre de Random Forest (RF), un algoritmo de clasificación supervisado, y un método de remuestreo (bootstrapping) para controlar la estabilidad de los resultados. Para la validación de los miARNs seleccionados, se llevó a cabo una amplificación y secuenciación en una cohorte externa mediante qRT-PCR. Una vez estimada la cuantificación relativa (RQ) de cada miARN, se ajustó modelos lineales generalizados de efectos mixtos para estudiar si había diferencias en la expresión de cada miARN entre los grupos de estudio. Finalmente, realizamos una predicción in silico de las dianas génicas de este perfil de miARNs e identificamos las redes de interacción proteína-proteína asociadas a estos genes para encontrar las subredes ontológicas más representativas. Un total de 313 participantes (108 HC y 205 pacientes) fueron incluidos durante los tres sub-estudios de este proyecto y, después de realizar la tarea de clasificación, identificamos un total de 23 DE miARNs (5 sobre-expresados y 18 infra-expresados) entre los controles y los pacientes. El modelo de clasificación presentó una precisión del 87,8% (IC95%, 73,8% - 95,9%). Para llevar a cabo la validación, decidimos cuantificar los 3 mejores miARNs: miR-DOWN006, miR-DOWN002 y miR-DOWN017. Aun así, solo obtuvimos la expresión del miR-DOWN006, que se expresó en el 97,2% de las muestras y del miR-DOWN002, que se expresó en el 87,3%; ambos transcritos infra-expresados en individuos diagnosticados de migraña. El análisis funcional de estos 2 miARNs enriqueció vías ontológicas que se han asociado previamente con la migraña: vías de señalización hormonal, vías de neurotransmisión sináptica, vías asociadas a la inflamación, vías asociadas a la homeostasis energética, vías asociadas al desarrollo neuronal y mecanismos de adaptación al ambiente. Los datos aportados por esta tesis nos pueden ayudar a desarrollar el primer kit para diagnosticar la migraña mediante el uso de indicadores cuantitativos. De este modo, podremos (i) reducir el estigma asociado a la enfermedad, (ii) diagnosticar mejor a los pacientes y (iii) a tratarlos precozmente. Diversos estudis pilot han estudiat el rol dels microARNs (miARNs) en la migranya. No obstant això, els estudis són poc comparables i reproduïbles degut a l'alt grau d'heterogeneïtat que existeix entre els grups. Així doncs, l'objectiu de la present tesi és identificar i validar un patró d'expressió diferencial de miARNs en cèl·lules mononuclears de la sang perifèrica (PBMC) en una cohort molt ben controlada d'individus amb migranya i controls sans (HC). Per dur a terme aquest objectiu, s'ha plantejat un disseny experimental basat en tres estudis seqüencials: (a) una prova exploratòria per a estimar l'efecte de l'anàlisi i ajustar la grandària de la mostra en (b) l'estudi principal i, finalment, (c) replicar les troballes en una tercera etapa en un estudi de validació. Es va reclutar individus diagnosticats de migranya i HC que negaven tenir antecedents familiars de primer o segon grau amb cefalea recurrent. Tots els participants van completar també un qüestionari molt exhaustiu per a controlar els factors ambientals. Es va realitzar l'extracció de l'ARN i es va estudiar el perfil d'expressió diferencial de miARNs mitjançant microxips específics durant els estudis exploratori i principal. Vam realitzar una anàlisi de les components principals i una anàlisis de clusterització jeràrquica per a estudiar la distribució de les mostres i detectar un possible efecte Batch. L'anàlisi diferencial es va realitzar mitjançant l'ajustament de models lineals basats en un enfocament Bayesià de la prova t de Student moderada. La selecció dels millors transcrits es va dur a terme mitjançant una tasca de classificació basada en arbres de decisió que rep el nom de Random Forest (RF), un algoritme de classificació supervisat, i un mètode de remostreig (bootstrapping) per a controlar l'estabilitat dels resultats. Per a la validació dels miARNs seleccionats, es va dur a terme una amplificació i seqüenciació en una cohort externa mitjançant qRT-PCR. Una vegada estimada la quantificació relativa (RQ) de cada transcrit, es va ajustar models lineals generalitzats d'efectes mixtes per a estudiar si hi havia diferències en l'expressió de cada miARN entre els grups d'estudi. Finalment, vam predir in silico els gens diana d'aquest perfil de miARN i vam identificar les xarxes d'interacció proteïna-proteïna associades a aquests gens per tal de trobar les subxarxes ontològiques més representatives. Un total de 313 participants (108 HC i 205 pacients) van ser inclosos durant els tres sub-estudis d'aquest projecte i, després de realitzar la tasca de classificació, vam identificar un total de 23 DE miARNs (5 sobre-expressats i 18 infra-expressats) entre els controls i els pacients. El model de classificació va presentar una precisió del 87,8% (IC95%, 73,8% - 95,9%). Per a dur a terme la validació, vam decidir quantificar els 3 millors miARNs de la signatura dels 23: miR-DOWN006, miR-DOWN002 i el miR-DOWN017. Tanmateix, només vam obtenir la RQ del miR-DOWN006, que es va expressar en el 97,2% de les mostres i del miR-DOWN002, que es va expressar en el 87,3%; tots dos transcrits infra-expressats en individus diagnosticats de migranya. L'anàlisi funcional d'aquests 2 miARNs va enriquir vies ontològiques que s'han associat prèviament amb la migranya: vies de senyalització hormonal, vies de neurotransmissió sinàptica, vies associades a la inflamació, vies associades a l'homeòstasi energètica , vies associades al desenvolupament neuronal i mecanismes d'adaptació a l'ambient. Les dades aportades per aquesta tesi ens poden ajudar a desenvolupar el primer kit per a fenotipar la migranya mitjançant l'ús d'indicadors quantitatius. D'aquesta manera, podrem (i) reduir l'estigma associat a la malaltia, (ii) diagnosticar millor als pacients i (iii) a tractar-los precoçment.
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handle: 10803/687499 , 20.500.11797/TDX4022
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handle: 2158/1359572
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handle: 10481/87660
A partir de los sistemas de organización interna y de las estructuras de poder en las comunidades rurales del mundo nazarí y en las aljamas castellanas se intenta explicar cómo la sociedad islámica andalusí se va apartando de sus precedentes medievales, integrándose cada vez más en los nuevos sistemas estatales del mundo moderno. Un contexto temporal suficientemente amplio y dilatado sirve para identificar elementos de continuidad e indicios de desvinculación progresiva, en definitiva, trazar los perfiles de un proceso tan complejo como la disolución de un sistema social y su reconversión, o su integración, en otro radicalmente diferente. Por eso el contenido de esta obra abarca el mundo nazarí, las prolongaciones de su organización estatal en el conjunto de la sociedad, y la situación de las minorías islámicas en el reino de Castilla, estudiadas a través de sus espacios de convivencia y de su forma de articular el poder político y económico.
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This dissertation studies the formation of the 1959 Obscene Publications Act with specific focus on the role of the publisher in its development. The 1950s contained a spate of obscenity trials alongside a five year struggle by the Society of Authors and MP Roy Jenkins to reform the Act and pass it through Parliament. A central question of this dissertation is with what power were publishers able to make an impact on the Act? Utilising the theories of Bourdieu and Foucault, this study aims to show that cultural and political capital were absolutely necessary to be able to be qualified to make significant societal contributions, particularly in the law. With Bourdieu’s theory of cultural capital and symbolic power, it is clear that publishers in the 1950s were able to use their positions as cultural agents to influence the public’s and the press’ attitudes to obscenity and censorship. Publishers studied in this dissertation are Alexander Stuart Frere, Fredric Warburg, Stanley Unwin, George Weidenfeld, and Nigel Nicolson. Using archival material like letters, court transcripts, articles and memoranda, this dissertation shows how they used their power and relationships with other important figures to both influence the outcome of their obscenity trials and change opinions on obscenity. Through exploring the material and analysing the role of the publisher—as well as the politician—this dissertation places these figures within the context of the 1950s and the development of the 1959 Obscene Publications Act, and looks forward to how their work against censorship and for freedom of expression manifested in its final form and influenced the 1960 trial of Lady Chatterley’s Lover.
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Reportatge en memòria del filòsof Josep Ferrater Mora. Recull la inauguració de la Càtedra Ferrater Mora de Pensament Contemporani, lliçò a càrrec del professor W.V.Quine, i les cròniques de la mort del filòsof a la televisió autonòmica, TV3, i a l'estatal, TV2. Finalitza amb una entrevista al director de la càtedra, el professor Josep Mª Terricabras que parla del llegat de Ferrater 7.mp4 7.mp3
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Polymer networks featuring dynamic bonds offer a promising solution to address key challenges in polymer materials, especially in terms of sustainability. The dynamic covalent chemistries enhance processability, enable recycling of the reversibly cross- linked thermosets and elastomers, and offer repairability. Over the last 10 years, the maturity of these networks has exponentially increased. However, they still haven’t made it into the mass consumer market. To stand a chance in the competitive polymer industry, dynamic polymer networks need to further exploit their advantageous properties and enhanced performance in combination with a focus on sustainability. This overarching objective has been divided in three stages, where the results of each stage were sequentially added to the next one. The first main focus is sustainability in all its aspects. Dynamic covalent polymer networks with self-healing properties were developed using the 12 principles of green chemistry as the designing driving force. The sustainability was tackled from a holistic point of view, taking into account the whole life cycle of the material (circular economy). The developed self-healing elastomers account for the sustainability of (1) the raw materials and (2) solvents, (3) the reusability, (4) the (re)processability, and (5) the end of life of the material. They are synthesized by a simple one-pot, solventless synthesis from commercially available reagents, they can be reprocessed and recycled, they autonomously heal at room temperature, and they can be hydrolytically degraded at the end of their service life. The objective of the second stage was to enhance the performance of dynamic covalent networks. Starting from the materials and the synthesis processes developed in the first stage, the trade-off between self-healing/reprocessability and creep resistance was minimized. Solving this trade-off has been a priority in the field, and it is currently the main performance challenge of dynamic polymer networks. This was achieved by combining a dissociative and an associative dynamic covalent chemistry in the same polymer network. This combination enables the fabrication of materials where the timescale at which they relax can be designed independently of its mechanical properties and tailored for the application and lifetime of the material. Finally, these materials were used in several applications where the dynamics bonds bring a performance advantage over traditional polymers. The materials were used in soft robotics, smart textiles, electronic skin, and 3D printing, proving its applicability in key technologies for the future without giving up on sustainability.
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handle: 10481/86920
This article proposes a hierarchy of translator and reviser competences in prototypical scenarios in legal translation with a view to determining the most appropriate revision foci to ensure translation quality. Built on a prior characterisation of the most common professional translator profiles in legal translation, the proposal for a hierarchy of competences derives from two premises: (1) The professional profile of those who translate and revise legal documents is very diverse in terms of competence and qualifications (training and experience), and (2) translation competence and specialist knowledge in legal fields (i.e., domain competence) are fundamental when revising to guarantee the quality of legal translations. The proposal is framed by quality assurance in legal translation through a revision process based on (1) the coherent management of the work of the translators and revisers involved in the translation project, and (2) the appropriate methodology for revision applied to legal translation by adapting the revision mode’s focus to ensure its effectiveness. Six common scenarios are identified in light of the translators’ profiles, for which revisers’ profiles are then proposed in order to detect any legal translation competence deficiencies among translators, and thus ensure quality. Firmé un contrato de cesión de derechos de autor a la editorial John Benjamins Publishing Company. No obstante, el artículo está disponible utilizando la VPN. Espero poder obtener el handle de los metadatos del artículo o de la copia off-print que he subido porque lo necesito para indicarlo en la solicitud de un sexenio de investigación. El archivo con la copia off-print del artículo que he subido indica lo siguiente: This electronic file is a contribution from Target 33:2 and may not be altered in any way. The author of this article is permitted to use this PDF file to generate printed copies to be used by way of offprints, for their personal use only. Permission is granted by the publishers to post this file on a closed server which is accessible onlty to members (students and faculty) of the author's institute. It is not permitted to post this PDF on teh internet, or to share it on sites such as Medeley, ResearchGate, Academia.edu. For any other use of this material prior written permission should be obtained from the publishers (John Benjamins Publishing Company) or through the Copyright Clearance Center. Gracias por su colaboración.
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Several studies have described the potential role of microRNA (miRNAs) in migraine but studies are scarcely reproducible primarily due to the heterogeneous variability of participants. Hence, the main objective of this project is to identify and validate differentially expressed miRNAs (DE miRNAs) in peripheral blood mononuclear cells (PBMCs) of patients with migraine compared to healthy controls (HC) in a well-controlled homogeneous cohort of participants using a three-step methodology: (a) exploratory study, (b) main study and (c) validation study. During the exploratory study, effect size was estimated in order to calculate the required sample size for the main and validation studies Patients diagnosed with migraine according to the International Classification of Headache Disorders and HC without family history of headache disorders were recruited. All participants completed a very thorough questionnaire and structured-interview in order to control environmental factors. RNA was extracted from PBMC and a microarray system was used to determine differential miRNA profiles between study groups in both exploratory and main studies. Principal components analysis and hierarchical clustering analysis were performed to study samples' distribution and to identify potential Batch effect. DE miRNAs were identified adjusting linear model with empirical Bayes moderated t-statistic and a classification task was performed for predictors selection using a supervised machine learning method widely used in the microarray analysis (RF, random forest models), with a bootstrapping algorithm in order to evaluate the stability of classification's results. During the validation study, a quantitative RT-PCR (qRT-PCR) procedure was performed of the DE miARNs identified during the main study and their relative quantification (RQ) was computed in terms of the difference between DE miRNAs and the housekeeping transcript (hsa-miR-93-5p) between study groups. Then, multivariate generalized mixed-effect regression models were estimated in order to study whether there were differences in the RQ of each DE miRNA between study groups, adjusted by covariates. Finally, a functional enrichment analysis of selected targets was computed through protein-protein interaction networks. A total of 313 participants (108 HC and 205 patients) were included during all the project and 23 DE miRNA (5 over-expressed and 18 under-expressed) were identified after the classification task as predictors which differentiate study groups (HC vs. patients with migraine). The RF model showed an accuracy [95% CI] of 87.8% [73.8% - 95.9%] with a 100.0% sensitivity, 84.4% specificity and kappa coefficient of 0.703. The area under ROC curve obtained was 0.821. Then, the top three miARNs (miR-DOWN006, miR-DOWN002 and miR-DOWN017) from this signature were validated through qRT-PCR in an external cohort of participants. Only miR-DOWN006 and miR-DOWN002 were under-expressed in the 97.2% and 87.3% of participants, respectively. Afterwards, multivariate mixed-effect regression models were fitted for each miRNA and we found that the group variable resulted statistically significant for both transcripts. Enrichment functional analysis showed that these 2 miARNs were closely associated with known migraine pathophysiology: hormone (estrogens, prolactin, oxytocin and cortisol) signaling pathways, dopaminergic and glutamatergic synapses, inflammatory signaling pathways (TGF-, chemokines, apelin, renin and Toll-like receptors), energy homeostasis (AMPK and mTOR signaling pathways), neuronal development (axon guidance and Notch signaling pathway) and environmental adaptation mechanisms (circadian rhythms). Hence, the results of this project will help us to develop the first kit for migraine diagnosis relying on quantitative indicators. This way we will be able (i) to reduce the stigma associated with the disease, (ii) to better phenotype patients and (iii) to treat them earlier. Moreover, the data gathered in this thesis will allow us not only to improve our understanding of migraine pathophysiology but also to identify new targets in order to develop future therapeutic approaches. Varios estudios piloto han estudiado el rol de los microARNs (miARNs) en la migraña. Sin embargo, los estudios son poco comparables y reproducibles debido al alto grado de heterogeneidad que existe entre los grupos. Así pues, el objetivo de la presente tesis es identificar y validar un patrón de expresión diferencial de miARNs en células mononucleares de la sangre periférica (PBMC) en una cohorte muy bien controlada de individuos con migraña y controles sanos (HC). Para llevar a cabo este objetivo, se ha planteado un diseño experimental basado en tres estudios secuenciales: (a) una prueba exploratoria para estimar el efecto del análisis y ajustar el tamaño de la muestra en (b) el estudio principal y, finalmente, (c) replicar los hallazgos en una tercera etapa, en un estudio de validación. Se reclutó individuos diagnosticados de migraña y HC que negaban tener antecedentes familiares de primer o segundo grado con cefalea recurrente. Todos los participantes completaron también un cuestionario muy exhaustivo para controlar los factores ambientales. Se realizó la extracción del ARN y se estudió el perfil de expresión diferencial de los miARNs mediante microchips específicos durante los estudios exploratorio y principal. Realizamos un análisis de las componentes principales y un análisis de agrupamiento jerárquico para estudiar la distribución de las muestras y detectar un posible efecto Batch. El análisis diferencial se realizó mediante el ajuste de modelos lineales basados en un enfoque Bayesiano de la prueba t de Student moderada. La selección de los mejores transcritos se llevó a cabo mediante una tarea de clasificación basada en árboles de decisión que recibe el nombre de Random Forest (RF), un algoritmo de clasificación supervisado, y un método de remuestreo (bootstrapping) para controlar la estabilidad de los resultados. Para la validación de los miARNs seleccionados, se llevó a cabo una amplificación y secuenciación en una cohorte externa mediante qRT-PCR. Una vez estimada la cuantificación relativa (RQ) de cada miARN, se ajustó modelos lineales generalizados de efectos mixtos para estudiar si había diferencias en la expresión de cada miARN entre los grupos de estudio. Finalmente, realizamos una predicción in silico de las dianas génicas de este perfil de miARNs e identificamos las redes de interacción proteína-proteína asociadas a estos genes para encontrar las subredes ontológicas más representativas. Un total de 313 participantes (108 HC y 205 pacientes) fueron incluidos durante los tres sub-estudios de este proyecto y, después de realizar la tarea de clasificación, identificamos un total de 23 DE miARNs (5 sobre-expresados y 18 infra-expresados) entre los controles y los pacientes. El modelo de clasificación presentó una precisión del 87,8% (IC95%, 73,8% - 95,9%). Para llevar a cabo la validación, decidimos cuantificar los 3 mejores miARNs: miR-DOWN006, miR-DOWN002 y miR-DOWN017. Aun así, solo obtuvimos la expresión del miR-DOWN006, que se expresó en el 97,2% de las muestras y del miR-DOWN002, que se expresó en el 87,3%; ambos transcritos infra-expresados en individuos diagnosticados de migraña. El análisis funcional de estos 2 miARNs enriqueció vías ontológicas que se han asociado previamente con la migraña: vías de señalización hormonal, vías de neurotransmisión sináptica, vías asociadas a la inflamación, vías asociadas a la homeostasis energética, vías asociadas al desarrollo neuronal y mecanismos de adaptación al ambiente. Los datos aportados por esta tesis nos pueden ayudar a desarrollar el primer kit para diagnosticar la migraña mediante el uso de indicadores cuantitativos. De este modo, podremos (i) reducir el estigma asociado a la enfermedad, (ii) diagnosticar mejor a los pacientes y (iii) a tratarlos precozmente. Diversos estudis pilot han estudiat el rol dels microARNs (miARNs) en la migranya. No obstant això, els estudis són poc comparables i reproduïbles degut a l'alt grau d'heterogeneïtat que existeix entre els grups. Així doncs, l'objectiu de la present tesi és identificar i validar un patró d'expressió diferencial de miARNs en cèl·lules mononuclears de la sang perifèrica (PBMC) en una cohort molt ben controlada d'individus amb migranya i controls sans (HC). Per dur a terme aquest objectiu, s'ha plantejat un disseny experimental basat en tres estudis seqüencials: (a) una prova exploratòria per a estimar l'efecte de l'anàlisi i ajustar la grandària de la mostra en (b) l'estudi principal i, finalment, (c) replicar les troballes en una tercera etapa en un estudi de validació. Es va reclutar individus diagnosticats de migranya i HC que negaven tenir antecedents familiars de primer o segon grau amb cefalea recurrent. Tots els participants van completar també un qüestionari molt exhaustiu per a controlar els factors ambientals. Es va realitzar l'extracció de l'ARN i es va estudiar el perfil d'expressió diferencial de miARNs mitjançant microxips específics durant els estudis exploratori i principal. Vam realitzar una anàlisi de les components principals i una anàlisis de clusterització jeràrquica per a estudiar la distribució de les mostres i detectar un possible efecte Batch. L'anàlisi diferencial es va realitzar mitjançant l'ajustament de models lineals basats en un enfocament Bayesià de la prova t de Student moderada. La selecció dels millors transcrits es va dur a terme mitjançant una tasca de classificació basada en arbres de decisió que rep el nom de Random Forest (RF), un algoritme de classificació supervisat, i un mètode de remostreig (bootstrapping) per a controlar l'estabilitat dels resultats. Per a la validació dels miARNs seleccionats, es va dur a terme una amplificació i seqüenciació en una cohort externa mitjançant qRT-PCR. Una vegada estimada la quantificació relativa (RQ) de cada transcrit, es va ajustar models lineals generalitzats d'efectes mixtes per a estudiar si hi havia diferències en l'expressió de cada miARN entre els grups d'estudi. Finalment, vam predir in silico els gens diana d'aquest perfil de miARN i vam identificar les xarxes d'interacció proteïna-proteïna associades a aquests gens per tal de trobar les subxarxes ontològiques més representatives. Un total de 313 participants (108 HC i 205 pacients) van ser inclosos durant els tres sub-estudis d'aquest projecte i, després de realitzar la tasca de classificació, vam identificar un total de 23 DE miARNs (5 sobre-expressats i 18 infra-expressats) entre els controls i els pacients. El model de classificació va presentar una precisió del 87,8% (IC95%, 73,8% - 95,9%). Per a dur a terme la validació, vam decidir quantificar els 3 millors miARNs de la signatura dels 23: miR-DOWN006, miR-DOWN002 i el miR-DOWN017. Tanmateix, només vam obtenir la RQ del miR-DOWN006, que es va expressar en el 97,2% de les mostres i del miR-DOWN002, que es va expressar en el 87,3%; tots dos transcrits infra-expressats en individus diagnosticats de migranya. L'anàlisi funcional d'aquests 2 miARNs va enriquir vies ontològiques que s'han associat prèviament amb la migranya: vies de senyalització hormonal, vies de neurotransmissió sinàptica, vies associades a la inflamació, vies associades a l'homeòstasi energètica , vies associades al desenvolupament neuronal i mecanismes d'adaptació a l'ambient. Les dades aportades per aquesta tesi ens poden ajudar a desenvolupar el primer kit per a fenotipar la migranya mitjançant l'ús d'indicadors quantitatius. D'aquesta manera, podrem (i) reduir l'estigma associat a la malaltia, (ii) diagnosticar millor als pacients i (iii) a tractar-los precoçment.
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