Lo que los personajes tienen que decir. La creación de diálogos mediante la metodología de William Layton

La creación de diálogos es una fase esencial de la escritura de guiones audiovisuales. Este artículo explica el sistema de análisis de textos dramáticos de William Layton, originalmente dirigido a la interpretación actoral, y lo aplica a la escritura de diálogos audiovisuales. También expone cómo se ha formado en este sistema a estudiantes de un Máster de guion y muestra su utilidad para la creación de diálogos a través de una encuesta a 85 alumnos de diferentes promociones. De ellos, el 59% tenía formación o experiencia previa en el audiovisual. Antes de conocer el método Layton, solo un 9% de la muestra manejaba otra técnica para crear diálogos y un 50% conocía consejos genéricos que guiaran el diseño de las conversaciones de sus personajes. Y es que las monografías técnicas o académicas sobre guion no ofrecen metodologías sistemáticas que permitan al aspirante a guionista iniciarse en la escritura de diálogos, solo presentan recomendaciones. Con la difusión de esta técnica para la creación de diálogos audiovisuales, este artículo pretende contribuir a evitar que esta carencia de metodologías sistemáticas se traslade a los programas formativos. Los encuestados confirman que este método favorece la síntesis (84%), la planificación previa de los diálogos y la funcionalidad de cada frase de los personajes (91%), permitiendo que estas se adecuen mejor a su caracterización psicosocial (94%) o momento del arco dramático (95%). Un 85% de los encuestados sigue empleándola, una vez terminada su formación, en su vida profesional como guionista o declara transmitirla en sus clases cuando trabaja como docente

Netflix Documentary Series in Spain. Challenging the Classical Seriality Norms

This research analyzes the impact of Netflix’s all-at-once premiere on the dynamics of seriality. The digitalization of television has profoundly modified the medium, and the binge watching, and the abandonment of the weekly premiere have completely changed the consumption and the creation of fictions. To address this issue, we focus on the documentary series produced by the company in Spain: Examen de conciencia (2019), El caso Alcàsser (2019), La Línea: La sombra del narco (2020) and Nevenka (2021). The main consequences generated by the elimination of the wait between episodes make the episodic plots overflow the chapter and serial storylines do not follow patterns linked to speculation. Consequently, the episodes lose their formal unity or, directly, seriality is abandoned as a narrative model. This paradigm reinforces the idea that, by modifying the distribution methodology, the syntactic characteristics of the products are also altered.Esta investigación analiza el impacto que ha tenido el estreno único de Netflix en las dinámicas propias de la serialidad. La digitalización de la televisión ha modificado profundamente el medio y el binge watching y el abandono del estreno semanal han cambiado por completo el consumo y la creación de ficciones. Para abordar esta cuestión, se analizan las cuatro series documentales que ha producido la compañía en España: Examen de conciencia (2019), El caso Alcàsser (2019), La Línea: La sombra del narco (2020) y Nevenka (2021). Al eliminar la espera entre los episodios las tramas capitulares desbordan la entrega y los seriales no siguen los patrones vinculados a la especulación. Consecuentemente, las entregas pierden su unidad formal o, directamente, se abandona la serialidad como modelo narrativo. Este cambio de paradigma refuerza la idea de que, al modificar la metodología de distribución, también se alteran las características sintácticas de los productos

What characters have to say. The creation of dialogues using William Layton's methodology

Dialogue creation is an essential phase of audiovisual scriptwriting. This article explains William Layton's dramatic texts analysis system originally aimed at acting performance, and applies it to the writing of audiovisual dialogues. It also explains how students of a Master's degree in screenwriting have been trained in this system and shows its usefulness for creating dialogues by means of a survey applied to 85 students from different classes. Of these, 59% had previous training or experience in audiovisuals. Before learning about the Layton method, only 9% of the sample used other techniques to create dialogues and 50% knew generic advice that would guide the design of their characters' conversations. The fact is that technical or academic monographs on screenwriting do not offer systematic methodologies that allow aspiring screenwriters to get started in dialogue writing, but they only provide recommendations. By disseminating this technique for the creation of audiovisual dialogues, this article aims to help prevent this lack of systematic methodologies from being transferred to training programs. Participants confirm that this method favors synthesis (84%) and pre-planning of the dialogues and the functionality of each sentence of the characters (91%), allowing them to better adapt to their psychosocial characterization (94%) or moment of the dramatic arc (95%). Eighty-five percent of the respondents continue to use it in their professional life as scriptwriters after completing their training or state that they pass it on in their classes when they work as teachers.La creación de diálogos es una fase esencial de la escritura de guiones audiovisuales. Este artículo explica el sistema de análisis de textos dramáticos de William Layton, originalmente dirigido a la interpretación actoral, y lo aplica a la escritura de diálogos audiovisuales. También expone cómo se ha formado en este sistema a estudiantes de un Máster de guion y muestra su utilidad para la creación de diálogos a través de una encuesta a 85 alumnos de diferentes promociones. De ellos, el 59% tenía formación o experiencia previa en el audiovisual. Antes de conocer el método Layton, solo un 9% de la muestra manejaba otra técnica para crear diálogos y un 50% conocía consejos genéricos que guiaran el diseño de las conversaciones de sus personajes. Y es que las monografías técnicas o académicas sobre guion no ofrecen metodologías sistemáticas que permitan al aspirante a guionista iniciarse en la escritura de diálogos, solo presentan recomendaciones. Con la difusión de esta técnica para la creación de diálogos audiovisuales, este artículo pretende contribuir a evitar que esta carencia de metodologías sistemáticas se traslade a los programas formativos. Los encuestados confirman que este método favorece la síntesis (84%), la planificación previa de los diálogos y la funcionalidad de cada frase de los personajes (91%), permitiendo que estas se adecuen mejor a su caracterización psicosocial (94%) o momento del arco dramático (95%). Un 85% de los encuestados sigue empleándola, una vez terminada su formación, en su vida profesional como guionista o declara transmitirla en sus clases cuando trabaja como docente

Coronary and carotid artery dysfunction and KV7 overexpression in a mouse model of Hutchinson-Gilford progeria syndrome.

Hutchinson-Gilford progeria syndrome (HGPS) is an extremely rare genetic disease caused by expression of progerin, a lamin A variant that is also expressed at low levels in non-HGPS individuals. Although HGPS patients die predominantly from myocardial infarction and stroke, the mechanisms that provoke pathological alterations in the coronary and cerebral arteries in HGPS remain ill defined. Here, we assessed vascular function in the coronary arteries (CorAs) and carotid arteries (CarAs) of progerin-expressing LmnaG609G/G609G mice (G609G), both in resting conditions and after hypoxic stimulus. Wire myography, pharmacological screening, and gene expression studies demonstrated vascular atony and stenosis, as well as other functional alterations in progeroid CorAs and CarAs and aorta. These defects were associated with loss of vascular smooth muscle cells and overexpression of the KV7 family of voltage-dependent potassium channels. Compared with wild-type controls, G609G mice showed reduced median survival upon chronic isoproterenol exposure, a baseline state of chronic cardiac hypoxia characterized by overexpression of hypoxia-inducible factor 1α and 3α genes, and increased cardiac vascularization. Our results shed light on the mechanisms underlying progerin-induced coronary and carotid artery disease and identify KV7 channels as a candidate target for the treatment of HGPS.Work supported by Ministerio de Ciencia e Innovación (MCIN) and Agencia Estatal de Investigación (AEI) (MCIN/AEI/https:// doi. org/ 10. 13039/ 50110 00110 33 grants SAF2016-79490-R and PID2019-108489RB-I00), with cofunding from Fondo Social Europeo (“El FSE invierte en tu futuro”), and a donation from Asociación Progeria Alexandra Peraut. Microscopy was conducted at the Microscopy & Dynamic Imaging Unit, CNIC, ICTS-ReDib, co-funded by MCIN/AEI/https:// doi. org/ 10. 13039/ 50110 00110 33. R.M.N was supported by the Ministerio de Educación, Cultura y Deporte (pre-doctoral contract FPU16/05027), and I.B. is supported by the Comunidad Autónoma de Madrid (grants 2017- T1/BMD-5247 and 2021-5A/BMD-20944), and the Ramón y Cajal contract (RYC2021-033805-I) funded by MCIN/ AEI/10.13039/501100011033 and the European Union “Next- GenerationEU”/PRTR. The CNIC is supported by the MCIN, the Instituto de Salud Carlos III, the Pro-CNIC Foundation, and is a Severo Ochoa Center of Excellence (grant number CEX2020-001041-S funded by MCIN/AEI/https:// doi. org/ 10. 13039/ 50110 00110 33).N

Monitoring of bluetongue virus in zoo animals in Spain, 2007–2019

Bluetongue (BT) is an emerging and re-emerging communicable vector-borne disease of animal health concern. A serosurvey was performed to assess exposure to BT virus (BTV) in zoo animals in Spain and to determine the dynamics of seropositivity in longitudinally sampled individuals during the study period. Serum samples were collected from 241 zoo animals belonging to 71 different species in five urban zoos (A-E) in Spain between 2007 and 2019. Twenty-four of these animals were longitudinally surveyed at three of the sampled zoos (zoos B, C and E) during the study period. Anti-BTV antibodies were found in 46 (19.1%; 95% CI: 14.1-24.1) of the 241 captive animals analysed by commercial ELISA. A virus neutralization test confirmed specific antibodies against BTV-1 and BTV-4 in 25 (10.7%; 95% CI: 6.7-14.6) and five (3.0%; 95% CI: 0.3-4.0) animals, respectively. Two of the 24 longitudinally sampled individuals (one African elephant (Loxodanta africana) and one aoudad (Ammotragus lervia)) showed anti-BTV antibodies at all samplings, whereas seroconversions were detected in one mouflon (Ovis aries musimon) in 2016, and one Asian elephant (Elephas maximus) in 2019. To the best of the authors' knowledge, this is the first large-scale survey on BTV conducted in both artiodactyl and non-artiodactyl zoo species worldwide. The results confirm BTV exposure in urban zoo parks in Spain, which could be of animal health and conservation concern. Circulation of BTV was detected in yearling animals in years when there were no reports of BTV outbreaks in livestock. Surveillance in artiodactyl and non-artiodactyl zoo species could be a valuable tool for epidemiological monitoring of BTV.info:eu-repo/semantics/acceptedVersio

Overcoming PLK1 inhibitor resistance by targeting mevalonate pathway to impair AXL-TWIST axis in colorectal cancer

© 2021 The Author(s).New therapeutic targets are revolutionizing colorectal cancer clinical management, opening new horizons in metastatic patients’ outcome. Polo Like Kinase1 (PLK1) inhibitors have high potential as antitumoral agents, however, the emergence of drug resistance is a major challenge for their use in clinical practice. Overcoming this challenge represents a hot topic in current drug discovery research. BI2536-resistant colorectal cancer cell lines HT29R, RKOR, SW837R and HCT116R, were generated in vitro and validated by IG50 assays and xenografts models by the T/C ratio. Exons 1 and 2 of PLK1 gene were sequenced by Sanger method. AXL pathway, Epithelial-to-Mesenchymal transition (EMT) and Multidrug Resistance (MDR1) were studied by qPCR and western blot in resistant cells. Simvastatin as a re-sensitizer drug was tested in vitro and the drug combination strategies were validated in vitro and in vivo. PLK1 gene mutation R136G was found for RKOR. AXL pathway trough TWIST1 transcription factor was identified as one of the mechanisms involved in HT29R, SW837R and HCT116R lines, inducing EMT and upregulation of MDR1. Simvastatin was able to impair the mechanisms activated by adaptive resistance and its combination with BI2536 re-sensitized resistant cells in vitro and in vivo. Targeting the mevalonate pathway contributes to re-sensitizing BI2536-resistant cells in vitro and in vivo, raising as a new strategy for the clinical management of PLK1 inhibitors.This study has been funded by Instituto de Salud Carlos III (ISCIII) -Fondos FEDER proyects PI16/01468 and PI19/01231

Monitoring of hepatitis E virus in zoo animals from Spain, 2007–2021

Hepatitis E virus (HEV, family Hepeviridae) is an important emerging and zoonotic pathogen. In recent decades, the number of human cases of zoonotic hepatitis E has increased considerably in industrialized countries and HEV has been detected in an expanding range of mammal species. Although domestic pigs and wild boar are considered the main reservoirs of zoonotic HEV genotypes, the role of other susceptible animals in the epidemiology of the virus is still poorly understood. A large-scale, long-term study was carried out (1) to assess HEV exposure in captive zoo animals in Spain and (2) to determine the dynamics of seropositivity in individuals that were sampled longitudinally during the study period. Between 2007 and 2021, serum samples from 425 zoo animals belonging to 109 animal species (including artiodactyls, carnivores, perissodactyls, proboscideans and rodents) were collected from 11 different zoological parks in Spain. Forty-six of these animals at seven of these zoos were also longitudinally sampled. Anti-HEV antibodies were detected in 36 (8.5%; 95% CI: 5.8–11.1) of 425 sampled zoo animals. Specific antibodies against HEV-3 and HEV-C1 antigens were confirmed in ELISA-positive animals using western blot assay. Two of 46 longitudinally surveyed animals seroconverted during the study period. Seropositivity was significantly higher in carnivores and perissodactyls than in artiodactyls, and also during the period 2012–2016 compared with 2007–2011. HEV RNA was not detected in any of the 262 animals that could be tested by RT-PCR. To the best of the author's knowledge, this is the first large-scale, long-term surveillance on HEV in different orders of zoo mammals. Our results indicate exposure to HEV-3 and HEV-C1 in zoo animals in Spain and confirm a widespread but not homogeneous spatiotemporal circulation of HEV in captive species in this country. Further studies are required to determine the role of zoo species, particularly carnivores and perissodactyls, in the epidemiology of HEV and to clarify the origins of infection in zoological parks

Acquired and Innate Immunity Impairment and Severe Disseminated Mycobacterium genavense Infection in a Patient With a NF-κB1 Deficiency

Background: NF-κB1 is a master regulator of both acquired and innate responses. NFKB1 loss-of-function mutations elicit a wide clinical phenotype with asymptomatic individuals at one end of the spectrum and patients with common variable immunodeficiency, combined immunodeficiency or autoinflammation at the other. Impairment of acquired and innate immunity and disseminated Mycobacterium genavense infection expands the clinical and immunological phenotype of NF-κB1 deficiency.Objective: Functional and molecular characterization of a patient with a novel phenotype of NF-κB1 deficiency.Methods: Circulating T, B, dendritic cell subsets and innate or unconventional T-cells were quantified. The cytokine production in stimulated whole blood samples was assessed and molecular characterization by next generation sequencing and gene expression assays were also performed.Results: We report a patient presenting with features of combined immunodeficiency (CID) and disseminated Mycobacterium genavense infection. Sequencing of genomic DNA identified a novel synonymous mutation (c.705G > A) in NFKB1 gene which resulted in exon 8 skipping and haploinsufficiency of the NF-κB1 subunit p50. The susceptibility to atypical mycobacterial infection has not been previously reported and may be the result of a dendritic cell deficiency. A selective deficiency of circulating follicular helper T (cTFH) cells responsible for mediating the differentiation of naive B cells into memory and plasma cells was also present in the patient. It could affect the maturation of innate or unconventional T cells where NF-κB1 could also be involved.Conclusion: These findings showed that the role of NF-κB1 in humans could be critical for the development of acquired and innate immunity and further highlights the role of human T cells in anti-mycobacterial immunity

Extreme Phenotypes With Identical Mutations: Two Patients With Same Non-sense NHEJ1 Homozygous Mutation

Cernunnos/XLF deficiency is a rare primary immunodeficiency classified within the DNA repair defects. Patients present with severe growth retardation, microcephaly, lymphopenia and increased cellular sensitivity to ionizing radiation. Here, we describe two unrelated cases with the same non-sense mutation in the NHEJ1 gene showing significant differences in clinical presentation and immunological profile but a similar DNA repair defect

Exacerbated atherosclerosis in progeria is prevented by progerin elimination in vascular smooth muscle cells but not endothelial cells

3 p.-2 fig.Hutchinson-Gilford progeria syndrome (HGPS) is a rare disease caused by the expression of progerin, a mutant protein that accelerates aging and precipitates death. Given that atherosclerosis complications are the main cause of death in progeria, here, we investigated whether progerin-induced atherosclerosis is prevented in HGPSrev-Cdh5-CreERT2 and HGPSrev-SM22α-Cre mice with progerin suppression in endothelial cells (ECs) and vascular smooth muscle cells (VSMCs), respectively. HGPSrev-Cdh5-CreERT2 mice were undistinguishable from HGPSrev mice with ubiquitous progerin expression, in contrast with the ameliorated progeroid phenotype of HGPSrev-SM22α-Cre mice. To study atherosclerosis, we generated atheroprone mouse models by overexpressing a PCSK9 gain-of-function mutant. While HGPSrev-Cdh5-CreERT2 and HGPSrev mice developed a similar level of excessive atherosclerosis, plaque development in HGPSrev-SM22α-Cre mice was reduced to wild-type levels. Our studies demonstrate that progerin suppression in VSMCs, but not in ECs, prevents exacerbated atherosclerosis in progeroid mice.Work supported by grant PID2022- 141211OB- I00 funded by MICIU/AEI/10.13039/ 501100011033 and ERDF/EU. I.B. was supported by Comunidad de Madrid (2017- T1/BMD- 5247 and 2021- 5A/BMD- 20944) with cofunding from European Structural and Investment Fund, RYC2021- 033805- I (MICIU/AEI/10.13039/501100011033, European Union NextGenerationEU/PRTR), and PID2022- 137111OA- I00 (MICIU/AEI/10.13039/501100011033, ERDF/EU). Salary support to A.B. (BES- 2017- 079705, MICIU/AEI/10.13039/ 501100011033, ESF), C.E.- E. (Fundación “la Caixa”, LCF/BQ/DR19/1170012), R.M.N (Ministerio de Educación, Cultura y Deporte, FPU16/05027), and M.R.H. (MICIU, IJC2019- 040798- I). CNIC is supported by Instituto de Salud Carlos III (ISCIII), Ministerio de Ciencia, Innovación y Universidades (MICIU), Pro-CNIC Foundation and is a Severo Ochoa Center of Excellence (grant CEX2020- 001041- S funded by MICIU/AEI/10.13039/501100011033). Generation of anti- progerin antibody funded by Wellcome Trust (098291/Z/12/Z, 221699/Z/20/Z). Microscopy and Dynamic Imaging Unit- CNIC/ICTS- ReDib supported by ICTS- 2018- 04- CNIC- 16 funded by MICIU/AEI/10.13039/501100011033 and ERDF–A way to make EuropePeer reviewe