Gasto turístico en alimentación y la cocina local del cantón Patate

Ecuador is a country with the 4 worlds, full of legacies and intangible cultural heritage such as gastronomy, it is one of the main economic engines of tourist destinations. Tourists are enriched and learn from the traditional knowledge of the inhabitants through experience and history in the place. The objective of this research is to identify tourist spending on food and spread the local cuisine of the Patate canton. The methodology used is a mixed approach, therefore, three instruments were applied, the interview, the food atlas cards and the survey. For the qualitative part, experts of the different characteristics of the local cuisine were carried out in the field. On the other hand, a survey of tourists was carried out in order to know the tourist food expenditure in the canton and their motivation for visiting. There is a high percentage of tourist spending on food and the local cuisine of Patate is represented by products that are connected to the history of the canton. As a result, a proposal suitable for people with visual disabilities was obtained, it is a spoken magazine that fulfills the function of adapting to all visitors and thus achieving accessible tourism.   URL: https://revistas.uta.edu.ec/erevista/index.php/bcoyu/article/view/1961/version/2615Ecuador es un país con los 4 mundos, lleno de legados y patrimonios culturales inmateriales como lo es la gastronomía, es uno de los principales motores económicos de los destinos turísticos. Los turistas se enriquecen y aprenden de los conocimientos tradicionales de los habitantes mediante la experiencia y trayectoria en el lugar. El objetivo de esta investigación es identificar el gasto turístico en alimentación y difundir a la cocina local del cantón Patate. La metodología utilizada es un enfoque mixto, por lo tanto, se aplicó tres instrumentos, la entrevista, las fichas del atlas alimentario y encuesta. Para la parte cualitativa se realizó en campo a expertos de las distintas características de la cocina local. Por otra parte, se realizó una encuesta a turistas con el fin de conocer el gasto turístico alimentario en el cantón y su motivación de visita. Existe un alto porcentaje de gasto turístico en alimentación y la cocina local de Patate está representada por productos que tienen conexión con la historia del cantón. Como resultados se obtuvo una propuesta apta para personas con discapacidad visual, es una revista hablada que cumple como función adaptarse a todos los visitantes y así conseguir un turismo accesible.   URL: https://revistas.uta.edu.ec/erevista/index.php/bcoyu/article/view/1961/version/261

A simplified implementation of the stationary liquid mass balance method for on-line OUR monitoring in animal cell cultures

This is the peer reviewed version of the following article: [Fontova, A. , Lecina, M. , López‐Repullo, J. , Martínez‐Monge, I. , Comas, P. , Bragós, R. and Cairó, J. J. (2018), A simplified implementation of the stationary liquid mass balance method for on‐line OUR monitoring in animal cell cultures. J. Chem. Technol. Biotechnol. doi:10.1002/jctb.5551], which has been published in final form at [doi:10.1002/jctb.5551]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.BACKGROUND: Compared with other methods, the stationary liquid mass balance method for oxygen uptake rate (OUR) determination offers advantages in terms of estimation accuracy and reduction of stress. However, the need for sophisticated instrumentation, like mass flow controllers and gas analysers, has historically limited wider implementation of such a method. In this paper, a new simplified method based on inexpensive valves for the continuous estimation of OUR in animal cell cultures is evaluated. The determination of OUR values is based on accurate operation of the dissolved oxygen (DO) control loop and monitoring of its internal variables. RESULTS: The method developed was tested empirically in 2¿L bioreactor HEK293 batch cultures. OUR profiles obtained by a dynamic method, global mass balance method and the developed simplified method were monitored and compared. The results show how OUR profile obtained with the proposed method better follows the off-line cell density determination. The OUR estimation frequency was also increased, improving the method capabilities and applications. The theoretical rationale of the method was extended to the sensitivity analysis which was analytically and numerically approached. CONCLUSIONS: The results showed the proposed method to be not only cheap, but also a reliable alternative to monitor the metabolic activity in bioreactors in many biotechnological processes, being a useful tool for high cell density culture strategies implementation based on OUR monitoring.Peer ReviewedPostprint (published version

Handling and novel object recognition modulate fear response and endocannabinoid signaling in nucleus basalis magnocellularis

[EN] Storage of aversive memories is of utmost importance for survival, allowing animals to avoid upcoming similar stimuli. However, without reinforcement, the learned avoidance response gradually decreases over time. Although the molecular mechanisms controlling this extinction process are not well known, there is evidence that the endocannabinoid system plays a key role through CB1 receptor-mediated modulation of cholinergic signaling. In this study, we measured fear extinction throughout 7 months using naive rats, assessed in passive avoidance (PA) test in a non-reinforced manner. Then, we evaluated the effect of gentle handling and non-aversive novel object recognition test (NORT) on the extinction and expression of fear memories by measuring passive avoidance responses. Neurochemical correlates were analyzed by functional autoradiography for cannabinoid, cholinergic, and dopaminergic receptors. Despite results showing a gradual decrease of passive avoidance response, it did not fully disappear even after 7 months, indicating the robustness of this process. Meanwhile, in rats that received gentle handling or performed NORT after receiving the PA aversive stimulus, extinction occurred within a week. In contrast, gentle handling performed before receiving the aversive stimulus exacerbated fear expression and triggered escape response in PA. The neurochemical analysis showed increased cannabinoid and cholinergic activity in the nucleus basalis magnocellularis (NBM) in rats that had performed only PA, as opposed to rats that received gentle handling before PA. Additionally, a correlation between CB1 mediated-signaling in the NBM and freezing in PA was found, suggesting that the endocannabinoid system might be responsible for modulating fear response induced by aversive memories.Basque Government IT975-16 to the "Neurochemistry and Neurodegeneration" consolidated research group; Instituto de Salud Carlos III, Grant/Award Number: PI20/0015

Elimination of the Warburg effect in Chinese hamster ovary (CHO) cells improves cell phenotype as a protein production platform

Lactate is a common metabolite and is central to many important processes. One of its more prominent roles is in the Warburg effect, in which cancer cells exhibit high rates of glycolytic flux followed by secretion of lactate, even in the presence of oxygen. This fermentation of pyruvate to lactate via lactate dehydrogenase (Ldh) accompanies increased proliferation of cancer cells and several other types of rapidly proliferating cell types in immune cell activation and embryonic development. Aerobic glycolysis is also prominent in biotherapeutic protein production, where mammalian production cells often secrete high levels of lactate. The accumulation of lactate is deleterious for cell growth, viability, product formation, and quality, both directly via acidification of the media and indirectly through base addition to control culture pH. Despite a clear genetic target, efforts to eliminate lactate secretion via knockout of Ldh(s) in mammalian cells have been unsuccessful, pointing to the essentiality of Ldh mediated NAD regeneration. A wide variety of approaches have been utilized to limit lactate accumulation in culture, including knockdown or inhibition of Ldh, replacement of glucose with alternate sugars, controlled feeding strategies, and many others, however none have proven successful in eliminating the Warburg effect. We report the elimination of the Warburg effect in a CHO cell line by using CRISPR/Cas9-based engineering to simultaneously knockout enzymes responsible for lactate production and ancillary regulators. The resulting cell lines remain proliferative while consuming significantly less glucose and can be used to generate protein producing lines using standard industrial processes. In a pH-controlled fedbatch process, the Warburg null cells require minimal base addition to maintain a stable pH, allowing an extended growth phase. The knockout strategy was also successfully applied to a CHO cell line producing Rituximab, again resulting in a prolonged growth phase. Additionally, protein production was maintained, while product quality was improved with increased glycan galactosylation. Thus, CHO cells without the capacity of Warburg metabolism may be useful for engineering production cell lines with enhanced bioproduction traits

Relative vaccine effectiveness against COVID-19 hospitalisation in persons aged ≥ 65 years: results from a VEBIS network, Europe, October 2021 to July 2023

VEBIS-Lot 4 working group: James Humphreys, Alexis Sentís, Joris Van Loenhout, Pierre Hubin, Katrine Finderup Nielsen, Chiara Sacco, Daniele Petrone, Patrizio Pezzotti, Itziar Casado, Aitziber Echeverria, Camino Trobajo-Sanmartín, Stijn Andeweg, Anja Bråthen Kristoffersen, Irina Kislaya, Patricia Soares, Carlos Dias, Ausenda Machado.Since 2021, the Vaccine Effectiveness, Burden and Impact Studies of coronavirus disease 2019 (COVID-19) and influenza (VEBIS) project monitors vaccine effectiveness (VE) in real-world conditions to inform vaccination programmes in the European Union/European Economic Area (EU/EEA) countries [1]. One project aims to monitor real-time COVID-19 VE using electronic health registries (EHR) in multiple countries, with initial findings previously published [2-4]. We report pooled VE results against hospitalisation due to COVID-19 by number of doses received and time since vaccination in a community-dwelling resident population aged ≥ 65 years between October 2021 and July 2023.info:eu-repo/semantics/publishedVersio

Autoantibodies against the immunodominant sCha epitope discriminate the risk of sudden death in chronic Chagas cardiomyopathy

In Chagas disease (ChD) caused by Trypanosoma cruzi, new biomarkers to predict chronic cardiac pathology are urgently needed. Previous studies in chagasic patients with mild symptomatology showed that antibodies against the immunodominant R3 epitope of sCha, a fragment of the human basic helix-loop-helix transcription factor like 5, correlated with cardiac pathology. To validate sCha as a biomarker and to understand the origin of anti-sCha antibodies, we conducted a multicenter study with several cohorts of chagasic patients with severe cardiac symptomatology. We found that levels of antibodies against sCha discriminated the high risk of sudden death, indicating they could be useful for ChD prognosis. We investigated the origin of the antibodies and performed an alanine scan of the R3 epitope. We identified a minimal epitope MRQLD, and a BLAST search retrieved several T. cruzi antigens. Five of the hits had known or putative functions, of which phosphonopyruvate decarboxylase showed the highest cross-reactivity with sCha, confirming the role of molecular mimicry in the development of anti-sCha antibodies. Altogether, we demonstrate that the development of antibodies against sCha, which originated by molecular mimicry with T. cruzi antigens, could discriminate electrocardiographic alterations associated with a high risk of sudden death.Ministerio de Economía y competitividad and Fondo Europeo de Desarrollo Regional (SAF2015-63868-R (MINECO/FEDER) to N.G., and SAF2016-75988-R (MINECO/FEDER) to M.F.); Ministerio de Ciencia, Innovación y Universidades-Agencia Estatal de Investigación and Fondo Europeo de Desarrollo Regional (PGC2018-096132-BI00 (MICINN/FEDER) to N.G.); Universidad Autónoma de Madrid-Banco de Santander Inter-University Cooperation Grant with Latin América (CEAL-AL/2015-12 to N.G.); Red de Investigación de Centros de Enfermedades Tropicales (RICET RD12/0018/0004 to M.F.); and Comunidad de Madrid (S-2010/BMD-2332 to M.F.). CBMSO institutional grants from Fundación Ramón Areces and Banco de Santande

Precariedad, exclusión social y diversidad funcional (discapacidad): lógicas y efectos subjetivos del sufrimiento social contemporáneo (II). Innovación docente en Filosofía

El PIMCD "Precariedad, exclusión social y diversidad funcional (discapacidad): lógicas y efectos subjetivos del sufrimiento social contemporáneo (II). Innovación docente en Filosofía" se ocupa de conceptos generalmente eludidos por la tradición teórica (contando como núcleos aglutinantes los de la precariedad laboral, la exclusión social y diversidad funcional o discapacidad), cuyo análisis propicia nuevas prácticas en la enseñanza universitaria de filosofía, adoptando como meta principal el aprendizaje centrado en el estudiantado, el diseño de nuevas herramientas de enseñanza y el fomento de una universidad inclusiva. El proyecto cuenta con 26 docentes de la UCM y otros 28 docentes de otras 17 universidades españolas (UV, UNED, UGR, UNIZAR, UAH, UC3M, UCA, UNIOVI, ULL, EHU/UPV, UA, UAM, Deusto, IFS/CSIC, UCJC, URJC y Univ. Pontificia de Comillas), que permitirán dotar a las actividades programadas de un alcance idóneo para consolidar la adquisición de competencias argumentativas y dialécticas por parte de lxs estudiantes implicados en el marco de los seminarios previstos. Se integrarán en el PIMCD, aparte de PDI, al menos 26 estudiantes de máster y doctorado de la Facultad de Filosofía, a lxs que acompañarán durante el desarrollo del PIMCD 4 Alumni de la Facultad de Filosofía de la UCM, actualmente investigadores post-doc y profesorxs de IES, cuya experiencia será beneficiosa para su introducción en la investigación. Asimismo, el equipo cuenta con el apoyo de varixs profesorxs asociadxs, que en algunos casos son también profesores de IES. Varixs docentes externos a la UCM participantes en el PIMCD poseen una dilatada experiencia en la coordinación de proyectos de innovación de otras universidades, lo que redundará en beneficio de las actividades a desarrollar. La coordinadora y otrxs miembros del PIMCD pertenecen a la Red de Innovación Docente en Filosofia (RIEF), puesta en marcha desde la Universitat de València (http://rief.blogs.uv.es/encuentros-de-la-rief/), a la que mantendremos informada de las actividades realizadas en el proyecto. Asimismo, lxs 6 miembros del PAS permitirán difundir debidamente las actividades realizadas en el PIMCD entre lxs estudiantes Erasmus IN del curso 2019/20 en la Facultad de Filosofía, de la misma manera que orientar en las tareas de maquetación y edición que puedan ser necesarias de cara a la publicación de lxs resultados del PIMCD y en las tareas de pesquisa bibliográfica necesarias para el desarrollo de los objetivos propuestos. Han manifestado su interés en los resultados derivados del PIMCD editoriales especializadas en la difusión de investigaciones predoctorales como Ápeiron y CTK E-Books

Development of tools for monitoring and controlling cell cultures through the metabolic analysis

Les cèl·lules de mamífer esdevenen un dels principals sistemes per a la producció d'una àmplia gamma de proteïnes. Al voltant del 60-70% de tots els productes biofarmacèutics es produeixen en cèl·lules de mamífer gràcies a la capacitat d'aquestes cèl·lules de realitzar complexes modificacions post-translacionals per produir proteïnes biològicament actives. Una de les principals limitacions de les cèl·lules de mamífer és el seu ineficient metabolisme en cultiu, caracteritzat pel consum de grans quantitats de glucosa i la alta producció de subproductes no desitjats, com l'amoníac i el lactat, que afecten negativament el creixement cel·lular. En el treball presentat en aquesta tesi, s’observa que sota certes condicions de cultiu, les cèl·lules de mamífer son també capaces de consumir el lactat, mantenint-se en la fase de creixement exponencial. Els capítols 3 i 4 es centren en presentar les diferents fases del metabolisme de la glucosa i lactat en dues línies cel·lulars: HEK293 i CHO. Depenent de les condicions de cultiu, s'han obtingut tres metabolismes diferents: Fase 1: consum de glucosa i producció de lactat (creixement exponencial), Fase 2: consum simultani de glucosa i lactat (creixement exponencial) i Fase 3: consum de lactat com a única font de carboni (sense creixement cel·lular). L’obtenció d’aquestes diferents fases metabòliques depèn principalment de dues condicions de cultiu: el pH i la concentració extracel·lular de lactat. Per realitzar un estudi més profund de les diferents fases presentades, s'ha realitzat un anàlisi de la distribució de flux intracel·lular mitjançant un Flux Balance Analysis (FBA). Aquest anàlisi demostra que, a la Fase 1, la generació de lactat es deguda a que el piruvat es converteix en lactat per complir els requisits de regeneració de NADH en el citoplasma i només una petita quantitat de piruvat s'introdueix en el TCA a través del Acetyl-CoA. En el consum concomitant de glucosa i lactat (Fase 2), el consum de glucosa es redueix significativament i es s’aconsegueix un equilibri entre la glicòlisi i els fluxos del TCA, produint un consum de substrats molt més eficient. Un cop entès el metabolisme de les cèl·lules de mamífer en cultiu, el següent pas és aplicar aquest coneixement en l'àrea de l’enginyeria de bioprocessos. Amb aquesta finalitat, en el capítol 5 es presenta una nova eina de monitoratge en línia en Bioreactor basada en la mesura de l'addició de base per mantenir el pH constant. Aquesta nova eina és comparada amb una eina de monitoratge àmpliament utilitzada basada en la mesura de la velocitat de consum d’oxigen (O.U.R.), mitjançant l'aplicació del mètode dinàmic. Les dues alternatives presentades han demostrat avantatges clars pel que fa a la concentració de producte final i, especialment, a les productivitats volumètriques. No obstant això, s'han obtingut millors resultats amb l'estratègia basada en l'addició de base, augmentant la concentració total de cèl·lules viables i la concentració final de producte en un 178% i un 257%, respectivament, i obtenint un increment del 109% de la productivitat volumètrica del procés respecte al cultiu de referència en Batch. Aquesta diferència entre les dues estratègies aplicades es deu a les distorsions constants que pateix el cultiu en quant al pO2 i pH realitzades a cada mesura de la velocitat de consum d’oxigen amb el mètode dinàmic. Per tancar el treball realitzat, en el capítol 6 es presenta un mètode no invasiu per a la determinació de la O.U.R. com alternativa al mètode dinàmic. Els resultats demostren que el mètode desenvolupat és una alternativa fiable per monitoritzar l'activitat metabòlica, sent una eina útil per a la implementació d'estratègies de cultiu on la finalitat és l’obtenció d’altes densitats cel·lulars basades en el seguiment de la O.U.R..Mammalian cells are well established systems for the production of a wide range of high added value proteins. About 60-70% of all biopharmaceuticals are produced in mammalian cells due to the capacity of these cells to perform complex post-translational modifications to yield biologically active proteins. One of the most important limitations of mammalian cells is their inefficient metabolism, characterized by the consumption of large quantities of glucose and concomitant production of similar amounts of by-products, like ammonia and lactate, which can detrimentally affect the cell growth. Interestingly, we have observed that under certain culture conditions mammalian cells are able to co-consume both glucose and lactate during the exponential growth phase. Chapters 3 and 4 are focused on presenting the different glucose and lactate metabolisms in cultures of HEK293 and CHO. Three different glucose and lactate metabolisms have been obtained: Phase 1: glucose consumption and lactate production (exponentially growth), Phase 2: glucose and lactate simultaneous consumption (exponentially growth), and Phase 3: lactate consumption as a sole carbon source (no cell growth). These different metabolic phases were observed mainly depending on two cell culture conditions: the pH and the lactate concentration. In order to perform a deeper study of the different phases presented, an analysis of the intracellular flux distribution for the different phases have been performed for both cell lines by means of Flux Balance Analysis (FBA). FBA showed that, in Phase 1, lactate is produced because pyruvate is converted to lactate to fulfill the NADH regeneration requirements in the cytoplasm and only a small amount of pyruvate is introduced into TCA through Acetyl-CoA. In glucose-lactate concomitant consumption (Phase 2), glucose uptake was significantly reduced and a balance between glycolysis and TCA cycle fluxes was reached, yielding a more efficient substrate consumption. Once understood the metabolism of mammalian cells in culture, the next step is to apply this knowledge in the engineering bioprocess area. To this end, a new robust on-line monitoring tool based on the alkali buffer addition used to maintain the pH set-point is presented in Chapter 5. This new tool is compared with a widely used monitoring tool based on the Oxygen Uptake Rate (O.U.R.) determination, by means of application of the dynamic method. The two alternatives presented have shown clear advantages in respect to final product titer and, especially, volumetric productivities. But better results have been obtained with the alkali addition strategy, increasing the total viable cell concentration and product titer by 178% and 257% respectively, and obtaining a 109% increment of the process volumetric productivity in respect to the batch culture. This is due to the culture constant distortions of the pO2 and pH performed in every O.U.R. dynamic measurement. To close the work performed, a different non-invasive method for O.U.R. determination based on the stationary liquid mass balance was presented and tested in batch culture in Chapter 6. The results demonstrated to be a reliable alternative to monitor the metabolic activity in bioreactors, being a useful tool for high cell density culture strategies implementation based on O.U.R. monitoring