The stable carbon isotopic composition of Daphnia ephippia in small, temperate lakes reflects in-lake methane availability

Daphnia can ingest methane-oxidizing bacteria and incorporate methanogenic carbon into their biomass, leading to low stable carbon isotope ratios (expressed as δ13C values) of their tissue. Therefore, δ13C analysis of Daphnia resting eggs (ephippia) in lake sediment records can potentially be used to reconstruct past in-lake availability of methane (CH4). However, detailed multilake studies demonstrating that δ13C values of recently deposited Daphnia ephippia (δ13Cephippia) are systematically related to in-lake CH4 concentrations (CH4aq) are still missing. We measured δ13Cephippia from surface sediments of 15 small lakes in Europe, and compared these values with late-summer CH4aq. δ13Cephippia ranged from −51.6‰ to −25.9‰, and was strongly correlated with CH4aq in the surface water and above the sediment (r −0.73 and −0.77, respectively), whereas a negative rather than the expected positive correlation was found with δ13C values of carbon dioxide (CO2) (r −0.54), and no correlation was observed with CO2aq. At eight sites, offsets between δ13 CCO2 and δ13Cephippia exceeded offsets between δ13 CCO2 and δ13Calgae reported in literature. δ13Cephippia was positively correlated with δ13C values of sedimentary organic matter (r 0.54), but up to 20.7‰ lower in all except one of the lakes (average −6.1‰). We conclude that incorporation of methanogenic carbon prior to ephippia formation must have been widespread by Daphnia in our study lakes, especially those with high CH4aq. Our results suggest a systematic relationship between δ13Cephippia values and CH4aq in small temperate lakes, and that δ13Cephippia analysis on sediment records may provide insights into past changes in in-lake CH4aq.

Fatal steamer accident; blunt force injuries and drowning in post-mortem MSCT and MRI.

Steamer accidents, through contact with the bucket wheel, are very seldom today. No publication of such a kind of fatal accident could be found in literature. We present the case of a fatal steamer accident, in which the findings of a blunt traumatization of a person by the ship was completely documented by post-mortem combined multi-slice computed tomography (MSCT) and magnetic resonance imaging (MRI) examinations. A rupture of the aorta was detected using both radiological methods without use of radiopaque material. Radiological examination revealed a comminuted fracture of the thorax vertebrae at the same level as the aortic rupture. Injuries of the soft tissues of the back, caused by the bucket wheel of the steamer, were also diagnosed. In addition to the signs of blunt force trauma the findings of drowning such as an over inflation of the lungs, fluid in the stomach and duodenum were revealed. Furthermore, algological analysis detected diatoms in the lung tissue and blood from the left heart. Therefore, the cause of death was considered being a combination of fatal hemorrhage, caused by the aortic rupture, and drowning. We conclude that virtual autopsy using combined post-mortem MSCT and MRI is a useful tool for documentation, visualisation and analysis of the findings of blunt force trauma and drowning with a large potential in forensic medicine.

129I/127I ratios and 129I concentrations in a recent sea sediment core and in rainwater from Sevilla (Spain) by AMS

In spite of the environmental relevance of 129I, there is still a scarcity of data about its presence in the different natural compartments. In this work, results are presented on the concentration of 129I in rainwater samples taken in Sevilla (southwestern Spain) and in a sediment core taken near the Ringhals coast (Sweden). Typical concentrations of 108 and 109129I at/l are found in rainwater samples, similar to other values in literature. In the case of the sediment core, our results clearly show the impact of anthropogenic sources, with concentrations in the order of 1013129I at./kg and isotopic ratios 129I/127I in the order of 10−8 in the higher layers.

New Clinical Indications for (18)F/(11)C-choline, New Tracers for Positron Emission Tomography and a Promising Hybrid Device for Prostate Cancer Staging: A Systematic Review of the Literature

CONTEXT Radiolabelled choline positron emission tomography has changed the management of prostate cancer patients. However, new emerging radiopharmaceutical agents, like radiolabelled prostate specific membrane antigen, and new promising hybrid imaging will begin new challenges in the diagnostic field. OBJECTIVE The continuous evolution in nuclear medicine has led to the improvement in the detection of recurrent prostate cancer (PCa), particularly distant metastases. New horizons have been opened for radiolabelled choline positron emission tomography (PET)/computed tomography (CT) as a guide for salvage therapy or for the assessment of systemic therapies. In addition, new tracers and imaging tools have been recently tested, providing important information for the management of PCa patients. Herein we discuss: (1) the available evidence in literature on radiolabelled choline PET and their recent indications, (2) the role of alternative radiopharmaceutical agents, and (3) the advantages of a recent hybrid imaging device (PET/magnetic resonance imaging) in PCa. EVIDENCE ACQUISITION Data from recently published (2010-2015), original articles concerning the role of choline PET/CT, new emerging radiotracers, and a new imaging device are analysed. This review is reported according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. EVIDENCE SYNTHESIS In the restaging phase, the detection rate of choline PET varies between 4% and 97%, mainly depending on the site of recurrence and prostate-specific antigen levels. Both 68gallium (68Ga)-prostate specific membrane antigen and 18F-fluciclovine are shown to be more accurate in the detection of recurrent disease as compared with radiolabelled choline PET/CT. Particularly, Ga68-PSMA has a detection rate of 50% and 68%, respectively for prostate-specific antigen levels < 0.5ng/ml and 0.5-2ng/ml. Moreover, 68Ga- PSMA PET/magnetic resonance imaging demonstrated a particularly higher accuracy in detecting PCa than PET/CT. New tracers, such as radiolabelled bombesin or urokinase-type plasminogen activator receptor, are promising, but few data in clinical practice are available today. CONCLUSIONS Some limitations emerge from the published papers, both for radiolabelled choline PET/CT and also for new radiopharmaceutical agents. Efforts are still needed to enhance the impact of published data in the world of oncology, in particular when new radiopharmaceuticals are introduced into the clinical arena. PATIENT SUMMARY In the present review, the authors summarise the last evidences in clinical practice for the assessment of prostate cancer, by using nuclear medicine modalities, like positron emission tomography/computed tomography and positron emission tomography/magnetic resonance imaging.

Effectiveness of healthcare waste management interventions in developing countries

Inefficiencies during the management of healthcare waste can give rise to undesirable health effects such as transmission of infections and environmental pollution within and beyond the health facilities generating these wastes. Factors such as prevalence of diseases, conflicts, and the efflux of intellectual capacity make low income countries more susceptible to these adverse health effects. The purpose of this systematic review was to describe the effectiveness of interventions geared towards better managing the generation, collection, transport, treatment and disposal of medical waste, as they have been applied in lower and middle income countries.^ Using a systematic search strategy and evaluation of study quality, this study reviewed the literature for published studies on healthcare waste management interventions carried out in developing countries, specifically the low and lower middle income countries from year 2000 to the current year. From an initially identified set of 829 studies, only three studies ultimately met all inclusion, exclusion and high quality criteria. A multi component intervention in Syrian Arab Republic, conducted in 2007 was aimed at improving waste segregation practice in a hospital setting. There was an increased use of segregation boxes and reduced rates of sharps injury among staff as a result of the intervention. Another study, conducted in 2008, trained medical students as monitors of waste segregation practice in an Indian teaching hospital. There was improved practice in wards and laboratories but not in the intensive care units. The third study, performed in 2008 in China, consisted of modification of the components of a medical waste incinerator to improve efficiency and reduce stack emissions. Gaseous pollutants emitted, except polychlorodibenzofurans (PCDF) were below US EPA permissible exposure limits. Heavy metal residues in the fly ash remained unchanged.^ Due to the paucity of well-designed studies, there is insufficient evidence in literature to conclude on the effectiveness of interventions in low income settings. There is suggestive but insufficient evident that multi-component interventions aimed at improving waste segregation through behavior modification, provision of segregation tools and training of monitors are effective in low income settings.^

2005 ACIP recommendations effect on newborn hepatitis B vaccination rates and the factors affecting the successful implementation of the recommendations in the US

Background. Hepatitis B virus infection is one of major causes of acute and chronic hepatitis, cirrhosis of the liver, and primary hepatocellular carcinoma. Hepatitis B and its long term consequences are major health problems in the United States. Hepatitis B virus can be vertically transmitted from mother to infant during birth. Hepatitis B vaccination at birth is the most effective measure to prevent the newborn from HBV infection and its consequences, and is part of any robust perinatal hepatitis B prevention program following ACIP recommendations. Universal vaccination of the new born will prevent HBV infection during early childhood and, assuming that children receive the three dosages of the vaccine, it will also prevent adolescent and adult infections. Hepatitis B vaccination is now recommended as part of a comprehensive strategy to eliminate HBV transmission in the United States. ^ Objective. (1)To assess if the hepatitis B vaccination rates of newborn babies have improved after the 2005 ACIP recommendations. (2) To identify factors that affects the implementation of ACIP recommendation for hepatitis B vaccination in newborn babies. These factors will encourage ongoing improvement by identifying successful efforts and pinpointing areas that fall short and need attention. Additional focus areas may be identified to accelerate progress in eliminating perinatal HBV transmission.^ Methods. This review includes information from all pertinent articles, reviews, National immunization survey (NIS) surveys, reports, peer reviewed literature and web sources that were published after 1991.The key words to be used for selecting the articles are: "Perinatal Hepatitis B Prevention program", "Universal Hepatitis B vaccination of newborn babies", "ACIP Recommendations." The data gathered will be supplemented with an analysis of vaccination rates using the National Immunization Survey (NIS) birth dose coverage data.^ Results. The data collected in the NIS of 2009 reveals that the national coverage for birth dose of HepB increased to 60.8% from 50.1% in 2006. The largest increase observed for the birth dose in the past 5 years is from 2008 which increased from 55.3 % to 60.8% in 2009. By state, coverage ranged from 22.8% in Vermont to 80.7% in Michigan. %. Overall, in 2009 the estimated vaccination rates are in higher ranges for most states compared to the estimated vaccination rates in 2006. States vary widely in hepatitis B vaccination rates and in their compliance with the 2005 ACIP recommendation. There are many factors at various stages that might affect the successful implementation of the new ACIP recommendation as revealed in literature review. ^ Conclusions. HBV perinatal transmission can be eliminated, but it requires identifying the gaps and measures taken to increase the current vaccination coverage, ensuring timely administration of post exposure immunoprophylaxis and continued evaluations of the impact of immunization recommendations.^

Transmisión oral en la narrativa española contemporánea : Un recurso para la construcción de la memoria de la Guerra Civil y del franquismo

Se propone la descripción y el análisis del empleo de la transmisión oral en la narrativa española actual como un recurso para configurar la memoria de la Guerra Civil española y del franquismo. Para ello se atendió especialmente a las estrategias que intervienen en la representación de la oralidad en relación con los diálogos y los relatos orales, entre otros procedimientos a través de los cuales se transmite el pasado traumático dentro de la ficción. Ha sido analizada la representación de la oralidad en la producción narrativa ficcional de diversos autores españoles de finales del siglo XX y comienzos del siglo XXI que han abordado la temática de la Guerra Civil en una situación particular en cuanto al surgimiento de replanteos y debates acerca del tratamiento del conflicto bélico. Los objetivos generales que guiaron el trabajo han sido indagar la presencia y la función de la transmisión oral en la literatura, profundizar el estudio de la oralidad como voz emergente de colectivos sociales desplazados y analizar la funcionalidad del empleo de la oralidad en textos representativos de la narrativa española contemporánea. A su vez, más específicamente, se propuso profundizar en el estudio de la ficcionalización de la transmisión oral como un artificio para la construcción de la memoria en la narrativa española actual, vincular las posibilidades de transmisión de la historia de la Guerra Civil española con medios propios de la historia oral en la literatura -especialmente en la novela-, problematizar determinadas perspectivas sobre el testimonio en la medida en que son literaturizadas para una concepción de la memoria de la Guerra Civil, de la posguerra y de la dictadura, y estudiar algunas miradas contemporáneas acerca de la Guerra Civil y del franquismo desde el campo de la narrativa como un correlato de ciertos reclamos sociales de la actualidad respecto del pasado reciente. Para esto fueron analizadas diversas propuestas que surgieron esencialmente desde la literatura -con algún detenimiento en un texto fílmico- para abordar el pasado traumático y se consideraron las técnicas de transmisión empleadas para la narración ficcional de aspectos largamente silenciados por la historia oficial. El corpus central de textos literarios ha quedado conformado por O lapis do carpinteiro y Os libros arden mal, de Manuel Rivas, Guárdame bajo tierra, de Ramón Saizarbitoria, Soldados de Salamina, de Javier Cercas, Home sen nome, de Suso de Toro, y Mala gente que camina, de Benjamín Prado. Como corpus complementario, se tornó pertinente incorporar un capítulo que articula un texto fílmico (Muerte en El Valle, de Christina Hardt) con un texto literario (Las esquinas del aire. En busca de Ana María Martínez Sagi, de Juan Manuel de Prada). Esto, que en principio puede parecer un apartamiento del corpus central, favoreció el análisis del tratamiento del testimonio desde la estetización de un texto fílmico o literario. Al realizar la selección, se estudiaron, junto con autores que escriben y publican en castellano, algunos textos escritos originariamente en lenguas minorizadas (gallego y euskera)

Transmisión oral en la narrativa española contemporánea : Un recurso para la construcción de la memoria de la Guerra Civil y del franquismo

Se propone la descripción y el análisis del empleo de la transmisión oral en la narrativa española actual como un recurso para configurar la memoria de la Guerra Civil española y del franquismo. Para ello se atendió especialmente a las estrategias que intervienen en la representación de la oralidad en relación con los diálogos y los relatos orales, entre otros procedimientos a través de los cuales se transmite el pasado traumático dentro de la ficción. Ha sido analizada la representación de la oralidad en la producción narrativa ficcional de diversos autores españoles de finales del siglo XX y comienzos del siglo XXI que han abordado la temática de la Guerra Civil en una situación particular en cuanto al surgimiento de replanteos y debates acerca del tratamiento del conflicto bélico. Los objetivos generales que guiaron el trabajo han sido indagar la presencia y la función de la transmisión oral en la literatura, profundizar el estudio de la oralidad como voz emergente de colectivos sociales desplazados y analizar la funcionalidad del empleo de la oralidad en textos representativos de la narrativa española contemporánea. A su vez, más específicamente, se propuso profundizar en el estudio de la ficcionalización de la transmisión oral como un artificio para la construcción de la memoria en la narrativa española actual, vincular las posibilidades de transmisión de la historia de la Guerra Civil española con medios propios de la historia oral en la literatura -especialmente en la novela-, problematizar determinadas perspectivas sobre el testimonio en la medida en que son literaturizadas para una concepción de la memoria de la Guerra Civil, de la posguerra y de la dictadura, y estudiar algunas miradas contemporáneas acerca de la Guerra Civil y del franquismo desde el campo de la narrativa como un correlato de ciertos reclamos sociales de la actualidad respecto del pasado reciente. Para esto fueron analizadas diversas propuestas que surgieron esencialmente desde la literatura -con algún detenimiento en un texto fílmico- para abordar el pasado traumático y se consideraron las técnicas de transmisión empleadas para la narración ficcional de aspectos largamente silenciados por la historia oficial. El corpus central de textos literarios ha quedado conformado por O lapis do carpinteiro y Os libros arden mal, de Manuel Rivas, Guárdame bajo tierra, de Ramón Saizarbitoria, Soldados de Salamina, de Javier Cercas, Home sen nome, de Suso de Toro, y Mala gente que camina, de Benjamín Prado. Como corpus complementario, se tornó pertinente incorporar un capítulo que articula un texto fílmico (Muerte en El Valle, de Christina Hardt) con un texto literario (Las esquinas del aire. En busca de Ana María Martínez Sagi, de Juan Manuel de Prada). Esto, que en principio puede parecer un apartamiento del corpus central, favoreció el análisis del tratamiento del testimonio desde la estetización de un texto fílmico o literario. Al realizar la selección, se estudiaron, junto con autores que escriben y publican en castellano, algunos textos escritos originariamente en lenguas minorizadas (gallego y euskera)

Transmisión oral en la narrativa española contemporánea : Un recurso para la construcción de la memoria de la Guerra Civil y del franquismo

Se propone la descripción y el análisis del empleo de la transmisión oral en la narrativa española actual como un recurso para configurar la memoria de la Guerra Civil española y del franquismo. Para ello se atendió especialmente a las estrategias que intervienen en la representación de la oralidad en relación con los diálogos y los relatos orales, entre otros procedimientos a través de los cuales se transmite el pasado traumático dentro de la ficción. Ha sido analizada la representación de la oralidad en la producción narrativa ficcional de diversos autores españoles de finales del siglo XX y comienzos del siglo XXI que han abordado la temática de la Guerra Civil en una situación particular en cuanto al surgimiento de replanteos y debates acerca del tratamiento del conflicto bélico. Los objetivos generales que guiaron el trabajo han sido indagar la presencia y la función de la transmisión oral en la literatura, profundizar el estudio de la oralidad como voz emergente de colectivos sociales desplazados y analizar la funcionalidad del empleo de la oralidad en textos representativos de la narrativa española contemporánea. A su vez, más específicamente, se propuso profundizar en el estudio de la ficcionalización de la transmisión oral como un artificio para la construcción de la memoria en la narrativa española actual, vincular las posibilidades de transmisión de la historia de la Guerra Civil española con medios propios de la historia oral en la literatura -especialmente en la novela-, problematizar determinadas perspectivas sobre el testimonio en la medida en que son literaturizadas para una concepción de la memoria de la Guerra Civil, de la posguerra y de la dictadura, y estudiar algunas miradas contemporáneas acerca de la Guerra Civil y del franquismo desde el campo de la narrativa como un correlato de ciertos reclamos sociales de la actualidad respecto del pasado reciente. Para esto fueron analizadas diversas propuestas que surgieron esencialmente desde la literatura -con algún detenimiento en un texto fílmico- para abordar el pasado traumático y se consideraron las técnicas de transmisión empleadas para la narración ficcional de aspectos largamente silenciados por la historia oficial. El corpus central de textos literarios ha quedado conformado por O lapis do carpinteiro y Os libros arden mal, de Manuel Rivas, Guárdame bajo tierra, de Ramón Saizarbitoria, Soldados de Salamina, de Javier Cercas, Home sen nome, de Suso de Toro, y Mala gente que camina, de Benjamín Prado. Como corpus complementario, se tornó pertinente incorporar un capítulo que articula un texto fílmico (Muerte en El Valle, de Christina Hardt) con un texto literario (Las esquinas del aire. En busca de Ana María Martínez Sagi, de Juan Manuel de Prada). Esto, que en principio puede parecer un apartamiento del corpus central, favoreció el análisis del tratamiento del testimonio desde la estetización de un texto fílmico o literario. Al realizar la selección, se estudiaron, junto con autores que escriben y publican en castellano, algunos textos escritos originariamente en lenguas minorizadas (gallego y euskera)

Gaussian Multiscale Aggregation oriented to Hand Biometric Segmentation in Mobile Devices

New trends in biometrics are oriented to mobile devices in order to increase the overall security in daily actions like bank account access, e-commerce or even document protection within the mobile. However, applying biometrics to mobile devices imply challenging aspects in biometric data acquisition, feature extraction or private data storage. Concretely, this paper attempts to deal with the problem of hand segmentation given a picture of the hand in an unknown background, requiring an accurate result in terms of hand isolation. For the sake of user acceptability, no restrictions are done on background, and therefore, hand images can be taken without any constraint, resulting segmentation in an exigent task. Multiscale aggregation strategies are proposed in order to solve this problem due to their accurate results in unconstrained and complicated scenarios, together with their properties in time performance. This method is evaluated with a public synthetic database with 480000 images considering different backgrounds and illumination environments. The results obtained in terms of accuracy and time performance highlight their capability of being a suitable solution for the problem of hand segmentation in contact-less environments, outperforming competitive methods in literature like Lossy Data Compression image segmentation (LDC).

Gaussian Multiscale Aggregation Applied to Segmentation in Hand Biometrics

This paper presents an image segmentation algorithm based on Gaussian multiscale aggregation oriented to hand biometric applications. The method is able to isolate the hand from a wide variety of background textures such as carpets, fabric, glass, grass, soil or stones. The evaluation was carried out by using a publicly available synthetic database with 408,000 hand images in different backgrounds, comparing the performance in terms of accuracy and computational cost to two competitive segmentation methods existing in literature, namely Lossy Data Compression (LDC) and Normalized Cuts (NCuts). The results highlight that the proposed method outperforms current competitive segmentation methods with regard to computational cost, time performance, accuracy and memory usage.

Using modular neural networks to model self-consciousness and self-representation for artificial entities

Self-consciousness implies not only self or group recognition, but also real knowledge of one’s own identity. Self-consciousness is only possible if an individual is intelligent enough to formulate an abstract self-representation. Moreover, it necessarily entails the capability of referencing and using this elf-representation in connection with other cognitive features, such as inference, and the anticipation of the consequences of both one’s own and other individuals’ acts. In this paper, a cognitive architecture for self-consciousness is proposed. This cognitive architecture includes several modules: abstraction, self-representation, other individuals'representation, decision and action modules. It includes a learning process of self-representation by direct (self-experience based) and observational learning (based on the observation of other individuals). For model implementation a new approach is taken using Modular Artificial Neural Networks (MANN). For model testing, a virtual environment has been implemented. This virtual environment can be described as a holonic system or holarchy, meaning that it is composed of autonomous entities that behave both as a whole and as part of a greater whole. The system is composed of a certain number of holons interacting. These holons are equipped with cognitive features, such as sensory perception, and a simplified model of personality and self-representation. We explain holons’ cognitive architecture that enables dynamic self-representation. We analyse the effect of holon interaction, focusing on the evolution of the holon’s abstract self-representation. Finally, the results are explained and analysed and conclusions drawn.

Background foreground segmentation with RGB-D Kinect data: An efficient combination of classifiers

Low cost RGB-D cameras such as the Microsoft’s Kinect or the Asus’s Xtion Pro are completely changing the computer vision world, as they are being successfully used in several applications and research areas. Depth data are particularly attractive and suitable for applications based on moving objects detection through foreground/background segmentation approaches; the RGB-D applications proposed in literature employ, in general, state of the art foreground/background segmentation techniques based on the depth information without taking into account the color information. The novel approach that we propose is based on a combination of classifiers that allows improving background subtraction accuracy with respect to state of the art algorithms by jointly considering color and depth data. In particular, the combination of classifiers is based on a weighted average that allows to adaptively modifying the support of each classifier in the ensemble by considering foreground detections in the previous frames and the depth and color edges. In this way, it is possible to reduce false detections due to critical issues that can not be tackled by the individual classifiers such as: shadows and illumination changes, color and depth camouflage, moved background objects and noisy depth measurements. Moreover, we propose, for the best of the author’s knowledge, the first publicly available RGB-D benchmark dataset with hand-labeled ground truth of several challenging scenarios to test background/foreground segmentation algorithms.

Functional characterization of YODA, a mitogen-activated protein kinase kinase kinase (MAP3K) that regulates a novel innate immunity pathway in Arabidopsis thaliana

Las cascadas de señalización mediadas por proteína quinasas activadas por mitógeno (MAP quinasas) son capaces de integrar y transducir señales ambientales en respuestas celulares. Entre estas señales se encuentran los PAMPs/MAMPs (Pathogen/Microbe-Associated Molecular Patterns), que son moléculas de patógenos o microorganismos, o los DAMPs (Damaged-Associated Molecular Patterns), que son moléculas derivadas de las plantas producidas en respuesta a daño celular. Tras el reconocimiento de los PAMPs/DAMPs por receptores de membrana denominados PRRs (Pattern Recognition Receptors), como los receptores con dominio quinasa (RLKs) o los receptores sin dominio quinasa (RLPs), se activan respuestas moleculares, incluidas cascadas de MAP quinasas, que regulan la puesta en marcha de la inmunidad activada por PAMPs (PTI). Esta Tesis describe la caracterización funcional de la MAP quinasa quinasa quinasa (MAP3K) YODA (YDA), que actúa como un regulador clave de la PTI en Arabidopsis. Se ha descrito previamente que YDA controla varios procesos de desarrollo, como la regulación del patrón estomático, la elongación del zigoto y la arquitectura floral. Hemos caracterizado un alelo mutante hipomórfico de YDA (elk2 o yda11) que presenta una elevada susceptibilidad a patógenos biótrofos y necrótrofos. Notablemente, plantas que expresan una forma constitutivamente activa de YDA (CA-YDA), con una deleción en el dominio N-terminal, presentan una resistencia de amplio espectro frente a diferentes tipos de patógenos, incluyendo hongos, oomicetos y bacterias, lo que indica que YDA juega un papel importante en la regulación de la resistencia de las plantas a patógenos. Nuestros datos indican que esta función es independiente de las respuestas inmunes mediadas por los receptores previamente caracterizados FLS2 y CERK1, que reconocen los PAMPs flg22 y quitina, respectivamente, y que están implicados en la resistencia de Arabidopsis frente a bacterias y hongos. Hemos demostrado que YDA controla la resistencia frente al hongo necrótrofo Plectosphaerella cucumerina y el patrón estomático mediante su interacción genética con la RLK ERECTA (ER), un PRR implicado en la regulación de estos procesos. Por el contrario, la interacción genética entre ER y YDA en la regulación de otros procesos de desarrollo es aditiva en lugar de epistática. Análisis genéticos indicaron que MPK3, una MAP quinasa que funciona aguas abajo de YDA en el desarrollo estomático, es un componente de la ruta de señalización mediada por YDA para la resistencia frente a P. cucumerina, lo que sugiere que el desarrollo de las plantas y la PTI comparten el módulo de transducción de MAP quinasas asociado a YDA. Nuestros experimentos han revelado que la resistencia mediada por YDA es independiente de las rutas de señalización reguladas por las hormonas de defensa ácido salicílico, ácido jasmónico, ácido abscísico o etileno, y también es independiente de la ruta de metabolitos secundarios derivados del triptófano, que están implicados en inmunidad vegetal. Además, hemos demostrado que respuestas asociadas a PTI, como el aumento en la concentración de calcio citoplásmico, la producción de especies reactivas de oxígeno, la fosforilación de MAP quinasas y la expresión de genes de defensa, no están afectadas en el mutante yda11. La expresión constitutiva de la proteína CA-YDA en plantas de Arabidopsis no provoca un aumento de las respuestas PTI, lo que sugiere la existencia de mecanismos de resistencia adicionales regulados por YDA que son diferentes de los regulados por FLS2 y CERK1. En línea con estos resultados, nuestros datos transcriptómicos revelan una sobre-representación en plantas CA-YDA de genes de defensa que codifican, por ejemplo, péptidos antimicrobianos o reguladores de muerte celular, o proteínas implicadas en la biogénesis de la pared celular, lo que sugiere una conexión potencial entre la composición e integridad de la pared celular y la resistencia de amplio espectro mediada por YDA. Además, análisis de fosfoproteómica indican la fosforilación diferencial de proteínas relacionadas con la pared celular en plantas CA-YDA en comparación con plantas silvestres. El posible papel de la ruta ER-YDA en la regulación de la integridad de la pared celular está apoyado por análisis bioquímicos y glicómicos de las paredes celulares de plantas er, yda11 y CA-YDA, que revelaron cambios significativos en la composición de la pared celular de estos genotipos en comparación con la de plantas silvestres. En resumen, nuestros datos indican que ER y YDA forman parte de una nueva ruta de inmunidad que regula la integridad de la pared celular y respuestas defensivas, confiriendo una resistencia de amplio espectro frente a patógenos. ABSTRACT Plant mitogen-activated protein kinase (MAPK) cascades transduce environmental signals and developmental cues into cellular responses. Among these signals are the pathogen- or microbe-associated molecular patterns (PAMPs or MAMPs) and the damage-associated molecular patterns (DAMPs). These PAMPs/DAMPs, upon recognition by plant pattern recognition receptors (PRRs), such as Receptor-Like Kinases (RLKs) and Receptor-Like Proteins (RLPs), activate molecular responses, including MAPK cascades, which regulate the onset of PAMP-triggered immunity (PTI). This Thesis describes the functional characterization of the MAPK kinase kinase (MAP3K) YODA (YDA) as a key regulator of Arabidopsis PTI. YDA has been previously described to control several developmental processes, such as stomatal patterning, zygote elongation and inflorescence architecture. We characterized a hypomorphic, non-embryo lethal mutant allele of YDA (elk2 or yda11) that was found to be highly susceptible to biotrophic and necrotrophic pathogens. Remarkably, plants expressing a constitutive active form of YDA (CA-YDA), with a deletion in the N-terminal domain, showed broad-spectrum resistance to different types of pathogens, including fungi, oomycetes and bacteria, indicating that YDA plays a relevant function in plant resistance to pathogens. Our data indicated that this function is independent of the immune responses regulated by the well characterized FLS2 and CERK1 RLKs, which are the PRRs recognizing flg22 and chitin PAMPs, respectively, and are required for Arabidopsis resistance to bacteria and fungi. We demonstrate that YDA controls resistance to the necrotrophic fungus Plectosphaerella cucumerina and stomatal patterning by genetically interacting with ERECTA (ER) RLK, a PRR involved in regulating these processes. In contrast, the genetic interaction between ER and YDA in the regulation of other ER-associated developmental processes was additive, rather than epistatic. Genetic analyses indicated that MPK3, a MAP kinase that functions downstream of YDA in stomatal development, also regulates plant resistance to P. cucumerina in a YDA-dependent manner, suggesting that the YDA-associated MAPK transduction module is shared in plant development and PTI. Our experiments revealed that YDA-mediated resistance was independent of signalling pathways regulated by defensive hormones like salicylic acid, jasmonic acid, abscisic acid or ethylene, and of the tryptophan-derived metabolites pathway, which are involved in plant immunity. In addition, we showed that PAMP-mediated PTI responses, such as the increase of cytoplasmic Ca2+ concentration, reactive oxygen species (ROS) burst, MAPK phosphorylation, and expression of defense-related genes are not impaired in the yda11 mutant. Furthermore, the expression of CA-YDA protein does not result in enhanced PTI responses, further suggesting the existence of additional mechanisms of resistance regulated by YDA that differ from those regulated by the PTI receptors FLS2 and CERK1. In line with these observations, our transcriptomic data revealed the over-representation in CA-YDA plants of defensive genes, such as those encoding antimicrobial peptides and cell death regulators, and genes encoding cell wall-related proteins, suggesting a potential link between plant cell wall composition and integrity and broad spectrum resistance mediated by YDA. In addition, phosphoproteomic data revealed an over-representation of genes encoding wall-related proteins in CA-YDA plants in comparison with wild-type plants. The putative role of the ER-YDA pathway in regulating cell wall integrity was further supported by biochemical and glycomics analyses of er, yda11 and CA-YDA cell walls, which revealed significant changes in the cell wall composition of these genotypes compared with that of wild-type plants. In summary, our data indicate that ER and YDA are components of a novel immune pathway that regulates cell wall integrity and defensive responses, which confer broad-spectrum resistance to pathogens.

Automatic physical activity pattern recognition and meal intake modelling in type 1 diabetes for their inclusion in artificial pancreas systems

La diabetes comprende un conjunto de enfermedades metabólicas que se caracterizan por concentraciones de glucosa en sangre anormalmente altas. En el caso de la diabetes tipo 1 (T1D, por sus siglas en inglés), esta situación es debida a una ausencia total de secreción endógena de insulina, lo que impide a la mayoría de tejidos usar la glucosa. En tales circunstancias, se hace necesario el suministro exógeno de insulina para preservar la vida del paciente; no obstante, siempre con la precaución de evitar caídas agudas de la glucemia por debajo de los niveles recomendados de seguridad. Además de la administración de insulina, las ingestas y la actividad física son factores fundamentales que influyen en la homeostasis de la glucosa. En consecuencia, una gestión apropiada de la T1D debería incorporar estos dos fenómenos fisiológicos, en base a una identificación y un modelado apropiado de los mismos y de sus sorrespondientes efectos en el balance glucosa-insulina. En particular, los sistemas de páncreas artificial –ideados para llevar a cabo un control automático de los niveles de glucemia del paciente– podrían beneficiarse de la integración de esta clase de información. La primera parte de esta tesis doctoral cubre la caracterización del efecto agudo de la actividad física en los perfiles de glucosa. Con este objetivo se ha llevado a cabo una revisión sistemática de la literatura y meta-análisis que determinen las respuestas ante varias modalidades de ejercicio para pacientes con T1D, abordando esta caracterización mediante unas magnitudes que cuantifican las tasas de cambio en la glucemia a lo largo del tiempo. Por otro lado, una identificación fiable de los periodos con actividad física es un requisito imprescindible para poder proveer de esa información a los sistemas de páncreas artificial en condiciones libres y ambulatorias. Por esta razón, la segunda parte de esta tesis está enfocada a la propuesta y evaluación de un sistema automático diseñado para reconocer periodos de actividad física, clasificando su nivel de intensidad (ligera, moderada o vigorosa); así como, en el caso de periodos vigorosos, identificando también la modalidad de ejercicio (aeróbica, mixta o de fuerza). En este sentido, ambos aspectos tienen una influencia específica en el mecanismo metabólico que suministra la energía para llevar a cabo el ejercicio y, por tanto, en las respuestas glucémicas en T1D. En este trabajo se aplican varias combinaciones de técnicas de aprendizaje máquina y reconocimiento de patrones sobre la fusión multimodal de señales de acelerometría y ritmo cardíaco, las cuales describen tanto aspectos mecánicos del movimiento como la respuesta fisiológica del sistema cardiovascular ante el ejercicio. Después del reconocimiento de patrones se incorpora también un módulo de filtrado temporal para sacar partido a la considerable coherencia temporal presente en los datos, una redundancia que se origina en el hecho de que en la práctica, las tendencias en cuanto a actividad física suelen mantenerse estables a lo largo de cierto tiempo, sin fluctuaciones rápidas y repetitivas. El tercer bloque de esta tesis doctoral aborda el tema de las ingestas en el ámbito de la T1D. En concreto, se propone una serie de modelos compartimentales y se evalúan éstos en función de su capacidad para describir matemáticamente el efecto remoto de las concetraciones plasmáticas de insulina exógena sobre las tasas de eleiminación de la glucosa atribuible a la ingesta; un aspecto hasta ahora no incorporado en los principales modelos de paciente para T1D existentes en la literatura. Los datos aquí utilizados se obtuvieron gracias a un experimento realizado por el Institute of Metabolic Science (Universidad de Cambridge, Reino Unido) con 16 pacientes jóvenes. En el experimento, de tipo ‘clamp’ con objetivo variable, se replicaron los perfiles individuales de glucosa, según lo observado durante una visita preliminar tras la ingesta de una cena con o bien alta carga glucémica, o bien baja. Los seis modelos mecanísticos evaluados constaban de: a) submodelos de doble compartimento para las masas de trazadores de glucosa, b) un submodelo de único compartimento para reflejar el efecto remoto de la insulina, c) dos tipos de activación de este mismo efecto remoto (bien lineal, bien con un punto de corte), y d) diversas condiciones iniciales. ABSTRACT Diabetes encompasses a series of metabolic diseases characterized by abnormally high blood glucose concentrations. In the case of type 1 diabetes (T1D), this situation is caused by a total absence of endogenous insulin secretion, which impedes the use of glucose by most tissues. In these circumstances, exogenous insulin supplies are necessary to maintain patient’s life; although caution is always needed to avoid acute decays in glycaemia below safe levels. In addition to insulin administrations, meal intakes and physical activity are fundamental factors influencing glucose homoeostasis. Consequently, a successful management of T1D should incorporate these two physiological phenomena, based on an appropriate identification and modelling of these events and their corresponding effect on the glucose-insulin balance. In particular, artificial pancreas systems –designed to perform an automated control of patient’s glycaemia levels– may benefit from the integration of this type of information. The first part of this PhD thesis covers the characterization of the acute effect of physical activity on glucose profiles. With this aim, a systematic review of literature and metaanalyses are conduced to determine responses to various exercise modalities in patients with T1D, assessed via rates-of-change magnitudes to quantify temporal variations in glycaemia. On the other hand, a reliable identification of physical activity periods is an essential prerequisite to feed artificial pancreas systems with information concerning exercise in ambulatory, free-living conditions. For this reason, the second part of this thesis focuses on the proposal and evaluation of an automatic system devised to recognize physical activity, classifying its intensity level (light, moderate or vigorous) and for vigorous periods, identifying also its exercise modality (aerobic, mixed or resistance); since both aspects have a distinctive influence on the predominant metabolic pathway involved in fuelling exercise, and therefore, in the glycaemic responses in T1D. Various combinations of machine learning and pattern recognition techniques are applied on the fusion of multi-modal signal sources, namely: accelerometry and heart rate measurements, which describe both mechanical aspects of movement and the physiological response of the cardiovascular system to exercise. An additional temporal filtering module is incorporated after recognition in order to exploit the considerable temporal coherence (i.e. redundancy) present in data, which stems from the fact that in practice, physical activity trends are often maintained stable along time, instead of fluctuating rapid and repeatedly. The third block of this PhD thesis addresses meal intakes in the context of T1D. In particular, a number of compartmental models are proposed and compared in terms of their ability to describe mathematically the remote effect of exogenous plasma insulin concentrations on the disposal rates of meal-attributable glucose, an aspect which had not yet been incorporated to the prevailing T1D patient models in literature. Data were acquired in an experiment conduced at the Institute of Metabolic Science (University of Cambridge, UK) on 16 young patients. A variable-target glucose clamp replicated their individual glucose profiles, observed during a preliminary visit after ingesting either a high glycaemic-load or a low glycaemic-load evening meal. The six mechanistic models under evaluation here comprised: a) two-compartmental submodels for glucose tracer masses, b) a single-compartmental submodel for insulin’s remote effect, c) two types of activations for this remote effect (either linear or with a ‘cut-off’ point), and d) diverse forms of initial conditions.