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In this study we are going to use different omic-techniques to analyze fruits of three species of berries such as strawberry, raspberry and black currant. Berry fruit are well appreciated for their delicate flavor and nutraceutical properties, with consumer demand increasing over the last years. Furthermore, climate change and market globalization have made necessary to improve the production while maintaining fruit quality traits. Goodberry project is developping analytical platforms, covering from transcriptomic to metabolites and volatile compounds analysis, to find new factors controlling plant adaptation, fruit production and quality. In this study we implement the metabolomic analysis of strawberry, raspberry and black currant fruits from the 2017 harvest, as well as 2018 harvest during this year. To analyze and compare the data we use multiomic tools and bioinformatics to extract properly conclusion The analyses take different berry cultivars, adapted to diverse environments, were grown in 2017 and 2018 in different latitudes (Germany, France, Norway, Italy, Poland and Scotland). The data comes from a combination of gas-chromatography-mass spectrometry (GC-TOF-MS) and headspace solid phase micro extraction (HS-SPME) coupled with GC-MS was used to semi-quantify fruit primary metabolome and volatilome. Around 50 key primary metabolites, including sugars and acids, which are fundamental factors influencing fruit taste and 75 volatiles, responsible of the aroma, were identified across the different genotypes and climates. Multivariate statistical approaches allow us to point out the genetic and environmental factors underlying complex metabolic traits involved in fruit quality. Preliminary analysis showed that both climate and genetic factors influence primary metabolite and volatile content, even if the environment seems to have a stronger impact on the first one. Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Berries, such as strawberry, raspberry and black currant, are well appreciated for their delicate flavor and nutraceutical properties, with consumer demand increasing over the last years. However, climate change and market globalization have made necessary to improve the production while maintaining fruit quality traits. Among the EU GoodBerry project’s objetive are develop state-of-the-art analytical platforms, covering from transcriptomic to metabolites and volatile compounds analysis, to find new factors controlling plant adaptation, fruit production and quality and use the data to face climate changes. Here we present the metabolomic analysis of strawberry, raspberry and black currant fruits from the 2017 harvest. Different berry cultivars, adapted to diverse environments, were grown in 2017 and 2018 in different latitudes (Germany, France, Norway, Italy and Poland) combination of spectrometry techniques was used to semi-quantify fruit primary metabolome and volatilome. Around 50 key primary metabolites, including sugars and acids, which are fundamental factors influencing fruit taste and 75 volatiles, responsible of the aroma, were identified across the different genotypes and climates. Multivariate statistical approaches allow us to point out the genetic and environmental factors underlying complex metabolic traits involved in fruit quality. Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech.

This article examines the effect of assumption or rejection on the non-filing joint venture members when a joint venture’s operating agreement is characterized as an executory contract and subject to assumption or rejection under the Bankruptcy Code by the co-venturer that filed for bankruptcy protection. The article presents a hypothetical to analyze the effect of dissociation, the effect of bankruptcy on managerial rights, and liquidation of the joint venture. Finally, the article provides practical considerations for protecting joint venture members prior to a bankruptcy and what to do when a partner is in financial distress.

[spa] La presente tesis se basa en cuatro artículos originales y un artículo de revisión que pertenecen a una misma línea de trabajo: el uso de técnicas de resonancia magnética (RM) y electrofisiológicas para evaluar y predecir la discapacidad física o cognitiva en pacientes con esclerosis múltiple (EM). En primer lugar, a través de RM convencional se ha evaluado el comportamiento radiológico de determinados patrones de lesiones cerebrales y su asociación con una peor evolución de la discapacidad. El trabajo muestra que las lesiones con un borde hipointenso en secuencias T2 y las lesiones captantes de gadolinio en anillo aparecen en el 9 y 12% de los pacientes con EM seguidos según la práctica clínica habitual. Además, la presencia de lesiones captantes de gadolinio en forma de anillo incrementa el riesgo de alcanzar una discapacidad elevada en menor tiempo y por tanto predice una peor evolución de la enfermedad. En segundo lugar se ha realizado un estudio multimodal del cuerpo calloso utilizando de forma conjunta técnicas electrofisiológicas con RM avanzada para indagar en el papel de la alteración de dicha estructura sobre la discapacidad física y cognitiva. El cuerpo calloso en pacientes con EM presenta una alteración tanto de la estructura e integridad microestructural como en la función de inhibición transcallosa. La alteración estructural y funcional de esta comisura se asocia con un bajo rendimiento cognitivo y con mayor discapacidad física. En tercer lugar se han utilizado diversas técnicas de RM avanzada para evaluar la integridad del tejido cerebral tanto de la sustancia gris como de la sustancia blanca y su relación con el rendimiento de diversas funciones cognitivas. En los pacientes existe un daño global de la integridad microestructural que afecta la sustancia blanca y la sustancia gris desde fases iniciales de la enfermedad. Sin embargo, la relación entre el daño microestructural y la disfunción cognitiva es específica de algunas regiones concretas según el test cognitivo evaluado. La integridad microestructural en la sustancia blanca y la presencia de lesiones parecen ser los principales eventos que explican la varianza del rendimiento cognitivo en pacientes poco discapacitados. En cuarto lugar, se ha investigado la capacidad de marcadores de desmielinización, daño axonal y astrogliosis en RM avanzada para predecir la discapacidad y la atrofia cerebral a medio plazo. Este último trabajo, actualmente enviado para su revisión a una revista científica, fue seleccionado para ser presentado en la “Contemporary Clinical Issues Plennary Session” de la Academia Americana de Neurología celebrada en Nueva Orleans en 2012. Del trabajo se concluye que la astrogliosis y el daño axonal son eventos relevantes en la progresión de la discapacidad y se propone el ratio de mioinositol/N-acetilaspartato obtenido mediante espectroscopia es un robusto predictor de la evolución de la atrofia y la discapacidad en pacientes con esclerosis múltiple. Estos estudios han servido para profundizar en el conocimiento de la fisiopatología de los condicionantes de la discapacidad, y en el avance en la búsqueda de marcadores de RM y electrofisiológicos útiles para evaluar y predecir la discapacidad en la EM., [eng] This thesis is based in four original articles and one review from the same research topic: the use of different magnetic resonance (MR) and electrophysiological techniques to evaluate and predict the clinical and cognitive disability in patients with multiple sclerosis (MS). First, the radiological behaviour and clinical meaning of lesions with hipointense rim on T2 weighted sequences and lesions with ring enhancement on T1 were evaluated. The presence of ring enhancement increases the risk of disability worsening. On the second article, the corpus callosum was evaluated through electrophysiological and MR techniques to understand the role of this structure on physical and cognitive dysfunction in MS patients. The structure and function of the corpus callosum is impaired in MS and relates to worse cognitive performance in memory and executive tests. Third, we used several advanced MR techniques to evaluate the integrity of brain tissue, both in white matter and grey matter, in MS and their relationship with cognitive functions. The microstructural integrity is widespread impaired although the cognitive performance relates to the damage of specific regions. The impairment of white matter microstructural integrity and the presence of lesions are the main factors explaining the cognitive impairment in patients. Finally, the capacity of markers of demyelination, axonal damage and astrogliosis to predict the evolution of disabilty and brain atrophy along four years was investigated. This work shows that astrogliosis and axonal damage are cardinal events leading to disesase evolution. The ratio of myoinositol and N-acetylaspartate obtained through spectroscopy is a robust predictor of disability and brain atrophy progression. These works have improved the knowledge on the physiopathological events underlying disability in multiple sclerosis and can be useful in the search of biomarkers that predict disease evolution.

The optic radiation (OR) is one of the major components of the visual system and a key structure at risk in white matter diseases such as multiple sclerosis (MS). However, it is challenging to perform track reconstruction of the OR using diffusion MRI due to a sharp change of direction in the Meyer's loop and the presence of kissing and crossing fibers along the pathway. As such, we aimed to provide a highly precise and reproducible framework for tracking the OR from thalamic and visual cortex masks. The framework combined the generation of probabilistic streamlines by high order fiber orientation distributions estimated with constrained spherical deconvolution and an automatic post-processing based on anatomical exclusion criteria (AEC) to compensate for the presence of anatomically implausible streamlines. Specifically, those ending in the contralateral hemisphere, cerebrospinal fluid or grey matter outside the visual cortex were automatically excluded. We applied the framework to two distinct high angular resolution diffusion-weighted imaging (HARDI) acquisition protocols on one cohort, comprised of ten healthy volunteers and five MS patients. The OR was successfully delineated in both HARDI acquisitions in the healthy volunteers and MS patients. Quantitative evaluation of the OR position was done by comparing the results with histological reference data. Compared with histological mask, the OR reconstruction into a template (OR-TCT) was highly precise (percentage of voxels within the OR-TCT correctly defined as OR), ranging from 0.71 to 0.83. The sensitivity (percentage of voxels in histological reference mask correctly defined as OR in OR-TCT) ranged from 0.65 to 0.81 and the accuracy (measured by F1 score) was 0.73 to 0.77 in healthy volunteers. When AEC was not applied the precision and accuracy decreased. The absolute agreement between both HARDI datasets measured by the intraclass correlation coefficient was 0.73. This improved framework allowed us to reconstruct the OR with high reliability and accuracy independently of the acquisition parameters. Moreover, the reconstruction was possible even in the presence of tissue damage due to MS. This framework could also be applied to other tracts with complex configuration.

Three nanostructured vehicles are proposed as potential carriers for photosensitizers to be used in photodynamic therapy: spherical nanoparticles, hexahedral microparticles and cylindrical magnetic nanorods. A comparative study of their photodynamic properties was performed, and the influence of their size and the amount of loaded porphyrin was considered to discuss their effects in the observed photodynamic activity. All the vehicles have a gold surface, allowing functionalization with a disulphide-containing porphyrin as the photosensitizer, as well as with a PEG-containing thiol to improve their biocompatibility and water solubility. The activity of the porphyrin loaded in each vehicle was assessed through in vitro photocytotoxicity studies using HeLa cells. A synergic effect for the porphyrin toxicity was observed in all of the vehicles. The zinc-containing porphyrin showed better production of singlet oxygen, and proved more photocytotoxic both in solution and loaded in any of the vehicles. The magnetism of the nanorods allows targeting with a magnetic field, but causes their aggregation, hampering the porphyrin's activity. Microparticles showed lower cell internalization but their bigger size allowed a high porphyrin loading, which translated into high photocytotoxicity. The highest cell internalization and photocytotoxicity was observed for the porphyrin-loaded nanoparticles, suggesting that a smaller size is favored in cell uptake.

[spa] Esta memoria de tesis se presenta en formato de artículos originales que corresponden a una misma unidad temática: el desarrollo de una metodología de reconstrucción de fibras de sustancia blanca basada en técnicas de difusión por resonancia magnética y su implementación en el estudio de conectividad estructural en pacientes con esclerosis múltiple. Para el primer artículo, seleccionamos las radiaciones ópticas para validar nuestra metodología de reconstrucción. Para ello, empleamos modelos avanzados de tractografía y un postprocesado basado en criterios de exclusión anatómicos propios de cada sujeto. La reconstrucción final de las radiaciones ópticas fue evaluada a través de un atlas histológico validado por la comunidad científica, obteniendo una precisión y exactitud volumétrica muy cercana a la anatomía real tanto en voluntarios sanos como en pacientes con esclerosis múltiple. En el segundo artículo se empleó la misma metodología desarrollada en el artículo precedente, con el fin de proporcionar reconstrucciones más precisas de las conexiones de sustancia blanca cerebral y obtener datos cuantitativos y patológicamente más específicos. Esto permitió conocer los cambios acontecidos en la conectividad cerebral estructural en pacientes con esclerosis múltiple y sus consecuencias sobre el rendimiento cognitivo. Pudimos identificar un daño difuso generalizado de la red cerebral así como la presencia de fenómenos de desconexión en regiones frontoparietales, sustancia gris subcortical e ínsula, relacionados con el rendimiento en funciones atencionales y ejecutivas. Además, evidenciamos posibles mecanismos de reestructuración cerebral a través de incrementos en la conectividad en conexiones específicas. Dichos mecanismos de plasticidad podrían ser tanto, herramientas para compensar el daño tisular como cambios estructurales maladaptativos. Estos estudios han permitido desarrollar y aplicar una nueva metodología de reconstrucción de fibras de sustancia blanca más precisa y reproducible, así como detectar aquellas redes más vulnerables sobre el rendimiento cognitivo. La identificación de estas redes permite contar con un biomarcador sensible a la discapacidad cognitiva y podrían llegar a utilizarse como diana para evaluar el efecto de nuevas intervenciones terapéuticas., [eng] This thesis was submitted with original research articles belonging to the same subject area: the development of an improved framework for tractography reconstruction of white matter fibers based on diffusion-weighted magnetic resonance imaging and its implementation in the study of structural connectivity in patients with multiple sclerosis. The purpose of the first original paper was to introduce an improved tractography framework to reconstruct the optic radiations in healthy volunteers and multiple sclerosis patients. The framework combined advanced diffusion models and an automatic post-processing based on anatomical exclusion criteria. The quantitative evaluation of the optic radiation was done by comparing the final reconstruction with a histological reference data. The proposed methodology was capable to reconstruct the optic radiation with a high accuracy and reliability. Moreover, the reconstruction was possible even in the presence of tissue damage. This framework could also be applied to other tracts with complex configuration in order to reduce the presence of anatomically implausible fibers. The second article used the same methodology developed in the previous article, in order to provide more precisely reconstructions of white matter connections. We aimed to investigate changes in structural connectivity in multiple sclerosis patients and their association with attention and executive performance. The structural brain connectivity is disturbed in multiple sclerosis due to widespread impairment of white matter connections and gray matter structures. The worse attention and executive performance was related to the decrease of integrity in frontoparietal networks, deep gray nuclei and insula. Contrarily, the increased edge connectivity suggests the presence of reorganization mechanisms at the structural level. These changes could be compensatory mechanisms to the maintenance of the normal level of cognitive performance or reflect maladaptative changes. These studies developed and improved framework for tractography reconstruction, useful to evaluate the changes in brain connectivity in patients with multiple sclerosis. The strategic networks related to attention and executive performance could be used as biomarkers to detect cognitive disability and targets to assess the effect of new therapeutic interventions.