Analysis of the influence of epitope flanking regions on MHC class I restricted antigen presentation

Peptides presented by MHC class I molecules for CTL recognition are derived mainly from cytosolic proteins. For antigen presentation on the cell surface, epitopes require correct processing by cytosolic and ER proteases, efficient TAP transport and MHC class I binding affinity. The efficiency of epitope generation depends not only on the epitope itself, but also on its flanking regions. In this project, the influence of the C-terminal region of the model epitope SIINFEKL (S8L) from chicken ovalbumin (aa 257-264) on antigen processing has been investigated. S8L is a well characterized epitope presented on the murine MHC class I molecule, H-2Kb. The Flp-In 293Kb cell line was transfected with different constructs each enabling the expression of the S8L sequence with different defined C-terminal flanking regions. The constructs differed at the two first C-terminal positions after the S8L epitope, so called P1’ and P2’. At these sites, all 20 amino acids were exchanged consecutively and tested for their influence on H-2Kb/S8L presentation on the cell surface of the Flp-In 293Kb cells. The detection of this complex was performed by immunostaining and flow cytometry. The prevailing assumption is that proteasomal cleavages are exclusively responsible for the generation of the final C-termini of CTL epitopes. Nevertheless, recent publications showed that TPPII (tripeptidyl peptidase II) is required for the generation of the correct C-terminus of the HLA-A3-restricted HIV epitope Nef(73-82). With this background, the dependence of the S8L generation on proteasomal cleavage of the designed constructs was characterized using proteasomal inhibitors. The results obtained indicate that it is crucial for proteasomal cleavage, which amino acid is flanking the C-terminus of an epitope. Furthermore, partially proteasome independent S8L generation from specific S8L-precursor peptides was observed. Hence, the possibility of other existing endo- or carboxy-peptidases in the cytosol that could be involved in the correct trimming of the C-terminus of antigenic peptides for MHC class I presentation was investigated, performing specific knockdowns and using inhibitors against the target peptidases. In parallel, a purification strategy to identify the novel peptidase was established. The purified peaks showing an endopeptidase activity were further analyzed by mass spectrometry and some potential peptidases (like e.g. Lon) were identified, which have to be further characterized.

Role of Reactive Oxygen Species in signalling and oxidative stress

Results reported in this Thesis contribute to the comprehension of the complicated world of “redox biology”. ROS regulate signalling pathways both in physiological responses and in pathogenesis and progression of diseases. In cancer cells, the increase in ROS generation from metabolic abnormalities and oncogenic signalling may trigger a redox adaptation response, leading to an up-regulation of antioxidant capacity in order to maintain the ROS level below the toxic threshold. Thus, cancer cells would be more dependent on the antioxidant system and more vulnerable to further oxidative stress induced by exogenous ROS-generating agents or compounds that inhibit the antioxidant system. Results here reported indicate that the development of new drugs targeting specific Nox isoforms, responsible for intracellular ROS generation, or AQP isoforms, involved in the transport of extracellular H2O2 toward intracellular targets, might be an interesting novel anti-leukaemia strategy. Furthermore, also the use of CSD peptide, which simulate the VEGFR-2 segregation into caveolae in the inactive form, might be a strategy to stop the cellular response to VEGF signalling. As above stated, in the understanding of the redox biology, it is also important to identify and distinguish the molecular effectors that maintain normal biological and physiological responses, such as agents that stimulate our adaptation systems and elevate our endogenous antioxidant defences or other protective systems. Data here reported indicate that the nutraceutical compound sulforaphane and the Klotho protein are able to stimulate the HO-1 and Prx-1 expression, as well as the GSH levels, confirming their antioxidant and protective role. Finally, results here reported demonstrated that Stevia extracts are involved in insulin regulated glucose metabolism, suggesting that the use of these compounds goes beyond their sweetening power and may also offer therapeutic benefits hence improving the quality of life.

MULTIDIMENSIONAL OPTIMAL DROOP CONTROL FOR WIND RESOURCES IN DC MICROGRIDS

Two important and upcoming technologies, microgrids and electricity generation from wind resources, are increasingly being combined. Various control strategies can be implemented, and droop control provides a simple option without requiring communication between microgrid components. Eliminating the single source of potential failure around the communication system is especially important in remote, islanded microgrids, which are considered in this work. However, traditional droop control does not allow the microgrid to utilize much of the power available from the wind. This dissertation presents a novel droop control strategy, which implements a droop surface in higher dimension than the traditional strategy. The droop control relationship then depends on two variables: the dc microgrid bus voltage, and the wind speed at the current time. An approach for optimizing this droop control surface in order to meet a given objective, for example utilizing all of the power available from a wind resource, is proposed and demonstrated. Various cases are used to test the proposed optimal high dimension droop control method, and demonstrate its function. First, the use of linear multidimensional droop control without optimization is demonstrated through simulation. Next, an optimal high dimension droop control surface is implemented with a simple dc microgrid containing two sources and one load. Various cases for changing load and wind speed are investigated using simulation and hardware-in-the-loop techniques. Optimal multidimensional droop control is demonstrated with a wind resource in a full dc microgrid example, containing an energy storage device as well as multiple sources and loads. Finally, the optimal high dimension droop control method is applied with a solar resource, and using a load model developed for a military patrol base application. The operation of the proposed control is again investigated using simulation and hardware-in-the-loop techniques.

Lack of cathelicidin processing in Papillon-Lefèvre syndrome patients reveals essential role of LL-37 in periodontal homeostasis.

BACKGROUND Loss-of-function point mutations in the cathepsin C gene are the underlying genetic event in patients with Papillon-Lefèvre syndrome (PLS). PLS neutrophils lack serine protease activity essential for cathelicidin LL-37 generation from hCAP18 precursor. AIM We hypothesized that a local deficiency of LL-37 in the infected periodontium is mainly responsible for one of the clinical hallmark of PLS: severe periodontitis already in early childhood. METHODS To confirm this effect, we compared the level of neutrophil-derived enzymes and antimicrobial peptides in gingival crevicular fluid (GCF) and saliva from PLS, aggressive and chronic periodontitis patients. RESULTS Although neutrophil numbers in GCF were present at the same level in all periodontitis groups, LL-37 was totally absent in GCF from PLS patients despite the large amounts of its precursor, hCAP18. The absence of LL-37 in PLS patients coincided with the deficiency of both cathepsin C and protease 3 activities. The presence of other neutrophilic anti-microbial peptides in GCF from PLS patients, such as alpha-defensins, were comparable to that found in chronic periodontitis. In PLS microbial analysis revealed a high prevalence of Aggregatibacter actinomycetemcomitans infection. Most strains were susceptible to killing by LL-37. CONCLUSIONS Collectively, these findings imply that the lack of protease 3 activation by dysfunctional cathepsin C in PLS patients leads to the deficit of antimicrobial and immunomodulatory functions of LL-37 in the gingiva, allowing for infection with A. actinomycetemcomitans and the development of severe periodontal disease.

Modelización y generación de mapas del comportamiento espacial de Sporisorium reilianum ((Kühn) Langdon y Fullerton) en Estado de México, México

El presente estudio se realizó con la finalidad de modelizar la distribución espacial del carbón de la espiga del maíz causada por Sporisorium reilianum durante 2006 en el Estado de México y su visualización a través de la generación mapas de densidad. El muestreo se realizó en 100 parcelas georreferenciadas por cada localidad analizada. La incidencia de la enfermedad (porcentaje de plantas enfermas) se determinó al establecer cinco puntos parcela, en cada punto se contabilizaron 100 plantas. Se realizó el análisis geoestadístico para estimar el semivariograma experimental, una vez obtenido, se ajustó a un modelo teórico (esférico, exponencial o gaussiano) a través de los programas Variowin 2.2., su ajuste se validó a través de la validación cruzada. Posteriormente, se elaboraron mapas de agregación de la enfermedad con el método de interpolación geoestadística o krigeado. Los resultados indicaron que la enfermedad se presentó en 20 localidades de 19 municipios del Estado de México; todas las localidades presentaron un comportamiento espacial agregado de la enfermedad, 16 localidades se ajustaron al modelo esférico, dos al modelo exponencial y dos localidades se ajustaron al modelo gaussiano. En todos los modelos se lograron establecer mapas de agregación que permitirá adecuar las acciones de manejo en términos de puntos o sitios específicos.

Cómo leímos, aquí y entonces, la literatura española

Se trata de un testimonio, con un enfoque personal (y probablemente muy parcial) de los momentos y las vías por las cuales fue llegando la literatura española a los escritores argentinos de mi generación, desde los años sesenta hasta finales del siglo XX

Cómo leímos, aquí y entonces, la literatura española

Se trata de un testimonio, con un enfoque personal (y probablemente muy parcial) de los momentos y las vías por las cuales fue llegando la literatura española a los escritores argentinos de mi generación, desde los años sesenta hasta finales del siglo XX

Cómo leímos, aquí y entonces, la literatura española

Se trata de un testimonio, con un enfoque personal (y probablemente muy parcial) de los momentos y las vías por las cuales fue llegando la literatura española a los escritores argentinos de mi generación, desde los años sesenta hasta finales del siglo XX

Isotope composition of sulfur, oxygen and carbon in Black Sea sediments

Inversion of isotopic composition in the SO4(2-)-H2S system is shown to be universal in Neoeuxine sediments and an explanation of its occurrence is proposed. Change in isotopic composition of sulfate sulfur in Black Sea waters over last 10-15 thousand years is reconstructed. Periods of alteration between aerobic and anaerobic situations are identified, the beginning of entry of Mediterranean waters into the basin is dated, presence of authigenic carbonates in sediments of the sea is established and amounts are determined. Methane generation from carbon dioxide is shown to have been replaced by its generation from acetate in the paleo-Black Sea period.

n-Alkanes and lignin in bottom sediments of the Norwegian Sea

Investigations of bottom sediments from the central and northern parts of the Norwegian Sea including study regions at the Storegga landslide, the Haakon Mosby mud volcano, and Knipovich Ridge were carried out. Concentration of n-alkanes in bottom sediments from these regions ranges from 0.53 to 22.1 µg/g of dry sediments that corresponds to 0.02-1.97% of Corg. Molecular composition of hydrocarbons indicates mixed allochtonous-authochtonous genesis of total organic matter (TOC) formed by hydrobiota and residuals of terrestrial plants. Terrigenous organic mater dominates in bottom sediments. Active redox, microbial and thermolytic processes of organic matter transformation take place in the sedimentary mass. Special character of chromatographic spectra of n-alkane distribution in both low and high-molecular ranges, as well as increased naphtene contents can be interpreted as a sign of oil hydrocarbon generation from maternal organic matter as a result of thermocatalytic reactions within sedimentary mass and their displacement into the upper sedimentary layers. Molecular compositions and concentrations of phenols and lignin were determined in core samples from the Norwegian Sea. Total concentration of phenols in the cores ranges from 8.1 to 101.8 (µg/g of dry sediments that corresponds to 0.15-1.15% of TOC. Lignin concentration was estimated at 21.0-459.0 µg/g of dry sediments (0.59-7.9% of ?org. Phenol compounds of p-hydroxybenzoic, vanillin, syringyl and cinnamyl families as basic components of lignin macromolecules were identified. It was found that sea currents and aerosols are the main contributors of lignin into the abyssal part of the Norwegian Sea.

Wind pumps for irrigating greenhouse crops

Agriculture is a major consumer of energy in many countries of the world. Only a few of these countries are self-sufficient in conventional energy sources, which are also exhaustible. Fortunately, there are other sources of energy, such as wind, which has experienced recent developments in the area of wind power generation. From irrigation projects to power supply in remote farms, wind power generation can play a vital role. A simple methodology for technical evaluation of windmills for irrigation water pumping has been developed in this study to determine the feasibility per unit amount of water supplied and the levels of daily irrigation demand satisfied by windmill irrigation system at various levels of risk (probability of failure). For this purpose, a series of three hourly wind-speed data over a period of 38 years at Ciego de Ávila, Cuba, were analyzed to compute the diurnal wind pump discharge at varying levels of risk. The sizes of reservoirs required to modulate fluctuating discharge and to satisfy the levels of irrigation demand, on function of crop development dates, cultivated area and water elevation height, were computed by cumulative deficit water budgeting. An example is given illustrating the use of the methodology on tomato crop Licopersicon esculentum Mill) under greenhouse.

Generación procedimental de contenidos en videojuegos

Este proyecto consiste en el desarrollo de un sistema completo de generación procedimental de misiones para videojuegos. Buscamos crear, mediante un encadenamiento de algoritmos y un modelado del juego y sus componentes, secuencias de acciones y eventos de juego encadenados entre sí de forma lógica. La realización de estas secuencias de acciones lleva progresivamente hacia un objetivo final. Estas secuencias se conocen en el mundo de los juegos como misiones. Las dos fases principales del proceso son la generación de una misión a partir de un estado de juego inicial y la búsqueda de una misión óptima utilizando ciertos criterios que pueden estar ligados a las propiedades del jugador, dando lugar a misiones adaptativas. El proyecto contempla el desarrollo íntegro del sistema, lo que incluye tanto el sistema de generación y búsqueda como un videojuego donde integrar el resto del sistema para completarlo. El resultado final es plenamente funcional y jugable. La base teórica del proyecto proviene de la simbiosis de dos artes: la generación procedimental de contenido y la narración interactiva. This project involves the development of a complete procedural game quest generation system. We seek to build, by linking a series of algorithms, game and game component models, sequences of logically chained game actions and events. The ordered accomplishment of these sequences lead progressively to the fulfillment of a final objective. These sequences are known as quests in the videogame world. The two main parts of the process are quest generation from an initial game state and optimal quest search. This last is achieved by using certain criteria that can defined by the player properties, thus giving birth to adaptive quests. In this project. The system is comprehensively developed, including the quest generation and optimal search, as well as a full videogame, in which the rest of the system will be embedded so as to complete it. The final result is fully functional and playable. The theoretical basis of the project comes from the symbiosis of two different arts: procedural content generation and interactive storytelling.

Improving brain injury cognitive rehabilitation by personalized telerehabilitation services: Guttmann neuropersonal trainer

Cognitive rehabilitation aims to remediate or alleviate the cognitive deficits appearing after an episode of acquired brain injury (ABI). The purpose of this work is to describe the telerehabilitation platform called Guttmann Neuropersonal Trainer (GNPT) which provides new strategies for cognitive rehabilitation, improving efficiency and access to treatments, and to increase knowledge generation from the process. A cognitive rehabilitation process has been modeled to design and develop the system, which allows neuropsychologists to configure and schedule rehabilitation sessions, consisting of set of personalized computerized cognitive exercises grounded on neuroscience and plasticity principles. It provides remote continuous monitoring of patient's performance, by an asynchronous communication strategy. An automatic knowledge extraction method has been used to implement a decision support system, improving treatment customization. GNPT has been implemented in 27 rehabilitation centers and in 83 patients' homes, facilitating the access to the treatment. In total, 1660 patients have been treated. Usability and cost analysis methodologies have been applied to measure the efficiency in real clinical environments. The usability evaluation reveals a system usability score higher than 70 for all target users. The cost efficiency study results show a relation of 1-20 compared to face-to-face rehabilitation. GNPT enables brain-damaged patients to continue and further extend rehabilitation beyond the hospital, improving the efficiency of the rehabilitation process. It allows customized therapeutic plans, providing information to further development of clinical practice guidelines.

PRISMA: Model-driven development of aspect-oriented software architectures

This summary presents a methodology for supporting the development of AOSAs following the MDD paradigm. This new methodology is called PRISMA and allows the code generation from models which specify functional and non-functional requirements.

Methodology for developing an advanced communications system for the Deaf in a new domain

A methodology for developing an advanced communications system for the Deaf in a new domain is presented in this paper. This methodology is a user-centred design approach consisting of four main steps: requirement analysis, parallel corpus generation, technology adaptation to the new domain, and finally, system evaluation. During the requirement analysis, both the user and technical requirements are evaluated and defined. For generating the parallel corpus, it is necessary to collect Spanish sentences in the new domain and translate them into LSE (Lengua de Signos Española: Spanish Sign Language). LSE is represented by glosses and using video recordings. This corpus is used for training the two main modules of the advanced communications system to the new domain: the spoken Spanish into the LSE translation module and the Spanish generation from the LSE module. The main aspects to be generated are the vocabularies for both languages (Spanish words and signs), and the knowledge for translating in both directions. Finally, the field evaluation is carried out with deaf people using the advanced communications system to interact with hearing people in several scenarios. In this evaluation, the paper proposes several objective and subjective measurements for evaluating the performance. In this paper, the new considered domain is about dialogues in a hotel reception. Using this methodology, the system was developed in several months, obtaining very good performance: good translation rates (10% Sign Error Rate) with small processing times, allowing face-to-face dialogues.