METABOLIC PROCESSES

Author: BOREN CHAMORRO JUAN ANTONIO.
Year: 2003.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA.

Summary: The metaboloma represents the end point where they make up all the signals coming from both the genome and the proteome, in a way that defines what is the biochemical phenotype of a cell or tissue. Characterization metaboloma a particular cell type allows us to recognize what those changes relevant presents both genomic and proteomic and, thus, enable them to exploit as potential targets which lead to a treatment. In order to apply this knowledge is necessary to study the metaboloma of different pathological conditions. In tumor cells was observed both a high plasticity metabolomics in response to external factors, such as an increase in genomic plasticity due to the high rates of mutation that presents these cells in response to the selection are subjected to. This requires a better understanding of tumor metabolism in order to use metabolomics to assess the effect of different drugs and also to be able to make a rational investigation of new therapeutic targets aimed at the treatment of this condition. In this thesis has deepened in the understanding of tumor metabolism using different strategies to be able to understand what their behavior both in normal circumstances and in situations in which they try to alter the development of the tumor. To achieve this objective has been used metabolomics, ie has characterized the metabolism of these cells in the study by analyzing metabolic fluxes, the study of the levels of various metabolites, the behavior of these enzymes that control processes Analysis Metabolic Control and supramolecular organization of these enzymes. The results allow in-depth analysis of metaboloma of the tumor cell and major metabolic pathways that it consists. This knowledge may be used to develop new strategies antirumorales based design inhibitors those routes which have proved essential for tumor metabolism, avoiding metabolic adaptation of the tumor cell, and to facilitate the assessment of those at genomic alterations and proteomic relevant to tumor development.

Author: CONTRERAS MONLEON INMACULADA.
Year: 2005.
University: VALENCIA [www.uv.es].
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTAD DE FARMACIA.

Summary: In this thesis, we have characterized the proteins on the coat COPI and COPII in plant cells and have investigated signals classification of membrane proteins that determine their inclusion in a coated vesicles or other proteins. First, we found that the proteins on the coat COPI are distributed in membrane fractions and cytosol in plant cells and form a cytosolic complex similar to coatómero of animal cells, which are recruited membranes as a single unit. The coatómero plant cell binds specifically to the grounds of dilisina present in the position -3, -4 regard end carboxi-terminal protein type I membrane, acting as signals retention in the endoplasmic reticulum in cells animals in yeast and plant cells. One reason for difenilalanina at position -7, -8 regard end carboxi-terminal acting cooperatively with the reason for dilisina in the recruiting of coatómero plant cell. In plant cells union of coatómero to membranes is mediated factor ribosilación ADP (ARF1) as GTP. Likewise, ARF1 is recruited by the tail cytosolic protein family p24 as GDP, thanks to reasons dilisina and difenilalanina acting cooperatively. All proteins of the family p24 of plant cells contain reasons difenilalanina and dilisina acting cooperatively in the recruitment efficient coatómero and ARF1, suggesting that these proteins are also efficiently packaged in vesicles coated COPI in plant cells. The recruitment of ARF1 the membranes could happen through a direct interaction between ARF1-GDP and tails of cytosolic protein p24. In this interaction are key reasons for dilisina and difenilalanina. After the exchange of GTP / GDP, it would lead to the dissociation of ARF1 of the cytoplasmic tail of the protein p24. Thus, ARF1-GTP, which interacts with the cytosolic domain of the protein p24, is now able to interact with the coatómero. The coatómero can also interact with the cytosolic tail of the protein p24, again through their reasons for dilisina and difenilalanina. Therefore, the signals classification in the tails of cytosolic proteins p24 can be used both for the recruitment of ARF1 and for the recruitment of coatómero in cooperation with ARF1. The reason for difenilalanina at position -7, -8 (at the extent carboxi-terminal) in the queue cytosolic protein p24 plant is also involved in the interaction with proteins on the coat COPII, particularly with the dímero Sec23/24p , suggesting that this is dímero, not dímero Sec13/31p, which interacts with the signal classification. However, citosólicos domains of the protein p24 of plant cells bind much more affinity to COPI that COPII. The molecular characteristics of proteins COPI and COPII of plant cells could reflect the morphological differences exist in the initial stages of the road biosintética / secretory between plants (which do not possess intermediate compartment between the ER and Golgi complex) and other eukaryotes.

Author: MARÍN MARTÍNEZ SILVIA.
Year: 2005.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA, UNIVERSIDAD DE BARCELONA.

Summary: The Systems Biology is a new discipline that seeks to explain the workings of complex biological systems from a systemic perspective in which all system components interact among themselves, through the action of a network or network. Along with the Systems Biology, metabolomics and more specifically fluxómica are providing tools and information which they allow this systemic vision of the cell, tissue or organism. In addition, the information obtained from these "ómicas" is directly related to the phenotype. But these ómicas are still at an early stage of development so that there is an urgent need to develop useful tools and practices to measure reliably both metaboloma as fluxoma its biological system. Here, in this thesis has been developed tools to characterize the metabolic network central glucose and in combination have been applied to existing ones (such as the analysis of the distribution of masses of isotopómeros -MIDA-), characterize the metabolic network liver conditions under both glucolíticas as gluconeogénicas well as the role played by futile cycles in this network. In addition it has also been a hallmark of the control exercised by enzymes glucoquinasa (GK), and glucose-6-phosphatase (G6Pase) on the metabolism of pig sperm. From the results obtained in this doctoral thesis concludes that have successfully developed two tools useful for the calculation of metabolic fluxes, one based on the use of analytical formulas and the other in the integration of information obtained by Tracer-Based Metabolomics in mathematical programs. The results obtained from the use of the tools developed revealed that futile cycles had differing responses to the presence of some or other substrates gluconeogénicos, being able to minimize the impact caused by acute presence on the entire network. It also concludes that, for the three cycles futile involved in the ways glucolítica and gluconeogéncia, catalyzed by enzymes GK and G6Pase is the one who is most affected by the presence of substrates gluoneogénicos. In this regard it has been found that both enzymes control the synthesis of glucose, glycogen and lactate from glycerol. It has also been found that different phenotypic characteristic profiles of the various species of sperm correlated with their profiles metabolómicos. From a global point of view, all results shown in this dissertation is a prime example of the potential of metabolomics and fluxómica in the study of complex biological systems.

Author: GARCÍA CORDENTE ANTONIO FELIPE.
Year: 2005.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTAD DE FARMACIA.

Author: ORMAZABAL HERRERO AIDA.
Year: 2005.
University: BARCELONA [www.ub.es].
Place of defense: HOSPITAL SANT JOAN DE DEU.
Place of preparation: FACULTAD DE FARMACIA.

Summary: In the last decade have been described different inborn errors of metabolism (ECM) of neurotransmitters (NT), especially tracks and serotoninérgica dopaminergic and the pterinas. There have also been described flaws in the transport of glucose and 5-metiltetrahidrofolato (5-MTHF) through dela blood-brain barrier, where all these defects rare diseases, and whose diagnosis is necessary to study cerebrospinal fluid (CSF). Our goal has been to assess the results of the implementation of a protocol analysis CSF in Spain and Portugal over three years in pediatric patients with neurological disorder of unknown origin. We studied 127 individuals control and 283 patients with neurological disorders of unknown origin. The NT analysis was conducted by HPLC with electrochemical detection and analysis of pterinas and 5-MTHF by HPLC with fluorescence detection. Have been diagnosed 3 deficiencies tyrosine hydroxylase, 2 cases of dystonia sensitive to L-dopa, 14 deficiencies GTP-ciclohidrolasa dominant, 2 failures glucose transporter and 43 deficiencies in folate CSF. This study has enabled diagnosis of new patients, and most importantly, all of them in the establishment of a drug treatment or nutrition. Weak 5-MTHF have been the most frequent and have been detected in different patient groups.

Author: CUESTA MATARREDONA EDUARD.
Year: 2005.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD MEDICINA.
Place of preparation: FACULTAD DE MEDICINA (BELLUITGE).

Summary: The F1 6BP is a metabolite reduces cellular cytotoxicity and apoptosis induced by the toxic galactosamina in hepatocytes in vivo model and in primary cultures. The effect of F1 6BP is that avoids histaminemia and endotoxemia generated pro galactosamina rat. Instead not recovered from the transcriptional inhibition induced galctoamina in hepatocytes. By bull side, F1 ,6-Bisfosfato reduces the activation of macrophages in vivo and in primary cultures, the administration DELPS. The mechanism of action of the F1 6-Biosfato by which avoids the effects of galactosamina and the endotoxemia is to reduce the level of activation of potassium channels involved in these types of injury.

Author: Bernal Sánchez Vicente.
Year: 2006.
University: MURCIA [www.um.es].
Place of defense: Facultad de Química.
Place of preparation: Facultad de Química.

Summary: In this Doctoral Thesis has been performed optimization of the production of L (-) -carnitina using the metabolism of compounds trimetilamonio Escherichia coli from the views of bioproceso and metabolism. It has carried out a systematic analysis of the main factors affecting the process of production of L (-) -carnitina using strains of Escherichia coli, studied the effect of the availability of aceptores electronics (such as molecular oxygen or fumarate) both in terms of discontinuous operation as continuous. The analysis using flow cytometry effect on the cells allowed us to determine the answers in the levels of DNA, RNA and proteins, and the integration of these data with heterogeneous cell populations. This allowed us to analyze a novel way in which the configuration of the reactor affects the physiology of the cells. In addition, we studied the DNA content in cells of E. Coli in continuous cultivation, allowing us to identify the factors that determine the genetic stability of the immobilized strain. Furthermore, the expression of secondary metabolism was coordinated with route through central regulatory proteins general, allowing the development of pools of cofactors and metabolites union or integration of the two metabolisms. The analysis metabolic stress conditions showed an increase in productivity due to permeabilización cell and the activation of metabolic pathways of power generation and precursors. It clonó, sobreexpresó and partially characterized protein CaiC, revealed as a highly specific ligasa of CoA. In addition, overexpression of strains were constructed and deletion of activities CaiB (CoA transferase) and CaiC (CoA ligasa) studied the effect of these changes on the production of L (-) -carnitina and underlining the importance of activating substrate. Finally, the expression cycle acids tricarboxílicos, cycle glioxilato and metabolism of acetate, which is related to the biotransformation, limits the maximum productivity, and have been experimentally developed and tested new strategies for improved strains for the production of L (-) -carnitina. Overall, the cellular metabolism and physiology are closely related to the mode of operation of the reactor and to contribute jointly in determining the performance of the process. It has deepened in clarifying the role of cellular metabolic state in determining the production of L (-) -carnitina, especially in the levels of cofactors (ATP and acetil-CoA/CoA). Furthermore, it has been shown that the mode of operation of the bioreactor, which determines the physiological state of Escherichia coli, largely confined performance of the process. In addition, it has been determined the role of CaiC and, above all, it has established the importance of considering jointly by the central and secondary metabolism in improved strains biotechnological purposes.

Author: VIZÁN CARRALCÁZAR PEDRO.
Year: 2006.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE BIOLOGIA UNIVERSIDAD DE BARCELONA.
Place of preparation: FACULTAD DE BIOLOGIA - UNIVERSIDAD DE BARCELONA.

Summary: The objective of this dissertation is to deepen the knowledge of cellular metabolism, specifically the central carbon metabolism during various processes associated with tumor progression. The acquisition of a malignant tumor phenotype is considered today an evolutionary process of accumulating changes by the tumor cells and adjacent cell types. The metabolism, seen as the standard integrator many of the cellular signal transduction, can serve not only to understand how these changes occur, but it can offer a range of therapeutic targets based on specific adaptation to prevent the tumor cell to changes in the process of malignancy. This has been used in vitro models known cellular and molecular level, and have been studied metabolic changes that accompany them: in the first chapter revealed metabolic changes that occur when a cell is transformed by constitutively active copies the K-ras oncogene, proving that they depend not only on the presence or absence of the same, but the type of mutation present. Here is metabolically as endothelial cells respond to angiogenic stimuli, a key process that trigger tumors to meet their needs for nutrients and oxygen. In the third chapter, using both experimental tools such as bioinformatics tools developed for this purpose, are carrying out a detailed study of changes in metabolic fluxes who suffers a major cell leucémica when it tries to force its programmed cell death. In the fourth chapter elaborates on the changes to metabolic level, particularly changes in the route of pentosas phosphate, which occur during cell cycle progression in two cell lines of colon cancer. Finally, the fifth chapter describes a method put point during their doctoral thesis, for analysis quantification of phosphorylated intermediaries of the glucólisis and path of pentosas phosphate in cells in culture. From the results that flow to these studies concluded that the metabolic alterations introduced tumor characteristics during tumor progression and thus their study can be of great interest for the design of therapeutic strategy against cancer.

Author: SALAS MANI ANNA.
Year: 2006.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: UNIVERSIDAD DE BARCELONA.

Author: RAMOS MONTOYA ANTONIO FRANCISCO.
Year: 2006.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA.

Summary: This Doctoral Thesis deeper understanding of the connections between metabolism and cell proliferation in order to exploit it in the design of new anti-tumor therapies. The results presented in the various sections that compose address this issue from complementary perspectives. Thus, the chapters of the paragraph I propose a strategy based on antitumor inhibit metabolic adaptation that supports the proliferation of tumor cells. Thus, it prevents the synthesis of precursors required for the de novo synthesis of nucleic acids and, therefore, out of the cell phase G1 cell cycle and its subsequent entry into the S. Paragraph II, however, deals with this subject from an angle, although not opposed, it slightly different. In the two chapters in it describes the effects at metabolómico of specific inhibition of enzymes that control the output of the cell phase G1 to S phase, the protein kinase-dependent ciclinas 4 and 6 (CDK4 and CDK6 also known as generically and pooled CDK4 / 6). Finally, the chapters of section III pointing deep into the role that may have brokers precursors required for the de novo synthesis of nucleic acids. This capability signaling has proven to be very important for the cell and shows metabolic changes that can affect an adaptation gene level.

Author: RAMAJO ESCALERA BEATRIZ.
Year: 2006.
University: OVIEDO [www.uniovi.es].
Place of defense: FACULTAD DE QUMICAS.
Place of preparation: FACULATAD DE QUIMICAS.

Summary: Within the chemistry of new materials phosphates metal laminates occupy a porminente. Because of its structural characteristics and thermal properties, these compounds have properties that make them applicable in fields as interesting as catalysis, ion exchange, or ionic conductivity. In addition, many of them may be utlizados as fertilziatnes chemical, acting as a source of some elements essential for plant nutrition, with the advantage that typically low solubility in aqueous media favors slow release of nutrients to the environment by preventing soil their accumulation in groundwater levels. In general, the addition of any substance to the soil for agricultural purposes or not, produces changes in the metabolism of soil microbial population and consequently altering the nature of the organic matter, which may cause degradation soil quality and increasing their emissions of CO2. In this report, describes the synthesis and characterization (structural and phenomenological) three metal phosphates habit laminar: NH4FePO4 H2O (AIP), (NH4) Zn2 (PO4) (HPO4) (AZP) and NH4ZnPO4 NH3 (DZP). Two of them (AIP and AZP) have characteristics suitable for potential use as additives em agricultural environments, utlilizándose calorimetry heat flow in the evaluation of their influence on the microbial metabolism of the population of three selected soils (Luvisol-Harpenden, UK; Rhodic-Campinas, Brazil; Podsol-Oviedo, Spain)

Author: MARIÑO GARCIA GUILLERMO.
Year: 2006.
University: OVIEDO [www.uniovi.es].
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: DEPARTAMENTO DE BIOQUIMICA Y BIOLOGIA MOLECULAR. FACULTAD DE MEDICINA.

Summary: Since the breakthrough occurred in recent years in understanding the molecular basis of the autofagia, we have begun to understand the decisive participation of this process in the maintenance of integrity and cellular homeostasis. In addition, the possible role of anti-tumor and the autofagia have anti prompted a new interest in the detailed study of the molecular mechanisms responsible for regulation of this process. These considerations make it very interesting finding and characterization of potential ortólogos human genes IGT, whose role is critical to the development of autofagia in lower eukaryotes. These facts, together, and given the general interest of our group for the study of systems proteolíticos human, we identified the ortólogos human protease ATG4, the only proteolítico enzyme involved in regulating the autofagia in yeast. Subsequently, once identified ortólogos human ATG4, did a comprehensive analysis aimed at clarifying the biochemical and functional properties of the autogaina-3, ortólogo human ATG4 with a larger tissue distribution. To that end, we generate genetically modified animals cerantes gene that encodes this protease. Finally, the availability to our laboratory for an animal model of premature aging caused by defects in the metaloproteasa FACE-1 (Zmpste24), directed our attention to an analysis of the possible involvement of the autofagia in developing these syndromes progeroides. In doing so, we draw some keys that enabled us to move forward in establishing the connections between autofagia and molecular aging.