BIOCHEMISTRY (3)

Author: VILLAR GAREA ANA.
Year: 2004.
University: VALENCIA [www.uv.es].
Place of defense: FACULTAD DE QUIMICA.
Place of preparation: CENTRO NACIONAL DE INVESTIGACIÓN ONCOLÓGICAS C.N.I.O. MADRID.

Summary: Alterations genélicas and desregulaclón cough epigenetic mechanisms are collaborating in cancer initiation and progression. The epigenetic defects are potentially reversible, which has prompted the search for drugs that selectively cause changes in the patterns epigtmélicos of cells tumareles, oon the subsequent differentiation, death and / or stop growth of the same. We have studied especially inhibitors metillransferasas deDNA (DNMT) and inhibitors desacetilasas-dependent histone (HDAC) Zn (II). Inhibitors DNMTs allow the revival of ganessilonciados by hypermethylation of CpG island of its promoter and inhibitors of HDAC. The winning silancfados vla hipoacetilación of histone élsociadas his promoter. Despite its promising effects in cultured cells, many of these substances present disadvantages that limit their application in chemotherapy. Therefore. In this thesis: 1. There were determinadolos affections of procaine in the melitacion DNA yen proliferation of online cell MCF7. Procaine reduces the amount of 5-meltilcitosina in genomic DNA, reactive geneS silenced by hipermetilaCión (RARb2) and induces cell cycle arrest in mitosis. 2. We compared seven inhibitors of HDACs dependent gives Zn (II) acid butanoico valproic acid. MS.275. Tricostatina A (TSA), Saha, PXD101 and NVP-LAQ824. There is a relationship astructura-actividad for purposes of eslas substances in tests in Vitro and in the growth of MCF7.Además, eunque these inhibitors induce similar changes in histone acetylation overall, not all reaclivan the same genes. Expression levels of CDKN1A and GADD45b seem to determine the effects of these substantiate the cell cycle. The promoters of the six genes studied. The inhibition of HDACs increases acetylation of H4 and dimetilación of tisina 4 of H3, while decreases dimetilación lysine 9 of H3 and the presence of HDAC1 and HDAC2.

Author: POSTIGO MARISCAL FERNANDO.
Year: 2004.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE QUÍMICA.

Summary: At work are set out in the first place different synthetic routes for obtaining photosensitivity useful in photodynamic therapy. All of them are based on the structure of metilpirofeofórbido woe are obtained in the laboratory from the chlorophyll a. In a second phase establishes the requísitos structural required for efficient incorporation of fotosensibilizadores hodrofóbicos liposome. It standardize and optimize processes for obtaining stable lipid vesicles. This part of the work is performed using porphyrins commercial and clorinas summarized above. Then he studies the effect of the incorporation of fotosensibilizadores liposome in the optical properties and fluorescence, as well as the efficiency of singlet oxygen generation. Later describing the detention of liposomes containing photosensitizer in solid matrices in order to obtain useful systems for sterility fotodinámica of blood. Here evaluates the effect inactivador different fotosensibilizadores solution, incorporated into liposomes and immobilized on a solid matrix using liposomes. The models are used viral ECMV and BVDV, examples of viruses with and without wrapper. Finally testing and phototoxicity of cell proliferation in vitro human skin fibroblasts, Hela cells and other cell lines. In this sense, evaluate the toxicity of fotosensibilizadores in the absence of light and fotocitoxicidad of photodynamic therapy after d radiate with appropriate wavelength. Further testing methods for determining the process of cell death that occurs after irradiation. All this is done to study the possible application to photodynamic therapy of cancer of the clorinas derívadas of clorofilia to synthesized and have sído incorporated into liposomes efficiently.

Author: UBER GARCÍA GENOVEVA.
Year: 2005.
University: VALENCIA [www.uv.es].
Place of defense: INST. AGROQUÍMICA Y TECNOLOGÍA ALIMENTOS.
Place of preparation: FACULTAD DE MEDICINA.

Summary: The aroma of wine is one of the most important characteristics in assessing their quality wine, and very important in rating their quality. Its chemical nature is very complex and difficult to characterize. However, in response to its source from, it can be calsificar into three broad categories: i) the aroma from the grape variety, or flavoring primary ii) the scent produced by the yeast over the fermentation, and therefore, stems from its metabolism, or secondary flavor iii) the aroma tertiary, or "bouquet" produced by the transformation of flavor during the aging wine, and that depends on environmental conditions and physico that has been stored . The present project deals with the increase of secondary flavor created by the yeast cell as metabolites (secondary alcohols and esters) during alcoholic fermentation. It devised various strategies for metabolic engineering: 1.1 Interruption and regulated overexpression of alcohol acetiltransferasas the encoded by genes ATF1 and ATF2 in yeast industrial T73 and laboratory strains. 1.2 Interruption of the ester hydrolase encoded by the yeast gene IAH1 in industrial and laboratory strains. 1.3 Increased production of primary alcohols, in particular alcohol isoamílico for: i) Sobreexpresión of α -cetoácido decarboxylase encoded by the gene THI3. Ii) Increase metabolic flux for the synthesis of leucine by deletion of permeasas amino acid in combination with overexpression of the gene THI3 and / or ATF1. Of all the strategies tested, controlled overexpression of the gene ATF1, proved to be the most effective way to achieve greater accumulation of these aromatic compounds.

Author: NOVÁS MASEDA ANA.
Year: 2005.
University: SANTIAGO DE COMPOSTELA [www.usc.es].
Place of defense: FACULTAD DE VETERINARIA CAMPUS LUGO.
Place of preparation: FACULTADE DE VETERINARIA CAMPUS DE LUGO.

Summary: The bivalve Mytilus galloprovincialis Lmk. Introducing a single type of immunity, called innate, which is exclusively cellular and phylogenetically oldest learned that, with hemocytes cell populations present in the hemolymph of the mollusk, responsible for defending these agencies. The immune response is regulated by multiple and complex interactions between molecules such as LPS, TNF, IL-2, PDGF, TGF, NO, and so on. And also for proteins involved in signal transduction pathways of the immune response. This work studied the influence dellipopolisacárido (LPS), and some cytokines (TNF, IL-2, PDGF YTGF) in the resuesta of hemocytes of Mytilus galloprovincialis Lmk. To that end, and among others, was quantified the production of nitric oxide (NO) and protein expression in hemocytes of mussel. First, it was necessary to adapt a culture medium, supplemented ALS, allowing maintain cell viability and to be incolor not interfere with the reagents used Gries colorimetric method for the quantification of NO. Of all the employees inducers (LPS, TNF, IL-2, PDGF YTGF), the only IL-2 causes notable increases in production of NO; surprisingly, the LPS, known stimulator of this synthesis in vertebrates, does not cause such hemocytes induction. During the development of experimental work, was assessed the proportion of cell types detected in hemolymph of Mytilus galloprovincialis, SH cells and cells RH, considering that the proportion of SH is less during the summer, when it showed a greater production basal NO. In turn, the analysis of signaling pathways involved in the synthesis of NO through the use of inhibitors of PKA (H89) And PKC (bisindolilmaleimida), suggests the involvement of PKA on the road signaling used by the IL-2 for the synthesis of NO, but the involvement of PKC appears to be influenced by environmental factors. In hemocytes, two proteins were detected, recognized by monoclonal antibodies anti-iNOS human anti-eNOS bovine, and the production of NO independent of changes in the expression of both isoforms. In turn, the isoform endothelial already noticed the difference in vertebrate cells, is detected only in cytosol, never in membrane. In subsequent years one of the largest environmental disasters occurred on the coast of Galicia, ie, during the years 2003 and 2004, the pattern of behavior of hemocytes was interumpió by unknown causes. During this biennium, was not found for NO synthesis, in turn confirmed by the absence of the two isoforms (iNOS and eNOS), which have been identified previously. There is also during this period of change, a reduction in protein expression, which affects both cytosolic and membrane proteins and it is during those years (2003 and 2004) and not in previous years, when it is possible to induce apoptosis to be the hemocytes incubated with cicloheximida, TNF and TFR.

Author: SANCHEZ MAS JESÚS MANUEL.
Year: 2005.
University: MURCIA [www.um.es].
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA - UNIVERSIDAD DE MURCIA.

Summary: Laos melanocytes are highly specialized cells whose main function known in the skin is the production and distribution of melaninas the skin, and therefore the human pigmentation. The melanocortinas, and more specifically stimulating hormone melanocito (MSH), play a central role in regulating these processes, both in basal conditions and after exposure to UV radiation. The stimulating effect of MSH in the melanocytes is produced by binding to a specific receptor belongs to the superfamily of G protein coupled receptors (GPCRs), the recipient of melanocortinas 1 (MC1R). The union is triggering cascades of cAMP which in turn leads to the activation of the biosynthesis melaninas. Furthermore, a change in any of these steps can lead to disorders that result in diseases of different severity, ranging from riots pigmentarlos to various types of skin cancer, such as melanoma. Therefore, an important aspect in the regulation of melanogenesis and biology melanocito is the function of MC1R. Thus the aim of our research is focused on the study of regulatory mechanisms that modulate signaling receptor melanocortinas 1 in melanocytes and melanoma cells in human and murine (MC1R) (Mc1r). Such modulation can be exercised at various levels to regulate the expression of the receptor in the plasma membrane of melanocytes, as well as their signaling. The most important conclusions of this study have been. 1, - melanoma cells and normal melanocytes in the amount of molecules MC1R expressed in membrane determines the level of response to agonist, which identifies the recipient as the limiting factor in the path of signal transduction in these cells. 2 - The new variant natural L93R and MC1R found in a line of human melanoma cells, corresponds to a protein with loss of function total inability to unite agonists. 3 - The MC1R presents constitutive activity independent of agonist. The Mc1r also is constitutively active, although less by the human recipient. Mutations of MC1R decreasing signaling induced by agonists (R151C, I40T, V92M, V122M, L93R and R162P) also reduces the constitutive activity. 4 - The pentapéptido carboxilo-terminal involved in the expression of molecules MC1R in the plasma membrane. Within this pentapéptido, acitación residue Cys315 could contribute to traffic and / or stability of the protein. 5 - The MC1R dimeriza constitutively in heterologous systems que sobre receiver. This dismerización takes place in the endoplasmic reticulum and is produced by exchanging domains. 6 - Some mutant alleles of MCR1R as L93R, R162P and delección of pentapétido carboxilo-terminal exerts a dominant negative effect on the expression of wild receiver in the plasma membrane. The existence of this dominant negative effect together with the high polymorphism observed in MC1R, might help to understand the wide range of phototypes described so far. 7-MC1R and Mc1r suffer desensitization counterpart in response to natural or synthetic ligands. This desensitization is independent of PKA, MAPK, PKC, and the mobilization of Ca2 +. 8 - The kinases GRK2 and GRK6 are expressed in melanocytes and melanoma cells, and are able to inhibit signaling by MC1R and MC1r induced agonists. In addition, GRK6 inhibits basal constitutive signaling.

Author: TOUS MÁRQUEZ MÓNICA.
Year: 2005.
University: ROVIRA I VIRGILI [www.urv.cat].
Place of defense: FACULTAD DE MEDICINA I CIENCIAS DE LA SALUD.
Place of preparation: FACULTAD DE QUÍMICA.

Summary: The atherosclerosis is an inflammatory disease. It has been shown that a chronic inflammation, such as periodontal disease, lupus eritomatoso, rheumatoid arthritis or chronic bronchitis contribute to increased mortality and morbidity from cardiovascular disease. Despite these developments, there is no data to prove the long-term effects of a chronic inflammatory process and what role they might have some anti-inflammatory agents in the development and progression of atherosclerosis. Several studies suggest that the use of certain anti-inflammatory agents in the development and progression of atherosclerosis. Several studies suggest that the use of certain anti-inflammatory drugs could be an effective tool for the treatment of atherosclerosis. Previous studies have shown that the administration of high doses of aspirin had a beneficial effect on the short-term atherogenesis in mice deficient in apolipoproteina (apo) E. Despite these results, there is no data evidencing the effect of high doses of aspirin in the mouse apo E-deficiente. The results presented in this thesis show that the administration of high doses of aspirin have no significant effect on long-term atherogenesis in mice deficient in apo E. It is noted that the size of lesions arterioscleróticas evaluated within aortic of these animals is significantly correlated with the concentrations plásmaticas cholesterol, suggesting that the ongoing administration of aspirin facilitates atherogenesis. We suggest that the observed effect is not due to the administration of aspirin per se, but could be due to the presence of one or more factors present in our model, we have not been able to identify, and that is affecting us our results. We must conclude that our model is not free of artifacts and propose that the diet with which the mice were fed could be a factor that complicates the interpretation of the results obtained in this animal model. In study 2 of this thesis, we have demonstrated that mice apo E-deficientes fed a diet high in fat and cholesterol, developed severe liver damage consisting of the fat accumulation, the proliferation of macrophages and the presence of inflammatory cells . We have also noted the existence of a relationship between quantitative and chronological lesion size arterioscleróticas and the degree of hepatic impairment. These results suggest that cholesterol in the diet can cause significant alterations in the liver of these animals dificultándonos and interpretation of results. Furthermore, we believe that mice apo E-deficients can provide a good animal model for the study of non-alcoholic steatohepatitis. Finally, we found that the ongoing administration of an agent pro-inflamatorio, as turpentina produces an inflammatory response very similar to what takes place in certain chronic inflammatory processes, and it appears that is not affected by the administration of aspirin. Mice treated with rupentina presented a significant decrease in plasma concentrations of cholesterol, the size of the lesions arterioscleróticas as well as the hepatic expression of the gene múltiple-drug resistance (mdr1b) in reaction to the control mice. We suggest that mdr1b can play a very important role in regulating the concentrations of plasma cholesterol and / or progression of atherosclerosis.

Author: Sidrach de Cardona Paniagua Lara.
Year: 2005.
University: MURCIA [www.um.es].
Place of defense: Universidad de Murcia.
Place of preparation: Universidad de Murcia.

Summary: INTRODUCCIÓNLa artichoke is a perennial widely distributed in the regions around the Mediterranean. The Mediterranean region produces 90% of world output. Some countries where production and consumption is traditional as Italy, France and Spain produce 80%. Spain is the world's second largest producer. For example, 70% by weight of the flower of the artichoke is waste not intended for human consumption. These wastes are used for animal feed, but in finding new uses for debris has been found, surprisingly, that these products have a high concentration of proteases, and peroxidases polifenoloxidasas.De We have focused this study work the use of such waste as a raw material for obtaining substances with high added value, such as enzymes, through the development of efficient processing technologies and scalable industrialmente.Las proteases involved in the breakdown of marriages peptide through proteolysis, which is a the enzymatic changes more frequent and major proteins. The aspartic proteases (APs) are widely distributed, found in vertebrates, fungi, plants, protozoa and retroviruses. They are all endoproteasas and are characterized by acid pH optimum and be specifically inhibited by pepstatina A. Among the possible applications of proteases is the manufacture of cheese through clotting enzyme milk. The peroxidases are enzymes involved in the enzyme system for the protection of cells consisting of the elimination of harmful species which occur during the process of reduction of O2 to H2O, such as superoxide anion (O2-), hydrogen peroxide (H2O2) and the hydroxyl radical (HO). The peroxidases are enzymes that catalyze the oxidation of a wide variety of organic and inorganic compounds using oxidizing power of H2O2 or other hidroperóxidos (Dunford and Stillman, 1976). Recently, our research group has a purified peroxidase artichoke (AKP-C ) (López-Molina et al., 2003). From artichoke flower has been purified to homogeneity the AKP-C using FPLC. There have been changes in the activity of AKP-C with hydrogen peroxide, dependent enzyme calcio.La tirosinasa or polifenoloxidasa (PPO) (monofenol, o-difenol: oxygen óxido-reductasa, EC 1.14.18.1) is widely distributed in all the phylogenetic scale. This cuproproteína catalyzes two kinds of reactions coupled in which molecular oxygen (a) hydroxylation of monofenoles to o-difenoles (activity monofenolasa) and (b) oxidation o-difenoles to o-quinonas (activity difenolasa). This Doctoral Thesis raised with a General Purpose consisting of the purification and characterization of proteases, peroxidases and polifenoloxidasas from agro-industrial waste artichoke (Cynara scolymus L.). Purification. We have managed to clean proteases and polifenoloxidasas waste agribusiness artichoke (Cynara scolymus L. White variety) from the development of this preserved vegetable, especially abundant in Murcia.Propiedades molecular. It has been performed the molecular characterization of proteases and polifenoloxidasas artichoke, determining the molecular masses and points isoeléctricos of isozymes aisladas.Caracterización kinetics. The kinetic characterization of proteases, peroxidases and polifenoloxidasas artichoke has revealed the behavior of these enzymes with their respective substrates, as well as the regulation of its activity by modifying the conditions 8 of ensa 5bf me. The study modulators and protease inhibitors, peroxidases and polifenoloxidasas artichoke, has allowed optimal performance in regulating its catalytic activity, which will promote the further implementation of these enzymes in the industria.Estabilidad. Considering the importance of the stability of enzymes for industrial application has studied the effects of different factors on the activity of the enzyme, thereby determining the optimum performance of enzymes for different applications industriales.Aplicaciones biotechnology. Based on previous studies, have been established major potential applications of proteases, peroxidases and polifenoloxidasas waste agribusiness, which includes various sectors: health, agri-food and industrial sectors.

Author: GARCIA MANSO DIEGO.
Year: 2005.
University: OVIEDO [www.uniovi.es].
Place of defense: DPTO.DE BIOQ. Y BIOL. MOLEC. FAC.DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA.

Summary: This thesis has brought the realization that the particular features kinetic opening and closing HERG channel, which are essential for its important pathophysiological roles, are due to the combined presence in its amino-terminal end of two functionally distinct domains: domain eag and proximal domain. The work shows on the one hand, the existence of interactions between amino domain eag end and the intracellular S4-S5 body loop central canal, which serves as a link between the sensor voltage and machinery opening pore ion channel, interactions which are critical for the particular features kinetic functional HERG. As for the understanding of these interactions is essential to know the distances between some of the intracellular domains of the channel, as well as other aspects of its structural organization, which are intractable through classical studies relations estructura-función also has been studied by a novel approach experimental molecular architecture of the intracellular domains of HERG allowing the existence of these interactions. They have looked through the technique of FRET (Fluorescence Resonance Energy Transfer) distances between different molecular intracellular domains of the protein channel in vivo, using the functional expression of collections HERG channel fluorescent where fluoróforos YFP PIC and were inserted throughout the polypeptide sequence of HERG using the random behavior of transposons recombinants and hyperactive through transposition reactions in vitro. The molecular architecture of HERG responds to a scheme in which the amino terminal ends of the channel tetramérico functional are packed, close to the loop S4-S5 and in a position close to the inner surface of the membrane, while the extremes of carboxyl tetrámero are further away from the intracellular side of the membrane.

Author: SÁNCHEZ HIDALGO MARY CRUZ.
Year: 2005.
University: SEVILLA [www.us.es].
Place of defense: FACULTAD DE FARMACIA DE LA UNIVERSIDAD DE SEVILLA.
Place of preparation: FACULTAD DE FARMACIA.

Summary: THESIS ON THE ENTITLED "STUDY ON EFFECTS PRODUCED BY THE ADMINISTRATION OF CHRONIC MANGANESE: IMPLICATION OF IRON INGESTA" HAS BEEN EVALUATED THE EFFECT THAT THE TOXIC MANGANESE, CONSIDERED A TRACE ELEMENT, MAY CARRY OUT IN THE CNS, MAINLY IN THE SYSTEM DOPAMINÉRGICO NIGRO-ESTRIADO, AS WELL AS IN VARIOUS BODIES PERIPHERALS. THIS ADMINISTRATION IS REALIZÓ IN CHRONIC RATAS WISTAR USING MnCl2 BY INTRAPERITONEAL. ALSO ALSO BE ABORDÓ THE ROLE THAT THE MANGANESE IRON AND COULD HAVE IN THE PROCESS OF NEURODEGENERACIÓN And AFECTACIÓN ORGAN PERIPHERALS FOR WHAT IS DISEÑÓ AN EXPERIMENT IN WHICH EMPLEARON DIETAS SYNTHETIC WITH NORMAL DOSE OF IRON AND LACK THEREOF. WELL BECAUSE, HE ESTUDIÓ RESPONSE GLIAL, INDICATIVE OF BRAIN DAMAGE TO LEVEL, ARE VALORARON PARAMETERS OXIDATIVO STRESS AND EFECTUARON MISCELLANEOUS TINCIONES INMUNOCITOQUÍMICAS WELL AS HIBRIDACIÓN IN SITU. In this manner, OUR STUDY REFLEJA THAT OF TOXICITY IN MANGANESE SNC IS BASED ON THE RISE IN STRESS OXIDATIVO PRODUCIÉNDOSE GREATER EFFECT OF MANGANESE IN THE SYSTEM DOPAMINÉRGICO NIGRO-ESTRIADO, IF NOT BE SELECTIVE FORM OF SERVING AS ALL THE STRUCTURES CEREBRALES ESTUDIADAS MUESTRAN STRESS OXIDATIVO. ALSO ALSO PRODUCE AN INCREASE IN BODIES OF SYSTEMIC OXIDATIVO STRESS AMONG THOSE STRESSES THE LIVER, PUDIÉNDOSE ESPECULAR WITH THE POSSIBILITY OF DYSFUNCTION OF THIS BODY AND DEVELOPMENT OF ENCEFALOPATIA HEPÁTICA. ALSO IS IMPORTANT TO HIGHLIGHT THE IRON DEFICIENCY AND HIGH LEVELS OF MANGANESE IN THE PRESENT AN EFFECT SNC TOXIC SINÉRGICO DESENCADENANDO A GREAT RESPONSE GLIAL WELL AS AFECTACIÓN VASCULAR AND PARÉNQUIMA BRAIN THAT PRESUPONE PHENOMENA ON NECROSIS APOPTOSIS AND CONCLUDING WITH BRAIN DEATH.

Author: HENRÍQUEZ HERNÁNDEZ LUIS ALBERTO.
Year: 2005.
University: LAS PALMAS DE GRAN CANARIA [www.ulpgc.es].
Place of defense: FACULTAD DE VETERINARIA.
Place of preparation: FACULTAD DE VETERINARIA.

Summary: Analysis of gene expression patterns liver hormono-dependientes: application of the technology of DNA microarrays. The technology DNAmicroarrays has seen a revolution in the field of genomics. The verification hypothesis enriched with this expression profiles allowing accelerate the discovery of mechanisms of hormone action. In this Doctoral Thesis this technology has been used to characterize the response of the liver to the actions of estrogen, growth hormone (GH) and thyroid hormone (TH). Studies so far are concentrated on the analysis of the actions of a single hormone, and there were few examples to try to use a compilation of different expression profiles regulated by hormones. This thesis is illustrated as the meta-análisis de multiples expression profiles obtained from the liver, can be used to assess endocrine hypothesis. Built Database called EndoGED (www.cmm.ki.se / EndoGED), which contains a set of expression profiles generated from treatment with growth hormone, T3 and 17Æ Do -ethinylestradiol (EE) in various experimental models. The analysis of all the results show that the continuous administration of GH or estrogen  gfeminizan hours on the pattern of hepatic gene expression. The EE has been used to elucidate the molecular mechanisms involved in the pathogenesis of estrogen-induced cholestasis. We apply DNA microarray in male rats by comparing the effects of EE in the presence or absence of pituitary hormones. It identified a number of new genes regulated by EE and multiple mechanisms that provide new areas of research in the pathogenesis of this disease. GH and TH have a critical role in the metabolism of lipids and bile acids. Our results indicate that these hormones regulate the expression of genes involved in the synthesis of bile acids or in the transcription of the same. Some actions of GH are opposite to those of T3. In the absence of pituitary gland, liver remains extremely sensitive to pharmacological doses of estrogen. We explore the profile of gene expression induced by EE in primary hepatocytes cultured on Matrigel, identifying a number of genes regulated directly into this model. The privacion, or high exposure to thyroid hormone during the neonatal period causes changes in the physiology of some organs to endure during adult life of the individual. We have studied the hepatic response to T3 in adults or infants with congenital hypothyroidism not with the purpose of identifying whether the endocrine environment surrounding the newborn can cause metabolic changes in the physiology hepatica during his adult life. The results corroborate our hipotesis.DOCUMENTACION ATTACHED TO TD-2

Author: Cabrera Heredia Jorge rubén.
Year: 2005.
University: AUTÓNOMA DE MADRID [www.uam.es].
Place of defense: Centro Nacional de Biotecnología.
Place of preparation: Centro Nacional de Biotecnología.

Summary: The Growth Arrest Specific gene 1 (Gas1) protein has been proposed to function during development as an inhibitor of growth and a mediator of cell death, and is also re-expressed in adult neurons during excitotoxic stimuli. Here we demonstrate that the Gas1 protein shows high structural similarity to the GDNF (Glial cell-derived neurotrophic factor) family receptors alpha, which mediate GDNF responses through the receptor tyrosine kinase Ret. We found that Gas1 binds Ret in a ligand-independent manner and sequesters Ret in lipid rafts. Signalling downstream of Ret is thus modified through a mechanism that involves the adaptor protein Shc as well as Erk2, eventually blocking Akt activation. Consequently, when Gas1 is induced, Ret-mediated GDNF-dependent survival effects are compromised.

Author: Matías Román Salomón.
Year: 2005.
University: AUTÓNOMA DE MADRID [www.uam.es].
Place of defense: Facultad de Medicina.
Place of preparation: Facultad de Medicina.

Summary: The matrix metalloproteinase membrane MT1-MMP is involved in the migration of endothelial cells and tumor, but its possible role in leukocyte migration has not been well characterized. Here we show that the monoclonal antibody anti-MT1-MMP affects the migration of monocytes stimulated with human MCP-1 on fibronectin, VCAM-1 and ICAM-1. In addition, the transmigration of monocytes through activated endothelium with TNF is also inhibited by antibody anti-MT1-MMP. Therefore, we decided to investigate the regulation of MT1-MMP monocytes isolated from peripheral blood. First, we saw piles of MT1-MMP in protrusiones membrane-associated motility in monocytes stimulated with MCP-1 migrating on fibronectin, VCAM-1 and ICAM-1, and in the face of progress, colocalizando with profilina, monocytes transmigrando to through activated endothelial cells. In addition, we also see an induction in expressing MT1-MMP in human monocytes following their accession to fibronectin in a manner dependent on the integrins 4 1 and 5 1. The binding of monocytes to endothelial cells activated by TNF-as well as VCAM-1 and ICAM-1 also induced the expression of MT1-MMP. These data correlate with the increased fibrinolytic activity observed in monocytes coupled with fibronectin, VCAM-1 or ICAM-1. Our data show how required MT1-MMP during the migration of monocytes and human endothelial during his transmigration, in addition to regulating the location, expression and activity in monocytes into contact with fibronectin or endothelial ligands, demonstrating the important role that MT1-MMP can play in the recruitment monocítico during an inflammatory process.

Author: Prados Pinto Belén.
Year: 2005.
University: AUTÓNOMA DE MADRID [www.uam.es].
Place of defense: Facultad de Medicina (Dpto. Bioquímica).
Place of preparation: Facultad de Medicina (Universidad Autónoma de Madrid).

Summary: The O-manosilacion is a modification post-traduccional importente in eukaryotes. The gene encoding the protein O-manosil transferase 1 in humans is POMT1. The alfa-distroglicano, glycoprotein component of the complex of dystrophin in muscle and neurons, is O-manosilado by POMT1. There are several neuromuscular diseases whose patients have deficits in the glycosylation of alfa-distroglicano. The syndrome Walker-Warburg (WWS) is a muscular dystrophy congenital malformations associated with severe eye and brain, caused by mutations in proteins involved in the glycosylation of alfa-distroglicano as POMT1. During embryogenesis mouse, Pomt1 expressed in the tissues most affected in patients with WWS. In the adult mouse, Pomt1 is located in the IR The muscle cells and the acrosoma of male germ cells. In this study, we generated a knockout animals to study the role of O-manosilación in mice, which has proved to be embryonic lethality due to defects in the formation of the membrane Reichert. To avoid embryonic lethality, we have produced through the use of chimeric cells IS Pomt1- /-, and we are generating a conditional knockout mouse based system CRE-loxP, allowing inactivate the gene Pomt1 espacio-temporalmente using promoters iducibles or specific tissue.

Author: GIRALDO RAMOS ESTHER.
Year: 2005.
University: EXTREMADURA [www.unex.es].
Place of defense: FACULTAD DE CIENCIA.
Place of preparation: FACULTAD DE CIENCIAS.

Summary: The fig (Ficus carica) along with olives and vines, is one of the three classic fruit associated with the onset of horticulture in the Mediterranean basin, going back to grow for more than 6000 years. The preservation and study of the genetic resources of fig tree, as in the rest of fruit species, involving the establishment of collections in the field. The research center Final "The Order" Extremadura along with other autonomous regions inición in 1985, a prospecting cultivated varieties of fig. As a result of this exploration has collected 215 accessions in a genebank represents a very high percentage of the variability existing fig tree in Spain. The main problem with the maintenance of the collections in situ plant is the correct identification of existing material, as well as new entries. The antigà ¼ age of the fig tree cultivation along with the ease of vegetative propagation has allowed a great exchange of plant material between different regions throughout history, resulting in the presence of synonyms and momonimias. The assessment and characterization of the diversity in cultivated species is very important to preserve both varieties disappearing, as alleles with interest agronomic current or future. In this paper we have used a set of 81 variables and 132 qualitative variables descriptors fig published in 2003, to characterize morphologically 35 varieties conserved in the genebank, chosen according to their age, uniformity of the trees and representation different types of fig. Under separate statistical analysis has been carried out an analysis UPGMA revealing different morphological profiles. The results indicate that the variables morgológicas that explain most variability of fig, are referring to the type of fruit production and morphology (Brevas and figs). The first two chapters of this work are dedicated to the development of this goal. However, the traditional characterization based on phenotype may present problems because of the influence of environmental factors. Therefore, this characterization has been complemented with an approximate molecular level. It has developed a battery of microsatellite markers in fig tree that has been used to characterize and study the genetic diversity of 207 genotypes preserved in the gene bank. The construction of a similarity matrix and an analysis has enabled UPGMA classify accessions in large groups with common characteristics. These results in conjunction with the existing literature have allowed the selection of 100 genotypes of reference. Moreover, molecular markers have shown low genetic polymorphism in the fig tree and the contents of something synonyms and homonimias. These works are described in Chapters 3 and 4. In Chapter 5 compares the results obtained on 35 varieties of fig SSR markers through and through morphological markers to determine if both characterizations are complementary and they are correlated with each other. The statistical analysis of Mantel and the Spearman coefficient Person and revealed a correlation between low similarity matrices obtained, as in other species. However, clusters of varieties obtained with both characterizations are very similar. A genebank duty to be dynamic, which involves the introduction of new plant material. Thus, to optimize the introduction of new genotypes has been evaluated the usefulness of microsatellites with accessions from prospecting in the Canaries and Aragon. The results showed a low polymorphism and population structure similar shows 8 nd one or 22c governing common these genotypes and selected only 27% of the material prospectado for inclusion in the bank. Chapter 6 responds to this point.

Author: BONILLA VALVERDE DANIEL.
Year: 2005.
University: CÓRDOBA [www.uco.es].
Place of defense: FACULTAD DE CIENCIAS.
Place of preparation: FACULTAD DE CIENCIAS.

Summary: Doñana biosphere reserve for its diversity of species and ecosystems, is affected by agricultural, industrial and mining, which pose a risk. In its surroundings are conducted several crops: rice, strawberry crops and "new agriculture". To unite all discharges to the Guadiamar River aceiteras or wine industries. Another potential source of contamination is located in Los Polos Industrial Huelva. At No. Doñana are pyrite mine of Aznalcóllar, whose products were stored in a bag stériles. In April 1998, the raft of Azanlcóllar released to you Agrio rivers and Guadiamar 4 Hm3 water acidic and 2Hm3 mud with toxic metals. The measurement of various biomarkers of pollution (Enzymes antioxidativas, enzymes biotransformadoras and levels of glutathione) sensitive metals and organic compounds in two species bioindicadoras abundant and unprotected: a mammal, the mouse moruno (Mus spretus), and a bird, common sparrow (Passer domesticus), has served to assess the biological effects of the spill of Aznalcóllar and assess the environmental pollution Doñana in terrestrial ecosystems. New techniques proteómicas have been used parala search for new biomarkers of contamination specific and sensitive mouse moruno sparrow and common environment Doñana.

Author: ROMERO RUIZ ANTONIO.
Year: 2005.
University: CÓRDOBA [www.uco.es].
Place of defense: FACULTAD DE CIENCIAS.
Place of preparation: FACULTAD DE CIENCIAS.

Summary: The main theme is to study and monitor the environmental situation Doñana after the dumping of toxic sludge occurred in April 1998 through biomarkers of environmental pollution. This has been used coquina mud (Scrobicularia flat) as a species bioindicadora pollution in ambient esturáicos which has identified a number of biochemical parameters as tools for monitoring and recovery of the affected area. In the second chapter of this work has developed a new method to quantify a physiological parameter (metalotioneínas) very important as biomarcador of metal contamination. This method overcomes in specificity and sensitivity to those used so far in this type of environmental monitoring programs. In the third chapter uses the most innovative techniques in the field of proteomics, which have been identified by sequencing novo protein altered in different sampling points with metal pollution on the shore of Doñana's Gudalquivir (with the difficulty that the genome of the organism used is not present in the database), which has developed an effective methodology in the description of molecular biomarkers last generation. Fruit of the results obtained are the four scientific papers published in prestigious scientific journals and two book chapters and a large number of contributions to Congress.

Author: JEREZ CABELLO MARÍA JOSÉ.
Year: 2005.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD DE CC QUÍMICAS.
Place of preparation: FACULTAD DE CC. QUÍMICAS.

Summary: The phosphodiesterase 7 has been validated as a therapeutic target for the treatment of inflammatory diseases such as esclarosis multiple or rheumatoid arthritis, as their inhibition in T lymphocytes produces declining interleukins inflammatory and transcription factors NFAT and NFkB associated with these diseases. Therefore, in this paper we develop the following points: 1-Identification of a farmacóforo of PDE7. Within this context design indirect, there is a determination through programs and DISC Catalyst model farmacóforo from inhibitors known PDE7, allowing the search for new seeded. Subsequently, the results of this search will be refined through the program Volsurf to select only those with the most appropriate pharmacokinetic properties, and through the progrma MetaSiete to analyze atoms most likely to be metabolized. 2-Validation of new tools for screening based virtual eligando: CODES and FALP. This initial validation will be carried out on an experimental basis through the synthesis and biological evaluation of new products. Those who are active will undergo cell biological studies to analyze the ability of these compounds for the treatment of inflammatory diseases. 3-Development of a method for predicting toxicity. As a follow-up to candidate selection carried out in the preceding paragraphs, it raises the development of an effective method of predicting mutaggenicidad, because business models currently available do not provide a predictive capacity enough to be considered reliable.

Author: MARTÍNEZ CALATRAVA M. JOSÉ.
Year: 2005.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA U.C.M.

Summary: At present, the main cause of death in Spain-morbi is cardiovascular disease. The work carried out so far in genetic Spanish population, have focused on the analysis of polymorphisms in candidate genes for components classics Metabolic Syndrome (SM): obesity, hypertension, dyslipidemia. The importance of these changes from a cardiovascular standpoint is well established. But there are other components of the S, which also confer cardiovascular risk (hipofibrinolisis, endothelial dysfunction, inflammation) and two that do not have studies that we reveal the extent to which the genes involved in these alterations are associated with the pathological phenotype in our population. For this reason we have analyzed polymorphisms in genes coding for PAI - 1, angiotensin converting enzyme (ACE), nitric oxide synthase endotelila (eNOS), C-reactive protein (CRP), TNF-alpha, adiponectina. To carry out these studies were analyzed using our general population drawn from a stratified random cross-sectional study conducted in the province of Segovia. Our results indicate that genotype 4G/4G of polymorphism 4G/5G PAI - 1 could be associated with the development of impaired fibrinolysis, which is characterized by the presence of circulating levels d ePAI-1 high, and therefore with a greater risk of cardiovascular disease. No evidence has been found that the consumption of snuff have an effect on fibrinolysis dependent on the genotype polymorphism 4G/5G of PAI-1. Also genotype 4G/4G PAI - 1 is associated with a higher prevalence of hypertension in those subjects who show no signs of submitting altered fibrinolysis. This partnership s independent of other metabolic risk factors for hypertension, as well as genotype polymorphism L / R of ACE. Regarding the gene of eNOS, the men of the population analyzed, the C allele of SNP rs3800787 gene of eNOS is associated with a higher prevalence of low HDL cholesterol regardless of other variables related to circulating levels of HDL cholesterol . The molecular mechanism by which this occurs the partnership must still be escarificado. On the other hand, our results suggest that genetic, metabolic and lifestyle are associated with circulating levels of CRP. So the GG genotype polymorphism 1059 G / C, waist circumference, the levels of leptin, and consumption of snuff are positively associated with serious levels of this marker of inflammation. Finally, our results reveal the existence of a gene-gene interaction of genes coding for TNF-alpha and adiponectina, so that both genes might be involved in regulating the levels of serious adiponectina and its impact on glucose homeostasis. In summary, the analysis of polymorphisms of genes coding for the PAI - 1, ACE, eNOS, CRP, TNF-alpha and adiponectina, are associated with different components of the SM (alteration of fibrinolysis, hypertension, dyslipidaemia, inflammation and hyperglycemia), and thus should be considered as markers of risk for SM and disease caridovascular.

Author: SANTOS ETXEPARE MIKEL.
Year: 2005.
University: PAÍS VASCO [www.ehu.es].
Place of defense: FACULTAD DE QUÍMICA.
Place of preparation: FACULTAD DE CIENCIA Y TECNOLOGÍA.

Summary: Candida albicans is a microorganism diner whose natural habitat are the mucosa, the gastrointestinal tract and skin of human beings. In immunocompromised individuals, C.albicans behaves as a pathogen capable of causing infections that are scattered in many cases are fatal. The search for more potent drug combinations for the treatment of invasive fungal infections caused by C.albicans led to the identification of the phosphatase calcineurin as a new therapeutic target. Clacineurina is a component of a signal transduction channel that plays a crucial role in the virulence of C.albicans and contributes significantly to the antifungal drug tolerance of the family delos azole. The information available on other components of this pathway and the operation of the path dependent clacineurina in C.albicans was very low, so this thesis has helped to increase awareness on this path of compromise through the isolation and characterization of a diana of calcineurin which was still unidentified: the effect CaZrz1 serving as a transcription factor regulated by calcineurin during transucción of the calcium signal. Genetic analysis revealed that the effect CaCrz1 has an important role in the regulation of calcium homeostasis and response to agents that cause stress membrane. In addition, the lack of effect CaCrz1 increases the sensitivity of C.albicans to azoles. However, the absence of effector CaCrz1 not affect the virulence of Candida ablicans in a mouse model of disseminated fungal infection.

Author: FERNÁNDEZ CANCIO MÓNICA.
Year: 2005.
University: BARCELONA [www.ub.es].
Place of defense: HOSPITAL VALL'HEBRÓN.
Place of preparation: HOSPITAL VALL D'HEBRÓN.