LIFE SCIENCES, 9

Author: BLANCO CHAPINAL SOLEDAD.
Year: 2005.
University: AUTÓNOMA DE MADRID.
Place of defense: CENTRO NACIONAL DE BIOTECNOLOGÍA.
Place of preparation: CENTRO NACIONAL DE BIOTECNOLOGÍA.

Summary: Malaria is a major cause of efermedad and mortality in humans against which there is as yet no effective vaccine. In this study we have analyzed various strategies designed to modulate immune responses induced in front of the CS protein of Plasmodium yoelii (PyCS), one of the murine models of malaria. It has been observed that the coadministration of various cytokines enhances antigen-specific cellular responses. We have produced recombinant vaccinia virus (rVV) coexpresan cytokines IL-15 and GM-CSF together with the antigen PyCS or with the gene witnessed the luciferasa. In rVV expressing luciferasa and IL-15 we found that IL-15 attenuates replication VV "in vivo". We have also determined that the coexpresión of these cytokines power not significantly specific cellular response versus PyCS. However, when cytokines are co-administered together with the antigen PyCS through DNA vectors followed by a booster dose with a rVV expressing PyCS, significant improvements are achieved. With co-administration of GM-CSF is a greater focus specific cellular response between 4 and 7 times and this empowerment of cellular response correlates with a high level of protection (97%). On the other hand, with the coinoculación of IL-15 is achieved by maintaining this specific cellular response over time. Seeking to enhance the cellular response induced by the inoculation vector DNA, a vector was generated by a sequence of the protein PyCS amended form containing codons used in most mammals, lacked two glycosylation sites and domain binding membrane. Despite these changes, the specific cellular response was similar to that obtained with the plasmid containing the original sequence of the protein. To carry out another of the proposed approaches, we generated rVV multiepitópicos containing sequences encoding various epítopos as minigenes separated elements IRES (internal initiation sites) from different backgrounds. The presentation of epítopo CD8 of the protein PyCS (PyCD8) by antigen presenting cells infected with the various rVV was similar regardless of: i) the context in which it is expressed (within the protein PyCS and isolated immediately after viral promoter or behind a stream IRES), ii) the presence of other epítopos in rVV iii) the origin of the IRES sequence preceding it. Moreover, these rVV inoculated in mice are able to induce specific cellular responses against each of epítopos that are present in rVV: PyCD8 (haplotype H-2d), V3 of the Env protein of HIV (haplotype H-2d) and PfCD8 of the CS protein of P.falciparum (haplotype H-2k) and these cellular responses were similar to those induced by inoculation of rVV expressing complete the corresponding proteins (PyCS, Env, PfCS). This is a novel approach which might be useful for developing vaccines containing epítopos multivalent antigen from different pathogens and / or antigens present in the various stages of the life cycle of a pathogen, which has great significance for parasites with a life cycle complex as is the case with Plasmodium.

Author: Porlan Alonso Eva.
Year: 2005.
University: AUTÓNOMA DE MADRID.
Place of defense: Facultad de Medicina.
Place of preparation: Instituto de Investigaciones Biomédicas "Alberto Sols".

Summary: The majority thyroid hormone receptor (TR) alfa1 and beta1 presented characteristic patterns of expression. Thanks to the models of in vivo gene inactivation is noting the existence of specific isoform functions in some cellular processes. While the role of TRs in cell proliferation is still controversial, and its expression in mitotic cells, the effects of differentiating thyroid hormone (TH) has been extensively characterized in a variety of cell types. However, in most published papers it is not possible to correlate the effect of the hormone with TR isoform of expressing cells under study. Likewise, information on possible physiological functions that TRs exercised in the absence of its ligand is even more rare in the literature. In an attempt to clarify these points were expressed ectópicamente the majority TRs separately in fibroblasts Swiss 3T3, and cell proliferation was studied in the absence of TH, describing a specific physiological function of TR beta1 unknown so far. TR beta1 blocks the proliferation of fibroblasts isoform-specific manner, regardless of their ligand, affecting the main effectors of phase G1; TR beta1 represses transcription promoter of ciclina D1 and blocks the transcriptional activity of factor E2F-1 in response to serum levels decreasing messenger RNA and protein of ciclinas D1, E and A2, preventing activation of Cdk2 and avoiding the phosphorylation of Rb. Characterizing gene profile of the TR beta cells through DNA microarrays identified a group of genes whose expression in a position proliferativas is affected by the presence of this receptor, which highlights the importance of the roles played by TR beta in the absence of its ligand. Finally, we study the role of TR beta in the proliferation associated with the neurogenesis in mice with inactivation of products locus Thrb. Our results show that TR beta is expressed in the SVZ and the SGZ of the adult brain, exerting strong antiproliferative effects on neural precursors of these areas, confirming the effect observed in vitro and involving TR beta in the regulation of the processes that drive the neurogenesis in the CNS.

Author: CUEVAS LABRADOR YOLANDA.
Year: 2005.
University: AUTÓNOMA DE MADRID.
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA.

Summary: MUTACIONES IN PROTEIN SUPRESORA OF TUMOR OF VON HIPPEL - LINDAU (vhl) PROVOKE A SYNDROME TUMORAL HEREDITARIO. RECENTLY, HE HAS SEEN THAT WORKS AS VHL COMPONENT OF RECOGNITION OF SUSTRATO OF A FACILITY E3-UBIQUITININ-LIGASA TO LEADING FACTOR INDUCIBLE IN HIPOXIA (HIF) DEGRADATION VIA PROTEOSOMA. IN NORMAL CONDITIONS OF OXYGEN, HIF MEAN LIFE IS SHORT, DUE TO ITS DEGRADATION OF THE PROTEOSOMA. IN CELL FOR DEFICIENTES VHL, HIF IS NOT UBIQUITINADO FORM THAT IS ACTIVE IN A MANNER CONSTITUENT. LEADING TO A SPECIFIC THERAPY AGAINST TUMORS DEFICIENTES VHL TO HAVE GENERATED A ADENOVIRUS REPLICATIVO CONDITIONAL (CRAD), WHOSE REPLICACION IS UNDER HIF. THIS VIRUS IS CPAZ OF MATAR EFFICIENT WAY OF CELL VHL-DEFICIENTES BOTH IN VITRO AS IN VIVO, JUST AS THE NORMAL CELLS UNDER CONDITIONS OF HIPOXIA. THESE DATA SUGGEST THAT THE AD-9XHRE1A CAN BE VERY EFFICIENT USE OF FORM AS THERAPY OF TUMORS VHL-DEFICIENTES AND MANY OTHER TUMORS TO PRESENT AREAS HIPOXIA OR HIGH WITH ENHANCE THE ACTIVITY OF HIF AS A RESULT OF OTHER TYPES OF DIFFERENT GENETIC DISORDERS THE LOSS OF VHL

Author: Martínez García Ma.Ángeles.
Year: 2005.
University: AUTÓNOMA DE MADRID.
Place of defense: Fac. de Cien.- C.B.M. Severo Ochoa.
Place of preparation: Centro de Biol. Molecular Severo Ochoa.

Summary: The main objective of this work was to characterize the molecular basis of acidaemia metilmalónica isolated (MMA). For this purpose we have conducted the analysis of 16 patients mutacional MMA belonging to the complementation groups mut (13) and cblA (3), describing 11 genotypes different (8 mut and 3 cblA). We have identified a total of 19 variants allelic differences in the gene MUT (deficient in the group complementation mut), which includes 16 potential pathogenic mutations and 3 polymorphisms previously described. Twelve variants have been described for the first time in this study, of which one (A324T) probably corresponds to a mut mutation associated with milder phenotype of the disease and another (I69V) to a polymorphism, or SNP. The gene MMAA (deficient in the group cblA) we encountered 4 mutations new, all responsible for the appearance of stop codons translation premature. We have also studied the degree of conservation between MCM proteins counterparts in different agencies and performed structural analysis of some variants MUT identified in order to obtain more information on the mechanism causing disease. The effect of structural missense mutations has been studied by the location of the different waste affected in the structure 3D enzyme metilmalonil-CoA mutasa in bacteria (subunit of the MCM P.shermanii). Thus we have seen that most of the changes affecting waste highly conserved among organisms phylogenetically distant likely to exert a major role in the folding and maintenance of optimal conformation and stability of the protein. In this paper we have also carried out the different expression of mutant proteins in a cell-free in vitro transcription / translation coupled with the purpose of studying the feasibility of this expression system in the synthesis of proteins that carry out their function in different environments as the cell cytosol (peptides PAH) and mitochondria (subunits -PCC and MCM) and provide additional information on the effect of physiological changes analyzed. We analyzed the effect of nine structural mutations PCCA on the synthesis, import and stability intramitocondrial of subunits -PCC. To do this we measured the rate of degradation of the mutant proteins expressed by in vitro experiments pulso-caza. Our results indicate that most of the alternatives discussed are mutations that affect the structural stability of peptides, introducing accelerated degradation respect to the normal protein inside the mitochondria. Additionally, we also analyzed three PKU mutations associated with the in vivo response to tetrahidrobiopterina (BH4) in patients fenilcetonúricos in order to explore whether the enzyme cofactor PAH exerts a stabilizing effect on the mutant peptides. We have optimized the system expression in vitro in the absence and presence of BH4 and noticed that analyzed for mutations in the cofactor not specifically protects the peptides from degradation accelerated. However, in subsequent studies with other variants of folding PAHs has been that the BH4 it acts as a chaperone chemistry increasing the stability of the mutant proteins. Finally, these results provide important information to better understand the complex relationship between genotype and the phenotype of patients affected by these diseases. Inconsistencies genotipo - fenotipo described some mutations may be explained in part by different modulation in vivo cellular machinery control (chaperonas and systems proteolíticos) and other f 8 players, 328 both epigenetic and environmental, suggesting that in some cases there only genetics but other factors influence the clinical presentation and these metabolic diseases.

Author: MORENO RISUEÑO MIGUEL ÁNGEL.
Year: 2005.
University: POLITÉCNICA DE MADRID.
Place of defense: E.T.S. DE INGENIEROS AGRÓNOMOS.
Place of preparation: ESCUELA TÉCNICA SUPERIOR DE INGENIEROS AGRÓNOMOS.

Summary: SUMMARY: The factors transcripcionales in class DOF are a family of proteins involved in the regulation of gene expression of specific processes of plants join the stream 5Â'- (T / A) AAAG-3Â 'on the promoters of genes regulate. This thesis focuses on: 1) the identification and annotation of new genes DOF barley, 2) the molecular and functional characterization of two new DOF factors of this kind involved in the regulation of specific genes of seeds, and 3) the realization of a phylogenetic analysis of the evolutionary family DOF representing seven species in the evolution of plants, the green alga Chlamydomonas reinhardtii, the moss Physcomitrella patens, the fern Selaginella moellendorffii, gimnosperma Pinus taeda, angiosperma dicotiledónea Arabidopsis thaliana and the two angiosperm moncotiledóneas Oryza sativa and Hordeum vulgare. In C. Reinhardtii, we have identified a single family member DOF, in P. Patens nine, and eight in S. Moellendorffii and P. Taeda. In barley, our species of interest have been identified and recorded 18 new members of this family in databases, in addition to the eight previously noted in this laboratory. To delve into the origin and evolution of this family of factors transcripcionales, we conducted a phylogenetic analysis with DOFs previously identified and those of A. Thaliana and O. Sativa. All these genes can be grouped into six families of genes ortólogos and parálogos to be originated from a grade basal parafilético or subfamily A. The structure of introns and exons of the gene DOF C. Reinhardtii supports the formation of the first DOF by barajado exons in the ancestor of plants. Thus, duplication of DOF ancestral during the subsequent process of evolution would have resulted in the formation of a family of ninety-two members in vascular plants studied here. Our data suggest that the loss, and barajado acquisition of reasons besides the DOF preserved between the new gene, it would have conditioned the formation of the seven subfamilies described herein. Among the twenty-six genes in barley Scored DOF, HvDof17 and HvDof19 were selected for further characterization, as it is expressed preferentially in the aleurona in germination of seeds. In aleurona post-germinativa of cereals, summarizes a set of hidrolasas, whose expression is induced by ABA GA and repressed, which was subsequently secreted in the endosperm amylaceous releasing nutrients that will be mobilized towards the embryo. In the promoters of these genes have been identified in a matter tripartite cis necessary to fully answer GA (GARC: GA-Responsive Complex). By contrast, in cis-regulatory elements that mediate the response of these genes are unknown to the ABA. The tránscritos gene HvDof19 accumulate in the layer aleurona in response to the ABA, while his expression, as well as that of another gene selected HvDof17 suppressed by GA. In this tissue, HvDOF19 half repression by ABA gene CatB, which encodes a thiol protease type catepsina B, and HvDOF17 contributes to the negative regulation of this gene in a path independent of this hormone. Both factors are capable of reversing the trans-activación gene CatB mediated GAMYB, which belongs to the class of factors R2R3MYB and is involved in activation of gene expression of hidrolasas in response to GA; and interact with him in vivo . In the endosperm developing barley, we find that HvDOF17 and HvDOF19 also regulate gene expression Hor2, which encodes a protein reserve B-hordeína. The pattern of expression of HvDof19 during maturation of endosperm, was consistent with their behavior as transcriptional activator gene Hor2 already detected the interaction between it and BLZ2, a promoter for the expression of this gene previously characterized in the lab. By contrast, HvDOF19 represses gene expression Hor2. In conlusión, this thesis contributes to a better understanding of the transcriptional regulation of genes hidrol 8 handles in 341 response to the ABA, and shows the involvement of HvDOF19 and HvDOF17 in the activation and repression, respectively, of genes encoding proteins booking at the barley seed.

Author: EDO SOLSONA MARÍA DOLORES.
Year: 2005.
University: VALENCIA.
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: INSTITUTO DE BIOMEDICINA DE VALENCIA (CSIC).

Summary: The protein p27 is an important cell cycle inhibitor that plays a key role in the transition G1-Sy proliferation of the cell cycle. Its regulation is governed by a multitude of mechanisms, notably by protein degradation route ubicuitín-proteasoma through various interactions with other proteins. The main objective of the work undertaken in this thesis has been the identification of new targets for interaction of the protein p27. Our experimental approach has allowed us to identify the interaction between p27 and SNX6, a protein involved in protein trafficking through the route endolisosomal. By testing in double hybrid yeast and test trawls with GST we identified the domains responsible for such interaction. Studies of expression and subcellular localization by immunofluorescence in HeLa cells 3T3 and have enabled us to demonstrate that SNX6 and p27 colocalizan and that the two proteins are found in different organelles of the route endolisosomal. We have also analyzed the effects of inhibitors of lysosomal degradation on the expression of p27 and the effects of the gain and loss of function SNX6 on the expression of p27 and cell cycle progression. Our results allow us to propose a new path of degradation of p27 through the route endolisosomal going on and cytoplasm during G1.

Author: MUÑOZ DESCALZO SILVIA.
Year: 2005.
University: VALENCIA.
Place of defense: BIBLIOTECA DEL CAMPUS DE BURJASSOT.
Place of preparation: FACULTAD DE CIENCIAS BIOLÓGICAS-UNIVERSITAT DE VALÈNCIA.

Summary: The dorsal closure is one of the last processes morfogenéticos suffered by the Drosophila embryo, between stadiums 12 to 15. During the first half of embryogenesis, the epidermis of the Drosophila embryo has a hole to 10 throughout the piece back, covered only by the amnioserosa, a monolayer of epithelial cells big polyploids coupled with the epidermis and not contribute to the formation the larva. This hole is caused by the retraction of the germ band, which leaves only membrane lining the embryo dorsally to the amnioserosa. A coordinated changes that occur in the form of cells in the epidermis and the amnioserosa to close the gap and generate a continuous dorsal skin is known as dorsal closure. The dorsal closure involves: a polarization flat cells more lateral ridges of the epidermis, changes in the cells of the amnioserosa that contract and invaginan and the formation of a contractile apparatus of actomiosina at the interface between the amnioserosa cells and epidermicas. To close the hole, the cell epidermicas become longer, the contractile apparatus strip them and the cells of the amnioserosa contract and invaginan. Some genes involved in the closure dorsal tambjen are involved in the establishment of cell polarity (PCP) flat in adult tissues, such as June, basketball and flamingo. In the eye of Drosophila, the CFP is reflected in the orderly distribution of omatidios with respect to an imaginary equator (the equator). The polarity defects manifested by the loss of quiralidad of omatidios and a misreading of the same rotation regardless of their position. In Drosophila wing of the PCP is manifested because each cell produces a hair pointing to the distal part of it. Defects in this process will result in cells that produce wing hairs that point toward any party, or even cells that produce more than a hair. In this study we characterized cabut (cbt), a new gene that is involved in the closure anterior-dorsal the Drosophila embryo. This gene was isolated in a crawl genetic gain function using collection lines EP seeking genes that might be involved in the establishment of PCP in the eye and wing, we have also discussed the possible role of cbt in this process.

Author: MARTINEZ LLAMAS MARIA SOLEDAD.
Year: 2005.
University: GRANADA.
Place of defense: Facultad de Ciencias, Universidad de Granada.
Place of preparation: UNIVERSIDAD DE GRANADA.

Summary: The loss of the expression of MHC class I molecules is an escape mechanism of the immune system that occurs in many human tumors and phenotype with murine MHC class I positive. This gives rise to variants tumor phenotype with MHC class I negatives that are recognized by T cells It has been described numerous alterations in the expression of MHC class I molecules in experimental tumors induced in mice, and these changes affect tumor growth and metastatic ability. In our laboratory we have developed a model GR9, consisting of cell lines derived from a fibrosarcoma induced metilcolantreno, clones of the primary tumor and metastases from some clones in mice BALB / C immunocompetent and nude. The primary tumor has been extensively studied in our laboratory. The clones of the primary tumor presents different expression of molecules H-2, different immunogenicity and metastatic power. In trials metastasis expontanes to clone B9, in nude mice (depleted all metastases obtained. These findings present experimental evidence supporting the hypothesis that the phenotype H-2 of the different metastasis is influenced by the code of cells T host implying the existence of inmunoseleccion during development metastatic. In investigating the mechanism responsible for the lack of expression of class I molecules on the surface show that this system tumor is a lack of coordinated expression of transcriptional certain components of APM such as TAP1, TAP2, LMP2, LMP7, LMP10, calnexina and tapasina. The expression of these components from APM is recovered after treatment with IFN-g also recovering the expression of class I molecules in the tests conducted in vivo immunocompetent mice shows that there is a direct relationship between the expression of MHC class I molecules on the surface and locally. Asi metastases MHC class I are very inmunogénas in positive mice suggests a direct involvement of T cells was also carried out tests in vivo immunization, showing that the lines metástasicas H2 class I affirmative protect against the growth of tumor metastases H2 class I negative. construct various libraries theft in order to investigate the possible common regulatory mechanism to explain the low coordinated expression of certain components of the APM, and we found several genes that might be involved and where will focus future studies. Between them highlight two of them: the gene suppressor gene your mores Fhit and Glis I

Author: Escoté Miró Xavier.
Year: 2005.
University: POMPEU FABRA.
Place of defense: Ciencias Experimentales y de la Salud.
Place of preparation: Escuela de Postgrado y Doctorado de la Universidad Autónoma de Barcelona.

Summary: The activation of protein kinases of stress is essential for proper cellular adaptation to external stimuli. In the yeast S. Cerevisiae, an increase of extracellular osmolarity activates the route of HOG1, which produces a complex adaptive response. This work has been characterized various aspects of this response, discovering that one of the protective mechanisms induced HOG1 involves a stop at cell cycle progression in G1 and in G2. This paper demonstrates that the activation of Hog1 arrested cells in phase G1 through a dual mechanism that ends with inhibition of CDK (ciclinas dependent kinase, a key enzyme that allows cell cycle progression). On the one hand, Hog1 inhibits the expression of ciclinas (protein required for the activation of CDK) and also stabilizes Sic1 (inhibitory protein of CDK). The work focuses on the latter mechanism, showing that Hog1 interacts physically in vivo and in vitro with Sic1, fosforilándolo in a threonine end carboxi terminal. The phosphorylation of Sic1 lengthens its half-life, slowing the process of ubiquitinación to which it is subjected in the absence of stress. The proposed model predicts, therefore, that the phosphorylation of Sic1 is essential for adaptation to conditions hiperosmolaridad. Indeed, cells deficient in Sic1 or containing allele of Sic1 mutated on the site of phosphorylation of Hog1 are unable to stop in G1 in the presence of salt in the middle, which produces a number of serious defects among these is the emerging poliploidias. For its part, the MAPK is capable of regulating the transition from G2 to mitosis through a mechanism similar to G1, where MAPK through a dual mechanism decreases the activity CDK this stage. Specifically activation Hog1 leads falling levels of messenger of the main ciclina of G2, Clb2 and induces stabilization inhibor of CDK, Swe1. This stabilization, unlike what happens in G1 is done using an indirect effect, affecting inhibitors Swe1, Hsl1-Hsl7. What has been demonstrated in these results is a direct phosphorylation of Hog1 on Hsl1, particularly in the region of union with Hsl7, resulting in a relocation of the latter, and therefore, in the inactivation of the complex Hsl1-Hsl7 . Therefore, inactivation of the complex Hsl1-Hsl7 leads to the stabilization of the inhibitor of CDK Swe1. If in a position to stress there are no such conditions, the cell enters prematurely in G2, appearing cells osmosensibles with high cell binucleadas. Overall, the results show that inhibitors of CDK Sic1 and Swe1 are molecular targets of Hog1 (directly and indirectly), and in turn strengthen the role of the MAP kinases as controllers of cell cycle in eukaryotic cells

Author: MURGUI PÉREZ ENRIQUE.
Year: 2005.
University: VALENCIA.
Place of defense: FACULTAD DE MATEMÁTICAS - UNIVERSITAT DE VALÈNCIA.
Place of preparation: FACULTAD DE CIENCIAS BIOLÓGICAS - UNIVERSITAT DE VALÈNCIA.

Summary: The municipality of Valencia (76 km2 excluding natural park L'Albufera) was divided into 210 grid of 700 m * 700 m With respect to the birds, field work is desarró during the winter breeding seasons and the years 1997 and 1998 and in each of these periods grid was covered exhaustively birds seen were identified, counted and assigned to different habitats using the time of observation With regard wings landscape features in each grid are located all parcels of habitat, and each of them won their area, perimeter, shape and insulation. These data were obtained both the composition (amount, number and diversity of habitats) and structure (spatial location) of the landscape at each sampling unit. The community is rich in bird species (79-85 species depending on the season) and with high diversity, although there is a numerical predominance of three species abundance and low population of most species terms fenológico predominated temporary species (wintering 0 breeding summer, although most of the wealth was due to the contribution of sedentary species. sharp contrast There seasonal so that the communities were better structured birds during the winter, aspect With factors biogeograficos and with the distribution and types of habitats that tend to reach their maximum bid trophic for many species during the winter and instead are severely impaired its ability to host species breeding during spring. analysis of the relationship between the characteristics of the landscape and birds were observed using three approaches: analysis of gradients, relaci6n between the structure and the community of birds, and finally an approximation semiexperimental by analyzing the variation diachronic of birdlife in a process of urbanization, All approximations coincided in pointing out that the species richness and diversity decreases with the degree of urbanization. abundance and biomass of birds, however, shows more complex patterns, dependent both on the inclusion or otherwise of the three most abundant species As the seasonality. strictly urban landscape, the main conclusion is that both the characteristics of the communities of birds, as the abundance or distribution of the different species, were explained by variables concerning the composition of the landscape rather than its structure , which is attributed to the short distance between landscape elements, adaptation of the species prior to parcelados landscapes and the complex relations between quantity and spatial arrangement of habitat. urbanization process, involves the loss and rarefacción of many species both in the regional and local levels. Despite est a trend, the municipality of Valencia still maintains a diversified birds with a significant percentage of species important for the maintenance of biodiversity in different regions. At a local level, natural habitats and agricultural landscape Peripheral, especially the portion north of the municipality, are the most interesting from the point of view of biodiversity conservation. Maintain or increase the value that even exists, goes through several actions that should be placed in a regional context. in locally, this beyond doubt that, both for their intrinsic value as its supplier of species and populations of the urban landscape, the landscape peripheral agricultural, Huerta, should be actively pr 8 otegida 4c3 through the different figures d protection that the law provides autonomic for the purpose. part of the urban landscape are the two principal actions. (i) the consideration of solar, in terms of their richness and value conservation of the species, such as green areas, on an equal basis other urban parks, (ii) a substantial change in the design and management of urban parks with regard to its size, proportion of paved land, floristic composition and structure of vegetation.

Author: MORANTE REDOLAT JOSÉ MANUEL.
Year: 2005.
University: VALENCIA.
Place of defense: GEOGRAFÍA E HISTORIA.
Place of preparation: INSTITUTO DE BIOMEDICINA DE VALENCIA (C.S.I.C.).

Summary: In this work, using an approximation of the candidate gene in the critical region of chromosome 10 previously described, identifies LGI1 as the gene causing the Temporary Autosómica Controlling Epilepsy Side, a type of epilepsy characterized by partial seizures originating in the region lateral temporal lobe, with frequent symptoms hearing and secondary generalization. To expand the limited information available at that time on the biology of the gene and to obtain data from its involvement in the pathology epileptic analyzes their genomic structure describing a variant of alternative processing, polyclonal antibodies are generated with which characterized the pattern expression of mouse brain using immunohistochemistry and immunocytochemistry and lIeva out a crawl through the technique of Double Hybrid in yeast to identify molecules with the ability to interact physically with LGI1. From the results of the Dual Hybrid makes a hypothesis that relates to LGI1 the route signaling mediated triple receiver p75NTR, NgR/Lingo1. Translating these results to a model of mammalian cell lines was demonstrated by the physical interaction of LGI1 with the three members of the receiver and its ability to act as extracellular ligand of this complex. Finally, it shows the functional effect of this molecule on the path of growth inhibition caused by neuritas inhibitors present in the myelin leading receiver p75NTR/NgR/Lingo1, so that the presence of LGI1 antagonises the inhibitory effect allowing growth of neuritas and avoiding the collapse of axonal growth cone of primary cultures of neurons in the presence of myelin. With all this data suggests a role LGI1 in synaptic plasticity processes that occur in the central nervous system and requiring a local pressure attenuation of growth inhibitory exercising myelin. It hipotetiza that mutations in LGI1 which alter their function could lead to errors in the process of synaptic remodeling, a phenomenon that has been linked to the onset of epileptic seizures

Author: CANALS MERSEBURGER M. GORETTI.
Year: 2005.
University: BARCELONA.
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA - UNIVERSIDAD DE BARCELONA.

Summary: An understanding of nutrient cycling in river ecosystems is important given the growing influence of human activity on its eutrophication. However, the existing knowledge about nutrient cycling in rivers affected by inflow of nutrients from human activity is still low. The overall objective of this thesis is to evaluate the effects of a point source of nutrients from the effluent of a Wastewater Purifying Plant (WWTP) on the functioning of rivers receivers. To examine these effects, we selected two sections located upstream and downstream from a point source into two rivers that drenaban basins with different land uses, to have two contrasting scenarios studies. Specifically, the study was conducted in the river Tordera and Gurri (Catalonia), which drains a watershed largely forested and agricultural sectors, respectively. In the Tordera, is the entry point source majority of nutrients in the river. In Curri, however, the river receives diffuse sources of nutrients from agricultural fields adjacent to the stretches of study besides the point source. The concentrations of ammonium (NH4 +-N), nitrate (NO3-N), soluble reactive phosphorus (SRP) and dissolved organic carbon (DOC) was significantly higher in the stretch downstream in the upstream in the river afforested . The effect of the point source on the water chemistry was not clear in the river farm, where only the concentration of SRP increased significantly below the point source. These effects were reflected in the effects on functional parameters in the study of rivers. The efficiency of retention river NH4 + -Ny SRP decreased downstream (distances of the order of assimilation km) from the point source in relation to upstream (assimilation distances on the order of hundreds of m) in the river afforested. By contrast, the efficiency of retention of nutrients (NH4 +-N, NO3 -Ny SRP) was not affected by point source in the river farm, where these efficiencies were already low (distances of the order of assimilation km) above the point source. The denitrification rates (mg N m-2 h-1) and efficiencies denitrification (mg Ng AFDM-1 h-1; where AFDM corresponds to the biomass) were higher upstream to downstream in the river afforested. These efficiencies and rates did not differ significantly between the two sections on the river agriculture. The daily rates of metabolism (reproduction gross primary or PG, or R and respiration) tended to be higher below the point source in all cases studied in the two rivers. However, these increases were significant only in the case of R in the river afforested. On the same stretch of the two rivers, the daily rates of net ecosystem production were negative in most cases, indicating that these ecosystems were predominantly heterotrophic. In the two rivers see a potential negative relationship between demand biological PO4 3--Py concentration SRP, and a potential positive relationship between rates and the concentration of PG SR, which remained consistent when we combine our results with those the literature of rivers in different biomes. These relationships indicate that the availability of SRP is a common factor in regulating demand biological PO4 3--Py of activity fotoautotrófica in a very different rivers. In conclusion, the sources of nutrients saturaban the local effect of the point source on the operation of agricultural River. Instead, the river was afforested very vulnerable to the point source.

Author: SOTO SÁNCHEZ CRISTINA.
Year: 2005.
University: SANTIAGO DE COMPOSTELA.
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: VIGO.

Summary: The subnúcleo lattice dorsal (SRD) is located in the reticular formation medial flow to óbex. Studies in rodents indicate that receives information nociceptiva and increases its transmission to both levels supraespinales as to the spinal cord. In rodents, transmission upward travel the lemnisco medial to the core ventral-medial talámico, and the transmission down the specification dorsolateral. Since the data from this nucleus were obtained in histological and behavioral studies conducted in rodents, we decided to determine the location of the SRD neurons in the cat and examine their electrophysiological properties. The results show that: * In the cat, the SRD is located in the reticular formation in the bone marrow oblong flow to óbex, but unlike what happens in rodents, there is a certain distribution somatotópica of neurons in the nucleus, so that they receive information through the dorsal columns are in the region of the medial reticular formation. * As in rodents and primates, neurons of the SRD are activated only by stimuli nociceptivos. * 62% (n = 102/165) of the cells registered send an axon to the spinal cord. * 44% (n = 24/55) of the neurons examined were activated antidrómicamente by stimulation of the medial bulbar reticular formation. * The 9% (9 / 103) of the neurons examined were antidrómicas to stimulation lemnisco medial, unlike what happens in rodents where the rostral projection of these neurons is done through lemnisco medial. * 82% (n = 27/33) of the cells activated antidrómicamente by stimulation rostral were also antidrómica the spinal stimulation, so that the information is sent as feedback to the marrow is the same as that sent rostralmente. The results show that in the cat, as in the rat cells SRD painful process information. However, the cat, these cells projected levels supraespinales more rostrales primarily through training lattice medial and to a lesser extent through lemnisco medial. Although we do not know the place of termination of these projections through the reticular formation, histological data indicate that at the end reticular more rostrales from where they may also be transmitted to the nucleus intralaminares talámicos to reach the cerebral cortex. Since the reticular formation bulbar houses circuits remotores related motor involuntary actions intending directly to motoneuronas the flagpole ventral spinal cord, the SRD can participate in reactions associated with motor processes nociceptivos and to prevent or reduce the pain generated by stimuli nociceptivos external and internal. This can be important for postural reactions defense being taken in response to the presence of an outbreak painful and involving the modification of the tone of the muscle groups aimed at immobilizes the region and away from painful stimuli that may enhance this situation. In addition, if the information is transmitted rostralmente reaches the cerebral cortex through the nuclei intralaminares, the crust could control postural reactions acting at the level of cells SRD intending to training lattice and the spinal cord.

Author: ALVAREZ FARIÑA PABLO.
Year: 2005.
University: SANTIAGO DE COMPOSTELA.
Place of defense: INSTITUTO DE ACUICULTURA.
Place of preparation: DPTO. DE FISIOLOGÍA - FACULTAD DE FARAMCIA.

Summary: The thesis examines the reproduction of two species of native clams in Galicia commercial interest: the clam Bicuda Venerupis aurea (Gmelin, 1791) and clam Babosa Venerupis pullastra (Montagà ¼, 1803). Both shares clams inhabiting player behavior and have different depending on the ability of resource management bionergéticos underpinning gametogenesis, the glycogen and lipid, in this aspect vesicular cells, tissue key constituent reserves, are capable of storing both bioenergy components. Note that the clam Bicuda, no vesicular cells, storing glycogen in the muscle intergonadal while the clam Babosa has a great ability to develop cell is, reaching about 60% shaping space gonadal. These Venéridos have the ability to invest in print up to 75% of their body (soma or soft) and its temporal evolution tell us, on the clam Bicuda put occur in the spring and the clam Babosa starting in late winter to early fall, the difference in these profiles is due to the capacity of the clam Babosa to store more resources bioenergy, which makes it more independent of environmental conditions, primarily food. The fecundity and fertility potential relative few parameters are studied in shellfish, so we made the estimate in sexually mature specimens, where the clam Babosa presented much of oocytes to be able to cast that Bicuda clam, and the number of oocytes by copy (fertility potential) not presenting significant differences, extracting it is possible that a species has increased fertility work, although the number of oocytes per unit volume (relative fertility) does not differ significantly. For this study we use qualitative methods -histología conventional and histoquímica- to observe aspects on the formation of the gonad and locate reservations bioenergéticas and quantitative methods -distribución size oocytes and estereología- to describe cycles gonadales and calculation of fertility in these mollusks.

Author: MOURIÑO LÓPEZ SUSANA.
Year: 2005.
University: SANTIAGO DE COMPOSTELA.
Place of defense: INSTITUTO DE ACUICULTURA.
Place of preparation: INSTITUTO DE ACUICULTURA.

Summary: Vibrio anguillarum is one of the principal agents of vibriosis of fish. Among the various mechanisms of virulence possessed, underlines its ability to obtain iron from different sources present in the host, including groups hemo. In this PhD thesis has been carried out the structural and functional characterization of a gene cluster that encodes the capture and transport of the molecule hemo from means to the extracellular bacterial cytoplasm, exploring the possible role played by this mechanism for obtaining iron in survival and multiplication of microorganism in the infective process. The system tangénico described is composed of nine genes organized into three units transcripcionales (gene huvA and operons huvXZ and tonB1exbBD1-huvBCD (), whose expression is regulated by the concentration of intracellular iron, through the transcriptional regulator Fur. Addition, the detection cluster gene sequence described in different strains V.anguillarum serotypes O1 to O10, confirms that the usability of hemo is a defining characteristic of the species. however, has shown a marked intraspecific genetic variation in the hemo use system, since only the genes tonB1, exbB1 and exbD1 are present in all strains tested. identification of two different external membrane receptors, HuvA and HuvS, with a low similarity in amino acid sequence, although functionally interchangeable , also suggests differences phylogenetic within V.anguillarum, or the presence of events transfernecia horizontal gene Finally, characterization and functional analysis of two complex TonB (TonB1 and TonB2) V.anguillarum, have proved the involvement of both systems in different mechanisms for obtaining iron, being functionally interchangeable for the use of hemo. virulence tests have revealed the relationship of the two systems with the virulence of this microorganism, as well as the role that the mechanism of uptake and transport of hemo play developments in the evolution of the infection caused by V.anguillarum.

Author: JULIÀ SAPÉ Ma. MARGARITA.
Year: 2005.
University: AUTÓNOMA DE BARCELONA.
Place of defense: FACULTAT DE CIÈNCIES.
Place of preparation: UNIVERSIDAD DE BARCELONA.

Summary: OBJECTIVE: To improve the characterization of the non-invasive brain tumors through the technique of magnetic resonance spectroscopy of proton. To achieve this goal, three studies were conducted with the following objectives: Study 1) Generate a database accessible through the Internet, containing clinical data and spectroscopic completely validated patients affected by brain tumors. Study 2) To determine the influence of time used in the acquisition reflected in the subsequent calsificación the spectra of human brain tumors. It compared the wisdom gained in the classification of tumor response times using two different (30 ms and 136 ms). Study 3) estimate the wisdom in interpreting the images resonance for the classification of tumroes cerebales, both in terms of type and degree of tumor malignancy. METHODS: Study 1) All data of patients who entered the draft INTERPRET (International Network for Pattern Recognition of tumors using Magnetic Resonance, http://azizu.uab.es/INTERPRET) will be kept in a database accessible via Internet (iDB). These data were selected through the interface questions of the database. same region which subsequently took the diagnostic biopsy, and the spectrum of reflected short time has not been discarded by purchasing artifacts or other reasons, and that a committee had reached consensus neuropathologists diagnosis. When spectra were obtained from normal volunteers, or were abscesses or clinically proven metastases were not required biopsy. Study 2) were selected retrospectively hundred and fifty-one studies of patients with brain tumors (37 meningiomas, 12 astrocytomas low grade, 16 astrocytomas grade, 54 glioblastomas and 32 metastases) in a series of three hundred and seventy-eight exams abnormal brain masses carried out under the project MEDIVO. Volume spectroscopy was performed only in time to echo 30 ms and 136 ms in all cases. It normalized areas composed of nine resonances of interest. The tumroes were classified into four groups using discriminant analysis: meningioma, astrocytoma low-grade, high-grade astrocytoma and gliobalstomas conjunction with metastases. We evaluated the outcome of discriminant analysis at each time echo through the method of leaving a case outside. Study 3) were retrospectively evaluated the agreement between the classification radiological and histopathological diagnosis, using data back three hundred and ninety-three patients of brain tumors that were stored in the database ITNERPRET (iDB). It created an ontology to define agreement between diagnoses. Each category tmoral compared bilaterally with the diagnoses histopatológcos. We calculated the values of sensitivity, specificity, positive and negative predictive values, and confidence intervals of 95% of Wilson. RESULTS: Survey 1) There was a subgroup of three hundred and four patients (twenty-two normal volunteers and two hundred and eighty two patients). These cases will be migrated to another searchable database (database validated). The meningiomas were the only group for which the grading was more sensitive and specific to echo long time. Results improved significantly when both echo times were considered simultaneously: the suggested diagnosis was corecto in 105 (94%) of 112 cases when the result predicted by both echo times was the same. It was suggested the correct diagnosis at least one of the two times in 136 (90%) cases of 151. Study 3) When radiologists reported and 8 studios bc2 magnetic resonance imaging, are highly specific (85.2-100%) in the characterization of the degree and type of tumor. Its sensitivity is variable depending on the type and degree, separately and in combination. Within the broad categories (neuroepithelial, meningeal strain) were very sensitive while when considered ways to inform the detailed type, the sensitivity was variable, being the highest for meningiomas low grade (sensitivity 100% confidence interval, 96.2-100.0%) and lowest in meningiomas high grade (sensitivity 0.0, confidence interval, 0.0-65.8%) and the low-grade oligodendrogliomas (sensitivity, 15% confidence interval, 5.2% -36.0% ). The sensitivity in detecting the origin of neuroepithelial tumors was correlated inversely with the accuracy in describing the extent and origin of the tumor cell. "Glioma was classified radiation frequently associated with a higher sensitivity in its category. The positive predictive value ranged between categories, reaching values generally higher than the prevalence of the respective tumors in this database. The negative predictive value was high for all categories analyzed (69.8-100%). CONCLUSIONS: The database validated complies with the regulations and ethical represents the population studied. It is accessible to neuroradiólogos around the world wishing to use the information espectroscópica in the non-invasive diagnosis of brain tumors. The short echo time provides a classification slightly better than echo long time, and the results improve when considering the two times echo simultaneously. The meningiomas are the only group for which the tumor echo long time is slightly better than the short echo time. The positive and negative predictive values for diagnostic imaging resonance were obtained can be used as the likelihood estimators post for the characterization of brain tumors in the database INTERPRET. In addition, it highlighted the need for greater sensitivity in the categorization of most brain tumors, while maintaining high specificity, especially through techniques such as magnetic resonance spectroscopy of proton.

Author: DOMENZAIN REYNA CLELIA.
Year: 2005.
University: AUTÓNOMA DE BARCELONA.
Place of defense: D.DE BIOQUIMICA Y BIOLOGIA MOLECULAR.
Place of preparation: UNIVERSIDAD AUTONOMA DE BARCELONA.

Summary: The composition of the extracellular matrix (ECM) identifies some crucial aspects of tumor behavior. Among its components highlights glycoproteins and proteoglycans (PGs), some of which have receptors on the cell membrane. Our research group is interested in the biology of PGs in human melanoma. The VS is a PG matrix of the family of hialectanos. It is composed of three domains: G1, G2 and G3. The alternative splicing domain Central G2 generates four isoforms called V0, V1, V2 and V3. In human melanoma cells has been described as the expression of proteoglycans the proteoglycan specific melanoma (mel-PG), CD44 or versicano (VS), whose presence was described in our laboratory. Also, we describe its ability to increase the proliferation and migration in human melanoma cells, in addition to reducing its commitment. In the first part of this work we determine the pattern of expression of the isoforms of VS and mel-PG during the process desdiferenciación of melanoma cells, using cell lines with different degrees of differentiation (early, intermediate and late). Our results show that the degree of differentiation of cells determines the differential expression of VS and mel-PG, so that more cell lines expressing indiferenciadas all isoforms of versicano and more differentiated either. De ello concluimos que la pérdida del estado diferenciado se acompaña de la expresión de las diferentes isoformas de VS, hecho que parece ser común en tumores derivados de células con mismo origen embrionario, como muestran nuestros análisis preliminares realizados en células de neuroblastoma y astrocitoma. During the process of differentiation in vitro cell lines SK-mel-1.36-1-5 and SK-mel-3.44 these PGs disappear, so we propose that the production of versicano is a sign of regression to an undifferentiated state during the tumor progression, and that there is a time for the expression pattern of each isoform of versicano. In the second part we analyze the regulation of the promoter of VS in human melanoma cell lines with varying degrees of differentiation early and late. Aislamos, secuenciamos and clone human promoter VS. The in silico analysis identified the same three areas of regulation: the first in the extreme 5 ', presents two boxes of recognition for TCF-4, the second in the central zone, 6 boxes union to Sp1, 7 for AP-2 and one for Smad3 / 4, and the third in the extreme 3 ', a box of union for AP-1. We build various deletion mutants designed to isolate the role of each element regulators promoter VS and transfectamos so transient in cell lines. The results show that TCF-4 is an activator responsible for more than 50% of the total activity of this promoter, as the sobrexpresión some of the participants of the road signaling TCF-4 has a direct impact on activity . Moreover, by EMSA tests determine the union's specific complex TCF-4 / -catenina. In the central region lies approximately 10% of the total activity of the promoter and we have given the union specifically 3 of potential binding sites for Sp1. We propose therefore that Sp1 is a modulator or perhaps regulator rapid response VS. Finally, in the region 3 'promoter, which contains the sequence of recognition for AP-1 is the remaining 40% of the activity and we find that this sequence is responsible for the basal activity of VS.

Author: LAZZARO ALEJANDRA SILVIA.
Year: 2005.
University: LA LAGUNA.
Place of defense: I.U.DE BIO ORGANICA.
Place of preparation: INSTITUTO UNIVERSITARIO DE BIO ORGANICA ANTONIO GONZALEZ.

Summary: This thesis explores various aspects of the molecules and mechanisms involved in the Evil of Panama. During the development of this disease, M. Acuminata and F. Oxysporum interact using various strategies developed during the coevolución between them. Given the presence of F. Oxysporum, M. Acuminata triggers the synthesis of various phytoalexins fenilfenalenónicas and other derivatives fenalenona, some with antifungal activity. The phytoalexins not only have antifungal activity directly on the pathogen, but also frees the species reactive singlet oxygen (1O2) in the presence of light. The antifungal activity of phytoalexins fenilfenalenónicas in the presence of light correlates with the intensity of the production of 1O2. This reactive oxygen species is highly damaging to cells. On the one hand is toxic to the pathogen, and secondly, active in the hypersensitive response leaf discs M. Acuminata. The ability of fenilfenalenonas to produce 1O2 is reflected in its greater capacity to damage the DNA plasmid in experiments in vitro. Moreover, it is described that the 1O2 is able to induce lipid peroxidation of membrane, causing the cells lose their integrity. In this cell damage enzymes involved are lipoxigenasas. M. Acuminata naturally produces the compound naproxen, which together with some structural synthetic analogues has been shown as an inhibitor of lipoxigenasas, besides possessing antifungal activity. Naproxen, an acid derivative naftalenacético influences on the growth of seedlings in vitro M. Acuminata and calluses of Bupleurum salycifolium. In turn, F. Oxysporum reacts to fenilfenalenonas M. Acuminata, detoxificando in vitro one of the most aggressive in darkness and methylation by converting it into another fenilfenalenona much weaker as antifungal. Both race pathogenic (FOC4) as a non-pathogenic (FOC1) are able to carry out detoxification, so it can be assumed that the difference in pathogenicity observed for both races would not be linked to the capacity of detoxification. In in vitro production of FOC4 in the presence of fenilfenalenona PN3, the fungus produces natural antioxidant tirosol. It would be interesting to explore whether the tirosol occurs in the interaction in vivo and whether it was involved in the relationship M. Acuminata - FOC4. The control of Evil Panama is quite complex for a variety of reasons. A control strategy is to use whole organisms, or exometabolitos produced by them to counter the growth of a pathogen. This study has investigated the production of antifungal compounds in a strain of Penicillium melinii found four compounds, two of them with antifungal activity against FOC4 and two inactive. Compounds isolated, and whose structures have been elucidadas relate acid 2,3-dihidro-2-metil-4-benzofuran carboxylic (PM1) and acid 3-hidroxi-2-metil-4-benzofuran carboxylic (FP4).

Author: CERVERÓ SANZ ANA CRISTINA.
Year: 2005.
University: VALENCIA.
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA.

Summary: The leptin system is involved in regulating body weight yen reproductive function acting at endocrine and paracrine. There is evidence that this system is essential for embryo implantation in mice. This paper describes the expression of leptin receptor and its different isoforms varies to 10 throughout the menstrual cycle in the human endometrium. The leptin receptor is present to 10 throughout the embryonic development preimplantatorio while leptin is expressed only in the blastocyst stage. There were no significant differences in the levels of expression nor the location of the leptin receptor in the endometrium of infertile women with endometriosis moderate / severe and fertile women during the window of implantation. In addition, both types of endometrios express leptin at very low levels. The blockade in vitro expression of the leptin receptor is not declining rates embryonic membership, indicating that this system is not involved at that stage. However, it does not rule out that it could be involved in the embryonic stage of invasion through the activation of metaloproteasas and angiogenic factors.

Author: DE ANDRADE BASTOS AFRANIO.
Year: 2005.
University: LEÓN.
Place of defense: ESCUELA DE VETERINARIA.
Place of preparation: UNIVERSIDAD DE LEÓN.

Summary: The practice of sports can be considered one of the biggest social phenomena of the contemporary world. In the sporting perspective, it is vitally important that every coach knows the reasons for the participation of athletes in their training, how the athlete perceives its level of physical ability, and that this measure aimed at achieving any goal. And among the great variety of existing sports, swimming is one of the most practiced throughout the world. For all these reasons, the main objective of this research was to analyze the relationships between the main reasons leading to the practice of competitive swimming with the perceived level of physical ability, and the prospects for achieving goals in Brazilian athletes. The sample consisted of 425 athletes (138 nadadoras and 287 swimmers), with a mean age of 14.7 years, from different clubs in five Brazilian states. To collect the necessary information was used a battery of tests. These tests or questionnaires suffered a treatment process and conforms to the characteristics of the study and the sample. Data analysis was performed using the statistical package SPSS version 11.0 for Windows (SPSS Inc. At Chicago, IL). For the study of the validity and reliability of the questionnaire was used factor analysis with varimax rotation, the study of correlations inter-item and calculating Cronbach alpha values, and the test-retest. Our results indicate that: The main reasons for the practice of competitive swimming in the population studied relate to the need to develop new motor skills and improve health, the need for socialization and cooperation among equals, all of intrinsic nature . The perceived physical ability is puntuada with specific values higher swimmers in the lower categories, perhaps because they overestimate their ability to provide even limited information on the techniques of swimming. Subjects participating in the study showed a motivation for achieving more task oriented toward the ego. This result can be attributed to the specificity of sport practiced. While there are gender differences, the data suggest a higher association of guidance to the task with motivational factors intrinsic and guidance to ego with extrinsic factors motivational.