LIFE SCIENCES, 6

Author: ALONSO CANTABRANA HUGO.
Year: 2005.
University: MIGUEL HERNÁNDEZ DE ELCHE.
Place of defense: UNIVERSIDAD MIGUEL HERNANDEZ CAMPUS DE SAN JUAN.
Place of preparation: UNIVERSIDAD MIGUEL HERNANDEZ.

Summary: In order to identify new gene functions involved in the development of gineceo in Arabidopsis thaliana, were two independent jobs. On the one hand, a chemical mutagenesis on the lineage cer6, allowing identification of the mutant zeppelin (zpl). The fruits of these plants are shorter and thicker than those of the wild strain, apparently due to an alteration in the direction of cell growth. It characterized the phenotype of these plants, and simultaneously began the study of genetic interactions with other mutant alleles, as fruitfull (ful) and claw crabs (crc). It also initiated a study transcriptómico this mutant. The mutation zpl affecting the region 3 'untranslated gene At5g16220, unknown function. Moreover, we studied the role of gene ASYMMETRIC LEAVES1 (AS1) in the development of gineceo. This gene is involved in the repression of genes KNOX class I lateral bodies, but so far had not been described his role during the formation of fruit. It characterized the phenotype of mutant fruit in simple as1, as well as in various combinations and in a mutant line overexpression of KNAT1. Also, we studied the expression of AS1, KNAT1, REPLUMLESS (RPL) and FUL in gineceos wild and as1. Our results indicate that KNAT1 and AS1 involved in the regionalization of the ovary. KNAT1 confers identity replum and activates the expression of RPL, while AS1 and other products that are expressed in the valva oppose its role.

Author: CASTAÑO CARDOSO JULIO.
Year: 2005.
University: AUTÓNOMA DE BARCELONA.
Place of defense: UNIVERSIDAD AUTÓNOMA DE BARCELONA.
Place of preparation: UNIVERSIDAD AUTÓNOMA DE BARCELONA.

Author: DIESTRA VILLANUEVA ELIA.
Year: 2005.
University: AUTÓNOMA DE BARCELONA.
Place of defense: FACULTAD DE CIENCIAS 103.
Place of preparation: UNIVERSIDAD AUTÓNOMA DE BARCELONA.

Author: Farré Martí Ricard.
Year: 2005.
University: AUTÓNOMA DE BARCELONA.
Place of defense: Facultad de Veterinaria.
Place of preparation: Facultad de Veterinaria.

Author: TAZÓN VEGA BÁRBARA YMAINE.
Year: 2005.
University: AUTÓNOMA DE BARCELONA.
Place of defense: FUNDACIÓ PUIGVERT.
Place of preparation: UNIVERSITAT AUTÒNOMA DE BARCELONA.

Summary: This thesis is a contribution toward understanding the molecular basis of the syndrome of Alport (SA), nephropathy hereditary responsible for 1-2% of cases of chronic renal failure (IRCT) terminal in western countries. The promise inherent in molecular genetics is that the revelation of these mutations could allow establish correlations genotipo - fenotipo and explain the clinical variability of the disease, as well as providing a more solid basis for genetic counseling and prognosis of these patients. Being HS a genetically heterogeneous disease and with different patterns of inheritance, have been addressed separately its various forms and with specific targets and novel set out below: Â investigate the possible relationship between molecular syndrome Alport autosomal recessive (SAAR) and the benign family Haematuria (HFB). Â Conduct a screening mutacional in gene COL4A3 and COL4A4 in patients with syndrome Alport linked to the X chromosome (SALX) * Analysis of linkage in the region Xq22-q23. * Screening mutacional gene COL4A5. . Studying the feasibility of genetic diagnosis preimplantacional specific syndrome Alport linked to the X chromosome (SALX). . To characterize a family syndrome Alport linked to the X chromosome (SALX) presenting a transmission varión-varón dela disease. The first chapter results of this paper elaborates on the controversy that existed before starting this study on the possible relationship between molecular HS and HFB, kidney disease, which presents some traits clinically identical to those in the early stage of HS or Holders of the SAAR. These two kidney disease that presents some traits clinically identical to those of genetic Stadium beginning at the start of this thesis, it is believed interesting identify molecular defects in patients who suffer and try to understand the parallels between these two entities clinics. We mutacionales two studies of genes COL4A3 and COL4A4. First, in 54 patients belonging to 11 families unrelated to HFB (Badenas et al. J Am Soc Nephrol 2002) and subsequently, 71 patients from 14 families with SAAR and 2 with HFB (Tazón-Vega B, et al., Am J Kidney Dis 2003). The second chapter is devoted to the shape of SA linked to the X chromosome: SALX. This teipo SA is the most common and therefore the best characterized. However, its molecular diagnosis was not well established at the start of this study. This chapter presents two works being published and a third already published. The first focuses on improving the diagnostic molecular SALX through refinement of the analysis deligamiento in the region chromosome Zq22-q23 and the development of a methodology for finding mutations in the gene COL4A5 cause of the disease. This allows advance in the molecular diagnosis of SALX previously based only on the analysis of linkage, exteniéndolo those cases in which this technique is not applicable: sporadic cases of the disease and those cases where families are available on a small number of individuals. This technique is the first method simultaneously simple, fast and efficient for direct molecular study of this syndrome from RNA. This makes it possible to determine the effect that casuan mutations in the processing of mRNA COL4A5. The technique used is based on an analysis of the entire coding region of the gene COL4A5, which in turn allows to determine the effect that casua mutation in the processing of mRNA of this gene. The second chapter of this work is the development of genetic diagnosis preimplantacional specific SALX and indpendiente sex of the embryo. Finally, during this work was marked the first case of concurrency HS and Klinefelter's syndrome in a family that had a transmission varón-varó 8 No HS 1275 linked to the X chromosome (Ars et al. Eur J Hum Genet 2005). The conclusions of this study are: 1 .- The HFB is casuada by mutations in the genes CIL4A3 and COL4A4, which are also responsible for the SAAR. We have identified 8 mutations not previously known in these type IV collagen genes in families with HFB, with the gene COL4A3 the first mutations described in this disease. 2 .- It has carried out the third full screening, as described so far, the gene COL4A3 and COL4A3 in families with SAAR, which has identified 9 new mutations responsible for the pathogenesis of this disease. This study confirms that the type of missense mutations affecting glycine residues are most often seen in these genes. 3 .- It has been demonstrated in 5 of the families studied, the HFB and state deportador the SAAR could be considered a single disease because these patients share the same molecular defect in the gene COL4A3 and COL4A4 and its clinical evolution is tantamount . 4 .- The HFB, the carrier status SAAR and SAAD, regarded hitherto distinct clinical entities are all caused by a single mutation of the gene COL4A3 or CIL4A4. Therefore, they could move within the term "Nefropatía collagen type IV (chains a3/a4)" resulting in a spectrum ranging from phenotypic microhematuria isolated until IRCT. 5 .- The technique SSCP / HAS applied to the analysis of genetic genes cIL4A3 and COL4A4 has demonstrated a sensitivity of 62%, so it can be considered suitable for the study of these genes. 6 .- It has improved the diagnosis indirect SALX with the use of microsatellite markers that define a region of 1.13-1.23 Mb about gene COL4A5. This has diminished the possibility of recombination between markers and gene ends of a 6% to 1.2%, which greatly reduces the possibility of errors in diagnosis. 7 .- It has developed a simple and quick method of finding mutations in the gene COL4A5, which causes SALX based on the screening of its coding region. This technique has a high efficiency of detection of mutations (76%). Accordingly, it is a methodology that enables direct diagnosis of this syndrome and it is applicable to the study of sporadic cases or small families. 8 .- 41% of patients with SALX to which he has identified the mutation show alterations in the normal splicing of mRNA COL4A5. Of the total of 22 mutations identified in the gene COL4A5, 5 of them (23%) were not as ad mutations desplicing having done a screening of gene-based gDNA. 9 .- The genetic diagnosis preimplantado specific SALX is feasible using linkage analysis in a single cell markers DXS1120, COL4A5-2B6 and DXS456 flanqueantes gene COL4A5. 10 .- It has described the first case of concurrency HS and Klinefelter's syndrome, demonstrating that in a family with SA transmission varón-varón of the disease is not always due to a pattern of inheritance autosóico but can also be caused by mutations in the gene CIL4A5. 11 .- It has been ruled out of the pattern of X chromosome inactivation, detecting that is random, as the cause of severe phenotype SA male who introduced the occurrence of this disease and Klinefelter's syndrome. It has identified a preferential expression of a transcribed aberrant splicing in this patient (50%) compared with that of her grandmother carrier (8%). This finding may explain why the demonstrations of SA in the patient with a karyotype 47. XXY are more similar to those of a man with SALX alongside a woman carrying. Finally, in the annex to the thesis presents three additional jobs also issued during this work: Bowl et al., Nephrology 2003; Prague et al., Nephrology 2003; and torra ycols. Nephrol Dial Translant 2004.

Author: Navarro Langa Juan Antonio.
Year: 2005.
University: VALENCIA.
Place of defense: Facultat de Química.
Place of preparation: Departamento de Genética.

Summary: Friedreich Ataxia of human disease is a neurodegenerative and autosomal recessive inheritance. It is the most common hereditary ataxia in the Caucasian population. The disease is caused by a deficiency of the protein frataxina. The keys on the role of frataxina come from the different studies carried out in several model organisms. We have isolated genes ortólogos in a large number of organisms from bacteria to mice and also in plants. Our laboratory has isolated the gene that encodes the Drosophila protein frataxina, the gene fh. Drosophila has proven to be an excellent model for studying human diseases because many biological processes are conserved between flies and humans, such as the formation and degeneration of the nervous system, the generation and function of the heart, mitochondrial metabolism , etcâ | This paper has approached the study of gene function in D. fh Melanogaster using three of the many strategies in this body oriented mutant phenotypes: mobilizing elements transponibles P, the system ARNi and overexpression. With experiments mutagenesis insercional did not obtain the expected results because the gene is a gene fh small located in a region rich in genes and flanked by two or even three points hot insertion. It then approached a second approximation consisting of the use of the system UAS-GAL4 for the development of overexpression experiments and interference. Interference or overexpression of the gene fh in target tissues in patients and in the embryonic development of Drosophila, induce the appearance of fatalities, in any phenotypic alteration or some behavior modification. The defects found were concentrated by a party in the SNP, both in neurons and axones of sensory component, as in axones component of the engine (in this case the motor neurons do not seem concerned). On the other hand, there have been major changes to several of the derivatives mesodermo embryonic, such as somatic muscle cells and cardiac and pericardiacas. All these defects lead to the appearance of 100% lethality before the rise of adult or a decrease in survivability and escalation in cases you get offspring. The interference and overexpression in muscle have proved effective only if the alteration occurs in the early stages of development of structures affected. For its part, the SNP only have occurred when changes have affected joint precursors sensory or just sensory neurons differentiated involved in the formation of sensory organs in the embryo and the adult. But apparently, the role of fh is not equally important in all sensory organs, and that changing its expression in organs cordotonales not produce any significant change. All results described permit to establish a model, in D. Melanogaster, for the study of Friedreich ataxia.

Author: ARAN CORBELLA BEGOÑA.
Year: 2005.
University: AUTÓNOMA DE BARCELONA.
Place of defense: INSTITUT UNIVERSITARI DEXEUS.
Place of preparation: SERVICIO MEDICINA DE LA REPRODUCCION. INSTITUT UNIVERSITARI DEXEUS.

Author: SANTIAGO GONZALVO NÉSTOR.
Year: 2005.
University: AUTÓNOMA DE BARCELONA.
Place of defense: INST.DE BIOL. MOL. DE BARCELONA (CSIC).
Place of preparation: INST. DE BIOL. MOL. DE BARCELONA (CSIC).

Summary: The work presented in this thesis focuses on the study of the impact of the elements transponibles on genes and genomes plant and the control mechanisms of transposition in these genomes. This has been discussed on the one hand, the dynamics that has followed the family Emigrant of MITEs in the Arabidopsis genome. And, on the other hand, he examines the control of transcription retrotransposón Tnt1 of snuff. 1. STUDY OF IMPACT OF ELEMENTS TRANSPONIBLES ON GENOMAS PLANT: Analysis of the family Emigrant of MITEs Arabidopsis thaliana. The MITEs are small DNA transposons without coding ability, present in a large number of copies of most genomes. It postulates that are mobilized by autonomous related elements present in the same genome. The first group of MITEs described in Arabidopsis was the family Emigrant (Casacuberta et al., 1998). The purpose of our study is to understand the dynamics of these elements and their effect on the evolution of genes and genome of Arabidopsis. Has designed a computer program to identify all copies of Emigrant elements contained in the Arabidopsis genome. In order to find the maximum number of elements Emigrant, the search was based on the terminal repeats (TIRs) preserved that define this family of MITEs. In this way can be found subfamilies represented by a small number of copies or those old copies, and therefore more divergent. Have also developed other software tools that have enabled evolutionary and phylogenetic analysis of these elements. In turn, using molecular techniques has been studied polymorphism of these MITEs between different Arabidopsis ecotypes. Through these tests have identified 151 copies Emigrant distributed throughout the genome of Arabidopsis. Most of these can be grouped into three families, each of which can be subdivided, in turn, subfamilies variability different. Analysis of these subfamilies has revealed that while the most recent copies are located far from gene sequences, those older appear frequently associated with genes. This suggests that the elements Emigrant have been able to play an important role in the evolution of genes from Arabidopsis. Moreover the existence of polymorphisms of these transposons integration between different ecotypes of Arabidopsis indicates the existence of a recent mobilization of these elements. 2. STUDY OF CONTROL TRANSPOSITION: Analysis of controlling the expression of retrotransposón Tnt1 of snuff. Tnt1 was the first retrotransposón active plant described (Grandbastien et al., 1989). It is present in hundreds of copies in the genome of snuff and it is expressed in certain stress conditions (Casacuberta and Grandbastien, 1993; Mhiri et al., 1997). Moreover, gene silencing seems to be the main mechanism by which different agencies controlling the proliferation of strange elements transponibles and sequences within their genomes (Hammond et al., 2001; Rudenko et al., 2003). The work presented in this report discusses the control of the expression of retrotransposón Tnt1 at the end of the transcript and polyadenylation of mRNA level and the formation of chromatin, by studying changes in the silencing of Tnt1 conditions stress. We analyze the existence and variation intermediaries silencing, as siRNAs and modification of chromatin. In this work we have designed various strategies to induce transcriptional silencing of transgene expression which is promoted 35S promoter of CaMV. Our results indicate that in terms of stress relaxes the silencing of retrotransposón Tnt1 without being committed in other transgene silencing studied. For this reason it was decided molecularly characterized the silencing of Tnt1. We have identified siRNAs that can be mediating repression of Tnt1 leaf of snuff, cu 8 ando is 4d7 and evidence was not transcribed sequences and their promoters are associated with inactive chromatin. Moreover, as long as knowing how Tnt1 exceeds his silence and expressed in stressful situations, it has analyzed the influence of the region U3 of the LTRs of this retrotransposón (where items are located promoters of their transcript) that process. These results suggest that the region U3 is not sufficient to overcome the silencing of Tnt1 and perhaps other sequences of the LTR or structural factors are those that allow the expression of retrotransposón.

Author: ALCARAZ SEGURA DOMINGO.
Year: 2005.
University: ALMERÍA.
Place of defense: DEPARTAMENTO DE BIOLOGÍA VEGETAL Y ECOLOGÍA, UNIVERSIDAD DE ALMERÍA.
Place of preparation: UNIVERSIDAD DE ALMERÍA.

Summary: The overall change will affect an important role in biodiversity and ecosystem services. The use of functional attributes of the ecosystem on a regional scale in the assessment of the impact of these changes is advantageous in terms of connecting local and global studies and expensive to establish monitoring programs. However, there are many jobs at regional assessing the effects of global change on ecosystem functioning of the Iberian Peninsula or provide a reference situation compared to compare the effect of land use and the climate and atmospheric changes on ecosystems. In this thesis documented the spatial and temporal patterns of the fraction of photosynthetically active radiation (fPAR) intercepted by vegetation present on the Iberian Peninsula, as well as trends in the same for ecosystems protected under the National Parks Spanish Network. Also explored climate controls and land uses underlying behind the patterns found. For this purpose, was used as a measure of operating integrated ecosystem an estimate of fPAR and, therefore, net primary productivity: Green Standard Index (NDVI) obtained from NOAA images / A VHRR between 1982 and 1999. We used the following functional attributes of the seasonal curves of this index: the comprehensive annual variability intraanual or relative range, maximum and minimum values of NDVI, the era in which these values are achieved, and interannual variability of them all . We identified Types Functional Ecosystem collected from that summarizes the spatial heterogeneity of fPAR on the Peninsula and whose patterns suggest that land use change on a regional scale carbon cycles. The rainfall, temperature, vegetation potential and the type of use for which it replaces, controlled the magnitude and direction of differences in the dynamics of NDVI. The characterization of functional attributes in natural areas, and semi-protected Spain revealed the various controls that determine the dynamics of NDVI along environmental gradients between the Mediterranean region and Eurosiberiana. The analysis on national parks showed some holes in the network and allowed to differentiate those types of parks that represent unique ecosystem functioning in the context Iberian tros those landscapes which are representative of the functioning of much of the Iberian vegetation. In addition, our results revealed how the Spanish national parks, areas with high levels of protection, tended to increase productivity and reduce the gap between growing seasons and rest in the short term. Overall, this thesis provides a baseline characterization compared to evaluate the effects of global change on the functioning of ecosystems and the Iberian that incorporate functional aspects of ecosystems to the political nature conservation and maintenance of Ecosystem services, in particular those related to carbon sequestration.

Author: PORRAS MILLAN PABLO.
Year: 2005.
University: CÓRDOBA.
Place of defense: FACULTAD DE VETERINARIA.
Place of preparation: FACULTAD DE VETERINARIA.

Summary: The glutarredoxinas (Grxs) are small proteins (10-12 kDa) that catalyze reactions oxidorredución of thiol groups of proteins and other biomolecules, to perform various functions (maintenance of homeostaiss redox cell catalysis of reactions glutationilación-desglutationilación protein). These processes are particularly important in the behavior of the cell mitochondrial eucariota, which produces the majority of reactive oxygen species (EROs). In this thesis has been studied glutarredoxina 2 (Grx2p) from the yeast Saccharomyces cerevisiae. We have found a new activity for Grx in which it can catalyze the reduction of glutathione by lipoamida reduced compound is located just inside the mitochondria. The Grx2p S. Cerevisiae proved to be particularly efficient in this process, compared to other glutarredoxinas, signaling a possible physiological significance yet to be determined. It has also been found several isoforms of Grx2p, which are split between the cytosol, the endoplasmic reticulum and the outer membrane and the mitochondrial matrix, all from the gene grx2, resulting in two products fde translation differentiated by the presence or not a presecuencia signal to mitocondría and reticle endoplásmatico. By techniques directed mutagenesis studies and significant in vivo and in vitro of Grx2p and its mutant forms, it has been determined that a phenomenon of dual nature (sweep imperfecto-traslocación inefficient), has not been described so far, is responsible for the peculiar subcellular distribution of the products of gene grx2 in S. Cerevisiae. It thus opens an interesting field for future research in which it is determined what role or physiological functions justify the evolutionary selection of this complex mechanism that extends the scope of action of the Grx2p several subcellular localizations.

Author: PIÑEIRO CID ROBERTO.
Year: 2005.
University: SANTIAGO DE COMPOSTELA.
Place of defense: FACULTAD DE MEDICINA, UNIVERSIDAD DE SANTIAGO DE COMPMOSTELA.
Place of preparation: HOSPITAL CLÍNICO UNIVERSITARIO DE SANTIAGO DE COMPOSTELA.

Summary: Growth hormone (GH) and adiponectina hormones are two closely related to cardiovascular function. This relationship is demonstrated by the fact that alterations in the secretion of both, are associated with the development of cardiovascular complications. For this reason, this thesis has focused on studying the effects of hormones on both aspects of the biology of cardiomiocitos such as the viability and metabolism. GH exerts beneficial effects on the heart, and is also a regulator death by apoptosis in various cellular models. For these reasons, the work tried to explore a possible regulatory role of GH on apoptosis in cardiomiocitos. Two models were used cell cardiomiocitos, the primary culture of cardiomiocitos neonatal rat (COP), and the line of mouse HL-1. The results showed the expression of GH receptor mRNA in cardiomiocitos and that the hormone was able to activate it through phosphorylation as well as protein JAK2 and STAT5. Treatment with GH exerted a protective effect on cardiomiocitos compared to apoptosis induced by serum deprivation (COP), or by treatment with cytostatic AraC (HL-1). This effect anti-apoptótico of the hormone was direct and not mediated by the action of IGF-1, because it did not change the levels of mRNA and protein IGF-1 after treatment with growth hormone, or the presence of an antibody anti - IGF-1 reversed protective action of GH. The analysis of this effect showed that the protein phosphatase calcineurin acts as a mediator of the actions anti-apoptóticas of GH in cardiomiocitos, since the combination of two specific inhibitors of this reversed the effect of GH. It was also found that treatment with AraC inducia the activation of p38 MAPK, facts which was partially prevented by growth hormone. These results suggest that GH exerts a protective effect against apoptosis of cardiomiocitos a direct and not mediated by IGF-1, and that this effect is mediated by the action of calcineurin. The adiponectina is a adipocitoquina produced abundantly by adipocitos and is believed to play a decisive role in the relationship between obesity, insulin resistance and cardiovascular risk. This work investigated the synthesis and secretion of adiponectina by cardiomiocitos, as well as potential functional effects on the metabolism of the same. Mobile phones were used as models the two mentioned above, and the primary culture of cardiomiocitos adult humans. The analysis of gene expression showed the expression of mRNA adiponectina by cardiomiocitos. Similarly, we determined the expression of the mRNA of the two receivers adiponectina, AdipoR1 and AdipoR2. The use of specific antibodies revealed adiponectina of protein synthesis and both receivers, and also determine the secretion adiponectina the culture medium by the cardiomiocitos. Addressing cardiomiocitos rat and HL-1 with adiponectina stimulated basal glucose uptake and the already-induced insulin. The hormone was also able to activate the kinase phosphorylation by AMP or AMPK, which could be the mediating effect of adiponectina on glucose uptake, as determined using a specific inhibitor of this (araA). Treatment with adiponectina also stimulated uptake of free fatty acids in cardiomiocitos HL-1, but had no effect on apoptosis induced serodeprivación or on the proliferation of basal cardiomiocitos rat and HL-1 respectively. Moreover, it was shown that adiponectina was able to encourage the accumulation of nitrite in the culture medium by cardiomiocitos rat, ef 8 ecto showed that 505 were mediated by inducible nitric oxide synthase (iNOS) or NOStipo 11. The presence of the inhibitor iNOS aminoguanidina, inhibit the action of adiponectina on the accumulation of nitrites. The stimulatory action of iNOS on adiponectina also found at the level of protein, with an induction of the expression of the synthase. These results demonstrate that cardiomiocitos expressed, synthesize and secrete adiponectina, and that this hormone presents functional effects on the metabolism of cardiomiocitos.

Author: LEIS MARTINEZ HUGO.
Year: 2005.
University: SANTIAGO DE COMPOSTELA.
Place of defense: FACULTAD DE VETERINARIA LUGO.
Place of preparation: FACULTAD DE VETERINARIA.

Summary: NF-kappaB, P13K1Akt AND MO essential molecules are involved in a joint and coordinated way, in controlling various aspects of homeostasis of the keratinocytes, including resistance to apoptosis, proliferation and differentiation epidermal as well as the cutaneous inflammatory process. The IKK complex is a central point in the regulation of NF-kB activity and numerous signals converge on him. The subunit regulatory IKKgamma is essential to the function of IKK complex and its lack of full or partial function correlates with various human patologias which are characterized by defects in one or more primary epithelia. In this paper we have addressed the functional study of IKKg in keratinocytes in culture and in mouse skin in vivo, using the mutant delección IKKg-DN97, which acts as a dominant negative role of NF-kB. IKKg-DN97 leads the decoupling of the complex, resulting in a partial inhibition and dependent doses of inducible NF-kB activity. Moreover produce morphological alterations and increased apoptosis of keratinocytes without stimulating, blocking the degradation IkappaBalpha and tracing nuclear p65 increasing levels of PTEN and reducing the phosphorylation of Akt. We generated transgenic mice expressing IKKg-DN97 in keratinocytes in the epidermis builds them and other stratified epithelia (K5-IKKg-DN97), which show changes in the follicle morphogenesis and cutaneous inflammatory response. By approximations of tumorigenesis in vivo, we have shown that Akt contributes to tumor progression of keratinocytes through partial inhibition of GSK3beta, allowing tracing nuclear CycD1 through various mechanisms that increase the expression and stability of the ciclina. We have also shown that some of the anti-tumor effects of GCs during carcinogenesis epidermal are mediated by the interaction of GR with PI3K/Akl. Such interaction leads to a decrease in activity PI3K/Akt, IKK activity and the activity of NF-kB binding to DNA due at least in part to a decrease in levels of IKKg.

Author: ABELENDA VILA JOSÉ ANTONIO.
Year: 2005.
University: SANTIAGO DE COMPOSTELA.
Place of defense: FACULTAD DE BIOLOXÍA.
Place of preparation: FACULTADE DE BIOLOXÍA.

Author: OTERO ADRÁN MIGUEL.
Year: 2005.
University: SANTIAGO DE COMPOSTELA.
Place of defense: FACULTAD DE MEDICINA. UNIVERSIDAD DE SANTIAGO DE COMPOSTELA.
Place of preparation: HOSPITAL CLÍNICO UNIVERSITARIO DE SANTIAGO DE COMPOSTELA.

Summary: The leptin, a hormone 16KDa discovered in the year 1994, is capable of uniting the energy homeostasis and the immune system by integrating various factors orexigénicos and anorexigénicos well as through its direct action on the thymus and the secretion of cytokines. The leptin is synthesized primarily by the adipose tissue, thus belong to the large group of adipocitoquinas. Structurally, the leptin belongs to the family of cytokines interleukin (IL) -6. The levels of leptin circulating maintain a direct relationship with the mass of body fat. The first function of laleptina was described their action as adipostatina: acting at the central level, leptin increases energy expenditure and minimizing the intake so maintaining optimal energy balance and a proper amount of body fat. In addition to this central role, leptin exerts a large number of peripheral functions, all of which are mediated by a specific receptor (Ob-R), as codified in the gene db, with six different isoforms of which only the long isoform, Ob-Rb , is fully functional. Within the peripheral functions of leptin emphasizes its role as regulator of the immune system: the alterations in circulating levels of leptin involve changes in the immune response due to changes in the function of T cells and changes in the secretion of cytokines, among others . Furthermore, it has been shown that leptin levels are elevated in rheumatoid arthritis, an autoimmune disease systemic involvement involving alterations in homeostasis articular cartilage. It has recently been discovered that chondrocytes, the cells that form the cartílago and maintain homeostasis, synthesize and secrete leptin. This secretion is increased during osteoarthritis, a primary pathology of cartilage degradation, and after administration of exogenous leptin, which suggested a role dírecto and pro-inflamatorio of leptin in articular cartilage. Given the importancía of nitrico oxide (NO) in mediating the response of cytokines in chondrocytes, considering the role of Nitróxido Sintasa (NOS) type 11en production of NO in immune and inflammatory processes, and addressing the ability of leptin to induce the formation of NO in macrophages, might action pro-inflamatoria direct this adipocitoquina in articular cartilage through the NOS type 11. The objectives of this work were:, 1. Studying the effect of leptin, in combination with interferon-gamma and interleukin-1 on the induction of Nitróxido Sintasa type 11en cultured chondrocytes in vitro. 2. To study the molecular mechanisms involved in the transduction path that leads to the activation of the Nitróxído Sintasa type I1inducida synergistically by leptin and interferon-gamma or interleukin-1 in chondrocytes in culture. To achieve this study has used the line condrogénica murine ATDC5, which has allowed us to work with chondrocytes in different states of maturation. They also have been used primary cultures of human chondrocytes. The activity of the type 11se has given us through the accumulation of nitrite in the supernatant culture. For the study of molecules involved in signaling via various inhibitors have been used for JAK-2 (Tyrphostin AG490 and Tkip) PI-3K (Wortmannina and LY294002), MEK-1 (PD098059) and p38K (SB203580). We also study the expression of mRNA and protein NOS type 11mediante RT-PCR and real-time Western Blot respectively. Our results showed that leptin itself is not capable of inducír the NOS type 11en chondrocytes, but in synergy with IL-1 or interferon-gamma, acts as a cytokine pro-in 8 flamator 4b6 ia, implying the induction of the enzyme via a route that involves the signaling kinase JAK-2, PI-3K, MEK-1 and p38 MAPK. In conclusion: 1. The leptin induces, in a synergistic manner with the interferon-gamma and interleukin-1, the expression of NOS type 11en chondrocytes. 2. The synergistic induction of Nitróxido Sintasa type 11en chondrocytes stimulated by leptin, in combination with interleukin-1 yel interferon-gamma, implies a common signaling channel, which involved the kinases PI-3K, MEK-1 and p38 MAPK .

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 travels upward by 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 óbez, 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 marrow. * 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 antidrómicas 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 in the SRD process information dolorsa. However, the cat, these cells projected levels supraespinales more rostrales fundametnalmente 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 promoters associated with motor actions involuntary intending directly to motoneuronas of astra ventral spinal cord, the SRD can participate in reactions associated with motor process 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 designed to immobilize 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: Mariñas Pardo Luis Antonio.
Year: 2005.
University: A CORUÑA.
Place of defense: Facultad de Ciencias.
Place of preparation: Facultad de Ciencias.

Summary: The reproductive system of males and females of the species that are gender Mytilus and Ensis presents no sign of sexual differentiation structural level. This paper deals with the study of sexual dimorphism in these genres, using different molecular markers. The mussels cover species dioicas of this kind with which most studies of all kinds have been made to date. Moreover, the knives are agencies whose scientific interest is growing because of its growing economic potential. The mechanisms that encourage sexual differentiation in both cases still are beginning to be clarified, they Mytilus and Ensis are used in this paper as a starting point for dealing with the little-documented sexual dimorphism of bivalves at the molecular level. The analysis of the behavior of the five paternal mitochondria in the sperm of M. Galloprovincialis shows that these penetrate inside the egg during fertilization. Following the same, remain viable inside the 56.4% of zigotos analyzed, aggregated and divided up without at least the third mitotic division, which can be seen in the micrómero 1d. The latter will result in the formation of the gonad in adult individuals, thus paternal mitochondria are confined to individual cells gonadales male. The polymorphisms generated RAPD showed a high sensitivity in the identification of molecular differences between sexes in M. Galloprovincialis, where the loci called MG-ABA5-750 presented effectiveness rates close to 80%. In E. Arcuatus four of the loci studied (EA-ABA2-500, EA-ABA3-700, EA-ABA7-750 and EA-ABA11-750) had rates close to 70% effectiveness in the presence and absence in males and females. The ISSR markers generated confirmed the potential of the technique to form an alternative to the isolation and characterization of micro-and generate markers especie-específicos. The microsatellite called MG-ISSR20-250, isolated from one of the sequences allows differentiate individuals of the species M. Galloprovincialis of M. Edulis. Another of them identifies individuals of E. Arcuatus and not ever amplified copies of E. Siliqua. Both markers might form a good method for the differentiation of these molecular species. The abundance of tandem repeats of the sequence (GT) n is greater than that of n (CT) in all species studied. In M. Galloprovincialis and E. Arcuatus there is a molecular basis for sexual dimorphism, under which males have a higher abundance of repeats (WP) No than females. Conducting a genoteca subtractive gender M. Galloprovincialis characterize allowed five pieces of messenger RNA present in gonads of adult males and absent in the hermbras, showing the existence of genes with differential expression between the sexes. The amplification with primers designed to amplify DNA sequences belonging to enzyme activity carboxileesterasa yielded a marker that differentiates males and females of M. Galloprovincialis regardless of the stage in the player they are, as amplified in 100% of the males surveyed and never makes it in females. It also allowed characterize specific sequences similar male in E. Arcuatus and E. Siliqua, corroborándose its presence in other species as Donax trunculus and Venerupis rhomboides.

Author: ARIAS ESTEBAN ARMANDO.
Year: 2005.
University: AUTÓNOMA DE MADRID.
Place of defense: SALA RAMON ARECES.
Place of preparation: FACULTAD DE CIENCIAS.

Summary: The populations of RNA viruses are complex distributions genomes highly related but not identical are called cuasiespecies. The distribution of a cuasiespecie viral depends on the continuous emergence of new mutants and competition and selection processes that act on them. The high genetic heterogeneity observed in the cuasiespecie is due to the low fidelity copying by the RNA-polimerasas viral activities and the absence of corrective error during the process replicativo. These distributions mutants give the cuasiespecie a great capacity adaptive front of selective pressures. That is why the populations of RNA viruses behave as an adaptive system, as they are able to respond to changes in the environment. It has been described that some adaptive systems, such as the immune system, have a "memory" of past events for those who have been exposed. The main aim of this is to deepen Doctoral Thesis on the molecular mechanisms involved in the replication of RNA viruses and the implications of that evolutionary process in the dynamics of cuasiespecies viral. We have characterized the presence of genomes "memory" in line C9.22 of the FMD virus (VFA). These genomes presented as a genetic marker insertion oligo-A internal preceding the second AUG codon start of the translation of the viral polyprotein. When C9.22 underwent passes at high multiplicity of infection genomes revertant who were not moved quickly this integration with the genomes insertion. This insertion leave if detactable in the consensus sequence of the population of the pass 20 (C9.22p20). However, the insertion was held in memory genomes of this storyline, as evidenced by the analysis of the peoples of the passes 50 and 150 (C9.22p50 and C9.22p150). The genomes memory increased their biological effectiveness during the passes, despite being displaced by the revertant, according to the hypothesis of Queen Cross. We analyzed the spectrum of mutants of cuasiespecie C9.22p50 by biological cloning and molecular cloning of three different genomic regions. The results have provided detailed information on types and frequency of mutation, as well as the presence of "hotspots" or "hot spots" for genomic alterations in the viral population. This analysis has shown a characterization indistinguishable from the specter of mutants of a cuasispecie through cloning biological and molecular cloning. We have expressed and purified polymerase VFA (3D) and we studied the basic requirements for its enzymatic activity in tests in vitro. The characterization of this enzyme is critical to the study and understanding of the nature erroneous replication VFA. Polymerase 3D purified has enabled the group Dr. Nuria Verdaguer obtain X-ray diffraction structures of 3D single three-dimensional and 3D complex with a molecule that acts as molde-cebador (1,9-Ay 3,0-A resolution, respectively). These structures have identified critical residues involved in the interaction with the RNA molecule that acts as molde-cebador. Have been identified in RNA virus populations subjected to mutagenesis numerous lethal mutations in polymerase 3D genomes present in this population. Some of them, especially the residue mutants Gly 118, causing the lack of enzymatic activity was detectable, suggesting a critical role for this residue in the activity RNA-polimerasa.

Author: AGIRREZABALA COLMENERO XABIER.
Year: 2005.
University: AUTÓNOMA DE MADRID.
Place of defense: CENTRO NACIONAL DE BIOTECNOLOGIA.
Place of preparation: CENTRO NACIONAL DE BIOTECNOLOGIA.

Summary: The bacteriophages DNA double-stranded model systems are suitable for the study of basic biological processes such as molecular interactions that occur during the assembly and viral maturation, or the behavior of molecular motors acting for the packaging of viral DNA. Through the use of electronic cryomicroscopy and processing of individual particles have been determined assembled from bacteriófago T7 produced during his route morfogenéteca. The structure of the precabeza reveals the presence of the protein scaffolding, the kernel and the connector, the latter involved in the packaging of DNA, which are located in one of the vertices pentaméricos. The reconstruction of virión mature shows drastic changes in cápsida, desaparación of protein scaffolding, and drastic structural changes in the kernel, which in this case stands for the cápsida to interact with the tail. Some of these changes would be caused by the pressure exerted by DNA packaging. Moreover, the structure of the connector to 8 A resolution obtained from recombinant particles, has shown the presence of a plea alfa-beta retained their dominance in the distal zone of the structure involved in the packaging of DNA.

Author: RIQUELME GABRIEL CRISTINA.
Year: 2005.
University: AUTÓNOMA DE MADRID.
Place of defense: CENTRO NACIONAL DE BIOTECNOLOGIA.
Place of preparation: FACULTAD DE CIENCIAS.

Summary: Throughout this report describes the generation of viral vectors based on the genome of the coronavirus TGEV in order to answer basic questions about the replication of the virus as well as their use as a system of heterologous gene expression. We have built genomes TGEV with all the genes separated sites restrictions endonucleasa. The separation of genes TGEV meant duplication sequence of extreme 5 'of each gene, which this is a domain sequence associated with the regulation of transcription, which affects the growth of the virus in vivo and its virulence. In this thesis, it provides the first demonstration that a gene 7 of TGEV is a gene is not essential for the viability of TGEV. Furthermore, it demonstrates that the deletion of the gene 7 affects replication TGEV and dilutes its virulence in piglets, its natural host. Likewise, it has been shown that the introduction of one or more changes in the genome of TGEV led to the creation of a collection of recombiantes of TGEV with a variable degree of virulence. Moreover, it has been determined that the E gene is essential to develop a genome TGEV deficient in the gene E genes in non-essential 3a and 3b. For the generation of virions in the defective gene E, it has been supplemented in trans by packing cells expressing the protein E. In the absence of the protein E blocked the formation of mature virions during morphogenesis virus obtained virions immature not infective that accumulate in the ERGIC. Also, vectors were generated tropic addressed to the canine species by replacing the globular domain of the S protein of TGEV by the canine coronavirus surpassing the first barrier barring infection, the entry of the virus. We have developed vectors tropic canine competent replication and spread. These skills would enable the generation of vectors tropic and virulence default, some caracterísitcas which are essential for the development of systems that allow expression using these vectors to obtain safe recombinant vaccines and gene therapy. They are written expression systems for specific species swine and canine with which they have expressed interest heterologous antigens to immunize these species in order to determine the degree of protection from frequent infections of the same.

Author: MOLNAR MURO CRISTINA.
Year: 2005.
University: AUTÓNOMA DE MADRID.
Place of defense: CENTRO DE BIOLOGIA MOLECULAR SEVERO OCHOA.
Place of preparation: CENTRO DE BIOLOGIA MOLECULAR SEVERO OCHOA.

Summary: The formation of the veins in the Drosophila wing requires the activity of the Notch signaling pathways, EGFR and Dpp. To identify new members of these routes have done a mutagenesis gain function and we have selected some 13,000 new insertions P-UAS. In combination with a line Gal4 specific veins have selected and mapped 500 insertions P-UAS identified approximately 300 genes that when sobre in the veins pupales affect the formation of veins, cell differentiation or morphology wing. We have identified in this mutagenesis for 70% of the known members of the routes Notch, EGFR and Dpp. In this work I present the results of mutagenesis and analysis of three of the selected lines, affecting genes Moesina, LanB1 and CG9056. Moesina belongs to the family MRA, a group of proteins that bind citoplasmáticas the cytoskeleton of transmembrane proteins actin sub-cortical to specific. We have created a new allele of a lack of role moe (moeC858) and we studied their phenotypes in the adult wing, finding two major flaws: 1) tail wing, defects in the armpit and presence of vesicles as differentiated tissue between the wing two superdicies dorsal and ventral wing, and 2) changes in the pattern of the vein L3. The analysis of the relevant disks imaginales reveal profound alterations in the morphology of the wing imaginal disc, as well as changes in the expression of several genes target in the path of Hedgehog signaling. The activity of other signaling pathways such as Notch, Dpp, Wg and EGFR is unchanged with disc wing moeC858. We suggest that Moe has a new requirement on signaling hh which is independent of its role in epithelial morphogenesis. The second analyzed gene encodes one of the three subunits of Laminina: LanB1. Laminina is a component of the basement membrane. The overexpression of lanB1 cause defects in the pattern of veins similar to those observed when the actividaad of Notch route is reduced. We have characterized the phenotypes gain function LanB1 and have prompted two alleles of a lack of funciín of this gene. The lack of LanB1 affects adherence between the surface of the dorsal and ventral wing. Finally, we analyzed the gene CG9056, whose overexpression causes lack of differentiation veins. We analyzed the function of lack of CG9056, finding that this gene is required to prevent formción vein antagonizando routes EGFR signaling and / or Dpp.