LIFE SCIENCES (7)

Author: LÓPEZ MOLINA DOROTEA.
Year: 2005.
University: MURCIA [www.um.es].
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA.

Summary: The canning of the artichoke originates a large amount of agricultural or industrial waste, composed mainly of leaves, stem and the outside of the flower (bracts) from the artichoke, unfit for human consumption. For example, 70% by weight of the flower of the artichoke is waste not intended for human consumption. However, such wastes are used for animal feed and are intended for the production of forage silage. This thesis deals with the search for new uses for such wastes to develop and characterize new products. The extracts of artichoke include enzymes with polyphenol oxidase and peroxidase activity, and can be obtained from waste (stems, leaves and bracts) through homogenization of such wastes in an aqueous medium, and can be used to decontaminate means liquids, solids and semi contaminated phenols, aromatic amines, organic halides and / or heavy metals, by treatment with peroxide in the presence of that artichoke extract. Another byproduct characterized in this thesis is a peroxidase (AKPC). Enzymes with peroxidase activity (EC 1.11.1.7) are óxidorreductasas that are widely distributed throughout the phylogenetic scale and that catalyze the oxidation of a wide range of organic and inorganic substrates, using power oxidant hydrogen peroxide. In addition to their academic interest and physiological these enzymes are widely used in clinical laboratories and industry. It has been characterized peroxidase isozyme of the bract of artichoke and some of their applications have been discussed. The final product obtained and analyzed in this report is the inulin. The insulin is being included today in numerous human and animal food products for its positive effect as prebiótico, stimulating the growth of intestinal flora non-pathogenic. It has found application in nutraceuticos and dietetics, dairy products, pet food, and to a lesser extent in animal production.

Author: ARNAN VIADIU XAVIER.
Year: 2005.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: FACULTAD DE CIENCIAS..
Place of preparation: UNIDAD DE ECOLOGÍA. UNIVERSIDAD AUTÓNOMA DE BARCELONA.

Author: Cuñé Castellana Jordi.
Year: 2005.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: Facultad de Ciencias de la U.A.B..
Place of preparation: Facultad de Ciencias.

Summary: The SOS system is a network multigénica controlled negative repressor LexA, vital for cell survival, as their desrepresión occurs when the cell damage introduced in the genetic material that can be transmitted to the offspring. This fact, coupled with its broad distribution in the domain Bacteria, present us with the knowledge of their operation in different groups, as a tool phylogenetic ideal. To that end, and as the unifying theme of this work was to study regulón LexA to Dehalococcoides ethenogenes, not green sulfur bacteria; Magnetococcus sp. Strain MC-1, a bacterium magnetotáctica and Leptospira interrogans serovar Lai, the first espiroqueta with a gene lexA identified. To that end, we proceeded to the cloning of the gene lexA of each and the purification of its protein product. In the first, the box was defined union of LexA (AGAACN (4) GTTCT), checking from it, their links with the Gram-positive bacteria, as well as non-regulated gene recA, regarded as canonical in the system SOS. In Magnetococcus sp. Strain MC-1, the box described SOS was CCTN (10) AGG. claralmente independent. As concerns Leptospira interrogans serovar Lai, the result was surprising, since the SOS box of this bacterium, it had to be identified in the promoter region of recA, because it is not present in the lexA, going well, the first example where LexA not autoregula his speech. This, as well as being unable to detect any gene under their control, we show the gender Leptospira, as an intermediate step in the evolutionary trend followed by spirochetes of losing their gene lexA.

Author: Ghirardi Juan José.
Year: 2005.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: Facultat de veterinaria.
Place of preparation: Facultad de Veterinaria.

Summary: This thesis focuses on the development of bowling for electronic identification (FDI) in sheep and cattle, in the estimation of a law to predict biological retention and finally in evaluating their use for the implementation of a traceability system based on IDE and DNA in lambs and calves.

Author: LOPEZ VIADO MARIA DEL CARMEN.
Year: 2005.
University: OVIEDO [www.uniovi.es].
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: UNIVERSIDAD DE OVIEDO.

Summary: One of the main interests in developmental biology today is to clarify the processes that are taking place, so that the cells initially indiferenciadas organized in the space and time to form complex organs. Our initial objective has been to identify new genes involved in the development of the limb, because the tip is a very classic model employee. In this paper we have cloned from a genoteca of cDNA chicken, cDNA corresponding to ps20 chicken (cps20). Proteins ps20 belong to the superfamily of proteins WAP. This includes superfamily proteins that have at least one domain WAP. The WAP domain is a domain that showed little structural homology in the peptide sequence between different proteins, except in 8 residues that are highly conserved cysteine. Between 8 cisteínas formed 4 bridges disulfide, and the 4 bridges are responsible for the domain disulfide WAP. Although they have no role, some domains WAP have inhibitory function of serinproteasas. The study of the pattern of expression of cps20 showed that expressed in a dynamic way on the tip of the chicken embryo, adopting a pattern posterodistal. This pattern was so interesting envisage a role during development, however overexpression of cps20 in chicken embryo using retroviral RCAS not vectors result in any alteration in the formation of the tip, obtaining extremities completely normal. The study of cells in culture resulted in an inhibition of cell growth that sobreexpresaban this protein. Also considered for cps20 the inhibitory function of serinproteasas which had been described in other proteins WAP. This was used recombinant protein cps20his and explored the role of trypsin inhibitor. It was obtained that cps20 was capable of inhibiting the proteolytic activity of trypsin. This feature is consistent with the previously described because it has been found that different protease inhibitors such as Pefabloc inhibit cell growth. Lastly was analyzed by inmunoflluorescencia tracing cellular cps20. This at first were obtained polyclonal antibodies in the lab that were purified by affinity to ensure its specificity. It was noted that, in different cell lines transfected with cps20, it was located in the plasma membrane. So has identified a new gene implicated in the development of limb, which through its inhibitory function could be regulating proliferation processes that occur during development. Furthermore, since several proteins involved in the development of the limb are activated by proteolísis, cps20 might be involved in controlling some of these pathways of activation.

Author: DUQUE ALVAREZ PALOMA.
Year: 2005.
University: OVIEDO [www.uniovi.es].
Place of defense: FACULTAD DE BIOLOGIA.
Place of preparation: UNIVERSIDAD DE OVIEDO.

Summary: This paper has been developed to improve the methods of production of bovine embryos in vitro. We have managed to identify factors likely to improve both the quality and the development of the embryo, thus contributing to the achievement of a system to produce embryos chemically defined features and free from substances of animal origin. The thesis is divided into four work papers published in journals in the area of Reproductive Biology ISI Journal Citation Reports Of: Effects of acetoacetate on in vitro development of bovine embryos in medium containing citrate and myo-inositol. Gomez E, Duke P, E and Diaz Diez C. Reprod Domest Anim 2001 Aug; 36 (3-4): 189-94. Effects of acetoacetate and D-beta-hydroxybutyrate on bovine embryo development in vitro in serum-free medium. Theriogenology 2002 Mar 15, 57 (5): 1551-62. Enhancement of development capacity of meiotically inhibited bovine oocytes by retinoic acid. Duke P, Diez C, L Royo, Lorenzo PL, Carneiro G Hidalgo P, N and Facal E. Gomez Hum Reprod 2002 Oct 17 (10): 2706-14. Use of two replacements of serum during bovine embryo cultire in vitro. Duke P, Gomez E, E Diaz, Hidalgo CO and Diez C. Theriogenology 2003 Feb, 59 (3-4): 889-99.

Author: Espadaler Mazo Jordi.
Year: 2005.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: Facultad de ciencias.
Place of preparation: Universidad Autonoma de Barcelona.

Summary: A genome alone can't make a person, because we are also influenced by where we live, what do we eat, and hundreds of other aspects of our environment. But you can't make a person without a genome. The Human Genome Project was just the first step in understanding humans at the molecular level. The words of Winston Churchill, spoken in 1942 after three years of war, capture well the post Human Genome Project era: "Now this is not the end. It is not even the beginning of the end. But it is, perhaps, the end of the beginning". Understanding the function of genes and their products - proteins - and other parts of the genome is known as functional genomics. Functional genomics relies extensively on the use of computers to store, organize and analyze data. Moreover, computers can be used to predict the structure or the function of a novel protein, using knowledge obtained from other proteins. This new research field is usually referred to as bioinformatics.

Author: RUIZ GARZÓN MARÍA AMPARO.
Year: 2005.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: DEPT. BIOQUÍMICA I BIOLOGIA MOLECULAR.
Place of preparation: FACULTAT DE VETERINÀRIA.

Summary: The phosphorylation / desfosforilación protein is the primary mechanism for regulating post-presenting all eukaryotic cells. The state of phosphorylation of a specific protein depends on the balance between the activities protein kinase and protein phosphatase acting upon it. In turn, these protein kinases and phosphatases are heavily regulated, making the phosphorylation mechanism in a complex and highly controlled. The fact that eukaryotic cells have a considerably larger number of protein kinases that protein phosphatases implies that these phosphatases are involved in many cellular processes and thus presenting often more complex regulatory mechanisms, such as the existence of regulatory subunits can adjust its enzymatic activity, or intracellular localization ability to interact with other proteins. In this regard, the yeast Saccharomyces cerevisiae represents a very useful tool for studying the process of phosphorylation and its regulation because, although it is a body eucariota is unicellular. Moreover, this yeast has advantages over others because it knows its chromosome sequence some years now and have a range of tools that make it suitable for basic research of any cellular process eucariota. The protein phosphatases Ppz1 and Ppz2, desfosforilan waste serine / threonine other proteins are structurally related to phosphatase PP1. In Saccharomyces cerevisiae this is a single phosphatase catalytic subunit (Glc7), which is essential, and a variety of regulatory subunits that confer the specific function. Unlike Glc7, which is highly conserved evolutionarily, phosphatases Ppz only found in fungi, and at the time of initiating this work was only described a regulatory subunit which modulates its phosphatase activity in all processes in which they being involved: protein Hal3. The phosphatases Ppz involved in regulating functions such as maintaining the integrity of cells, the transition G1 / S cell cycle, homeostasis saline and fidelity traduccional. Although little was known of the elements through which they exercise these functions during the conduct of this paper described the existence of a relationship between phosphatases Ppz and transporters potassium Trk1 and Trk2. The defosforilación of these transporters per share of phosphatases Ppz would result in a lower corner of potassium inside cells, which can explain in large measure the functions they perform these phosphatases. In this paper we present new targets of biological phosphatases Ppz, such as signaling via calcium / calcineurin and the transport system potassium low affinity. Analysis of the salt tolerance of cells lacking Ppz1 and Ppz2 shows that these cells are tolerant lithium and sodium because they have an increased expression of the ATPase Ena1, the main system detoxificador of yeast S. Cerevisiae. This increase in the expression of ENA1 is independent of the transport system potassium high affinity (Trks), but totally dependent on the path of the phosphatase calcineurin signaling. Our results suggest that phosphatases Ppz, and specifically Ppz1, negatively regulates calcium transporters to the plasma membrane, altering calcium homeostasis and thus inhibiting the activity of the phosphatase calcineurin. On the other hand, analysis of the requirements of potassium in cells that lack of transporters potassium Trk1 and Trk2 and phosphatases Ppz1 and Ppz2, has allowed us to identify these phosphatases as positive regulators of the transport system potassium low affinity, called NSC1. Moreover, we describe a new regulatory subunit refusal phosphatases Ppz, protein Vhs3. Unlike phosphatases Ppz1 and Ppz2, which are unique to fungi, Hal3 and Vhs3 presented counterparts in higher eukaryotes. In both yeast proteins act by inhibiting the activid 8 Ad-Pp 96d z1 and Ppz2 in all its functions described, although Hal3 appears to be more efficient than Vhs3. They also demonstrate the involvement of proteins Hal3 and Vhs3 in the cellular process of flocculation. Hal3 and Vhs3 would govern in a negative way towards signaling the PKA, which regulates the transcription factor Flo8 and expression of floculina Flo11, so that a double mutant conditional hal3 vhs3 presents a phenotype Flocculation accompanied by an increase in expressing FLO11. This effect appears to be dependent phosphatases Ppz and could be explained by the negative control exerted on these transporters potassium. Furthermore, we demonstrate the existence of a new role for proteins Hal3 and Vhs3 that does not involve phosphatases Ppz, since the double mutant hal3 vhs3 is lethal and this is not due to an excess of activity phosphatase Ppz. For this new feature is important residue His90 described in AtHal3a Arabidopsis thaliana as essential to catalyze the in vitro descarboxilación of 4'-fosfopantotenoilcisteína to fosfopanteteína, a key step in the synthesis of coenzyme A. Although Hal3 and Vhs3 conserved histidine residue, not conserved residue equivalent to Cys175, also necessary in eukaryotes and eubacterias to take place this reaction in vitro. Therefore, it is not clear that this new feature is described participate in the synthesis of coenzyme A. Even so, the lethality of the double mutant hal3 vhs3 is suppressed by expression in yeast counterparts Hal3 of higher eukaryotes, such as plant, mouse and human, so whatever its nature, this essential function is preserved along evolution.

Author: RUIZ GARZÓN MARÍA AMPARO.
Year: 2005.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: DEPT. BIOQUÍMICA I BIOLOGIA MOLECULAR.
Place of preparation: FACULTAT DE VETERINÀRIA.

Summary: The phosphorylation / desfosforilación protein is the primary mechanism for regulating post-presenting all eukaryotic cells. The state of phosphorylation of a specific protein depends on the balance between the activities protein kinase and protein phosphatase acting upon it. In turn, these protein kinases and phosphatases are heavily regulated, making the phosphorylation mechanism in a complex and highly controlled. The fact that eukaryotic cells have a considerably larger number of protein kinases that protein phosphatases implies that these phosphatases are involved in many cellular processes and thus presenting often more complex regulatory mechanisms, such as the existence of regulatory subunits can adjust its enzymatic activity, or intracellular localization ability to interact with other proteins. In this regard, the yeast Saccharomyces cerevisiae represents a very useful tool for studying the process of phosphorylation and its regulation because, although it is a body eucariota is unicellular. Moreover, this yeast has advantages over others because it knows its chromosome sequence some years now and have a range of tools that make it suitable for basic research of any cellular process eucariota. The protein phosphatases Ppz1 and Ppz2, desfosforilan waste serine / threonine other proteins are structurally related to phosphatase PP1. In Saccharomyces cerevisiae this is a single phosphatase catalytic subunit (Glc7), which is essential, and a variety of regulatory subunits that confer the specific function. Unlike Glc7, which is highly conserved evolutionarily, phosphatases Ppz only found in fungi, and at the time of initiating this work was only described a regulatory subunit which modulates its phosphatase activity in all processes in which they being involved: protein Hal3. The phosphatases Ppz involved in regulating functions such as maintaining the integrity of cells, the transition G1 / S cell cycle, homeostasis saline and fidelity traduccional. Although little was known of the elements through which they exercise these functions during the conduct of this paper described the existence of a relationship between phosphatases Ppz and transporters potassium Trk1 and Trk2. The defosforilación of these transporters per share of phosphatases Ppz would result in a lower corner of potassium inside cells, which can explain in large measure the functions they perform these phosphatases. In this paper we present new targets of biological phosphatases Ppz, such as signaling via calcium / calcineurin and the transport system potassium low affinity. Analysis of the salt tolerance of cells lacking Ppz1 and Ppz2 shows that these cells are tolerant lithium and sodium because they have an increased expression of the ATPase Ena1, the main system detoxificador of yeast S. Cerevisiae. This increase in the expression of ENA1 is independent of the transport system potassium high affinity (Trks), but totally dependent on the path of the phosphatase calcineurin signaling. Our results suggest that phosphatases Ppz, and specifically Ppz1, negatively regulates calcium transporters to the plasma membrane, altering calcium homeostasis and thus inhibiting the activity of the phosphatase calcineurin. On the other hand, analysis of the requirements of potassium in cells that lack of transporters potassium Trk1 and Trk2 and phosphatases Ppz1 and Ppz2, has allowed us to identify these phosphatases as positive regulators of the transport system potassium low affinity, called NSC1. Moreover, we describe a new regulatory subunit refusal phosphatases Ppz, protein Vhs3. Unlike phosphatases Ppz1 and Ppz2, which are unique to fungi, Hal3 and Vhs3 presented counterparts in higher eukaryotes. In both yeast proteins act by inhibiting the activi 8 dad P 96e pz1 and Ppz2 in all its functions described, although Hal3 appears to be more efficient than Vhs3. They also demonstrate the involvement of proteins Hal3 and Vhs3 in the cellular process of flocculation. Hal3 and Vhs3 would govern in a negative way towards signaling the PKA, which regulates the transcription factor Flo8 and expression of floculina Flo11, so that a double mutant conditional hal3 vhs3 presents a phenotype Flocculation accompanied by an increase in expressing FLO11. This effect appears to be dependent phosphatases Ppz and could be explained by the negative control exerted on these transporters potassium. Furthermore, we demonstrate the existence of a new role for proteins Hal3 and Vhs3 that does not involve phosphatases Ppz, since the double mutant hal3 vhs3 is lethal and this is not due to an excess of activity phosphatase Ppz. For this new feature is important residue His90 described in AtHal3a Arabidopsis thaliana as essential to catalyze the in vitro descarboxilación of 4'-fosfopantotenoilcisteína to fosfopanteteína, a key step in the synthesis of coenzyme A. Although Hal3 and Vhs3 conserved histidine residue, not conserved residue equivalent to Cys175, also necessary in eukaryotes and eubacterias to take place this reaction in vitro. Therefore, it is not clear that this new feature is described participate in the synthesis of coenzyme A. Even so, the lethality of the double mutant hal3 vhs3 is suppressed by expression in yeast counterparts Hal3 of higher eukaryotes, such as plant, mouse and human, so whatever its nature, this essential function is preserved along evolution.

Author: TOMAS ZAPICO CRISTINA.
Year: 2005.
University: OVIEDO [www.uniovi.es].
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA. DPTO.BIOQUMICA Y BIOLOGA MOLECULAR.

Summary: The Harderian gland of the Syrian hamster (Mesocricetus auratus) as a main feature shows a high metabolísmo porfirinogénico, even more than the liver. The low activity of the last enzyme in the path porfiriogénica, ferfroquelatasa, leads to the accumulation of porphyrins, mainly protoporphyrin IX, the Harderian gland in females. Due to the location of this blándula, these metabolites are exposed to light, generating reactive oxygen species. Therefore, the Harderian gland presents a physiological oxidative stress, with a large number of signs of degeneration, but that the integrity of the gland is compromised. The occurrence of abnormal morphology in this gland has been described in the past, but had never done a interpertación of them so far. Studies in this Doctoral Thesis part of the analysis of the physiological factors that mark the metabolism characteristic of the gland Hader, that is the main enzyme porfirinogénicos, linking both the antioxidant enzyme glandular system, with the cellular damage caused by species reactive oxygen, as with the presence of molécdula antioxidant melatonin, which also plays other roles in the gland Hader as modular certain processes both morphological and physiological. The results of these studies led to the description of processes autofágicos as the first barrier against high metabolism porfirinogénico, observed in both sexes. This mechanism is concerned, in the Harderian gland, as a system of constant renewal suitable to continue with the normal glandular activity. Besides, there is a second mechanism, consisting of invasive processes to the connective tissue, and may even reach blood vessels resulting in a process of intravasation material glandular damaged. This effect is the result of an environment with high oxidative stress, which is seen primarily in females, recalling tumor progression. Both mechanisms, autofagia and invasive procedures, should be involved in maintaining the intgridad cell.

Author: Fernandez Tajes Juan.
Year: 2005.
University: A CORUÑA [www.udc.es].
Place of defense: Facultad de Ciencias.
Place of preparation: Facultad de Ciencias.

Summary: The knives are a group of molluscs bivalbos grouping species of the genus Ensis and Solen. Recently there have initiated a number of studies on the biology and ecology in order to improve the production and exploitation of marine resources of large national and commercial interest galician. So far, genetic studies are virtually non-existent so in this work have been raised doctoral thesis the following objectives; Analyze the cytogenetic characteristics of Ensis arcuatus, E. Siliqua and Solen marginatus, carrying out the molecular analysis of ribosomal genes nuclear rDNA 18S-5, 8S-28S and rDNA 5S in different European populations of knives and establish the genetic variation and population structure in E.arcuaturs, E. Siliqua and E. Directurs of Galicia, Portugal, Ireland and the Netherlands through molecular markers of nuclear anonymous regions (RAPDs). Cytogenetic characterization has revealed that the three species analyzed presented a budget diploid chromosome of 2n = 38. The species E. Arcuatur and E. Siliqua accessories B chromosomes present in varying numbers as both intra interindividual. In the species E. Arcuatus and E. Siliqua have found a large number of genomic regions rich in GC. It is located 13 regions CA3 in E. Siliqua, nine in E. Arcuatus and two in S. Marginatus. Different regions Ag-NORs and position of the gene ribosomal mendiante FISH. The molecular characterization of the gene ribosomal 5S has revealed that all species have a size of 120 bp coding region, the region is not being transcribed variable size. We must highlight the existence of at least three types of NTS, the type I characteristic of E. Arcuatus and E. Siliqua, NTS Type II characteristic of E. Directus and NTS Type III characteristic of S. Marginatus. Besides the species E. Directus showed other NTS characterized by the insertion into the interior of a microsatellite compound. The ITS region showed some divergence of values very similar between the two species and the region ITS2 the most divergent. The phylogenetic analysis allowed to separate into two distinct clusters clones of E. Arcuatus of E. Siliqua. For the analysis of the regions anonymous DNA in different populations of E. Arcuatus, E. Siliqua and E. Directus have selected a total of 62 loci generated from five primers, using these loci RAPDs has been observed that the species E. Directus is shown far more respect to other species and populations studied. Refieriéndose only to the populations of E. Siliqua populations Galicia showed less differentiation reflecting the existence of a structure panmictica, with respect to other populations of E. Siliqua data supports that there is an influence of the geographic distance on individual stocks showing the most far greater differentiation. In the case of the people of Strangford Lough analysis shows the major components separated from the rest of populations.

Author: Fernandez Moreno Mercedes.
Year: 2005.
University: A CORUÑA [www.udc.es].
Place of defense: Facultad de Ciencias.
Place of preparation: Facultad de Ciencias.

Summary: The ADNmt is a useful genetic marker studies genético-poblacionales and in the development of filogenias. In the case of different species of mollusks (both belonging to the subclass Heterodonta as Pteriomorphia) amplification patterns were generated from -420 bp for the gene ARNr 12S and -710 bp for the IOC. For the gene ARNr 16S consisted of a fragment -550 bp (I band) and one -610 bp (Banda h). The nucleotide composition of the three genes revealed a higher content in AT (-60%) than in GC. The values of genetic distances are generally higher among species subclass Heterodonta that among the subclass Pteriomorphia. The values for the ratio Ts / TV close to 10 were obtained from gene ARNr 12S. In the case of ARNr 16S, these values were =, 756 to 8168 between species of the genus Ensis, and less than 1 in the case of clones of E. Banda h Siliqua and E. Arcuatus. Among other species analyzed value was -1. The values obtained from gene IOC were below or close to 1 for all species. These results indicate that, in general, the sequences analyzed transitions are saturated or close to saturation, and that levels of genetic divergence between species are quite high in the case of genes ARNr 16S and IOC. The phylogenetic analysis made in the three mitochondrial genes show the grouping in a cluster of species belonging to the families Pharidae and Solenidae, reflecting on the same two distinct groups, one comprising species of the genus Ensis and another to S. Marginatus. However, in the case of ARNr 16S grouping gender Ensis varies. Thus, individuals with pattern amplification and single band clones band 1 appear in a cluster near the species subclass Heterodonta, while clones Banda h do with another species belonging to the subclass Pteriomorphia. In the case of the species studied pectinidae phylogenetic analysis indicates a possible polyphyletic origin for this group. Moreover, the order shows a possible Veneroida polyphyletic origin in the light of trees generated from the three genes. The values of D, D and F * * obtained indicate that the three regions analyzed still neutralista theory. However, clones of E. Siliqua band h seem to be under the action of purifying selection, or have recently experienced a bottleneck that has eliminated the variation. The heteroplasmia detected from the analysis of gene RNA 16S could be due to the existence of three types of mitochondrial chromosomes: chromosomes with the gene ARNr 16S type 1, with the chromosome gene ARNr 16S type hy chromosomes with both. Thus, the mitochondria may be formed by any of the three types of chromosomes or mixture of them. This heteroplasmia could be generated and maintained or because the process of eliminating lasd mitochondria male was inefficient, by the existence in itself of great intraindividual variability and / or the presence of homologous recombination mechanisms homologous or not. The analysis of the patterns of restriction generated from gene ARNr 16S allow establish 50 haplotypes different and shows that a higher polymorphism in the Galician towns. It did not deliver haplotypes specific kind but if locality. The distribution of haplotypes in different localities Galician suggests that they are not genetically isolated. However, the towns of Galicia and the Irish if 8 are. E 500 l low polymorphism detected in the towns Irish and Portuguese may be the result of the geographical features of those areas. Moreover, we must also consider the fact the downsizing of natural populations caused, in the case of Portugal, by overfishing, which could cause a bottleneck transitional affect mitochondrial diversity. The results of the AMOVAs reveal that most of the genetic variation is distributed within the localities (intraindividual variation) rather than between groups.

Author: Acerete Rodríguez Laura.
Year: 2005.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: Univ.Autónoma de Barcelona.
Place of preparation: Universidad Autónoma de Barcelona.

Summary: The cortisol is the primary glucocorticoid in fish teleósteos, assuming also functions mineralocorticoideas. This is the main indicator of the response to stress, is involved in various metabolic pathways, is immunosuppressive and anti-inflammatory and is the main hormone osmorreguladora in fish. In the liver, glucocorticoids increase transcription of genes involved in gluconeogenesis, in the catabolism of amino acids and the acute phase response. The secretion of cortisol in fish is regulated by the shaft hipotalámico-pituitario-interrenal (HPI), equivalent to the axis hipotalámico-pituitario-adrenal (HPA) in mammals. Hormones corticosteroideas in mammals, acting through specific intracellular receptor, mineralocorticoid receptor (MR) and glucocorticoid receptor (GR), which act as transcription factors dependent ligand. Lately it has been shown that some species of fish express a MR and / or more than one MO. In this paper we have cloned, for the first time, the golden MO (Sparus aurata). The coding region is translated into a protein of 784 amino acids and shows high homology with the DNA binding domains and the hormone, other glucocorticoid receptors, including those of mammals. The expression in the golden MO is ubiquitous, as has been detected in all tissues studied: heart, liver, spleen, kidney and back, intestines, gonad, adipose tissue, branquia, brain and muscle. The overall response to stress requires interaction neuro-inmuno-endocrina through communications paracrinas bidirectional. This communication is important for maintaining homeostasis in various stressful conditions, including endotoxemia. The bacteria endotoxin of gram-negative bacteria or LPS, in vertebrates, triggers an immune reaction complex comprises the production of proinflammatory cytokines by macrophages, mainly TNFa (tumor necrosis factor alpha) and IL1b (interleukin 1 beta). The cytokines produced in response to a statement of LPS are also involved in the activation of the HPA axis and activate, therefore, the release of cortisol. Our results show that the administration of LPS stimulated saw in the shaft HPI in the golden triggering the release of cortisol, following a pattern of acute response. This hormone modulates the inflammatory response by inhibiting the production of cytokines induced by LPS. Our study shows that this regulation could be given through the CR because it interferes with the transcription factors responsible for inducing the transcription of genes involved in the inflammatory response. The expression of the GR in the golden after an injection of LPS is regulated in a specific manner depending on the time and the fabric. Overall, we see that there is an inverse correlation between plasma cortisol levels and the levels of expression of the GR in the tissues. The intraperitoneal injection of LPS also increases the expression of other genes immune (TNFa, IL1b, catepsinaD, Mx protein, gamma PPAR) in the major tissues of the immune gold, specifically in previous kidney, spleen, intestines and gills earlier. Changes in the expression of these genes is different depending on the fabric and time. In the primary culture of hepatocytes of golden, low doses of immune stimulation (TNFa recombinant), or endocrine (cortisol) modify the differential expression of genes immune (TNFa, IL1b) and endocrine (MO, catD), respectively. At higher doses of cortisol decreases the expression of GR and TNFa 12 hours after treatment. Changes in the expression of both genes follow a pattern with these temporary because high doses of cortisol, increases the expression of GR and decreases of TNFa 3 hours after treatment. This response shows that in fish, the regulation of the inflammatory response may also be made travé 8 s of SW 3cd. Finally, the European perch (Perca fluviatilis) subjected to stressors typical aquaculture practices (shipping and handling) shows the physiological response primary release of cortisol, but the low level of activity in the responses secondary release of glucose and lactate.

Author: Meghelli Nacima.
Year: 2005.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: Facultad de ciencias.
Place of preparation: Unidad de Ecología. Universidad Autónoma de Barcelona.

Summary: This thesis aims to assess different methodologies for optimizing production viverística and the establishment in different fields Mediterranean forest species with different morphological features. During the establishment phase field seedlings may experience significant periods of stress, especially water, which may limit their survival and development. For this reason, quality morfo-fisiológica of plants is a factor that plays a vital role in the success of a reforestation project. This grade represents a compendium of different features designed to obtain a higher strength material once it is in the bush, so as to promote the reduction of stress suffered by the handling, improved the ability of attachment, and their optimum growth, therefore, ensuring the repopulation of the area in a long time. Most of the research on improving the quality of plant nursery (increase in temperature, application atmospheres enriched CO2 different levels of water stress, the use of different substrates) have especially focused on the intensive production of horticultural species, where there is very little information in its application in the case of forest species. In this context, this theory provides new insights into the use of different treatments in pre nursery on 4 species of Mediterranean forest P. Nigra, P. Pinaster, Q. Ilex and Q. Humilis. This thesis also provides news on the use of some techniques for improving the establishment of seedlings in the field, such as the use of sludge drying heat of purification, waste generated in large quantities, which are reusable a priori to improve the physical characteristics chemical degraded soils, but which has very little information. In this thesis project envisages the research conducted all key processes with respect to the use of plant material for the restoration of degraded areas, from production in nursery until the establishment in the field.

Author: Oliva Paterna Francisco José.
Year: 2005.
University: MURCIA [www.um.es].
Place of defense: Facultad de Biología.
Place of preparation: Facultad de Biología (Universidad de Murcia).

Summary: The purpose was to establish the scientific basis for the recovery and conservation of Aphanius iberus in the region. In the context of the axioms of Conservation Biology, the main results have been obtained: 1) Identification and quantification of the distribution of species in the study area, with the establishment of units Ecogeográficas management. In turn, provides a list of potential habitats for rebuilding stocks of the species in the region. 2) It presents a first approach to the genetic variability of the species in the region and the establishment of Operating Units Conservation of the species. 3) It has analyzed its strategy of living in a coastal wetland with saline, with the aim of improving the management focused on the kind of these systems. 4) have been characterized types of local populations of the species present in the Mar Menor and its environment and proposes a model of dynamics and structure metapoblacional. 5) It establishes the status of conservation of the species in the study area with the implementation of IUCN criteria applied at the regional level. In order to submit a recommendation from the academic context for the management of the species: 6) gather experience and sets out recommendations for the management of the species. 7) It produces a document with guidelines to be followed in the proceedings on the species until it is developed its recovery plan.

Author: GALLEGO DEL SOL FRANCISCA.
Year: 2005.
University: VALENCIA [www.uv.es].
Place of defense: FACULTAD DE MEDICINA DE LA UNIVERSITAT DE VALÈNCIA.
Place of preparation: INSTITUTO DE BIOMEDICINA DE VALENCIA.

Summary: The lectin derived proteins are not immune, which specifically recognize carbohydrates but exerted no enzymatic action on them. They are ubiquitous in organizations throughout the evolutionary scale, being involved in many biological processes, both physiological and pathological. In addition to being essential for the normal functioning of living organisms, are important tools in biotechnology. The aim of the thesis was the study of plant lectin belonging to the legume family. On the one hand, it has been resolved and the first primary structure of a crystalline lectina of the subfamily of Mimosoideas. The lectina of Parkia platycephaia presents three domains of type jacalina arranged in tandem. Each subunit has a site joined by sugar with glucose and manosa. This lectina form dimeros regardless of pH. Moreover has deepened in the study of the structural foundations of equillibrio dimero-tetrámero in lectin gender Diocleinae. To this have been solved crystal structures lectin seed Dioclea guianensis. Dioclea violacea Cratylia floribunda and to be able to compare between them.

Author: Porras Pardo Mònica.
Year: 2005.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: Facultad de Veterinaria.
Place of preparation: Unidad Fisiología. facultad de Veterinaria.

Summary: The bowel inflammation is one of the most common causes of gastrointestinal pathology. Under the name of inflammatory bowel disease (IBD) are grouped two and relapsing chronic inflammatory diseases of unknown etiology: Crohn's disease and ulcerative colitis The relatively high incidence of these diseases makes studying its pathogenesis is an important objective, as currently there is no cure and / or prevent recurrence Despite that IBD patients suffering from symptoms related to alterations of the gastrointestinal motility unknown implication that these disorders can have in the pathogenesis of the disease. Our group has focused on the characterization of an experimental model of chronic bowel inflammation, with the initial aim of assessing the changes that causes inflammation in the intestinal motility and to explore mechanisms that are involved in these changes. The study has been carried out in an experimental model in rats induced by indomethacin, which have succeeded in reproducing relapses that occur spontaneously in patients human Thus, animals which were given indomethacin presented alternate stages of active inflammation, characterized by leukocytosis, increased serum TNF and increased activity MPO, other apparent recovery of normal. Studies in this model clearly demonstrate that the different phases are accompanied by changes in the intestinal flora and alterations of motility. While during the active phase showed a hipomotilidad generally accompanied by an overgrowth of bacteria luminal, the recovery phase is associated with an increase in engine parameters that help with the restoration of microbial flora regulation of the synthesis of NO plays a role key in the course of motor impairment associated with the inflammatory process while increasing motility observed during the recovery appears to be due to reduced inhibitory tone derived from intestinal inhibition of the expression of nNOS, lower motility and overgrowth bacterial associated with the active phases appear to be due to an increase in the synthesis of NO produced by overexpression of iNOS. Recently we have also demonstrated that animals with induced inflammation present a sustained increase in intestinal permeability, suggesting that the vulnerability of the epithelial barrier function is one of the factors involved in the perpetuation of the inflammatory process. So together, these results give rise to the following hypothesis: while the burden antigenic stays within certain limits, because of a normal or increased motility, no inflammatory response in spite of increased intestinal permeability. However, when the motor activity decreases, increased interaction antígeno-mucosa facilitates the passage of molecules through the epithelium, resulting in the activation of the immune system and the release of inflammatory mediators

Author: Peluffo Zavala Hugo.
Year: 2005.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: Escuela de Postgrado, UAB.
Place of preparation: Unidad de Histología, Fac.de Med. UAB.

Summary: Acute central nervous system (CNS) damage consists of a multitude of inter-related and complex events, playing excitotoxicity, oxidative stress and inflammation important roles in the initial and secondary injury. In particular, the immature brain displays several characteristics that make it special in its reactions against acute injuries. In this context, this Thesis contributes to the basic understanding of the oxidative stress and inflammation occurring after an NMDA-mediated excitotoxic lesion to the immature brain, and to the development of neuroprotective gene therapy strategies for reducing immature brain damage by inhibiting oxidative stress. This Thesis shows the in vivo expression and cell localization of one of the most important antioxidant proteins, Cu/Zn superoxide dismutase (SOD), whose expression was observed mainly in neuronal cells, glia limitans and ependyma in the normal immature brain. However, after an excitotoxic lesion, the expression of this enzyme rapidly disappeared (at 4 hours) from the acutely affected neurons before they underwent cell death. One day after, expression of SOD begun to increase in reactive astrocytes, present in the lesion for up to 7 days, the last time studied. Moreover, in the normal brain, the peroxynitrite product nitrotyrosine was observed in neurons and scattered astrocytes, however, after the excitotoxic lesion it was early increased in neurons of the lesion core, and after at 1 day post-lesion it increased also in astrocytes. Interestingly, the nitrated hypertrophied astrocytes from 3 days post-lesion onward represented a separated population of cells sharing several markers such as vimentin, metallothionein, SOD, and high GFAP content and hypertrophy, being always phenotypically the most reactive astrocytes. In addition, although being heavily nitrated and showing activated caspase 3 in their nuclei, nitrated astrocytes did not display any morphological sign of cell death nor TUNEL staining at any time-point studied. The next step of this Thesis was to explore the putative beneficial effects of the overexpression of SOD after the NMDA excitotoxic lesion. This Thesis focused on the development of a new gene therapy strategy based on a non-viral modular recombinant protein vector. These vectors could deliver a transgene to the whole excitotoxically lesioned zone of the immature brain when injected 4 hours after the lesion. Moreover, they could transfect neurons, astrocytes and microglial cells, without generating additional inflammation. With these promising results, one of these protein vectors was used for overexpressing SOD 2 hours after the excitotoxic lesion. SOD overexpressing animals displayed decreased nitrotyrosine formation, reduced lesion volume, increased neuronal survival, and a complete functional recovery after 3 days in relation to NMDA+saline injected animals. Surprisingly control lesioned animals injected with the protein vector overexpressing the transgene for the green fluorescent protein or with the naked protein vector without any DNA, showed also a reduced lesion volume. Finally this Thesis showed that the neuroprotective potential of the protein vectors was mediated by the vectors integrin-interacting Arg-Gly-Asp (RGD) motif, as a cyclic RGD peptide was sufficient to induce this neuroprotection. Accordingly, both the protein vector and the cyclic RGD peptide were neuroprotective against a NMDA mediated injury to mixed cortical cultures. However, none of these molecules were neuroprotective under the same treatment conditions in cortical neuron purified cultures, suggesting that the neuroprotective mechanisms include triggering of a glial derived neurotrophic phenotype. This Thesis concludes that oxidative stress, and in particular the O2-./ONOO- pathway is a mayor contributor to lesion expansion in the acutely injured immature brain, and that the overexpression of SOD is an interesting neuroprotective strategy. In addition, it shows that modular recombinant protein vectors are efficient gene therapy vectors that can be applied to therapeutic interventio 8 ns to th 2b9 e acutely lesioned immature brain.

Author: Ribas Cabezas Laia.
Year: 2005.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: Universitat Autònoma de Barcelona.
Place of preparation: Universitat Autònoma de Barcelona.

Author: FORMENT DASCA JOSEP VICENT.
Year: 2005.
University: VALENCIA [www.uv.es].
Place of defense: INSTITUTO DE AGRICULTURA Y TECNOLOGÍA DE ALIMENTOS.
Place of preparation: FACULTAD DE CIENCIAS BIOLÓGICAS-UNIVERSITAT DE VALÈNCIA.

Summary: The filamentous fungus Aspergillus nidulans is a model organism for the study of metabolic pathways given the wide variety of substrates on which it can grow. Although much is known about the type and the regulation of enzyme activity that expresses the fungus depending on the available carbon source, very little is known about the mechanisms for attracting key metabolites such as monosaccharides. This paper describes the cloning of six genes (mstA, mstB, mstC, mstD, mstE and mstF) hypothetical whose protein products might be related to the transport of sugars. Two of these six genes are characterized functionally, discovered that the gene mstC encodes a protein transport glucose high affinity is expressed preferentially in conidia in germination of A. Nidulans, while the gene mstE encodes a permease manosa glucose and low affinity. The expression of the gene mstC is suppressed by the presence of carbon sources repressive in the middle through the action of transcription factor CreA and is induced under conditions of pH alkaline environment thanks to the action of transcription factor PacC. The elimination of the gene mstC leads to loss of glucose transport of high affinity similar to what happens in the mutant sorA3 A. Nidulans. The gene mstC is able to complement the mutation sorA3, by implying that mstC and sorA are the same gene. The expression of the gene mstE this induced by the presence of carbon sources repressive in the middle of a dependent transcription factor CreA. A fusion of the protein MstE the fluorescent protein GFP let the subcellular localization of the transporter in the plasma membrane of the fungus. The elimination of the gene mstE causes glucose transport in low affinity conidia in germination of A. Nidulans. Given the large number of genes in A. Nidulans which a priori could present transport capacity of sugars (with the consequent problem in the search for functional phenotypes by possible redundancy), this work also describes the development of a molecular tool for the elimination consecutive genes in a single strain A. Nidulans. This strategy is based on the system of recombinasa Cre and sites 10XP of bacteriofago P1, which allows the recycling of a selective marker selection bidirectional (ie, which can be selected either positively his absence as their presence). In this case, we use the gene pyr-4 of Neurospora crassa, supplementing the inability of growth media without uracilo and uridina of the strain pyrG89 A. Nidulans, which in his absence was very resistant to the toxic effects of acid 5-fluoroorotico. To demonstrate the effectiveness of the system is removed from consecutively genes and A and wA involved in coloración of spores of the fungus.