LIFE SCIENCES

Author: ULLOA DARQUEA FAUSTO ALEXANDER.
Year: 2001.
University: AUTÓNOMA DE BARCELONA.
Place of defense: Fac.de Medicina, UAB. Hosp. del Mar..
Place of preparation: INSTITUT MUNICIPAL D'INVESTIGACIÓ MÈDICA.

Summary: The benzyl GalNAc was initially described as an inhibitor of O-glicosilación type mucin (Kuan et al., 1989). The chronic treatment of cells mucosecretoras HT-29 M6 with the drug produced a series of effects including: i) reduction of the proliferation ii) inhibition of secretion mucinas iii) accumulation of glycoproteins in apical vesicles citoplasmáticas electronlúcidas and iv) reduction in sialilación of glycoproteins (Huet et al., 1998; (Hennebicq-Reig et al., 1998). benzyl GalNAc also induce these effects on the population HT-29 parental and other cell populations derived from it regardless of their phenotype. benzyl GalNAc is metabolized to benzyl GalNAc-Gal, which in turn competitively inhibits the activity a2, 3 sialiltransferasa cell HT-29, activity sialiltransferasa predominantly expressed in these cells. As a result of this, synthesized compounds sialilados as benzyl GalNAc-Gala2, 3Neu5Ac (huet et al., 1995); Delannoy et al., 1996). Moreover, treatment with benzyl GalNAc not sensitive to the cells Caco-2, which express essentially 2.6 sialliltransferasas, enzymes that are not inhibited by metabolites derviados benzyl - GalNAc (Huet et al, 1998). The aim of this study was to identify the factor (s) responsible (s) of origin of the cell phenotype HT-29 treated chronically with benzyl GalNAc. More specifically, we are interested in determining how a glycosylation inhibitor produces different effects and therefore whether the effect of this inhibitor was not restricted or cells derived from HT-29. The hypothesis that was handled initially proposed that the overall effects of treatment with benzyl GalNAc could be caused by lahiposialilación of glycoproteins. This hypothesis was based on the assumption that acid siálico be required for the transportation / apical sorting of glycoproteins. In the absence of these glycoproteins are siálico acid accumulate in trasponrtadores exocíticos arising from the TGN, which correspond to the vesicles citoplasmáticas electronlúcidas (vesículas-BG). In a manner consistent with this hypothesis, we found through analysis lectin, which siálico acid linked in a position to 2.3 was distributed primarily in the apical membrane of epithelial cells in culture and in tissues. Notariamente, acid siálico linked in a position to 2.6 showed a wider distribution, possibly located at the apical membrane and in the basolateral. The study of the effects of benzyl GalNAc on a panel of cell lines showed that the presence and severity of the same dependent deltipo cell but were not restricted to cells HT-29. Thus, in cells IMIM-PC-1 treated with benzyl GalNAc effects were observed similar to those described in cells HT-29. Notariamente in these cells accumulated intracellularly apical glycoproteins, as MUC1 and basolaterales as b1 integrina. MUC1 and b1 integrina colocalizan part in the same vesicles, suggesting that they may not correspond to structures exocíticas, since glycoproteins apical and baslaterales transported in different vesicles (Kellelr et al., 2001). The analysis of glycoprotein processing cells in HT-29 M6, showed that the benzyl GalNAc affecting the processing of glycoproteins apical and lisomales.Así, cells were treated with the inhibitor induced a delay in the maturation of the catepsina D block lamaduración the a-glucosidasa lysosomal (AAG) in a behavior acidic post-TGN. In cells tratatas with benzyl GalNAc, AAG is processed into a form that migraba between aberrant forms intermediate and mature this enzyme. Analysis at different concentrations and time of treatment with the inhibitor, showed that experimental conditions resulting in the inhibition of sialilación not meant necesarimente the appearance of the remaining effects observed in these cells, indicating against what initially postulated that lahiposialilación 8 of glico c3d protein is not the calusa the overall phenotype of cells treated with benzyl GalNAc. Treatment with benzyl GalNAc affected the urta endosomal cell IMIM-PC-1 and HT-29 M6. Thus, benzyl GalNAc induced a decrease in endocytosis and the accumulation of material internalized from the plasma membrane in cells IMIM-PC-1. In cells IMIM-PC-1 this material colocalizaba with vesicles positive for MUC1 or b1 integrina. Markers endosomas late as Rab7 and LBPA, but not endosomas primary (Rab5, EEA1) and Golgi (GRASP65, TGN46) colocalizaban with b1 integrina of vesicles. All the results indicate that the vesículas-BG correspond to a heterogeneous population of ensomas aberrant, related endosomas late. The phenotypic characteristics of the cells treated with benzyl GalNAc reproduced partially defects described in some cells of lysosomal diseases deposit (EDL), specifically those produced by the accumulation of saccharide compartments degradativos. Thus, the morphology of vesículas-BG is similar to those seoriginan in cells treated with sucrose (sacarosomas) and several EDL as sialidosis or infantile sialic acid storage disease (ISSD). Analysis of processing the AAG in fibroblasts of patients with ISSD showed defects similar to those found in cells HT-29 treated with benzyl GalNAc. In addition, cells treated with benzyl GalNAc also accumulate LBPA and cholesterol, findings characteristic of some EDL. Confirming the idea that the benzyl GalNAc induces a phenotype of lysosomal deposit, treatment with sucrose cell IMIM-PC-1 produced changes sobrela morphology of the cells and the intracellular accumulation of MUC1 and b1 integrina, in a similar way to what he had observed in cells treated with benzyl GalNAc. The sialilación of glycoproteins in cells IMIM-PC treated with sucrose was not altered, resulting queindica that hiposialilación is not a requirement pra intracellular accumulation of membrane glycoproteins. Since cells treated with benzyl GalNAc accumulate a large amount of oligosaccharides derived from the drug, a situation similar to that which occurs in cells treated with sucrose and some EDL, we propose that the accumulation of these metabolites, and not hiposialilación of glycoproteins is the that causes the cell phenotype overall HT-29 and IMIM-PC-1 treated with the drug.

Author: PASCUAL PEÑA EMILIANA ESTHER.
Year: 2003.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTAD DE MEDICINA - UCM.

Summary: Chronic ulcers of the lower limbs pose a major challenge to social welfare because its high prevalence, the difficulty of treatment and its high social and economic cost. Decommissioning cures through topical and postulares is rarely spontaneous, and usually require grafts or flaps that do not always have good results. The crops querantinocitos are presented as an alternative therapy. In this paper, two kinds of value for carriers epithelium cultivated: gauze vaselinaza and fibrin gel. HYPOTHESES Because the fibrin gel provides growth factors involved in the regeneration of injured and has greater consistency, its use as a carrier matrix of growing keratinocytes should improve the quality of cobrtura of chronic ulcers in lower extremities. OBJECTIVES 1, - compare clinical outcomes of empelo sheets of cultured keratinocytes, transported in gauze vaslinadas and fibrin gel, for coverage of chronic ulcers in lower extremities. 2, - Analysis of time handling with the two types of carriers. 3, cost-analysis of the two carriers means. RESULTS The results in the group of fibrin indicate that requires significantly less (p less 0001) that the group of gauze to get healing comppleta of injury to a patient. The index 1 (I1) links the closing time with the surface, to say how much time is needed to heal 1cm2 of the ulcer. The index 2 (I2) relates the number of grafts with the surface, and expresses cúantos grafts are ncesarios for complete healing of 1cm2 of injury. The results for both indexes have shown statistically significant differences between the two types of treatment (p less 0002 in the I1 p lower 0001 in I2), being the group fibrin which requires less time and fewer grafts to heal 1cm2 . The cost-effectiveness analysis demonstrates our population is needed fewer grafts and less time to cover 1cm2 injury in the group treated with fibrin. In addition, production costs are lower in the group of fibrin. But the difference between the two groups was not statistically significant. Conclusions The results show that the use of fibrin gel as a transporter sheeting cultured keratinocytes to heal ulcers, chronic lower extremity: 1, - Increases cellular proliferation, obtaining prior grafts ready for cultivation. 2, - Remove enzymatic treatment during the process of separation of the blades from the bottle cultivation. 3, significantly reduces the time-handling of the crop. 4-significantly diminishes the time to heal 1cm2 of chronic ulcer. 5, - significantly reduces the cost of production of the plates. 6-Aumetna efficiency, as required less graft skin to cover the defects.

Author: GONZÁLEZ HOYUELA ALONSO MARITZA CLARA.
Year: 2003.
University: AUTÓNOMA DE MADRID.
Place of defense: FACULTAD DE CIENCIAS.
Place of preparation: FACULTAD DE CIENCIAS, UNIVERSIDAD AUTÓNOMA DE MADRID.

Author: CASES GONZÁLEZ CLARA ELISA.
Year: 2003.
University: AUTÓNOMA DE MADRID.
Place of defense: FACULTAD DE CIENCIAS.
Place of preparation: FACULTAD DE CIENCIAS.

Summary: The retrotranscriptasa (RT) of the virus from inmunodeficienciahumanatipo 1 (HIV-1) is a enzimamultifuncionalcon activities DNA polymerase dependent ADNy RNA, RNasa H, and transfer and despazamiento Hebra. It is a heterodímero composed of two subunits, p66 and p51, which contain the subdomains called "fingers," thumb "," palm "and" connection ", and a domain RNasa Hen largest subunit. Resíduos Ala-114 and Tyr-115 along with other debris as Asp - 113 and Gln-151, form the pocket which accommodates the group 3'-OHdel nucleotide incoming. residue Met-230se located in a highly conserved region called "first grip," held liable for maintaining the end of the initiator in a suitable position to be carried out the attack nucleofílico on an incoming dNTP. Doctoral Thesis This has studied the effect of mutations at positions 114, 115 Y230 on the accuracy synthesis ADNy the specific nucleotide under different conditions, using biochemical testing of nucleotide incorporation based gel as a new genetic test to measure the frequency of mutation G / A for the synthesis of ARNdependiente DNA. &% / Replacements not preserved in the Tyr- 115 causing a decline in the loyalty of ADNen presence synthesis of Mgcomo cofactor.'s presence Mntiene a mutagenic effect on the polymerization of DNA, which operates primarily at the extent of extreme desapareados. is needed aromatic amino acid at position 115 to discriminate in front of nucleotides that contain a group 2'-OH participating the Tyr-115 as a barrier estérica. substitutions of Ala-114 did not appear to influence on the accuracy of the enzyme, although this aminácido it contributes to discriminate against ddNTPs. Mutants in the region of "first grip," as M2301y M230L, behave as highly mutagenic in the presence of unbalanced concentrations of dNTPs, to use a test that measures the genetic fidelity frequency of mutation G / A.

Author: MORÁN GRACÍA JOSÉ M..
Year: 2003.
University: EXTREMADURA.
Place of defense: FACULTAD DE VETERINARIA.
Place of preparation: FACULTAD DE VETERINARIA.

Author: TAMAYO HURTADO MANUEL GABRIEL.
Year: 2003.
University: GRANADA.
Place of defense: FACULTAD DE CIENCIAS DE LA EDUCACIÓN.
Place of preparation: CATÓLICA DEL MAULE, TALCA, CHILE..

Summary: The biological evolution gives meaning to the Biology and helps to explain the characteristics of the world around us. However, one of the areas most complex and most difficult courses in Biology, and in different countries, there are movements that have unleashed a source opposed to his teaching, pressure has led to progressively reduce or tergiversrla. It examna this situation in the Chilean education. It provides a review of background about the origin of evolutionary biology and their education, and religious conflict that generated the most important milestones in the history of education in Chile. We reviewed the information in order to form a clear picture about three aspects interrelacinados: Chile, for the last high school or middle assessed the conflicting positions between evolutionists and antievoucionistas reflected in the contents, related ideology of qutores with its position versus evolutionism, it determines the relative importance ing to the theme of evolution in the schools, the contribution of textbooks Chileans to the development of evolutionary ideas, the advance of scientific knowledge and its manifestation in the teaching of evolution. We compared various edicinos of texts you to analyze the changes ne content, it was a review of the topics considered in Evolutionary Biology texts published since the late period of the synthesis and upheld the veracity and the afimaciones. Scientists mentioned were classified according to their relevance and relationship with schools evolutionary offered. In some textbooks were found deterministic arguments and explanations teleológicas and antropocéntrisa. Sometimes are scarce or missing connections between evolutionary biology and might explain the phenomena. There are analyzed, or discussed superficially, the concepts of scientific facts and theories and the relationship between religion and evolutionary biology. The language can help reinforce ideas lamarquistas or on the alleged survival of the strongest. Other aspects dificitarios are not relsiconar the lamarquismo with concepcione of students or fijismo with creationism present not linking genetics with evolution, not to mention any ideas sinteticistas. In some cases are doubts as to whether the evolution embraces all living beings. Being reviewed didicultades pair compression of the theory of evolution, alternative conceptions of students, the confusion between the scientific language and common language, the rejection of biological evolution worst religious grounds and the reasons that sometimes in doubt Teória the scientific nature of the developments. According to the tests, the textbooks on the subject of biological evolution may be required compliance with certain aspects, such as biological evolution adequately define and clarify its scope, to clarify the scope of témino "theory" in connection with the evolving adequately explain the main evidence of evolution, discuss various types of "adaptation", explain and demonstrate proper lamarquismo and darvinimo, etc.. We present a proposal for teaching Evolution Biologíca from analysis of the historical development, according to the published texts and the different aspects reviewed.

Author: López Lasala Beatriz.
Year: 2003.
University: AUTÓNOMA DE MADRID.
Place of defense: Facultad de Ciencias.
Place of preparation: Facultad de Ciencias, Universidad Autónoma de Madrid / Facultad de Ciencias, Universidad de Chile.

Summary: The main objective of this thesis was to assess whether there was variation in behavior between individuals chemical recognition of the same sex of the species island LZ Ater and explain variations in a context filogeográfico. To do so: 1) identified the phylogenetic relationships of the group Nigromaculatus through analysis of sequences of cytochrome b mitochondrial DNA; 2) were established relations filogeográficas of island populations of Lz Atrer of the Fourth Region of Chile, using the same marker as above; 3) were analyzed as part of semioquímicos secretions precloacales of Lz Ater from the islands Tilgo, Totoralillo and Chungungo, when individuals from each population were confronted with chemically individuals of the opposite sex, their own people and other peoples. The analysis of mitochondrial DNA sequences confirm that the species L. Platei does not belong to the group Nigromaculatus. The origin of this group is established in the Miocene, but the processes leading to the current species would have occurred during the Pleistocene. The group Nigromaculatus would consist of the L. Atacamentsis, L. Bisignatus, L. Copiapoensis, L. Silvai, L. Nigromaculatus, L. Kuhlmani and two nevas species: sp. Nova 1 sp. Nova 2. Likewise, the taxonomic status of the species LZ Zapallarensis and LZ Ater needs to be revised. Based on the analysis of cytochrome b both subspecies are two different species. The geographical area of contact between the two species would be established around the 29.5  ° latitude south to the mainland populations, and the island Tilgo for forms island. Both species have originated from two independent ancestral cores during the Pliocene half, which would have colonized the islands during interglacial periods when the masses pleistocénicos glaciers led to a decline in sea level which enabled the connection of these islands to the mainland. It has not been possible to establish direct relationship between history filogeográfica complex island Tilgo, Totoralillo Chungungo and with the results obtained from analysis of the secretions precloacales and conduct reconnaissance chemical. The chemical composition of secretions precloacales is similar to that in other species characterized Liolaemus but differs in the absence of low molecular weight compounds in the presence of compounds metilados. These differences would be related to the environmental conditions of habitat that would allow better detection of the signal chemical. The comparison of interpoblacional secretions precloacales suggests that we are in initial stages of divergence chemistry. These initial data are caracterizarián by introducing a large number of chemical compounds and low levels of chemical differentiation. Among the populations of the islands Tilgo, Totoralillo Chungungo and there concordance between the results obtained from analysis of  º secretions precloacales and analysis of the behavior. Low levels of chemicals detected differentiation suggest that the drift is more that the selection, the main force responsible for evolutionary change, both in composition and in the detection of chemical signals. Under certain weather conditions, such as high relative humidity and reduced solar radiation, the lizards could increase efficiency in attracting the chemical signals simultaneously making use of the systems and WNV MO.

Author: OUARDANI FATIMA EL.
Year: 2003.
University: GRANADA.
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTAD DE FARMAICA UNIVERSIDAD DE GRANADA.

Summary: As a point of techniques for the diagnosis through anatomical characters of 90 medicinal species belonging to 9 families botánicaS: Apiaceae (3 species), Asteraceae (29 species). Elerberidaceae (1 species), Caryophytlaceae (6 species), Ericaceae (1 species), Fabaceae (6 species), Lamniaceae (37 species), Rhamnaceae (3 espeices) and Rosaceae (4 species). Bring to identify and categorize the elements anatomical considered key to the diagnosis of taxa and on the basis of these elements have tabled a proposal for keys to the systematic identification of plant material irrespective of the state in which it finds itself. The memory of 733 pages is illustrated with 90 figures and 37 sheets in color.

Author: JIMENEZ SEGOVIA EDUARDO TERENCIO.
Year: 2003.
University: SEVILLA.
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA.

Summary: In the Mediterranean area, with high temperatures and high evapotranspiration during the summer, sugar beet (B.vulgaris) is planted in the fall (autumn sowing) and harvested at the end of the period of Spring and early summer. In temperate regions, the cultivation of sugar beet is planted in the spring (spring planting) and harvested in the autumn. With this autumn beet farming practice avoids the high temperatures during an early stage of development. By contrast, suffers from water shortage and high temperatures during the harvest. This fact leads to the plant in response to stress that is often associated with high levels of compounds such as glucose, fructose, alfa-amonio, Na, K, which involves a decrease in the quality of the root. In sta thesis has been studied physiological aspects related to the loss of quality beet planting autumn. There is an increase in levels deprolina at harvest time, this being indicative of the water stress situation in which the plant is located. During this period also produces glucose accumulation that is associated with an increase in activity Invertase acid (an enzyme hydrolyzed sucrose). The harvest period is noted that high levels of nitrogen in plant coincide with increases in the levels of glucose and decrease the sucrose content in the root. This fact could be related to an increase in activity Invertase acid and sucrose synthase respectively. Thus, these data show that the stress suffered by the beet seed in autumn reduces the quality and production of sugar in the root. Excess nitrogen increases the surface of the leaf transpiration and hence the stress suffered by the plant. This stress has a dramatic impact on the quality and production of sugar in the autumn crop of beet. This work shows that a variety of "road close" with a smaller transpiration, suffer less stress introducing proline levels of reducing sugars and lower during the period of data collection, which improves the quality of the root. Then he studies the metabolic state in which the beet planting autumn compared with the spring planting, comparing levels adenilatos that we find in the wake of the autumn sowing compared with levels adenilatos determined in spring planting. The results show a more active metabolic status in the beets harvested in the autumn sowing (summer), with high temperatures and long photoperiods, compared to less active metabolic status in planting beets harvested in spring (October-November), where the plant is to start the Dormition in response to environmental signals as short photoperiods and cooler temperatures. Finally, we present a number of new treatments applied to sugar beet planting autumn aimed at interfering in the responses to stress the plant, mainly proline accumulation and reducing sugars, which occur in the data collection period. These innovative treatments could reduce the loss of quality of the beet root crops planted in autumn.

Author: PRATS RUIZ M. TERESA.
Year: 2003.
University: BARCELONA.
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA.

Summary: It has investigated the influence of major effectors alostéricos (Ins-P5, ATP 2,3-BPG, CO2, H +, Cl-and TÂ th) on the affinity hemoglobina-oxígeno (Hb - O2) in two types of vertebrates that use models alostéricos individuals: 1 - The quail (Coturnix coturnix japonica) as a model of bird, characterized by use as a regulator of the erythrocyte affinity Hb - O2 a phosphate disconnected from the main channels of energy cells: Ins-P5. 2 - The Goat (Capra Hispanic pyrenaica), ruminants and unique case in mammalian erythrocytes as their lack of organic phosphates regulators affinity Hb - O2 and modulate the affinity for C1- introducing its beta chains of amino acid substitutions make it less sensitive to phosphates. Both groups are of interest because presentna demands metabolic relatively high and have been little studied in terms of the molecular regulation of the transport of O2 in two different functional models, such as birds and mammals, allows contrasting aspects differentials two and expand existing information in order to resolve some outstanding issues to clarify in the modulation alostérica these types of Hbs. For the realization of this work have been developed, a series and chromatography techniques with the aim of: isolate, purify and separate the different Hbs, and isolate and purify Ins-P5 for subsequent use in experiments modulation alostérica ( Prats and Riera, 1994. Anal. Biochem. 221:335-339). Studies affinity Hb - O2 has been made using two different methodologies; method polarográfico (Neville, 1974; Hughes et al. 1988) and methods tonométrico (Giardina and Amiconi, 1981), the latter used in a placement at the University of Tor Vergata of Rome with Drs. S. Condó and B. Giardina. It has also had the opportunity to characterize the transport of oxygen in the blood of two wild species and compare them with their domestic counterparts: * We have studied the possible adaptations in the transport of O2 in populations of native wild quail (coturnix coturnix coturnix) during his nomadism areas from sea level to about 1,200 m above sea level (Prats et al. 1995. Physiol. Zool. 69 (4): 912-929), comparing them with those of the domestic quail (Coturnix coturnix japonica). * We have studied the possible adaptations in the transport of O2 in populations of native wild goats (Capra Hispanic pyrenaica), comparing them with those of domestic goats (Capra hircus sp.). This paper provides a detailed knowledge of the regulation and novel molecular affinity Hb - O2, which will allow a better understanding of the observed changes in the transport of oxygen at the molecular level in different physiological situations. Evidence also adaptations in the transport of oxygen populations of wild animals that are subjected to a higher level of activity and influence of different environmental factors and behavior.

Author: Armengol Barnils MMa. del Pilar.
Year: 2003.
University: AUTÓNOMA DE BARCELONA.
Place of defense: UNIVERSITAT AUTÃ’NOMA DE BARCELONA.
Place of preparation: Unitat Docent UAB Hosp. Germans Trias i Pujol.

Author: GARCÍA VILLADA LIBERTAD.
Year: 2004.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE CIENCIAS BIOLÓGICAS.
Place of preparation: FACULTAD DE VETERINARIA.

Summary: Currently, the level of environmental pollution is a threat to the maintenance of ecosystems. The microalgae are a group of basic commodities in the functioning and structure of aquatic ecosystems and soil. Previous studies have shown that contaminants pose a risk to these microorganisms. However, it was not known what the capacity of microalgae to adapt to current levels of pollution. In this thesis has been investigated capacity and coping mechanisms of populations microalgales phenomena of acute pollution. As experimental models used three laboratory strains, belonging to the species Dictyosphaerium chlorelloides (Cloroficeae) (DcG1wt), Scenedesmus intermedius (cloroficaea) (Si31Mwt) and Microcystis aeruginosa (Clanobacteira) (MaM1), and four toxic: herbicide 3-( 3.4-diclorofenil) -1.1 -dimetilurea (DCMU), a complex mixture of heavy metals discharged from the Aznalcóllar mine, the explosive 2,4,6-trinitrotolueno (TNT), and alguicida copper sulphate (CuSO4.5H2O). The basic tool used in this study was the analysis of fluctuation. Moreover, this approach also investigated the usefulness of the analysis of fluctuation to provide specificity to biosensors microalgales. Our results indicate that the microalgae are capable of withstanding the effects of lethal concentrations of pollutants through spontaneous mutations of large effect. The estimated frequency of resistant mutants in balance, coupled with the size typical of the populations microalgales suggests that these populations puedne persitir after a toxic spill despite the cost of the mutations. Although the problem addressed in this paper should be investigaco in greater depth and through in-situ experiments, the results suggest that contributed Environmental pollutants, they can be atlerando the evolutionary rate of populations microalgales. On the other hand, the use of resistant and susceptible strains of a particular contaminant has proved a useful way in the development of specific biosensors. KEYWORDS: Adaptation, fluctuation analysis, Aznalcóllar, bionsensor, DCMU, Dictyosphaerium chlorelloides, micro - algae, Microcystis aeruginosa, Scendesmus intermedius, rate of mutation, TNT.

Author: GENESCÀ FERRER MERITXELL.
Year: 2004.
University: BARCELONA.
Place of defense: FACULTAD DE BIOLOGIA.
Place of preparation: IIBB-CSIC (IDIBAPS).

Summary: In the thesis, have studied the mechanisms that trigger the activation of apoptosis during ischemia syndrome and reperfusión.Durante ischemia occur a series of biochemical events that can modulate the development of apoptosis.Uno of the first events that take place is degradation ATP.Esto induces accumulation products catobólicos substances quese changed depending on the time of ischemia, such as adenosine and xanthine, which through the nitric oxide and superoxide respectively may induce apoptosis.Por elsewhere Another major consequence of ischemia is the alteration of citoesqueleto.Recientemente, it has been observed that these alterations may induce the development of apoptosis in certain cell types. Thus, the overall objective of this thesis has been to characterize the role of certain molecules in relation to the development of apoptosis during the preservation of organs, to modify and pharmacologically predict. With the work done in this thesis we have shown that the development of apoptosis during ischemia and reperfusion is modulated by adenosine and xanthine and, in the case dela adenosine, this is mediated induction oxide nítrico.La cell death by apoptosis can be minimized by the addition of theophylline and fructose - 1, 6-difosfato to the preservation solution, which not only prevents him develop apoptosis, but also prevents bacterial translocation to trasplante.Por Furthermore, the development of apoptosis in reperfusion depends on the modificaciónes in the actin cytoskeleton during preservación.Finalmete, in connection with the study of predictive markers of organs can be a very helpful in this respect, since it allows for the preservation discriminate between bodies that have undergone a period of warm ischemia and those no.Además, one of the parameters derived from this measure parameters related to alterations of the cytoskeleton produced prior to the transplant.

Author: ESPARZA GORDILLO JORGE.
Year: 2004.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE BIOLOGÍA.

Summary: The complement system is a set of proteins lmpbcadas in detensa dél agency ftente infections. The effectiveness of this mechanism lies in its ability to activate specific pathogens on limiting the harm associated with the uncontrolled activation of complement on the tissues themselves. Such specificity is based on the existence of a set of regulatory proteins, many of which are encoded by genes located. In the genomic region Iq32 within the grouping gene RCA (Regulators ofComplement Activation). In this dissertation I applied the method of decomposition of the varianza.para analysis of the genetic factors responsible for the variability in the plasma levels of two regulators supplement to the RCA (C4BP and factor H). Also evaluated the possible implications associated with pathological variability in these phenotypes. The protein C4b-Binding Protein (C4BP) has the ability to interfere with the clotting system through its interaction with the protein S (PS). This work has applied the model threshold or threshold in order to analyze the contribution of phenotypes PS plasma C4BP and the susceptibility to suffer from thrombosis. Moreover has studied the role of the complement regulatory proteins in the RCA predisposition syndrome hemólítico uremic atypical (aHUS), a disease caused by the uncontrolled activation of complement on the endothelium kidney as a result of alterations in functional proteins of RCA. In this context, our study has been carried out considering changes. Quantity (of expression) and qualitative (functional deficit) genetically determined proteins RCA.

Author: TORRE MARTINEZ IRATXE.
Year: 2004.
University: PAÍS VASCO.
Place of defense: FACULTAD DE MEDICINA Y ODONTOLOGIA.
Place of preparation: FACULTAD DE MEDICINA Y ODONTOLOGIA.

Summary: The objective in this work Doctoral Thesis, is to deepen the understanding of the mechanisms leading to the death of oligodendrocytes, following the activation of receptor glutamtérgicos and purinérgicos in the optic nerve of adult rats. To address this study have been used Immunohistochemical techniques, biochemical and molecular biology. Our results indicate that this tissue oligodendrocytes present recipients ionotrópicos of glutamic acid AMPA and kainato, which produces activated forward apoptotic cell death, through a mechanism that requires the activation of the caspasa-3. These conditions excitotoxicidad decrease the expression of the phosphatase nuclear MKP-1 and increase the expression of the phosphatase cytoplasmic MKP-3. It was found that a brief stimulus excitotóxico also is involved in cell death, as it sensitizes cells in the optic nerve to attack the supplement. Under these conditions it was observed that reduces the expression of key molecules in the path of MAP kinase, as well as their regulators. Our results indicate that the oligodendroglial apoptotic death is not only mediated by receptors glutamic acid, but it is also mediated by the activation of the prolonged subclass receptor purinérgicos P2X7. Under these conditions, it was found that the death oligodendroglial also requires the activation of the caspasa-3. Therefore, these results suggest that knowledge of the intracellular components involved in these mechanisms of cell death and the role played by each of them, it is important to develop new therapeutic agents for the treatment of neurological diseases that involve the loss of oligodendrocytes .

Author: Núñez Naveira Laura.
Year: 2004.
University: A CORUÑA.
Place of defense: Facultad de Ciencias.
Place of preparation: Facultad de Ciencias.

Summary: Kluyveromyces lactis is a leaven of high interest to biotech differs from traditional yeast Saccharomyces cerevisiae to study various aspects of metabolism. While in terms of aerobic and glucose as a source of carbon K. Lactis has a metabolism fundamentally respirativo, S. Cervisiae has fermentative in the same context. These metabolic differences might reflect a different transcriptional regulation of those genes associated with breathing in these bodies so close evolutionarily. In this thesis explored a number of genes that are considered essential in this regard. ScHEM1 and ScHEM12 coded respectively delta-aminolevulinato synthase and uroprofirinógeno decarboxylase catalyzing the first and fifth step of the route biosynthesis hemo. This group structural hemoproteínas as cytochromes, which participate in the processes of breathing. ScHAP1 encodes a factor trascripcional activated hemo regulating the expression of genes related to breathing, and also activates the expression of ScROX1 that represses gene hipóxicos in aerobic conditions. ScSRB10 encodes a kinase involved in processes both activation and repression trascripcional. The latter is characterized by complex interactions with the repressor Ssn6-Tup1p after being recruited to the promoter regions of genes repressors by several factors, including protein Rox1p. In this paper we have cloned genes K1HEM12, K1SRB10 and K1HAP1 demonstrating for the first two complementing its functionality through mutations in genes counterparts in S. Cerevisiae. K1HAP1 has not proved to be functional in S.cerevisiae at least in control of its own promoter. The expression K1HEM12 has proven dependent strain, in addition to sugrir activation by non-fermentable source of carbon through participation factor regulator Hap2/3/4/5p. The developer can be seen as TATA-less with multiple sites beginning of the transcript without definite preference for any of them. Through analysis of the first extension have located several starting points for the promoter K1HEM1; the position -48 predominant. It has verified the importance of the element-rich pirimidinas by testing deletions and mutagenesis directed toward specific regions of the promoter fused to lacZ. This element is responsible for determining high levels of expression in both aerobic and anaerobic conditions. About cis to the union of Gcr1p has an effect repressor Media glucose 2% and consensus for the union of Buf1p, that the promoter gene counterpart S. Cerevisiae has a repressive effect, does not show a clear regulatory role in either aerobic conditions or hipóxicas. In the area between the positions -558 to -443 is located a signal required for the expression and response to hypoxic hemo. By alignments of K1Srb10p with other proteín-quinasas related revealed that the yeast there were a number of reasons highly conserved. The functionality of these reasons was found by deletions of each of them, both on the protein Srb10p K. Lactis on the S. Cerevisiae. The reasons DC-Iy DC-II and the reason for ATP binding are essential to the function of Srb10p in vivo as demonstrated by the effects of its delección on associated phenotypes and the expression trancripcional of genes FLO11, CYC7 and SPI1 . Also, it has proven its direct or indirect participation in the interaction between kinase and its ciclina Srb1p, as well as for interaction with the repressor Tup1p. Another source also maintained DC-Ala, near the end carboxyl terminalk, lacks relavancia functional interaction with Srb11p, but has some influence on the interaction with Tup1p. The reason WRKY only present in the protein K. Lactis and near the amino end, it does not seem essential in the phenotypes and interactions analyzed.

Author: FERIA BOURRELLIER ANA BELEN.
Year: 2004.
University: SEVILLA.
Place of defense: FACULTAD DE BIOLOGIA.
Place of preparation: FACULTAD DE BIOLOGÍA.

Summary: The phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31) is a cytosolic enzyme widely distributed both in photosynthetic organisms such as plants and green algae, as in most of the non-photosynthetic bacteria and protozoa, however is absent from fungi, yeast and animals (Chollet et al., 1996). Catalysing beta-carboxilación irreversible fofoenolpiruvato (PEP) in the presence of HCO3-and Mg2 +, to produce oxaloacetato (OAA) and Pi (Chollet et al., 1996), which is quickly transformed L-Malato, participating in this great metabolic variety of contexts. Previous work carried out in our laboratory showed an increase in activity PEPC during seed germination and regulation of the enzyme by reversible phosphorylation (Osuna et al., 1996; Gonzalez et al., 1998, 2002). Unlike the PEPC-quinasas studied so far, the activation of the PEPC-quinasa involved in the germination is not dependent on protein synthesis, but it was already present in the dry seed (Osuna et al., 1999), suggesting that the synthesis of PEPC-quinasa was conducted during the development and maturation of the seed. The results of this thesis show that in the course of barley seed two polypeptides of PEPC of 103 and 108 kDa of whom, of 103 kDa subunit is a constituent while the 108 kD is induced in the final stage of development . The results show that both versions polypeptides are complete and do not contain the N-and C-terminal ends, including the last 4 aa ANTG the C-terminal. On the other hand, showed that the PEPC-quinasa accumulates in the barley seed from 30 PDA and is accumulated in the seed dry, but during this period and dry seed, PEPC is desfosforilada. Further results suggest that the relationship Glucosa-6P/L-Malato rather than isolated the concentration of these metabolites or quantity of PEPC-quinasa accumulated in the tissues, which determines the in vivo phosphorylation dela PEPC during development seed germination and barley. In this sense, this is the first physiological context to show a metabolic regulation of the PEPC-quinasa in vivo contrast to the synthesis / activation classical described for the regulation of PEPC-quinasa in all plants and organs studied to date. All this suggests that the aleurona / endosperm the PEPC compensates inhibition caused by high levels of malato accumulated during development, with increased activity up to 12 times, based on the total activity. This heightened activity could respond to the accumulation of L-Malato without recourse to the phosphorylation. This is according to a stadium development of low metabolic activity, high levels of ADP and a Glucosa-6P/L-Malato low that keeps the PEPc-quinasa, though abundant, essentially dormant in vivo. However, during germination, PEPC accumulated in the seed is phosphorylated rapidly being completely fosforilada to 48h of imbibition. The amount of a high number of PEPC accumulated in the seed, but a high level of phosphorylation, made possible by the abundance of PEPC-quinasa cumulada in the seed during development and for the high regard Glucosa-6P/L-Malato suggest that during germination the PEPC should reach its highest levels in vivo activity which would justified by the ubiquity, versatility and usefulness of its final product the L-Malato. In an attempt to clarify the possible role of the ABA in the regulation of PEPC-quinasa and in the phosphorylation of PEPC, we have seen that the ABA does not stimulate the synthesis of PEPC-quinasa but regulate their degradation impact on the activity of a PEPC-quinasa protease that is already present in the dry seed. This work has also been carried out cloning and expression of the PEPC C3 wheat (WPPC1) and the PEPC C4 leaves of sorghum (PEPC311) in Escherichia coli as a fusion protein with a etiquet 8 to de His 5cb. The immunological characterization of their properties, and kinetic s regulation of the phosphorylation of the enzyme, it can be concluded that are substrates suitable for any type of studies both kinetic and regulation by phosphorylation. The MS analysis of peptides of PEPCs obtained from different tissues of wheat to conclude that; 1, - peptides from WPPC1 have high levels of coverage in all tissues studied, but he noted the presence of the sequence MALSAPGGGGK specific the roots of WPPC1 in mature. 2-In wheat the PEPC is also encoded by a family multigénica from which we have identified at least 4 representatives: WPPC1, WCP21, WCP28 and WSOLTU. 3 - The existence of an isoenzyme counterpart to the PEPC4 (PEPC bacterial A.thaliana) in wheat. Moreover, they represent the first evidence that the bacterial protein is translated, differentially accumulated in certain tissues and in response to a pattern of development as in the case of escutelos during germination.

Author: CASAÑAS ACOSTA NIEVES.
Year: 2004.
University: LA LAGUNA.
Place of defense: INST.UNIVERSITARIO DE BIORGANICA ANTONIO GLEZ.
Place of preparation: FACULTAD DE BIOLOGIA.

Summary: It has analyzed the distribution, characteristics and possible functions of the cells astrogliales in the retina, nerve, optic chiasm and tract of Gallotia galloti, and have been used to identify these proteins glial populations, including S-100, protein gliofibrilar acidic (GFAP), vimentina and glutamine synthetase (GS), and to mark the population in proliferation has been used nuclear proliferation cell antigen (PCNA). It has been shown that this kind of the retinal ganglion cells are able to re-establish connections to the roof after acute injuries optical nerve. Taking into account the important role of glial cells in the maintenance of central nervous system (CNS) and the repair of the system in other groups such as fish and amphibians, the results are important to know the characteristics of these glial cells in the SNC-Gallotia galloti and resolve issues raised in the mock injured.

Author: ROIG NAVARRO IGNASI.
Year: 2004.
University: AUTÓNOMA DE BARCELONA.
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA.

Summary: The meiosis is the cell division that occurs during gametogenesis in which the chromosome strength is halved. The meiosis in mammals introduced gender differences, while the male is ongoing and begins in adult life, the female begins during fetal life and is aborted before birth. Thus, the mating and sinpasis of counterparts in human oocytes can only be studied in fetal ovaries. Thus, most of what is known about meiosis comes from work performed in male samples. In this study, we have used 33 samples to conduct an exhaustive analysis of mating and synapses of counterparts in human oocytes. The results have allowed for the first time to link the progression of meiotic prophase over fetal development in women. It shows that during the mating begins stadium leptoteno when oocytes presented morphology bouquet. In addition, the involvement of the dynamics in the synapses of telomeric counterparts. The results suggest that the mating processes and synapses are very efficient in oocytes euploides, as in oocytes cultured, as in oocytes aneuploids, introducing error rates on the order of 0.13%. The results presented also indicate that the involvement of the process of mating in the origin of aneuploidies is not as important as other factors. The oocytes euploides human analysis has revealed significant differences with regard to the progression of meiosis between men and women might be responsible for the high maternal origin of aneuploidies in humans. Furthermore, significant differences were found in meiotic progression among humans and mice that might suggest the search for another animal model to study meiosis in humans.

Author: AYLLON GOMEZ FERNANDO.
Year: 2004.
University: OVIEDO.
Place of defense: FACULTAD DE BIOLOGIA.
Place of preparation: FACULTAD MEDICINA.

Summary: This thesis arose to study the various patterns of gene flow in salmonid populations, especially the brown trout and Atlantic salmon. We studied the population structure of the salmon populations of northern Spain and southern France determined the presence of a single population panmíctica. We analyze the contribution of stocking allochthonous in determining and maintaining this genetic structure and the relative success of different stocks used in reforestation. For brown trout explored the role of gene flow in maintaining populations affected seriously by the fragmentation of habitat. It was determined the existence of high levels of gene flow between populations adjacent. Thanks to the existence of experimental populations in the Kerguelen Archipelago were studied patterns of gene flow and colonization of various stocks of brown trout, both wild origin as captive bred. Finally, it investigates the possibility of gene flow between the interspecific brown trout and Atlantic salmon, as well as the symmetry of the reproductive barriers between species.