LIFE SCIENCES, 4

Author: EL ALLALI ABDESLAM.
Year: 2004.
University: GRANADA.
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTAD DE FARMACIA.

Summary: It introduced digital mapping at 1:10.000 c of the southern slope of the Sierra Nevada Natural Park (Andalusia, Spain), accompanied by a fitosociológico study of vegetation in that territory, providing further data physical environment, biogeografícos, bioclimatic , geological and edáficos. The methodology followed in the study of vegetation has been the school sigmatista of Braun-Blanquet. The methodology seguída at work cartografíco has been using Information Systems Geografíca (GIS) with their corresponding databases. It has submitted a memory on paper, with annexes cartographic paper A1, and a caróm of the digital cartographic coverage.

Author: GONZÁLEZ GUERRERO MANUEL.
Year: 2004.
University: GRANADA.
Place of defense: ESTACIÓN EXPERIMENTAL DEL ZAIDÍN.
Place of preparation: ESTACIÓN EXPERIMENTAL DEL ZAIDÍN (CSIC).

Summary: This thesis examines the effects of exposure to heavy metals in the fungus micorrícico level morphological, and the organelles involved in the accumulation of the metal. Once these objectives are met proceeded to describe the mechanisms involved in the detoxification of Zn, Cu and Cd or its side effects, with special emphasis on carriers family CDF and MRP, metalotioneínas and Cu, Zn SOD. Finally, we evaluated how modifying gene expression of these genes between ecotype tolerant and another who was, in order to elucidate mechanisms of tolerance.

Author: PÉREZ CONTRERAS TOMÁS.
Year: 2004.
University: GRANADA.
Place of defense: FACULTAD DE CIENCIAS.
Place of preparation: FAUCLTAD DE CIENCIAS.

Summary: We have studied various aspects of ecológia of procesionaria pine (Thaumetopoea pityocampa), a lepidopteron heterócero, family Thaumetopoidaes, which is the principal scourge defoliadora of different species of pine in the Mediterranean area. The field work was conducted during the period 1993-1996, in a pine reforestation composed of Aleppo pine (Pinus halepensis) and maritime pine (Pinus pinaster), located east of the province of Granada. Due to the different characteristics of the leaves of two species of pine, we study the selection of species of pine, found, at least for the variables considered (infection rate and number of put attacked by pine), a preferential selection by female procesionaria, of Aleppo pine. Moreover, the selection of this species carries a most successful hatching and pupation of the set due to a lower incidence of parasites. Through experiments transposing larvae between the two pine species in the study area, we found a certain adaptability of larvae exploiting hospoedadores subóptimos. The selection of host subóptimos (maritime pine) can be explained by the reduction of competition for larval food asiciada to this selection. Among the species of pine optimal (Aleppo pine), we found that procesionaria selects those copies that have a higher rate of growth in height, acículas larger, asymmetric addition, due to an inverse relationship with the defensive compounds pines. This preference for copies of pine largest tamño seems hide, due to stronger growth of these pines, the negative impact of defoliation caused by the porcesionaria. As parasitism delas puttings procesionaria demonstrate that it is an important determinant of failure in the hatching of eggs procesionaria. To be covered by these flakes seems that the function of these is to prevent the exact location of the parasite eggs in the host. Regarding the gregarismo introducing this species in larval state, we found that the sizes of large group promotes growth and larval survival and a size of 32 individuals marks a turning point in the advantages and benefits of being in a group higher or lower for that number.

Author: ENGUITA MARTÍNEZ MARTA.
Year: 2004.
University: BARCELONA.
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA.

Summary: The fate of a neuron depends on the extracellular signals it receives and its inherent ability to respond. The depolarization of the plasma membrane is one of the major incentives that promote the survival of a neuron, both during embryonic development and in their adult life. It has been postulated that K + could act activating the intracellular signaling pathways of life and also blocking the tracks of death. However, the mechanism that has depolarization induces cell survival has not been established with precision and might depend on the cell type. Previous studies conducted in our laboratory have shown that, in the granular cells of the cerebellum in culture, the program of death that is induced when withdrew concentrations despolarizantes K + depended on the overexpression of NP1, a glycoprotein that is expressed only in the nervous system . Based on these data, work goals were: 1-To study the influence of intracellular signaling pathways in the process of deprivation-induced death of K + in the cerebellum granule cells in culture. 2, discussing the signal transduction pathways that regulate the overexpression of NP1 that occurs by reducing the activity of neurons in the model of deprivation of K + in granular cells of the cerebellum. 3-Identify a strategy to protect long-term death induced reduction of activity. Our results show that activation of the path of PI3K-Akt by IGF-1 protects so transient granular cells of the cerebellum of death induced reduction of activity, but does not block the overexpression of NP1 that occurs in cells closed K +. The signaling pathways of death that were studied were those of the stress-activated kinases (p38MAPK and JNK) and the way in which it participates GSK-3. Our results show that the route of p38MAPK not involved in the death process that takes place in the deprivation of K + in the cerebellum granule cells in culture. Inhibition of JNK route through a pharmacological inhibitor, CEP-11004-2, blocking a complete, but transient, the process of neuronal death that occurs when reduced neuronal activity. Despite being neuroprotective, the inhibitor of JNK route does not preclude the overexpression of NP1 that occurs when deprive the granular cell cultures of concentrations despolarizantes K +. Regarding GSK-2, the experiments show that the treatment of granule cell cultures with SB-415286, inhibitor of GSK-3 is also transient. However, when dealing with private cultivation of K + with the inhibitor route and JNK inhibitor of GSK-3 in conjunction triggering long-term survival. These data suggest that the signaling pathways of JNK and GSK-3/NP1 are the main channels responsible for the death induced by reducing activity and participating in the process independently.

Author: GONZALEZ RUBIO JUANA MARIA.
Year: 2004.
University: AUTÓNOMA DE MADRID.
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA.

Summary: The nicotinic acetylcholine receptor is a structure widely distributed at the central and peripheral nervous system, which is involved in many physiological processes important to the agency. However, in últomos years have proved their involvement in different patotogía charged particularly important in Western society such as dementia (Alzheimer's, Parkinson's, etc.). In this thesis is a study of one of the most striking features of these receptors, as is its permeability to Ca2 +, using as a model cell receptors nicotinicos that are expressed in the cell cromafin bovine, through its expression in Xenopus laevis oocytes . Based on the results obtendos, one might conclude that bovine nicotinic receptors expressed in oocytes are highly permeable to Ca2 +. In addition, different concentrations of this cation regulate the activity of this nicotinic receptor in a positive way, increasing their peak flow and cargo that passes through the receiver after activation. The incorporation of the subunit a5 receptor heteromérico a3b4 causes significant changes in the permeability of Ca2 + described on this receiver.

Author: CLAVERO VILLARRUBIA SONIA.
Year: 2004.
University: AUTÓNOMA DE MADRID.
Place of defense: DEPARTAMENTO DE BIOLOGIA MOLECULAR.
Place of preparation: CX 210.

Summary: The main objective of this work was to characterize the molecular basis of acidaemia propiónica. To this end, we genotipado patients acidemia propiónica identifying new allelic variants associated with this disease. Also, there has been a structural and functional analysis of the different allelic variants to know the molecular mechanism responsible for its pathogenesis. The analysis has enabled mutacional characterize a total of twenty-three allelic variants in patients analyzed. The analysis protein in fibroblasts of patients provided additional information on the physiological effect. The effect structural and functional eleven variants allelic PCCA has been analyzed by modeling and structural expression in eukaryotic systems. The protein PCCA human has two major domains: the biotin carboxylase domain and the domain of biotinilización with mutations localized in both domains. Due to the sequence homology in the polypeptide and the reaction catalysed, biotin carboxylase and the subunit - subunit carries carboxibiotina (BCCP) of the Acetil-CoA carboxylase of E.coli were chosen as models structural domains PCCA. The molecular modeling domain of biotin carboxylase protein PCCA shows that all waste affected by mutations PCCA (with the exception of the residue M229) are located in the same region of the protein, as opposed to the active site, defining a domain probably important from the viewpoint structural and / or functional protein. The mutation M229K affects a residue located in a hydrophobic pocket near the ATP binding site, so it can be expected that the effect can be catalytic. In a eukaryotic expression system, all variants allelic PCCA have an enzymatic activity on PCC void, with the exception of A75P, A138T and I164T, who have a moderate activity PCC. The change nucleotídico G668R, located in the domain biotinilización affects the union of biotin. The effect of several structural changes PCCB has been studied by the different location of waste involved in the structure 3D subunit 12S of the transcarboxilasa P. Shermanii, revealing most of the changes nucleotídicos alters interactions intra-and intertriméricas ring hexamérico. In the latter part of this study has analyzed the potential impact of the alternatives on PCCB allelic PCCA and the splicing mechanism. To do so, it has done an in silico analysis of the presence of ESEs and its affectation by mutations exónicas punctual. The results predict that this program were validated experimentally by minigenes, revealing no missense mutation analyzed appear to affect the processing of the messenger. The prediction of the secondary structure of mRNA through thermodynamic parameters is an approximation to explain the possible effect of some allelic variants on the processing of messenger, as is the case with the alternatives PCCA c.184-17delTG and c.231 +44 del4. The methodology of real-time PCR using specific primers has enabled the quantification of tránscritos normal in patients homozygous for mutations splicing c.1899 +3 del4 and c.2041-2A> G. Furthermore, this methodology and the use of probes alelo-específicas have been identified tránscritos normal from the mutation c.1091-11del6 in a heterozygous patient. The results previously described for mutations PCCA and PCCB, provide important information about the complexity of the correlation genotipo - fenotipo, and suggest that, in some cases, regulation of the splicing mechanism could also influence the parameters of phenotypic acidemia propiónica.

Author: MARTIN DE LARA FERNANDO.
Year: 2004.
University: AUTÓNOMA DE MADRID.
Place of defense: FACULTAD DE CIENCIAS.
Place of preparation: FACULTAD CIENCIAS.

Summary: The main aim of this thesis has been the study of the entire party related to the possible biological function of FoxJ2, a transcription factor, transactivador of the transcript of the family Fork Head. The transcription factors Fork Head of the family, are characterized by a domain binding to the DNA, called domain Fork Head. The previous work conducted in the laboratory led to the cloning and characterization of this novel protein, as well as the determination of both the levels of regulation and the pattern of expression of the same. The question that still has not been given an answer, was that it might be associated with the biological function of this transcription factor. We used different approaches to attempt to answer this question. The first one was to try to determine the pattern of expression of FoxJ3, protein, which showed a greater degree of homology at the level of amino acid sequence with FoxJ2, on the assumption that if both shared the same pattern of expression, probably with potential partners or redundant functions . The second and third approach was used respectively, to generate transgenic mice of overexpression of FoxJ2, and by means of analytical seek to identify genes that could be regulated by FoxJ2, containing in its regulatory regions (promoters and enhancers) binding sites of this transcription factor. Through these approaches succeed somehow suggest that the function could be carried out by FoxJ2 during embryonic development and in the adult. Thus, the role seems to be carrying out FoxJ2 in adults, is a regulatory function quite widespread, and that it would be carried out by regulating genes, presumably Conexina 43 and Cadherina-Ey in a provable in major organs such as the heart or gonads. Thanks to the experimental approaches described above, we see a number of phenotypes that lead us to propose which could be the functions FoxJ2 conducting both during development pre-implantación that this probably related functions compaction morular and formation of the blastocyst. As in post-implantation stage where it is suggested an important regulation FoxJ2 on Conexina-43 in the heart in the embryonic stage of E10, 5. These are thus the final outcome of a trial that was proposed as an eventual goal try and determine what you are doing, what is the role of FoxJ2.

Author: QUESADA NAVIDAD ANTONIO JESUS.
Year: 2004.
University: AUTÓNOMA DE MADRID.
Place of defense: CENTRO DE BIOLOGIA MOLECULAR SEVERO OCHOA.
Place of preparation: CENTRO DE BIOLOGIA MOLECULAR SEVERO OCHOA.

Summary: The ability of tumor cells to induce and maintain vasculature is critical for the development of tumors. Thus, tumors promote the development of a vascular network within them, a process called angiogenesis. The doxorubicin (DXR) is the antineoplastic choice in a wide variety of processes both solid and liquid tumors. The problem of DXR, like that of similar agents, are the side effects that its use causes in the long term. Previous work in our group described that DXR induced apoptosis in cells primaras human from the umbilical cord vein (HUVEC), also described as causing an increase in surface expression of CD95, a receptor death belongs to the superfamily of TNF . Trombospondina-1 (TSP1) is an endogenous protein, which many have described biological actions depending on the recipient and which is a type of cell on the act. TSP1 capability antiangiogénica, because it is capable of inhibiting the migration, adhesion, proliferation and morphogenesis of the endothelial cells. It has been shown activity antiangiogénica depends on its binding to CD36, a membrane protein that is expressed, among others, in human cells microvasculature. When activated CD36, triggering a gene that causes the expression on the surface of CD95L, ligand natural CD95. The ability antiangiogénica of TSP1 stems from the fact that in endothelial cell proliferation is in the presence of factors proangiogénicos, producing a change in the subcellular localization of CD95, was carried from the Golgi apparatus to the cell membrane. The binding of ligand and receptor on the surface of endothelial cell apoptosis occurs. However, if the concentration of proangiogénicos exceeds a certain threshold, the effects antiapoptóticos of them will prevail. The aim of this study was to increase the susceptibility of activated endothelial to DI-TSP a nonapéptido derivative TSP1. DI-TSP seems to be more suitable for use in the clinic because it is only capable of joining activate CD36. We investigate whether low doses of DXR, which had no apparent side effects, in combination with subtherapeutic doses of DI-TSP were capable of increasing endothelial apoptosis, inhibit neovascularization and delay tumor progression in a synergistic manner. The combination of these agents in vitro caused an induction of apoptotic events in cultured human endothelial cells. As we move to study in vivo models, we chose an administration metronómica (very low doses of agent administered at regular intervals, resulting in a lower toxicity) DXR and DI-TSP. As doses of DXR use a dose choose between 25-100 times lower than the therapeutic traditional DI-TSP was administered daily at a concentration 60 times lower than that described as effective in vivo. When combined the two substances, reduced microvascular density and tumor progression in a synergistic manner. In addition, the synergistic effect was due to the induction of apoptotic signals triggered by the activation of the path of CD95/CD95L cells endoteliales.Las therapies antiangiogénicas represent a new option for the treatment of tumors in which attacking the endothelial cells in growth in a tumor. This is a type of cell genetically more stable than the tumor cells, so it is less likely to acquire resistance to treatment. In addition, since virtually all tumors depend on angiogenic processes, is a therapeutic scheme that can be used in a wide variety of treatments. With this paper we have demonstrated that the rational combination of substances based on their effects on the endothelium can be an attractive therapeutic option for the development of improved anti-tumor therapies.

Author: MOLINA ARRANZ SUSANA.
Year: 2004.
University: AUTÓNOMA DE MADRID.
Place of defense: CENTRO DE BIOLOGIA MOLECULAR SEVERO OCHOA.
Place of preparation: CENTRO DE BIOLOGIA MOLECULAR.

Summary: Gender Alfavirus, which includes the virus Sindbis, comprises virus genome RNA chain simple and positive polarity. The genomic RNA (49S) serves as a mold for the synthesis of RNA polaridd negative is utiizado to generate more copies of the genome. Using an internal promoter, the RNA can be transcribed creating a RNA subgenómico (26S), from which summarizes the structural proteins. At times late in the infection, only RNA 26S translates, while RNA 49S and protein synthesis is inhibited host. At this very moment, there is an increase in the phosphorylation of the alpha subunit factor initiation of translation eIF2. Despite this, the synthesis of protein structural continues for hours. The Brefeldina A (BFA) is an antibiotic that disrupts the system of vesicular eukaryotic cells. The BFA has been used as an antiviral agent with virus wrapped in preventing the maturation of glycoproteins. This compound can also act on some viruses without wrapped, as it interferes with the replication of the genome, which requires a vesicular system intact. In this paper, we analyzed the effect of BFA on the translation of RNAs of BHK Cells - 21 and the virus Sindbis. Moreover, it has compared the action of the BFA with other compounds, such as Ditiotritol (DCT), an inhibitor of protein folding in the endoplasmic reticulum. Both treatments exert an inhibitory effect diferencual on the synthesis of cellular and viral proteins, the latter being more resistant. Both the BFA as the DCT induce phosphorylation of subinidad alpha factor eIF2. Furthermore, we have built a series of replicones with mutations that alter the initiator AUG codon or the secondary structure of RNA 26S. These replicones have been used to examine the dependency factor eIF2 the translation of RNA subgenómico, as well as to try to clarify the significance of the secondary structure or sequence of this RNA leader in the resistance to treatment with BFA and DCT.

Author: BLANCO FUENTES RAQUEL.
Year: 2004.
University: AUTÓNOMA DE MADRID.
Place of defense: CENTRO DE BIOLOGIA MOLECULAR SEVERO OCHOA.
Place of preparation: CENTRO DE BIOLOGIA MOLECULAR SEVERO OCHOA.

Author: JIMENEZ PALACIOS FRANCISCO JOAQUIN.
Year: 2004.
University: SEVILLA.
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTAD DE FARMACIA.

Summary: After conducting an exhaustive review of the literature on the cefepime antibiotic, were on the verge various analytical methods for determination specialty pharmaceutical and biological fluids. Thus, we propose a method voltamperométrico by Polarografía differential pulse that is applied to the identification and quantification of cefepime in especialidaes pharmaceutical and urine and serum samples from patients who had been treated with this drug, a method by voltammetry of redisolución adsorptive Cathodic and its application to real samples in urine and cerebrospinal fluid obtaining high sensitivity, a method by high pressure liquid chromatography effectiveness and its application to samples of urine, serum and cerebrospinal fluid, and a method by ultraviolet spectrophotometry for the quantification of cefepime in proprietary products and the determination of dissociation constant. Finally, with the data obtained in the quantification of cefepime in real samples in different biological fluids, a pharmacokinetic and pharmacodynamic study of that drug for results of the clinical behavior of cefepime in hospitalized patients in the Intensive Care Unit of Hospital Universitario Virgen del Rocío Seville, and to establish dosage regimens dosing after computer simulations.

Author: Mulero Cánovas Juana.
Year: 2004.
University: MURCIA.
Place of defense: Facultad de Biología.
Place of preparation: de Murcia.

Summary: Wine is a product of millennial tradition around which has developed a culture of its own, but beyond the pleasures and the popularity of wine there are studies that attributed to moderate consumption prevention of cardiovascular diseases. Moreover, the spectacular development of organic farming in recent years has also come to the field of viticulture and thus to the emergence of wines produced from grapes ecological, but the question arises whether there's growing influence on the nutritional quality eco the grape and wine both made from it. Therefore the objective of this work is to study the influence of organic farming in the composition and phenolic antioxidant activity of red wine grapes and the variety Monastrell and their comparison with grapes and red wine of the same variety obtained from techniques traditional cultivation and the study of the influence of different vinification techniques (traditional red wine, wine with maderación prolonged fermentation with the addition of tannin wine and wine with added enzymes ecological) on the antioxidant activity and phenol compounds wine tiento made from two types of grapes. After the study results indicate that in the case of the grapes are not significant differences in the concentration of phenolic compounds and antioxidant activity of grape traditional ecological and on the date of the harvest. There were no significant differences in the composition and phenolic antioxidant activity in the wine made from grapes and wine ecological produced from grapes in the traditional winemaking traditional red as in the rest of vinificaciones made, despite the fact that both parameters are higher in organic wines.

Author: LEÓN LÓPEZ ANA MARÍA.
Year: 2004.
University: GRANADA.
Place of defense: ESTACIÓN EXPERIMENTAL DEL ZAIDÍN (CSIC).
Place of preparation: ESTACIÓN EXPERIMENTAL DEL ZAIDÍN (CSIC).

Summary: Nitric oxide (NO) is a gaseous molecule and a free radical that, in animals, it acts as a signal molecule in the regulation of various physiological process and patofisiológicos in vascular systems, immune and nervous. It was synthesized by a group of enzymes called óxidonítrico sintasas (NOS). In plants, much less is known about this species radical, but it seems that participates in processes such as the regulation of growth and the response of the plant compared to different stress. However, the identification of potential sources of NO enzyme has been very controversial. In this thesis, has been studied in pea plants modulation of endogenous NO in the development postgerminativo in roots, stems and leaves, and to different stress situations, to elucidate their physiological implications. Thus, it has been found that NO is synthesized primarily in the shoots of pea plants during the first days postegerminación and was the first reference that exists on NO production in a plant stalks higher. NO also could intervene in the plant's response to stress by extreme temperatures or high light intensity. Moreover, it has tried to identify source of NO enzyme in the leaves. It has been shown that this kind of radical synthesized form enzyme in the peroxisome leaves pea through a type NOS activity. These organelles present an essentially oxidative metabolism, and contain various antioxidant enzymes, as well as being a source of reactive oxygen species. The NOS activity of peroxisome pea leaves depends on the same substrate that NOS animals, L-arginine and HADPH, and also needs the cofactors calcium, calmodulina and tetrahidrobiopterina. The ability of peroxisome NO synthesis of endogenously confirms the importance of these organelles as a source of signal molecules, which could be especially important in cases where it has been observed that there is an increase in the population peroxisomal, as during senescence and to various abiotic stress situations.

Author: HACKENBERG MICHAEL.
Year: 2004.
University: GRANADA.
Place of defense: UNIVERSIDAD DE GRANADA.
Place of preparation: UNIVERSIDAD DE GRANADA.

Summary: The retrotransposones Alu elements are transponibles more abundant in the human genome. Since its discovery, it has been suggested that may have played a role in the evolution of primates. A prominent feature of Alus is their differential distribution along the chromosomes according to their evolutionary age: Alus younger show greater density in regions rich in A & T, while the oldest are more dense in regions rich in G + C. The main objective of this thesis is to explore the evolutionary dynamics of Alus in the human genome and to identify the major factors involved in creating and maintaining their spatial distribution along the chromosomes. To achieve this goal have been implemented in the first place a number of methodological improvements: 1-Development of a collection of Perl programs to process the data generated first by RepeatMAsker and IsoFinder. 2-correction for multiple substitutions, applied here for the first time alignments of Alus, in order to estimate more accurately the evolutionary age. 3-Development of a technique for defragmentation of Alus to eliminate usual overestimation of these elements and better align their actual number in the genome. 4-Definition precise techniques in silico, the trímeros of Alus and the Alus lonely, which has led to trace the genome in search of the traces left by recombination. 5, - Development of a new measure, independent of density, for the study of clusterización of Alus. Then buying the density of elements with different evolutionary age at different isocoras, discarded some of the mechanisms proposed to explain the change in density: the interaction Alu/LINE1 (competition for retrotransposasa), the compositional adjustment, and the selection positive (preferential accumulation in the environment of genes). Subsequently, results are presented that demonstrate the involvement of recombination in changing density. This has been made possible by defining silicon in the trímero resulting from a process of unequal homologous recombination (RHD) between Alus, and the status of the process of integration into the computer, making it possible to trace the genome in search of the traces left by recombination and quantify its impact on the change in density. There has been a higher proportion of trímeros in isocoras L, which indicates greater activity RHD, or greater survival of their products in these isocoras. This suggests that the RHD could help change the density Classrooms. The analysis of the evolutionary age of the subfamilies of recent insertion and more frequent Alus solitary point to the recombination as the causative agent of change in density because it is the only mechanism whose business depends on the age of the elements. It has also tested the process aglomerción (or clusterización) of Alus along the chromosomes. It begins by proposing a new measure, independent of the density, to quantify the clusterización in the spatial distribution of Alus. Then, through simulation of the process of integration, are associated levels of statistical significance (valores-P) at frequencies clusterización observed. Thirdly, the incorporation into models of inserting different evolutionary mechanisms, has helped evaluate the impact of each of them in the generation and maintenance of clusterización. In parallel, it has also made an analysis of the clusterización of the goals of integration, having verified that this process can not explain the strong preferential accumulation in the environment Alu that can be seen in the isocoras H. However, the comparison between the frequencies of targets for integration and Alus in the sidewalls of other elements shows that the insertion preferential queue poli-A of Alus preexisting can contribute significantly to the strong clusterización observed in the isocoras H. Finally, the insertion into existing elements may also contribute to greater clusterización observed in these isocoras.

Author: CUERDA CORREA M. TERESA.
Year: 2004.
University: GRANADA.
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTAD DE FARMACIA.

Summary: The liposome vesicles are formed by one or more bicapas lipid concentric formed spontaneously. They are potential vehicles for drugs that are incorporated according to their solubility, at bicapas lipid or aqueous compartments. Moreover, archaea are microorganisms that have evolved under the conditions prevailing in the primitive Earth: high temperature and salinity and anaerobic atmosphere. They are divided into three groups: extreme halophytic (haloarqueas) metanógenas, termoacidófilas (extreme thermophilic). So far only been tested obtaining liposomes from the lipid membrane archaea metanógenas and have been called arqueosomas. The membrane lipids tested in this work for the development of arqueosomas come from halophilic archaea extreme. These microorganisms require very high salt concentrations (20-30g NaCl/100mL) and it is not possible to replace the salt by other solutes not ionizables as sugars. Only KCl can partially replace the NaCl. They also need abnormally high concentrations of magnesium, iron and other trace elements. The objectives of this report are: 1-To study the ability to form liposomes from different strains of collection haloarqueas and evaluate the properties of these arquesomas. 2-To develop and validate a method of valuation of the active subsequently encapsulará in arquesomas and liposomes. 3-Designing formulating arquesomas and liposomes allowing to optimize the properties of ether and ester liposomes, and the uptake of the drug. 4-Consider the influence of the composition of the arquesomas and liposomes in attracting corticosteroid and the ownership of these morphological vectors. 5-To assess the stability of the various formulations developed with both types of lipids at room temperature and 4-8Â ° C. The methodology highlights two distinct parts: 1-lipid extraction and precipitation of the polar fraction. 2 - The development of arqueosomas and liposomes. Of the 18 strains tested collection, 14 have shown ability forming arqueosomas, preseleccionando for four of them following studies: Haloarcula califomiae ATCC 33799, Halobacterium salinarum CECT 396, Halococcus morhuae NCMB 757 and Halorubrum coriense DSM 10284. The properties and has chosen Halobacterium salinarum CECT 396 to develop the following investigations raised at the beginning of this work. The design optimized formulations areuosomas and liposomes concluded that the proportion "lipids: active" best was 15:1 in arquesomas and 40:1 in liposomes. In both cases, the stirring time selected was the lowest. The results of the study prior to encapsulation of the drug has been quite satisfactory for both arqueosomas as liposomes, ranging between 42% and over 80% of the active withheld. The characterization of samples of the various formulations confirmed the blistering lipid multilaminares carriers active in all formulations tested. The mean diameter of the vesicles formed has been very small, less than 1 m in all cases. The analysis of the stability of arqueosomas and liposomes revealed that the formula of choice would be developed with lipids, cholesterol and the combination of antioxidants. He stressed that the long-term stability is greater in arqueosomas.

Author: ARCO PRADOS YOLANDA.
Year: 2004.
University: GRANADA.
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTAD DE FARMACIA.

Summary: Halomonas maura is a bacterium halophytic asylum status was moderate soil next to the salt of Asilah (Morocco). This bacterium is a Gram negative bacillus that is characterized by a diazotroforo living free. Its genome, with a size of 3,574 kb, consists of a circular chromosome and two plasmids of 619 and 70 kb. Halomonas maura S-30 summarizes an EPS called maurano with key properties that make it useful for use in various fields of industry. Among these functional properties highlights the viscous, and thixotropic pseudoplastic their preparations aqueous emulsifier and its activity on various phases oleasas, as well as the stability of aqueous such preparations under conditions of extreme pH and high concentration of solutes. In addition, the maurano is an anionic polymer in nature, it is capable of capturing heavy metals. In this Doctoral Thesis deals with the study of the determinants gene responsible for the biosynthesis of maurano. The cluster eps of Halomonas maura S-30 has been sequenced in the mutant defective in the synthesis of EPS, called TK71 and have so far been identified ten genes. Genes epsA, epsB, epsC and epsJ are involved in the transport and polymerization of maurano; protein EpsC regulates the length of the hydrocarbon chain. Genes epsD, epsE, epsF, epsG and epsH are responsible for the addition of sulfate substituents; protein EpsE is a regulatory process sulfation. It also has identified a glicosiltransferasa, encoded by the gene epsI, has been implicated in the synthesis of units oligosacarídicas. The genes described form a transcriptional unit headed by a promoter located in front of the gene epsA. By merging transcriptional gene lacZ gene epsA, seha found that the highest expression of genes eps comes to salt concentrations of 5% (w / v) and exponential phase of growth. In order to know the physiological function of maurano and drawing mutants deficient in the synthesis of it, has revealed the involvement in the formation of biofilms on inert surfaces.

Author: VARELA HERNÁNDEZ NIEVES YAQUELIN.
Year: 2004.
University: GRANADA.
Place of defense: INSTITUTO DE PARASITOLOGÍA Y BIOMEDICINA.
Place of preparation: INSTITUTO DE PARASITOLOGÍA Y BIOMEDICINA "LÓPEZ NEYRA", CSIC, GRANADA.

Author: Mora de Zayas Laura.
Year: 2004.
University: GRANADA.
Place of defense: Facultad de Farmacia.
Place of preparation: Facultad de Farmacia.

Summary: The production of bacteriocins (rizobiocinas) strains of Rhizobium genre has been repeatedly mentioned as a highly influential factor in competitiveness, as well as a feature susceptible to manipulation for the construction of improved strains. As part of a larger study on the interaction between microorganisms rhizosphere of common bean (Vicia faba L.), and within a project funded by the National Program for Agricultural Research (AGRI91-0714), the group Environmental Microbiology at the University Granada conducted a search strains of R. Leguminosarum bv. Viciae producing rizobiocinas. We isolated the strain Z25 (CECT 4585), which showed a potent activity bacteriocinogénica against all strains of R. Leguminosarum bv. Viciae used as indicators, with no inhibitory effects on other groups of microorganisms in the soil. These results suggest that the strain Z25 produces a rizobiocina with a spectrum of action restricted to the strains that can compete with directly by the nodulation, without affecting other saprophytes or beneficial microorganisms in the rhizosphere. Bacteriocina production by the strain Z25 of Rhizobium leguminosarum bv. Viciae is determined by genes rzcCADE, which shares homology with genes that are part of operons responsible for the production of toxins family RTX. Genes rzc are located in one of three megaplásmidos possessing strain Z25 of Rhizobium leguminosarum bv. Viciae, with an approximate molecular mass of 360 MDa. Rhizobium leguminosarum bv. Viciae strain Z25 carries counterparts genes cinR and cinI involved in mechanisms for collecting quorum mediated acil-homoserina lactones (AHLs). By the reaction of a primer extension, we have identified two regions flanking promoters in 5 'to the genes rzcC and rzcA respectively feature that has not been described previously in other strains of the genus Rhizobium producing bacteriocins the family of toxins RTX. The regulation of transcription of genes rzc responds to a complex model in influencing various elements. The extent of activity beta-galactosidasa mergers rzcC-lacZ and rzcA-lacZ has shown the presence of two possible regulatory regions, Region M R region, which act causing an effect repressor of transcription. The deletion of the region R, which is in 5Â 'regarding ORF3 causes deregulation of the activity beta-galactosidasa the merger rzcC-lacZ of plasmid pMAGR37 which, however, was not observed in the mutant Z25-cinI -, which shows a possible influence of the mechanism for collecting quorum in regulating the production of bacteriocina. Additionally, the mutation in the gene cinI significantly decreases production bacteriocina amid solid. Under conditions of the study, the production of bacteriocina is a determining factor in the competitiveness of the strain Z25 front of the strain M17 Spr (spectinomycin resistance) sensitive to bacteriocina, both in testing competition cocultivos in liquid medium in tests competition for the nodulation of plants pea (Pisum sativum cv. Frisson). The mutant in the gene rzcA is very aggrieved to compete with the strain M17 Spr sensitive to bacteriocina, compared with the parental strain. The mutation in the gene rzcA not affect the phenotype of nodulation of pea plants, but there has been a significant increase in activity nitrogenasa in symbiosis. In the mutant strain Z25 in the gene cinI and deletion of the region there is no significant difference R with the parental strain in 8 s paráme 2ae tions nodulation nor the symbiotic nitrogen fixation.

Author: GARCÍA FERNÁNDEZ MARÍA.
Year: 2004.
University: SEVILLA.
Place of defense: SALON DE ACTOS DEL EDIFICIO DE GOBIERNO DEL HOSPITAL UNIVERSITARIO VIRGEN DEL ROCÍO.
Place of preparation: UNIVERSIDAD DE SEVILLA.

Summary: The maintenance of a stable internal or homesotasia, is special for the living. Therefore there are systems that control changes in the variables that form to launch compensatory responses. Oxygen and glucose are two of the most important variables to be controlled. This thesis has been studied as cells cromafines of the adrenal medulla detect lower levels of oxygen during neonatal and adult rats and as cells glomicas body both carotid detect changes in oxygen tension as lower the concentration of glucose. For this study have been used various techniques including slices of tissue and cell cultures scattered. These preparations have been employed techniques of electrophysiology (amperometría, patch-clamp and fluorimetría), histological and molecular biology. The main results show that the ability of cells cromafines to detect directly the reduction in the partial pressure of oxygen decreases but does not disappear with the maturation of the rat. This does not appear to be due to a state of excitability in smaller cells cromafines adult animals. With regard to acute hypoxia sensor mechanism in the cells glomicales, we found that the secretory response to hypoxia is eliminated specifically when using the previously inhibitor complex i rotenora or MPP +. It seems, therefore, that a specific target of rotenone can this mechanism involved in this sensor. However, neither the nitric oxide nor the system of prolil hidroxialasas seem to be involved in detecting acute oxygen in the carotid body. Lastly, the reduction in the levels of glucose causes the secretion of catecholamines in the cells glomicas of rat carotid body following the same model quimiotransducción accepted for sensitivity to hypoxia. However, despite the observed inhibition of microscopic flow of potassium in some cells by removing glucose from the external solution, similar to what found during hipoxía seems that the hypoglycemia caused mainly the empowerment of a flow of sodium through a channel catiónico not selective. Experiment preliminary suggest the involvement of a TRP channel type. The mechanism of low glucose sensor cells glimicas requires glucose metabolism but not dependent on changes in the ATP generated via oxidative damage. In addition hypoxia and hypoglycemia not share the same way of detecting both stimuli but finally converge in the release of catecholamines. We did not find the glucose transporter of low affinity glut2, although the enzyme glucoquinasa which has been linked in other systems with the sensor mechanism hypoglycaemia. However, their presence in other nodes adjacent to the carotid body and in different cell types within the same (not only in the cells glimicas sensing of glucose) is necessary experiments to confirm his involvement in this mechanism sensor.

Author: LUQUE LAÓ M. ÁNGELES.
Year: 2004.
University: SEVILLA.
Place of defense: DEPARTAMENTO DE FISIOLOGÍA Y ZOOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA.

Summary: The biological significance of the movements of orientation is shifting eyes, head and / or body to stimuli that appear on the live animals, including man, for projecting this boost to fóvea. The neural circuit involved in these movements has been extensively studied in mammals, but not in other vertebrate species. In particular current knowledge on the formation of lattice mesencéfalo are restricted to different species of primates. This paper discussed the implication of this area in the generation of eye movements in teleósteos fish. Both were used electrophysiological techniques of electrical stimulation as morfológias plot routes. The most relevant results can be summarized as follows: 1 - The formation of lattice mesencéfalo is an area tectorecipiente major quantitative. 2 - The connection tectoreticular is mutual. 3 - The connection tectoreticulotectal is organized anatomic and functional. 4 - Electrical stimulation of the formation lattice mesencefálica shows the involvement of this area in the generation of eye movements guidance. 5-In training lattice mesencefálica can differentiate functional areas according to the characteristics of the movements they produce. 6, - Since the formation lattice mesencefálica can generate five different types of rapid eye movement or sacádicos. The metric and kinetic characteristics of these movements depend on the parameters of stimulation. 7 - The source output tectal towards the formation of lattice mesencéfalo comes from the layers of intermediate and deep. These same layers are reticulorecipientes. 8-Interconnections reticulotectales depend on the functional properties of the lattice area. Together these results suggest that the formation lattice mesencefálica is a structure crucial in the generation of eye movements and orientation of this feature appeared early in the filogenía vertebrate.