LIFE SCIENCES, 15

Author: GUILLEN FRETES ROSA MARIA.
Year: 2006.
University: LA LAGUNA.
Place of defense: INSTITUTO UNIVERSITARIO ANTONIO GONZALEZ.
Place of preparation: UNIVERSIDAD DE LA LAGUNA.

Summary: The assimilation of nitrate is one of the most important route for the acquisition of nitrogen in the biosphere and thus for the incorporation of inorganic nitrogen. H. Polymorpha yeast is able to assimilate nitrate as the sole source of nitrogen and has been used as an experimental model for studying the way. Our present knowledge of the rules of the road of assimilation of nitrate pose to the overall process is the result of the balance between positive signals (induction nitrate) and negative signals (suppression by reduced nitrogen sources). To date, researchers have isolated genes involved in the induction nitrate (YNA1 and YNA2) and the suppression / desrepresión by nitrogen sources preferential (URE2, GAT1, GLN3, GZF3, DAL80 and DEH1), in addition to indirect evidence indicates the involvement of signaling mediated via the Tor kinase.

Author: Ramírez Hidalgo Cristina Micaela.
Year: 2006.
University: LA LAGUNA.
Place of defense: Universidad de La Laguna.
Place of preparation: Facultad de Medicina.

Summary: In this dissertation work are identified and characterized a number of ways to estrogen receptor alpha in the plasma membrane of the cells SN56 and HT22 and brain areas particularly affected in degenerative processes such as the septum, the cortex and the hippocampus. These cellular models demonstrate the involvement of a membrane channel VDAC, traditionally known as porina mitochondrial, the preservation neuronal versus toxic effect of beta-amyloid peptide. Studies on the estrogen receptor membrane and VDAC, as well as previous data indicating a possible link between molecular, led to the first experimental evidence of the association between these two proteins on the neuronal plasma membrane. These new data on the association of estrogen receptor membrane and VDAC and on the location joint signaling domains such as caveolas, open up new avenues for study on the modulation and regulation of this protein complex in relation to the phenomena of estrogen initiated neuroprotection at the plasma membrane.

Author: SICILIA MARTÍN DESSIRE C..
Year: 2006.
University: LA LAGUNA.
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTA DE FARMACIA.

Summary: STUDY TAXONÓMICO AND SYSTEMATIC THE Biota LIQUÉNICA OF GARAJONAY WITH DATA TO CLOSE ITS DISTRIBUTION IN THE PARK AND THE CORRELATION TO VEGETATION CORMOFÍTICA. UTILIZATION OF LÍQUENES AS BIOSENSORS THE POTENTIAL OF POLLUTION FROM CENTRAL OF THERMAL GRANADILLA (SUR DE TENERIFE).

Author: CABALLER GUTIERREZ MANUEL.
Year: 2006.
University: CANTABRIA.
Place of defense: E.T.S. DE INGENIEROS DE CAMINOS, CANALES Y PUERTOS.
Place of preparation: E.T.S. DE INGENIEROS DE CAMINOS, CANALES Y PUERTOS.

Summary: In order to determine the identity of the cryptic species of sacoglosos and opistobranquios present in the bay of Santander, a revision of the 5 genera contain: Hermaea Lovèn  ¡, 1844; Hermaeopsis Coast, 1869, Placida Trinchese, 1879, Cuthona Alder and Hancock, 1855 and Eubranchus Forbes, 1838. It looks for this material from both sides of the North Atlantic. In gender hermaea, three species have proved to be new to science, one of them has already been published (Hermaea ghanensis Caballer, Ortea and Moro, 2006). A fourth, Hermaea Coirala Marcus, 1955, was redescribe citándola both sides of the Atlantic. Additionally, describes the first Atlantic species of the genus Alderiopsis Baba, 1968, Alderiopsis hook, Caballer, Ortea and Esponosa, 2006, mixed with the past. In gender Placida, establishing the diagnostic characters of all Atlantic species of the genus with the digestive green, clarifying the historical problem of its synonyms. It redescribe a species was known only apartir of its original description. Placida dakariensis (Pruvot-Fol, 1953). In gender Cuthona, it is shown that assumption to Cuthona genovae (O'Donoghue, 1926) - Cuthona folaita (Iforbes and Goodsir, 1839), is actually a single species. In gender Eubranchus, recovers the name Eubranchus capellinii (Trinchese, 1879), a sort traditionally considered synonymous with Eubranchus doriae (Trinchese, 1873), which itself remains uncertain. Additionally redescribe another kind cryptic Eubranchus cingulatus (Alder and Hancock, 1847). Also cited for the first time by the Iberian Peninsula recent description of a species, Eubranchus vascoi Ortea, Caballer and Moro, 2002. There will always be a catalog illustrated and commented on the 82 species of mollusks sacoglosos and opistobranquios collected in the Bay of Santander, giving details of their anatomy, biology and distribution. Seven of these species are cited for the first time for wildlife Iberian.

Author: TURON PLANELLA CLAUDIA.
Year: 2006.
University: GIRONA.
Place of defense: FACULTAD DE CIENCIAS..
Place of preparation: FACULTAD DE CIENCIAS.

Author: PRADO VILLEGAS PATRICIA.
Year: 2006.
University: BARCELONA.
Place of preparation: UNIVERSIDAD DE BARCELONA.

Summary: The reassessment large spatial scale (less than 300 kilometers) and by methods of quantifying direct impact herbivorísmo in ocean ecosystems Posidonia reveals losses of foliar biomass much higher than previous estimates based on indirect methods (ie, frequency of brands action). On an annual average, herbivores cause the loss of approximation for 57% of production foiliar, although there are big differences seasonal and increased pressure occur in the summertime. The fish Sarpa salpa stands as the main herbivore, with rates of defoliation of ca. 40% of production leaf cancels (70% of herbivorísmo) followed by the hedgehog sea Paracentrotus lividus (17% of the leaf production and 30% of herbivorísmo). However, it also detects the existence of a large spatial variability, with pressure values of herbivorísmo fluctuating between 23 and over 85% of production leaf depending on the locality. Among the factors behind these differences, fishing and the availability of P.oceanica as habitat and as a resource for food, were identified as the most influential on the population structure of S.salpa. The effect of fishing pressure resulting in a decrease in the number of individuals and hence the pressure herbivorísmo. By contrast, the effect on the availability of P.oceanica within the territorial extent of S.salpa, because its concentration on the appeal, and therefore, the increased pressure deherbivorísmo. As regards the sea urchin P.lividus, the contribution of new individuals to the population is produced by two fundamental process: the migration from the rocky substrate in contact with prairie and the availability of structure kills the rhizome of P. oceanica as an element associated with the shelter and / or feed the early stages of this kind and without which it is detected recruitment. Other factors such as variability of quantitative and qualitative changes induced by the availability of nutrients (eg biomass and composition of the communities epífitos leaf, content of nutrients in plant and epífitos), but capable of disrupting preferences s.salpa by P.oceanica, possess, the spatial scale study, a range of less than variability factors - type landscape and the influence of fishing. Therefore, the results of this thesis show that the pressure herbivorísmo can reach magnitudes locally relevant and very resulting from the interaction between human alterations in the environment (eg, habitat loss and reduction in the abundance of herbivores) and biology of the species to its particular habitat use.

Author: FERNÁNDEZ SANTIDRIÁN ANTONIO.
Year: 2006.
University: BARCELONA.
Place of preparation: UNIVERSITAT DE BARCELONA.

Summary: The doctoral thesis has studied the mechanisms of induction of drugs that directly affect the mitochondria, such as 2-metoxiestradiol and PK11195 in leukemia linfocíticacrónica. It has also been studying the mechanism of induction of apoptosis by acadesina in chronic lymphocytic leukemia, and has analyzed the effects of acadesina on lymphocytosis in vivo in mice Balb / c During the work we have shown that 2-ME induce apoptosis through a mechanism in which it could be implicated VDAC or chlorine mitochondrial channels. However, normal B cells are equal to sensitive cells LLC.2-ME induces genes NOXA and PUMA. An interesting observation is that the Ruthenium Network (RuR), an inhibitor of calcium channel unidirectional of the mitochondria, in a clear sensitizes the cells LLC to very low doses of 2-ME. Following studies with 2-ME, we have analyzed the effect of other drugs action mitochondrial target as having one of the proteins that form the PTPC to study the effect of altering this group of proteins in apoptosis cells LLC. PBR is a protein found sobreexpresada in the LLC. Note that PK11195, a ligand of PBR, induces apoptosis in B cells LLC and also in the B lymphocytes of healthy donors, whereas T lymphocytes are more resistant. Yet now, the target propaoptótica of PK11195 is unclear and the involvement of PBR is controversial. It is the first time described that PK11195 induce apoptosis, and not just awareness, in leukemic cells primaries. In addition, the study of the mechanism has shown us that PK11195 induce apoptosis independently of p53, point of interest for possible use in patients refractorios to chemotherapy today. The main aim of the thesis was to study the mechanisms of induction of apoptosis by acadesina in cells LLC. Before the start of the thesis observábamos that activation of AMPK always correlacionaba to induction of apoptosis. However, we have shown that activation of AMPK, other activators as fenformina or A-769662, is insufficient to induce apoptosis. We noticed that acadesina induces a decrease of intracellular ATP. Furthermore, we noticed that acadesina induce apoptosis through mitochondrial track, and induces changes in the family members of BCL-2. Lastly, acadesina is a drug that may come during the year 2007 in clinical trials. Because of the work done, we have shown that acadesina may also have effects on the in vivo mouse lymphocytosis, and although we must wait for testing in humans, this is a hopeful figure towards having an activity after systemic administration.

Author: BAO CUTRONA MERITXELL.
Year: 2006.
University: BARCELONA.
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA (UNVIERSIDAD DE BARCELONA).

Summary: The main aim of the thesis was to create a conditional system for in vivo study (transgenic mice), in which the expression of a protein (SOD1, carrier dela mutation G93A) could be controlled. So far Sclerosis Side Amiotrófica of the family has been analyzed using transgenic models classics. Since these models have been developed on different scenarios which could fuer relationship causa-consecuencia between the protein SOD1 and the molecular mechanisms that cause degeneration (death of motoneuronas) and therefore símptomas clinical manifested in mice models. But the mechanism / s true that triggers the disease is still unknown. For this reason, we decided that it would be interesting to be able to control the lapsus of time that the mutant protein was present in the cells of the body transgéncio, to be able to study the reversibility the fenótipo, and understand that key mechanisms were implicated in the pathology. To make this work, we set out to build a conditional system (based on tetracycline; tet-off) supported by successful examples published. In particular, the system tet-off is composed of two elements: an activator (of expression) and a responsive element (from which controls the expression of the gene cloned human and interest). Therefore, we selected this system, since it allows us to the cDNA clone of the SOD1 coding for the protein mutant known as G93A (indicated the mutation on the position amonoacídica) and control their expression in time and space (eg the nervous system, where the disease manifests itself). Thus generate eight lines viable mice trasngénicos "responsive" to be crossed with the strain of mice "activators" who are holders of transgen in which the expression of the "tTA" (which evokes the transcriptional activity of the promoter responsive) is controlled by the promoter of the gene to the prionic protein hamster. Unfortunately, it was observed that the system tet-off expressed clear difficulties in its operations in vivo. Thus, we concentrated on trying to dissect the issues affecting their potential role in trying to repair and restore the desired format. The main effect was to the expression restricted to a few cell types, basically the recipients of the olfactory mucosa. Analysis of the levels and distribution of the messenger RNA and protein for the tTA, SOD1 human and EGFP (cDNA its reporter was included in the responsive transgene), showed that tTA was present throughout the nervous system. Furthermore, the protein was detected in the nuclei of excerpts from anywhere on the same system. Despite this activation Responsive single case in specific types of olfactory epithelium, where it certainly manifested itself as a heterogeneous pattern, which traditionally has been linked to regulation and epigenetic phenomena of RNA interference. We aimed to analyze the levels of DNA methylation in the construct to see if the transcriptional response was suppressed by epigenéticas causes. In fact it was found that the sequences were mainly those metiladas home procariota (virus and bacteria), and that its distribution correlacionaba with the levels of expression observed. Anyway, this phenomenon did not explain why the restriction observed, and suggests that additional factors interfere in the regulation of transcriptional response of this system, which may be in relation to other comments as inversely proportional dependence between the number of copies and levels of expression and / or the robustness of the pattern of change despite the strain of mice.

Author: ESCALONA ARAS NOELIA.
Year: 2006.
University: BARCELONA.
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGIA. UNIVERSIDAD DE BARCELONA.

Summary: The cGMP is an intracellular messenger serving as a mediator of important actions of nitric oxide and natriurético atrial peptide. It has been described a noticeable reduction in the content of cGMP in the myocardium of rats and pigs subjected to a transient ischemic. In addition, it has been demonstrated that cGMP reduced by myocardial reperfusion injury. In this work we have investigated the mechanisms that modulate the concentration of cGMP during myocardial ischemia and reperfusion in coronary microvascular endothelial cells and caridomiocitos of adult rats. In microvascular endothelial cells, simulated ischemia (hypoxia in acidic conditions) drastically reduces the content of cGMP baseline and the ability of these cells to synthesize both cGMP in response to the stimulation of guanilil cyclase sensitive to NO (GC-NO) and the guanilil cyclase particulate (GCp). Acidosis depletion and energy involved in this reducicón. In cardiomiocitos rat, ischaemia reduces synthesis GMOc dependent on the activity guanilil cyclase particulate, but not significantly affect the synthesis mediated guanilil cyclase dependent NO. The effect of acidosis on cGMP synthesis seems to be a direct effect on the activity of enzymes synthesis (GC-NO and GCp) given that caused the pH dependence that is observed when it determines the in vitro activity. The simulated reperfusion completely reverses the effects of acidosis previous two cell types, the reduction was only partially induced by hypoxia. The fact that cardiomiocitos synthesis of cGMP-mediated GC-NO not be affected by conditions of intracellular acidosis, when the enzyme is extremely sensitive to pH in vitro, to think in a complex regulation of the enzyme during ischemia . In this sense, has been reported recently that the activity GC-NO can be found associated with the plasma membrane of myocardial tissue and platelets, where the translocation of the enzyme would take place during the process of activation of these and would be regulated by intracellular calcium. In the latter part of this paper has analyzed the potential importance of the functional activity GC-NO associated with the membrane of caridomiocitos. It was noted that one-third of the GC-NO was associated with the membrane fraction of these cells. The cytosolic enzymes and participated in vitro presents the same characteristics kinetics for the factors analyzed: GTP, NO, pH and calcium. It was noted that the influx of calcium into the cell leads to the translocation of GC-NO to the membrane, increasing the activity in this fraction and increases the response of the cell donor NO. This effect is powered by inhibitors of protein kinases and phosphatases by inhibitors, which suggests a mechanism for regulating complex via phosphorylation and desfosforilación protein (probably including the very GC-NO and regulatory protein). Finally, the effect on cGMP synthesis of various pharmacological agents used in caridomiocitos isolated correlates with the effect of these drugs on the activity of the enzyme associated with the membrane, suggesting that the membrane associated enzyme was determined to a large degree cGMP-dependent synthesis of NO in this cell type.

Author: POYATOS LÓPEZ RAFAEL.
Year: 2006.
University: BARCELONA.
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA.

Summary: Sweat two forest species representative of the average mountain Mediterranean (Pinus sylvestris and Quercus pubescens) was measured with methods of sap flow in the area pilot Vallcebre (Eastern Pyrenees). The regulation of perspiration on the part of both species was studied in relation to various meteorological variables, soil moisture and other physiological parameters of the plant. In addition, physiology of P.sylvestris regarding the evaporative demand and the availability of water was studied along its distribution area.

Author: MATEO LOZANO SILVIA.
Year: 2006.
University: AUTÓNOMA DE BARCELONA.
Place of defense: HOSPITAL UNIVERSITARIO VALL D'HEBRON.
Place of preparation: AUTONOMA DE BARCELONA.

Summary: The family of Ewing's sarcoma tumors (ESFT) includes a heterogeneous group of neoplasms composed of small round cells with minimal evidence of morphological differentiation, or by larger cells with different degrees of differentiation neuroectodérmica. The Ewing's sarcoma is the second most common bone tumors, affecting mainly children and adolescents. Although appearing primarily in bone, approximately 15% of patients have primary tumors in soft tissues. Current treatment combines high doses of chemotherapeutic agents for the control of systemic disease with surgery and / or radiation for local control. Despite the use of multimodal therapy and aggressive actions of local control, survival of patients with metastases is less than 30%, whereas in the absence of metastatic disease is approximately 50-70%. Because of this, we need new therapeutic approaches to reduce treatment-related morbidity and improve the survival rate. Fortunately, ESFT present a perfect target molecular resulting from the chromosomal translocation that defines this type of tumor, and involves the EWS gene and a member of the family of transcription factors STD (Erytroblastic Transforming Sequence), primarily FLI-1 or ERG. The most common translocation, t (11; 22) (q24, q12), generates the formation of the oncoproteína merger EWS/FLI-1 serving as a factor in regulating transcription and aberrant form of the expression of target genes, favoring this how the tumorigenic process. This inactivation of the fusion protein EWS/FLI-1 becomes an attractive strategy, not only because of its specificity in transformed cells, but also because of its key role in the tumorigenesis of ESFT. This study evaluated different strategies to reduce the expression levels of EWS/FLI-1 in vitro and in vivo. The first strategy used to inhibit the levels of expression of the fusion protein EWS/FLI-1 was based on the use of rapamycin, an immunosuppressant and antifungal properties anticancerígenas, resulting in inhibition of the channel signaling mTOR/p70s6K . The rapamycin inhibited cell proliferation of ESFT without causing apoptosis, suggesting the use of this drug as a cytostatic agent in the treatment of such tumors. The second approach was based on the therapeutic simultaneous inhibition of EWS/FLI-1 at transcriptional and post-through combination therapy of antisense oligonucleotides and rapamycin. The results showed an increase in cell death of ESFT, through a process that involves the restoration of the road signaling pro-apoptótica of TGFß1/TGFß-RII. In addition, in vivo testing immunosuppressed mice showed a delay in the growth of tumors. These data provide the basis for further exploration of the potential of combination therapy as a new strategy in the treatment of such tumors. Tumors of the family of Ewing's sarcoma show alterations in cell cycle regulatory proteins, including the overexpression of protein kinases ciclina-dependientes (CDK) and the loss or low expression of their inhibitors. Based on this, the third strategy was based on the reversal of some of these conditions through the use of roscovitina, a potent inhibitor of the kinase activity of the CDKs. The results showed that the roscovitina induce apoptosis in cells ESFT in vitro and in vivo, by a mechanism dependent on the activation of caspases. Numerous studies have identified genes whose expression is regulated by the expression of EWS/FLI-1, thus favoring the tumorigenic process. In order to identify and assess new proteins that interact 8 with EWS / 4e3 FLI-1 and thereby contribute to the understanding of the mechanisms of transformation, identified as a direct target of EWS/FLI-1 to caveolina-1, protein involved in a variety of cellular processes such as endocytosis, cholesterol homeostasis, signal transduction and tumorigenesis. The results of this study showed that caveolina-1 plays a decisive role in the tumorigenesis of cells ESFT and its inhibition could allow the development of new molecular therapeutic strategies aimed at improving the treatment of patients ESFT.

Author: Pérez Lluch Sílvia.
Year: 2006.
University: AUTÓNOMA DE BARCELONA.
Place of defense: Centre d'Investigació i Desenvolupament (CSIC).
Place of preparation: Institut de Biologia Molecular de Barcelona.

Author: MARTÍNEZ GALLO MÓNICA.
Year: 2006.
University: AUTÓNOMA DE BARCELONA.
Place of defense: HOSPITAL DE LA SANTA CREU I SANT PAU..
Place of preparation: DEPARTAMENTO DE INMUNOLOGÍA. HOSPITAL DE LA SANTA CREU I SANT PAU..

Summary: The primary immunodeficiencies or congenital (IDPs) are a group of diseases of diverse etiology they have in common a defect qualitative or quantitative immune system as a result of which alter their functions, compromising the defense against external aggression. At present, it has flexionado this definition dadno including a larger spectrum of diseases affecting the immune system, and that does not always imply a greater increase in the frequency of infections, such as those affecting the homeostasis of the immune system. In this paper we describe the clinical and immunological features of three patients with IDP monogenic, two patients with syndrome Hiper-IgM, several families with Syndrome Linfoproliferativo Autoimmune ALPS and a patient with linfoproliferación of double positive T cells, which led out molecular diagnosis. The oxidative capacity was analyzed by NBT in granulocyte stimulation with WFP and E.coli, in a patient who suffered from repeated infections since childhood by bacteria catalase positive, with the lack of production of superoxide. The study was conducted genetic subunit p47-phox of NADPH oxidase and found that the mutation responsible for the defect in phagocytosis was a deletion of dinucleótido GT at the beginning of exon 2 gene NCF1 being diagnosed with Chronic Granulomatous Disease (ECG ). We studied a patient with repeated infections in early life with lifnopenia Ty B. The diagnosis was made Deficit ADA through a trial of enzyme activity. Subsequently, we performed an allogeneic donor emperantedo haploidéntico. The system recovery inmunollógico patient after transplantation was evident. We studied a male patient with sospeccha clinic IDP humoral that showed a decreased number of B cells. We examined the presence of kinase Btk by intracellular staining and flow cytometry using western blot, the result showed a lack of expression of the protein in the patient. We performed genetic analysis of gene BTK, finding a new mutation Gln-103-STOP (Q 103X), which affects the PH domain of the protein, which was diagnosed Agammanglobulinemia linked to the X chromosome (XLA) with a complete correlation between genotype, protein expression and clinical phenotype. The aproximaicón for functional methods bioquímcios and provides a rapid diagnosis, which is essential to implement an early treatment, as has been done in patients with ECG deficit ADA and XLA. The syndrome Hyper Ig-M includes a series of molecular defects that directly or indirectly affect the lineage B. In both cases studied was ruled forms that were due to alterations in either of the two molecules main shaft CD40-CD40L. It was analyzed by flow cytometry the expression of HLA-DR, CD25 and ICOS without finding abnormalities. We studied the presence of memory B cells to analyze the expression of IgM and CD27 on B cells CD20 positive, finding missing this population in the two patients. The genetic study was conducted gene AID, as well as the regions of the constant heavy chains IgG1 and IgG2 of inmunogloblinas without encountering alterations in any of the patients. Although still unaware of the molecular bases in the two cases presented here Hyper IgM, clinical and immunological are well characterized and the failure to find cells memory B CD27 none of the cases shows that the defect that leads to a cambo of isotipo altered, could find itself in this process after the break and allows us to characterize DAN within the group HIGM4. Lastly, has studied a population double positive (DP) in a patient with patolgoía breathing. The characteristics of the population DP indicate that phenotypically 8 is 73c a population T CD3CD4Cd8ab, functionally belongs to the lineage of T cells CD4. There was an increase in the expression of integrina CD103 and recipient of chemokines CCR6 in the DP population, which together with the presence of these tissues in the body, to justify his tropism lung mucus and relates this population with respiratory pathology. Although the patient did not provide organomegaly / lymphadenopathy, lymphocytosis or other features of malignancy, the analysis of beta and gamma chains CT showed reordenamietnos clonal. The study showed cytogenetic alterations cromosómics surigiendo a state preleucémico, requiring additional genetic alterations to develop a frank neoplastic process. The results of this thesis show that the complexity of the diseases that affect the immune system, requires increasingly innovative techniques and specific for identifying defects responsible for a particular phenotype. The study genetic, molecular and functional systematic these diseases or can diagnose and classify entities already described as addressing the diagnosis of new diseases.

Author: Andreu Martín Nuria.
Year: 2006.
University: AUTÓNOMA DE BARCELONA.
Place of defense: Facultad de Biociencias.
Place of preparation: Instituto de Biotecnología y de Biomedicina.

Summary: Knowledge of the virulence factors of Mycobacterium tuberculosis is essential for the development of new drugs and vaccines, which are essential to combat this infectious disease that annually causes 1.7 million deaths. A distinctive feature of the virulent strains of M. Tuberculosis is the ability to set the red color neutral as acid. In previous work was refuted the relationship, widely accepted so far, between this behavior and sulfolípido (a component of the cell involved), and found that the gene Rv0577, cotranscrito with possible regulatory transcriptional Rv0576, gave M. Smegmatis (micobacteria saprófita) phenotype neutral red positive virulent strains of M. Tuberculosis. In this paper, through mergers transcripcionales with microchips and analysis has shown that the gene Rv0576 is, indeed, a transcriptional regulator that suppresses the expression of operon Rv0576-Rv0577. However, the study of mutant strains and complemented by these genes has shown that neither of these two genes is required for the neutral red staining of M. Tuberculosis, or virulence in the mouse model of infection. They have also shown that M. TB in vitro form heterogeneous cell populations for the red and neutral for the synthesis of dimicocerosatos of ftiocerol, some of the lipids wrapped cellular related virulence bacillus. The spontaneous mutant red neutral reverse negative, with some frequency, the phenotype wild, while it has not been detected in the mutant reversal in the synthesis of dimicocerosatos of ftiocerol, which accumulate with successive resiembras M. Tuberculosis, until result in the majority population. These observations, and the relationship of the neutral red staining and the dimicocerosatos of ftiocerol with the virulence of M. Tuberculosis, highlight the need to check these phenotypes before performing an analysis of the virulence of a particular or mutant strain of M. Tuberculosis.

Author: GOMEZ MORUNO ANTONIO.
Year: 2006.
University: AUTÓNOMA DE BARCELONA.
Place of defense: UNIVERSITAT AUTONOMA DE BARCELONA.
Place of preparation: INSTITUT DE BIOTECNOLOGIA I BIOMEDICINA (UNIVERSITAT AUTONOMA DE BARCELONA).

Summary: THE FORECAST BIOINFORMATICA OF ROLE FROM THE SEQUENCE PROTEICA TOOL IS A KEY TO SUCCESS FOR DEVELOPMENT OF EXISTING PROGRAMS ANALYSIS PROTEOMAS. IN THIS WORK IS USED A SERIES OF TOOLS BIOINFORMATICAS AVAILABLE TO DATE FOR PREDECIR THE ROLE FROM STREAM, AT A TIME TO DEVELOP NEW ALGORITMOS THAT INTENTAN IMPROVE AND / OR SUPPLEMENT TO THE EXISTING, WITH THE AIM OF THE MAXIMUM GET INFORMATION ON THE ROLE OF A PROTEIN IN BIOLOGICAL ANALYSIS PARTEN THAT ONLY ITS sequence.

Author: López Castejón Gloria.
Year: 2006.
University: MURCIA.
Place of defense: Facultad de Biología.
Place of preparation: Facultad de Biología.

Summary: During the development of this work have been cloned and characterized three key molecules in the system of interleuquina-1beta (IL-1beta) of gold (Sparus aurata L., Teleostei): receiver P2X7, caspase-1 and type receiver II to IL-1beta (IL-1RII). En primer lugar, se clonó el receptor P2X7 de dorada y se compararon sus perfiles de actividad en respuesta a diferentes agonistas y antagonistas con los de los receptores P2X7 de rata, xenopus y pez cebra expresados en células HEK, y también se estudió dicha respuesta en the fibroblast cell line of golden SAF-1. This information was used to investigate the mechanisms of release from IL-1beta induced receptor mammal and fish. Despite exposure fosfatidilserina in the external face of the membrane and cellular permeabilización observed in leukocytes of gold after activation with high concentrations of BzATP, IL-1beta remained inside the cell without being processed. However, the activation of receptor P2X7 rat expressed so Ectopic cell HEK293 with caspase-1 human produced the release of the precursor form of IL-1beta of gold. By contrast, neither the activation of receptor P2X7 of golden zebrafish or induced secretion of the IL-1beta or mammal or fish when expressed in cells HEK293. These data indicate that activation of caspase-1-mediated receptor P2X7 and release of the IL-1beta occur through different signaling pathways and suggest that, although the mechanisms involved in the secretion are conserved during evolution, different inflammatory signals, have been selected for the secretion of this cytokine in different vertebrates. Secondly, it clonó caspase-1 of gold, which presented a domain conserved amino terminal CARD (caspase recruitment domain) and a catalytic domain of caspase carboxi-terminal. Se comprobó que el gen de la caspasa-1 se expresaba en larvas de un día después de la eclosión y que los niveles de dicho mRNA aumentaban durante el desarrollo. In adults, caspase-1 is constitutively expressed in all tissues analyzed and that decreased expression both after infection with Vibrio anguillarum in all tissues examined, as after stimulation of phagocytes of gold with different molecules derived from pathogens (PAMPs ). It also showed that caspase-1 recombinant form of golden expressed Ectopic cell HEK293 was capable of processing a substrate fluorogénico specific caspase-1 and that this activity increased after activating receptor P2X7 with BzATP. In addition both cells SAF-1 as leukocytes of golden showed activity endogenous caspase-1, which was completely inhibited with a specific inhibitor of caspasa-1. Lastly, was isolated and characterized in a golden counterpart receptor type II to the IL-1 (IL-1RII) mammal. The IL-1RII of golden presented two Ig - like domains in the region and a small extracellular domain citoplasmático without mastering TIR signaling. The cDNA of gold showed a 3'UTR unexpectedly long compared with other species, which contained three reasons for instability ATTTA, which is probably responsible for its relatively short half-life (less than 2h). There was an increase in the expression of the receptor golden greater than the expression of IL-1beta in all tissues analyzed after an infection V. Anguillarum. The stimulation of leukocytes golden with bacterial DNA and flagelina in vitro increased mRNA levels of IL-1RII in macrophages, whereas only flagelina was able to induce an increase in gene expression in granulocyte acidophilous. Finally, it was observed that IL-1RII was located in the plasma membrane and was able to interact with the IL-1beta of gold.

Author: MARTÍN GARCÍA PALOMA.
Year: 2006.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE CC. BIOLÓGICAS.
Place of preparation: FACULTAD DE CC. BIOLÓGICAS.

Summary: This paper develops a new methodology for the management of the visit in protected natural areas, which is based on the account of the visit described as a system consisting of pieces iteradas. This makes it possible to formalize such systems in the so-called "Modular Description" and specify the information necessary to see how it works. From the description Modular System visit, the parameterization of its constituent elements and functions of impact and perception, it is possible to construct models Functional Flow Visitors using the tools integrated into the so-called "Monitoring System Functional De La Tour ( SSFV), which allow an understanding of the distribution and dynamics of visitors in the interior space as their associated consequences on the environment and perception of the quality of the visit. This methodology provides a methodological breakthrough in the development and application of new tools for the management of the visit, which in addition to providing a very detailed knowledge of the functioning of these systems, allows you to define a comprehensive monitoring program that optimizes data collection for the delas detect variations that can produce and manage enables the carrying capacity of visitors by the mere actions alternatives to limit the total number of visitors, thanks to the ability to easily simulate different scenarios of operation and know well its implications on the management objectives . Its application to all National Parks has to be understood as a first approximation knowledge of the operation of these spaces which demonstrates the flexibility and usefulness of the methodology outlined in the management of the visit in any protected natural space.

Author: Cerviño Gómez MMa. del Carmen.
Year: 2006.
University: SANTIAGO DE COMPOSTELA.
Place of defense: Facultad de Biología.
Place of preparation: Facultad de Biología.

Summary: The protein kinase C (PKC) is a family of serina-treonina kinases involved, among other processes, in the signal transduction activation in T lymphocytes This family consists of at least ten isoforms with expression varies according to the cell type. In order to analyze the specific role of PKC beta in the process of activation of T lymphocytes, his expression was inhibited by antisense technology, using as a model to study cell Jurkat (a line leucémica of human T cells). Using this technique, which consists in the introduction into the cell of a sequence of RNA complementary to the messenger RNA that encodes the protein under study, inhibimos the expression of this enzyme. For a permanent inhibition was introduced into cells Jurkat by electroporation, a vector episomal (plasmid pREP3) with an insert for a fragment of the gene coding for PKC beta in antisense orientation. In parallel, the cells Jurkat was introduced vector pREP3 without insert, with the aim of obtaining populations that could be used as a control. In order to obtain homogeneous populations in terms of their expression of this isoform of PKC, the cells were cloned by dilution limit, then selected those with lower expression of PKC beta with respect to controls analyzed. In these cells could be demonstrated selective inhibition of PKC beta between 40% and 60% and the decrease in the levels of messenger RNA that encodes. The reduction in the expression of PKC beta is associated in cells Jurkat with an alteration in the formation of aggregates, more numerous and consist of a greater number of cells with respect to the controls, but differences were observed with respect to the feasibility or cell proliferation in the absence of stimulation. After stimulation of the cells with an ester forbol (WFP) and a ionóforo calcium (ionomicina), the relationship was observed between the decrease in the expression of PKC beta and the increase in the expression of CD25 (chain alpha receptor IL-2) and the reduction of the expression of CD69, molecules marker of T cell activation Moreover, it was shown in the same cells inhibiting the production of IL-2 and IL-8, and the increase in production of TNFbeta. It was observed in cells deficient in PKC beta no alteration with regard to the phosphorylation in tirosinas nor the degree of activation of protein from the ERK MAP kinase cascades or p38. However, inhibition of the expression of PKC beta is associated in cells Jurkat with an increase in the degree of activation of the JNK route (reflected in increased activation of p46 and its target, the transcription factor ATF- 2). These findings help establish the differential involvement of two isoforms calcio-dependientes of PKC (PKC alpha and beta PKC) in the lymphocyte activation process, comparing the results with those of other work, using the same technique and the same cell type , demonstrated the relationship between inhibition of PKC alpha with the decline in the synthesis of IL-2 and its high affinity receptor and a decrease in the expression of the cytokine TNF-alpha. In these cells, the observed alterations could not be associated with a different degree of activation of any of the MAP kinase analyzed (ERK, JNK and p38) and the transcription factor ATF-2. The comparison of the results obtained in this work demonstrates that the functions of PKC alpha and beta PKC may be similar, different and even antagonistic in different aspects of cell activation Jurkat.

Author: NAVARRO AVILES GLORIA M.
Year: 2006.
University: MURCIA.
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA.

Summary: The bacteria are adapted to the fluctuating environmental conditions regulating the expression of specific genes. An external stimulus critical light, affects several metabolic processes, development and behavior both in bacteria and in higher organisms. In some species of bacteria light causes the synthesis of carotenoids, which are used to protect the injury fotooxidativo cell caused by singlet oxygen and other free radicals produced as a result of the lighting (Armstrong, 1997). Carotenoids also play a role as an intermediary and as a precursor of redox molecules necessary for the fotorrecepción and hormonal action (Frank and Brudvig, 2004). An important model for the study of carotenogénesis induced light is the Gram negative bacterium M. Xanthus, in which genetic and molecular analysis reveals a complex circuit that includes several genetic factors and regulatory mechanisms for fotoinducción of novel enzymes responsible for synthesis of carotenoids. All genes of M. Xanthus involved in carotenogénesis are grouped in the operon carB, with the exception of the gene crtlb (Fontes et al., 1993; Bottle et al., 1995). The promoter who directs the expression of operon carB, PB, depends on the binding of RNA polymerase (RNAP) A partner, the main factor sigma vegetative M. Xanthus. The induction of PB by light is closely linked to the activity of repressor CarA (Ruiz-Vázquez et al., 1993; Whitworth and Hodgson, 2001; Cervantes and Murillo, 2002; López-Rubio et al., 2002). Thus, in the dark, PB is suppressed by the union of CarA its operator bipartite design. The operator of a site consists of high affinity (pI), which corresponds to a palíndrome perfect interrupted located upstream of PB, and a low-affinity site (pII), a version of imperfect pI that overlaps with the region -35 - PB . CarA joins in a cooperative manner to its operator, first pI pII and then, in an effective way to prevent the access of RNAP to the promoter and suppress carB (López-Rubio et al., 2004). Furthermore, the activation of carB by light is due to the dismantling of complex CarA-operador. To that end, the light induces protein CarS a antirrepresor of CarA. CarS, which does not bind to DNA, physically interacts with CarA and prevents binding of repressor to its operator. CarS expressed from operon fotoinducible carQRS, which also produces CarQ, a type of alternative sigma factor ECF ( "ExtraCytoplasmic Function"), and factor antisigma, CarR, located in the membrane (McGowan et al., 1993) . The expression of carQRS (and also crtlb) requires CarQ (McGowan et al., 1993; Fontes et al., 1993). CarQ, sequestered in the dark by their union to CarR, is released when light inactive CarR (Browning et al., 2003). The mechanism for this inactivation is not known, but involves at least one constituent protein expression, CarF (Fontes et al., 2003), although the need for light and CarF may be ignored by the presence of copper (Moraleda-Muñoz et al., 2005). The protein CarQ free associated with RNAP to transcribe the operon carQRS. This requires, in addition, the action of three other factors expressed in a manner constituting: CarD, the only known example in bacteria that resembles architectural eukaryotic proteins of the family HMGA (Nicolas et al., 1994; Nicolas et al., 1996 ; Padmanabhan et al., 2001; Cayuela et al., 2003); IhfA, another architectural protein in bacteria similar to the histones (Moreno et al., 2001); and CarG, a novel transcription factor that is associated with zinc (Peñalver - Mellado et al., 2006). The suppression of carB in darkness and their activation in the light reflected a game between antagonistic interactions CarA your operator or CarS. Previously, we had shown that both interactions occur through its segment Nt 8 erminal 86c of 78 residues, which is an autonomous domain, monomer and stable CarA (Nter), which alone can suppress carB in vivo in a manner dependent the dose (Pérez-Marín et al., 2004). In this work has deepened in the structural foundations of molecular mechanism underlying this competitive binding. We have determined the three-dimensional structure by NMR CarA (Nter), and identified specific residues and structural elements that are crucial for the recognition of yourself and CarS and therefore for its role in vivo. CarA (Nter) takes the topology hélice-giro-hélice winged ( "winged helix") typical of the DNA binding domain (DUD) of proteins MerR, consistent with the predictions obtained by the in silico analysis of its sequence. Furthermore, we have shown that CarA (Nter) has the most contact with the DNA that are characteristic of DUD of proteins type MerR in their motives hélice-giro-hélice and "wings". A significant finding of this work is that the propeller 2 of the plea hélice-giro-hélice not only is the region-specific binding to the operator but also to protein antirrepresora CarS. This implies that the union of CarS and union operator CarA are two facts mutually exclusive. The occlusion of the region's repressive CarA involved in the union operator for the antirrepresor CarS thus provides the molecular basis of the mechanism that controls the induction of gene expression by light on M. Xanthus

Author: ALONSO PERAL MARIA MAGDALENA.
Year: 2006.
University: MIGUEL HERNÁNDEZ DE ELCHE.
Place of defense: DEPARTAMENTO DE BIOLOGÍA APLICADA E INSTITUTO DE BIOINGENIERIA.
Place of preparation: UNIVERSIDAD MIGUEL HERNANDEZ.

Summary: The vascular system of agencies pluricelulares part of a family of biological structures branched arborescentemente, ranked and three-dimensional, which also includes the respiratory and nervous systems of many animal species. For disecarlas rules generativas of these topologies can be used as a model venation patterns of the leaves of plants and wings of insects, which are two-dimensional and simple. In this thesis we studied Doctoral mutants hemivenata (hve), which have altered their pattern of leaf venation. We have cloned posicionalmente gene HVE and characterized three of its mutant alleles, hve-1, hve-2 and hve-3 at the molecular and phenotypic. These mutants show a pattern of leaf venation very simple, with a number of secondary and tertiary veins far below the wild, and the absence or shortening of the higher-order. The mutants hve also show a delayed flowering and senescence, decreased fertility, bushy habit and lack of ripples in the root. Some of these features resemble mutants alerados in the perception of auxin. We have established that mutants are slightly insensitive to hve auxin, as though responding to the wild type inhibitors of this hormone transport. The proceeds of Ben HVE is a protein CAND1 (CULLIN-ASSOCIATED AND NEDDYLATION-DISSOCIATED1) whose participation in the ubiquitinación has been demonstrated in mammalian cells. The allele spontaneous hve-1 alters the prosecution of transcribed gene HVE, and hve-2 and hve-3 are carrying inserts ADN-T. The size of the inflorescence and fertility are more severely curtailed by hve-2 and hve-3 that hve-1, suggesting that this allele is hipomorfo. The pattern of venation simple plants hve appears to be due to a defect in the early development of the pattern of venation. We have determined that in vitro protein HVE joins the CULINA1 and venation patterns of mutants axr1 and hve are similar. These results and our analysis of the mutant phenotype vascular hve show that the auxin signaling mediated by the ubiquitina involved in establishing the pattern of venation of cotyledons, vegetative leaves, petals and sepálos. Our analysis of double mutants and transgenic plants showed that the transportation and the perception of auxin involved in the training regardless of the pattern of venation, and that HVE acts before ATHB8 on a path of development in which it participates AXR1, but do not LOP1, PIN1, CVP1 nor CVP2.