CLINICAL CHEMISTRY

Author: MORENILLA PALAO M. CRUZ.
Year: 2003.
University: MIGUEL HERNÁNDEZ DE ELCHE [www.umh.es].
Place of defense: MEDICINA.
Place of preparation: FACULTAD DE MEDICINA. UNIVERSIDAD DE VALENCIA.

Summary: The receiver vanilloides (TRPV1) is an ion channel sensitive to capsaicin, protons and heat that is expressed in nociceptors, and which plays a major role in the peripheral pain and heat in inflammatory pain. The expression and activity of TRPV1 are positively modulated by the presence of agents pro-algésicos and pro-inflamatorios, although still not known what transduction pathways responsible for this phenomenon awareness of the channel. This study using the technique of double hybrid in yeast to identify proteins that interaccionaran with cellular domain amino-terminal of TRPV1. Two vesicular protein, SNAPIN and Sinaptotagmina 9, interact strongly with the domain amino-terminal of TRPV1 in studies both in vitro and in vivo. In addition neurons gánglio of dorsal TRPV1 co-localiza with root proteins vesicular membrane, as VAMP-2, Ni SNAPIN nor Sinaptotagmina 9 affect the activity of the channel under normal conditions, on the other hand if inhigen partially protenciación the response of TRPV1 - induced activation of the protein kinase C (PKC). The power of the blockade observed by these proteins was similar to that caused by botulinum toxin A, a potent blocker of exocytosis. The derived results of this study indicate that a mechanism led by the rapid mobilization of PKC channel TRPV1 housed in vesicles to the plasma membrane. This mechanism for regulating levels of the channel in the membrane could be implicated in the development and maintenance of hiper-algesia thermal associated with the inflammatory processes. The discovery of this new route of regulation highlights the complexity of the intracellular machinery of the nocicepción and its role in inflammatory pain. In turn, represents a new study to ascertain its validity as a therapeutic target for the development of new drugs more potent analgesics and specific.

Author: RODRÍGUEZ VARGAS ELISENDA.
Year: 2004.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: INSTITUTO DE CIÈNCIA DE MATERIALS DE BARCELONA.
Place of preparation: ESCUELA DE POSTGRADO.

Author: GARCERAN ORTEGA M. JOSE.
Year: 2004.
University: MÁLAGA [www.uma.es].
Place of defense: MEDICINA.
Place of preparation: MEDICINA.

Summary: APPROACH AND OBJECTIVES The primary knee arthroplasty (APR) may result in a substantial loss of blood and 31 30-50% of these patients received allogeneic blood transfusion (TSA). Since the TSA is a scarce resource therapeutic increasingly expensive and not exente risk has shown the effectiveness of two strategies that reduce the need for TSA in patients undergoing APR. PATIENTS AND METHODS In the HCU Our Lady of Victory, results in 200 patients who received blood recovered from drains postoperative (group A2) were compared with those from a control group retrospective (Group A1). In HU Servetus, the outcomes of 81 patients who received iron saccharate period (2x200 mg iv), but if erythropoietin preoperative Hb was less than 13 g / dL (1x40, 000 IU sc) (B2 group), it compared with a retrospective group of 100 patients (group B1). RESULTS The percentage of patients with TSA in the A2 group was lower than the A1 (56.5 vs 11%, respectively, p less 0.01). The same happened in the B2 group regarding the B1 group (30 vs. 3.7%, respectively, p less 0.01). However, 23% (17/74) anemic group A2 and 10% (2 / 19) of the B2 group needed TSA. Conclusions In patients APR not anemic both were equally effective strategies to eliminate the need for TSA. However, in anemic patients would need a combination of both to have a greater margin of safety.

Author: TRULLOLS SOLER ESTHER.
Year: 2005.
University: ROVIRA I VIRGILI [www.urv.cat].
Place of defense: FACULTAT DE QUIMICA.
Place of preparation: FACULTAT DE QUIMICA. URV.

Author: EGIDO GABÁS MERITXELL.
Year: 2005.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE QUÍMICAS.
Place of preparation: FACULTAD DE QUÍMICA - UNIVERSIDAD DE BARCELONA.

Summary: The biosynthesis of proteins, one of the key steps is the folding and aggregation. This process usually requires cofactors and other proteins known as molecular chaperonas. If the amino acid sequence there is a mutation, it may not occur on folding and / or aggregation of the protein, and the resulting protein is not fully active or is degraded and do not reach their place of action. The chemical therapy chaperonas described as a new therapeutic approach for treating various diseases sisosomales or neurodegenerative. This therapy is based on the interaction with small molecules dela protein low molecular weight attending the correct folding of the protein through a non-covalent interaction type ligando-proteína, with a mode of action equivalent to the molecular chaperonas. This thesis has been synthesized new aminociclitoles candidates chaperonas chemical to treat one of the glicoesfingolipidosis: Gaucher disease. The synthesis of these compounds has been carried out through the opening regio-and stereoselective epoxides and aziridinas whose synthesis have been widely described in the literature. They also discussed the results of in vitro activity of these compounds on various enzymes; glucosylceramide synthase microsomal, glucosylceramide lysosomal hydrolase, Imiglucerasa (Cerezyme ®, Genzyme), the alpha yeast, beta-glucosidasa almond and various glicosidasas involved in the metabolism of glicoesfingolípidos. The synthesized compounds showed a high specificity glucosylceramide hydrolase with competitive inhibition studies and the determination of enzyme activity in denaturing conditions indicate that these compounds could, in low concentrations, stabilize and increase their glucosylceramide hydrolase activity, ie , are candidates for chaperonas chemicals for the treatment of Gaucher disease.

Author: GELLA CONCUSTELL ALEJANDRO.
Year: 2005.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA.

Summary: The thesis is aimed ultimately to the development of immunoassay for the determination of choriogonadotropin (hCG) in samples of human serum or urine. The first methods used for the determination of hCG were "bioassay" and then "radioinmunoensayos." Both lost their validity for various reasons: laborious, insensitive and not very specific (bioassays), the need for regulated facilities, and so on. When purified hCG was affordable, could prepare specific antibodies and developed various types of immunoassay with better benefits as "bioassay" and ease of use "radioimmunoassay. En los casos en que la determinación se realiza como prueba rápida de embarazo, son especialmente útiles determinadas variantes de inmunoanáisis denominadas "pruebas rápidas", que no precisan de instrumenteal y pueden ser realizadas por personal no especilaizado. In this paper we have developed and validated two systems test for the determination of hCG. On the one hand enzimoinmunoanálisis in a microplate to perform quantitative determinations of serum samples in Blood and an inmunocromatografia in nitrocellulose to undergo qualitative urine sample oriented and fast as evidence of pregnancy. In addition they have developed a variety of technologies applicable to the procurement of raw materials of biological origin involved in the immunoassay as obtaining and purification of hCG, obtaining and purification of monoclonal antibodies against hCG, the preparation of colloidal gold nanoparticles large scale and the conjugation of antibodies against hCG peroxidase and colloidal gold.

Author: Camprubí Giménez Sandra.
Year: 2006.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: Facultat de Medicina.
Place of preparation: Facultat de Medicina - Unitat de Bioquímica - Universitat Autónoma de Barcelona.

Summary: Streptococcus pyogenes is a bacterium grampositiva responsible for infections estreptocócicas in humans, such as sore throat or impetígeno, but it is also the cause of development in the aftermath postinfecciosas com rheumatic fever or acute glomerulonephritis. Rheumatic fever is the leading cause of death from heart disease in people younger than 45 years in underdeveloped countries. As the human body immunological response to infections estreptocócicas with a plethora of antibodies against components of cellular and extracellular S. Pyogenes, the serologic diagnosis of these infections is based on the immune response against extracellular products as estreptolisina O (SLO), desoxiribonucleasa B, hyaluronidase, NAD-glicohidrolasa or streptokinase. The detection of antibodies antiestreptolisina O (ASO) is ámpliament acceptada com a prova for diagnosing infections estreptocócicas, because about 80% of infected patients show high degrees of these antibodies in blood. Currently, there are several immunoanálisis for the detection of these antibodies, but the most used is the immunoanálisi turbimétrico based on the agglutination with SLO latex sensibilitzado with purified apartir crop S. Pyogenes. However, cultius this bacterium presentacos because of its pathogenicity. In addition, the crops are difficult standardization, and the nutritional requirements of S. Pyogenes, culture media is expensive. For these reasons, the production of a recombinant SLO was raised as an alternative to resolve these difficulties. In the present work, it chose a system of Expression procariota, Escherichia coli, because it was biologically secure their crops were reproducible i relatively low cost. First, it got the sequence coding for the SLO apartir genomic DNA from S. Pyogenes and prepared two recombinant expression vector, pBAD-SLO and pJLA603-SLO-His. Then, studies were conducted for the Expression of these two vectors and identified best condicions for maximum production of SLO. Apartir of these studies, it chose the system induà ¯ sible by L-arabinosa, with the vector pBAD-SLO to perform scaling of the crop in the bioreactor due to increased production of the recombinant protein. Next, the SLO is purified recombinant by affinity chromatography of metal ions immobilizados as the recombinant protein has a queue histidinas merged in the C-terminal end. We optimized the condicions union of the SLO to the resin and climbed through the purification technology StreamlineTM. It marked molecular and antigenically i purified protein was confirmed both their identity as its antigenicity. It was finally purified recombinant SLO applied in the development of a immunoanálisis turbidimétrico. Based on the controls and the characterization made, it was felt that it was feasible using the purified recombinant SLO for the identification of antibodies in blood ASO.