BIOCHEMICAL ANALYSIS

Author: MUÑOZ MARTÍNEZ JOSÉ ANTONIO.
Year: 2004.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: FACULTAT DE CIÈNCIES.
Place of preparation: ESCUELA DE POSTGRADO.

Summary: The work presented in this thesis focuses on the study of lithiasis oxalocálcia like illness that affects a large segment of the population, which is characterized by a high prevalence and incidence, morbidity and high rates of relapse. Even in the currently unknown a significant number of aspects relating to the mechanisms of formation of the calculations and their causes, and thus does not have the diagnostic or therapeutic treatments efficient than desirable. This paper attempts to elaborate on a better understanding of the etiology litiásica by developing analytical tools for greater eficiacia in the diagnosis of urinary disorders and the prevention of this disease. These objectives have been carried out to work at different levels: 1-Development of a series of simple analytical methodologies applicable to the identification of markers of litogénesis. 2, - study the temporal variation (daily and seasonal) risk litógeno. 3-In vitro characterization of the properties inhidoras of different substances. 4-Evaluation of the adequacy of the urine specimen night as best suited to the study of the alterations responsible for litogénesis, compared to the inconvenience that presents the urine of 24 hours, normally used for this purpose.

Author: GONZÁLEZ GOMEZ DAVID.
Year: 2004.
University: EXTREMADURA [www.unex.es].
Place of defense: FACULTAD DE CIENCIAS.
Place of preparation: FACULTAD DE CIENCIAS.

Author: PEÑA LUQUE FERNANDO.
Year: 2004.
University: CÓRDOBA [www.uco.es].
Place of defense: FACULTAD DE CIENCIAS.
Place of preparation: DEPARTAMENTO DE QUÍMICA ANALÍTICA.

Summary: The purpose of this report was the development, validation and implementation of various analytical methodologies allowing direct analysis of samples of olive oil with a view to conduct a quality control of the same. This was carried out through the characterization of the oil analyzed through the overall signal provided in the instrument by the volatile profile of the same. To achieve this objective using an instrument consisting of a commercial space module head coupled to a mass spectrometer (HS / MS), which acted as a detector. The system also allowed employees the option of both modules include a gas chromatograph for information discriminated against oil analyzed. The quality of olive oil was studied in this report based on three different aspects: classification of the three main types of oil pollution with various compounds (such as volatile halogenated solvents and hydrocarbons) and adulteration with other types of inferior quality oil, either through olive inferior quality, or through seed oil. The use of various types of tools quimiométricas for the treatment of the data was instrumental in the development of these methodologies. We used three kinds of different techniques such as pattern recognition techniques (including visualization techniques, classification and modeling), regression techniques and techniques for selecting variables. It has followed the line of research of our group FQM-2l5 of the Junta de Andalucia oriented to the development of analytical strategies for vanguardia-retaguardia.

Author: BENASCO RODRIGUEZ JOSE PEDRO.
Year: 2004.
University: LA LAGUNA [www.ull.es].
Place of defense: FACULTAD DE QUIMICAS.
Place of preparation: LA LAGUNA.

Summary: The thesis is a study of maturation of the grape variety Malvasía, in three denominations of origin canary, Abona, Ycoden-Daute-Isora and Lanzarote for vintages of 1999, 2000 and 2001. This study explores the maturing weight of 100 berries, pH, total acidity, the degree brix, tartaric acid, malic, citric and easily assimilable nitrogen. Also, are performed microvinificaciones processing sweet wines in the three vintages and the three denominations of origin, and were analyzed five sets of parameters, one related to the acidity of wine, the second related to the body of the wine, with the third color, polyphenols and sulfur, the fourth with metals and conductivity, and the fifth related to the volatile majority.

Author: LOZANO PÉREZ CARLES.
Year: 2005.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGIA.

Summary: It has obtained a series of conjugates flavanoles from a plant extract through a process of despolimerización acid in the presence of derivatives cistenia. The monomers flavanoles derivatizados separated efficient oil extraction by ion exchange chromatography, due to the net burden introduced derivatizar. They were subsequently purified by chromatography on reversed phase C18 getting the corresponding derivatives epicatequina, catechin and epicatequin 3-O-galato conjugated with cysteine, O-etilcisteina and N-acetil-O-metilcisteina. These compounds showed a greater capacity antiradicalaria the corresponding epicatequina through testing radicals linbres DPPH (which is reduced via donation hidrógenos and / or by electronic transfer) and HNTTM (which is reduced only by electronic transfer). In general, compounds containing the group gálico were generating greater capacity antiradicalaria in both studies. Studies were also carried out in vitro with human melanoma tumor lines (A375 and M21), and colon (HT29) and with a line of non-tumor keratinocytes (HaCaT), a moderate inhibition observed in cell viability, and compounds containing a group galato which presented more effectively. At the level of cell cycle, only derivatives containing the group gálico submitted an alteration in the cicclo, increased phase S. They also noted a greater induction of apoptosis in linias tumor, but not in the line of non-tumor keratinocytes. This induction to the apopotosis be corroborated with other techniques in which it was observed chromatin condensation and DNA fragmentation. Surprisingly, a genetic derivatives containing a group galato did not change in the cilco cellular or induced apoptosis in tumor cells. Likewise, their power antiradicalario through electronic transfer (HNTTM) was insuitadamente under even containing gurpo galato. These results led us to hipotetizar the possible relationship between the electronic transfer and its effect on cell machinery. Similarly, the radical stable HNTTM would allow us to predict easily capacity por-apoptótoica of compounds according to their ability to transfer electronically.

Author: ABELLÁN SÁNCHEZ MARÍA DEL ROSARIO.
Year: 2005.
University: POMPEU FABRA [www.upf.edu].
Place of defense: DEPARTAMENTO DE CIENCIAS EXPERIMENTALES Y DE LA SALUD.
Place of preparation: DEPARTAMENTO DE CIENCIAS EXPERIMENTALES Y DE LA SALUD.

Summary: Hormones recombinant erythropoietin (EPO), growth hormone (GH), almost equal to the endogenous and short half-life in circulation, are difficult to detect directly in doping control. The values were determined populations of biomarkers indirect EPO, soluble transferrin receptor, insulin-like growth factor (IGF-I) and procolágeno type III peptide (P-III-P) in selected populations of athletes, and effect of exercise and different types of training on their serum. The comparison of results obtained through different trials showed the need for a comprehensive validation prior to their use. Except for P-III-P, serum biomarkers proposed for the detection of rhEPO and rhGH are not directly affected by the athletic level, exercise or different burden of training done over the sports season. Age is the main influence on the serum concentrations of IGF-Iy P-III-P.

Author: Dondapati Srujan Kumar.
Year: 2006.
University: ROVIRA I VIRGILI [www.urv.cat].
Place of defense: Escuela Técnica Superior de Ingeniería Química.
Place of preparation: ETSEQ.

Summary: There is a growing demand for devices analysis multianalito, with potential applications in the fields of biomedicine and biotechnology, as well as in the industrial and environmental fields. For the development of these devices is essential good control space during the immobilization of biomolecules of interest, each one of them must be placed in a precise manner on the surface of the sensor (for example, a transducer amperométrico), avoiding overlaps that the specificity may compromise the system. The objective of this thesis is to develop different methods of patterning for the selective immobilization of biomolecules. The first method is based on selective electroplating gold biofuncionalizadas for nanoparticles development of biochips. It is a method of patterning electroquímicamente controlled, in which gold nanoparticles amending first recubriéndolas with various enzymes and then electrodepositan selectively on the surface of an electrode. As part of this methodology, were prepared nanoparticles of gold biofuncionalizadas using three different strategies: via the link dativo oro-tiol by adsorption directly or through electrostatic interaction by following the technique layer-by-layer (layer by layer). For funcionalización of gold nanoparticles used different biomolecules, such as enzymes horseradish peroxidase (HRP), glucose oxidase (GOX) and bovine serum albumin (BSA), and finally modified oligonucleotides with fluorescent molecules and thiol groups. The nanoparticles biofuncionalizadas were characterized using techniques spectroscopy UV-visible, transmission electron microscopy (TEM) and measure the zeta potential. Through spectroscopy UV-visible there was a resonance peak of plasmón characteristic of the modified nanoparticles, linked to the stability of the preparation. The measure zeta potential allowed the characterization of gold nanoparticles modified with polymer layer by layer and redox enzymes. It also studied changes in the zeta potential of nanoparticles modified with BSA at different pH values. After preparing the particles biofuncionalizadas, studies were carried out fundamental electroplating gold nanoparticles modified with BSA and a redox polymer, in order to analyze the effect of various parameters: potential applied deposition time, distance between the electrodes , auxiliary electrode surface and pH of the medium. To study the behavior electrocatalítico of nanoparticles modified once electrodepositadas, experiments were carried out using modified gold colloids with HRP and GOX. Then using this methodology for the development of biochips using two different configurations. The first was electrodepositaron nanoparticles gold funcionalizadas with GOX and HRP and modified with a redox polymer on the surface of a chip electrodes interdigitados (SDI), and to the complete elimination of non-specific responses. In the second configuration, the particles were modified with an extra layer of polymer redox checking again the total absence of non-specific responses after the electroplating. This method of patterning is generic and can be used for the production of various biochips. The second method of patterning is also based on electrochemical control, and involves the modification of the electrodes with topcoat Self electroactivas whose functionality is modular in function of applied potential. In this methodology, the monolayer electroactiva contains acetal groups that can be selectively unprotected by applying a potential in specific areas of the surface of the electrode. Thus are exposed on the surface active groups aldehyde, which can be easily combined with primary amines present in the biomolecules of interest. The e 8 nzimas G db8 OX and HRP were used as a model protein to test the versatility of this technique. Its applicability to the manufacture of biochips demonstrated by measures amperométricas and action in real time through resonance plasmón surface combined with electrochemical (eSPR). The third methodology is a system of patterning electroquímicamente controlled, but in this case it is used for the immobilization of 4,4-bipiridil as a basis for the creation of biochips. It synthesized molecules 4,4-bipiridil funcionalizadas with carboxylic groups, which were characterized electroquímicamente and then conjugated with biomolecules of interest to the creation of biochips. The selectivity of these systems is demonstrated colorimetrically, obtaining minimum levels of nonspecific response. Finally, the fourth of the methods of patterning developed is based on the technique of photolithography. The enzyme glucose oxidase and sarcosina oxidase were deposited selectively along with a redox polymer on the surface of electrodes interdigitados using a lift off process, thereby completely eliminating signals crossed or cross-talk. As part of this methodology is optimized several procedures immobilization of biomolecules, in order to select the most appropriate strategy. It also carried out tests with different reagents to eliminate nonspecific adsorption. Finally, the optimized system was implemented on IDEs manufactured by photolithography. The glucose sensor and sarcosina responded selectively to their respective substrates, with total absence of cross-talk. This thesis is divided into 7 chapters. In Chapter I presents the foundations for the development of biochips, methods of patterning controlled electrochemical, other methods of selective patterning and photolithography techniques, as well as a summary of the thesis. Chapter 2 and 3 describes the synthesis of gold colloids, changing with biomolecules, stability studies and studies of the fundamental electroplating gold nanoparticles modified on the surface of the electrodes. In Chapter 4 shows the application of nanoparticles electroplating gold biofuncionalizadas for building biochips. Chapter 5 describes the selective immobilization of biomolecules through unprotected Electrochemical Self electroactivas topcoat. In Chapter 6 shows the synthesis, characterization and selective immobilization of derivatives 4.4-bipiridil funcionalizados with HRP. Chapter 7 describes the selective patterning in the micrometer scale two oxidasas on a chip electrode interdigitados through photolithography. Finally, Chapter 8 summarizes the findings and future work.