Tesi realitzada a l'Instut de Recerca en Energia de Catalunya – IREC / Thin film solar cells based on direct band gap semiconductors have attracted much research during last decades. Thin film technologies are currently commercial and display record power conversion efficiencies up to 20% at the laboratory scale. However, typical direct band gap semiconductors, CdTe and CuIn1-xGaxS2 (CIGS), content scarce and/or toxic elements such as In, Ga or Cd. An alternative to these materials is Cu2ZnSnS4 (CZTS), formed by abundant and non toxic elements. CZTS is a quaternary p-type semiconductor which presents great absorption coefficient (> 104 cm-1), has similar crystalline structure and optical properties to CIGS, and a suitable and tunable band gap (1.00-1.5 eV) by varying the S/Se ratio. An interesting strategy to develop thin film solar cells is the solution processing. Solution based approaches are especially interesting for their potential low production costs and their easy scalability. Among different solution processing techniques, the spraying of nanocrystals or metal salts is an especially interesting approach. The easy scalability of spraying techniques to prepare large-area panels in a non vacuum atmosphere, which is translated in a significant reduction of the production costs, renders the spraying very attractive for industrial implantation. A pulsed spray deposition system, which was custom made and operates in open air, is here used to produce CIGS and CZTS films from colloidal CIGS and CZTS NPs. This work is divided in 5 chapters. The 1st chapter is an introduction to the photovoltaic (PV) technology and in particular to thin film PV technology, with special focus to chalcopyrite CIGS and kesterite CZTS. In the 2nd chapter I review the work done towards solving one of the major challenges associated to NP-based PV technologies: the complexity to transform NPs into highly crystalline thin films by sintering processes. The 3rd chapter describes the experimental procedures used to prepare all the required materials and thin films and to fabricate solar cells. This chapter also describes the techniques used to characterize the morphological, compositional, structural and optoelectronical properties of the materials and films. Chapter 4 and 5 describe the work done regarding CIGS and CZTS technologies, respectively. Both chapters describe: NP colloidal synthesis, ligand exchange strategies to remove organic carbon surrounding the NP, subsequent thin film deposition techniques used, thermal treatments performed and final hetero-junction formation and device completion. Chapter 5 also describes a scale up method to produce large quantities of NPs using a continuous flow reactor. In summary, the goal of this thesis is to establish a non vacuum technology to produce CIGS and CZTS PV devices prepared by solution process of the absorber. Additionally, drawbacks involved in the solution processing of NP-based films, such as elimination of organic carbon present in NP and film crystallization are addressed. / Avui en dia, la major part de la industria fotovoltaica està basada en el silici. Aquesta és una tecnologia provada i robusta, però, a causa de l’alt cost dels wafers de silici, el seu potencial de reducció de costos sembla limitat. Així, s’ha desenvolupat una segona generació de cel.les solars, formada per capes primes de semiconductors inorgànics, que gràcies al consum reduït de material semiconductor permet la fabricació de cel·les solars de baix cost. Les tecnologies de capa prima són actualment comercials i presenten eficiències records de fins a 20%, a escala de laboratori. No obstant això, en general els semiconductors de banda prohibida directa, CdTe i CIGS, contenen elements tòxics i poc abundants, com In, Ga o Cd. Una excel.lent alternativa a aquests materials és el CZTS, ja que està format per elements no tòxics i abundants. Aquesta tesi explica el treball fet en la preparació i caracterització de capes primes de CIGS i CZTS mitjançant tècniques de processat en solució, utilitzant tintes de nanopartícules preparades prèviament mitjançant síntesis col·loïdal. Les etapes seguides i detallades en aquesta tesi són les següents: 1. Obtenció d’una solució de nanopartícules de CIGS i CZTS mitjançant síntesi col·loïdal. 2. Preparació de capes primes de CIGS i CZTS usant tècniques de processat en solució mitjançant les nanopartícules de CIGS i CZTS. S’ha establert la composició de la tinta amb la qual obtenim capes sense defectes superficials i lliures d’esquerdes. 3. Seguidament, s’han establert els paràmetres per realitzar el correcte tractament tèrmic per tal d’obtenir un material cristal·lí. 4. Finalment un cop establertes les condicions per obtenir capes primes amb les propietats òptimes per al funcionament d’una cel.la fotovoltaica, procedirem al muntatge de la mateixa i a la mesura dels paràmetres fotovoltaics aixi com la seva eficiència mitjançant simulador solar.

Tesi realitzada a l'Institut de Bioenginyeria de Catalunya (IBEC) / Natural vesicles produced from genetically engineered cells with tailored membrane receptor composition are promising building blocks for sensing biodevices. This is particularly true for the case of G-protein coupled receptors (GPCRs) present in many sensing processes in cells, whose functionality crucially depends on their lipid environment. Membrane receptors are involved in a variety of biochemical pathways and therefore constitute important targets for therapy and development of new drugs. Bioanalytical platforms and binding assays, using these transmembrane receptors, for drug screening or diagnostic require building well-characterized lipid membrane arrays, acting as supports to prevent protein denaturation during biochip processing. The controlled production of natural vesicles containing GPCRs, their characterization and their reproducible deposition on surfaces are among the outstanding challenges in the road map to realize practical biomolecular devices based on GPCRs. In addition, quantification of the protein receptors in such lipid membrane arrays is a key issue in order to produce reproducible and well-characterized chips. In this thesis we present the production and characterization of membrane nanovesicles (NV) from Saccaromyces Cerevisiae containing heterologously expressed olfactory receptors - a member of the family of GPCRs. We have demonstrated that membrane fractions from yeast cells spontaneously form closed spherical nanovesicles in solution. A simple method to homogenize the size of the nanovesicles to a diameter of around 100 nm at a concentration of more than 1010 nanovesicles mL-1 is also presented. It is also showed that after a genetic engineering process the olfactory receptors of interest were well expressed in the yeast membrane. Furthermore, we report for the first time a novel immunochemical analytical approach for the quantification of transmembrane proteins (i.e. GPCR) in their natural lipid environment. The procedure allows direct determination of tagged receptors (i.e. c-myc tag) without any previous protein purification or extraction steps. The proposed approach uses monoclonal antibodies addressed against the c-myc tag, frequently used in protein expression, on a microplate-based ELISA format with high detectability. The immunochemical method quantifies this tag on proteins or bioreceptors embedded in nanovesicles with detectability in the picomolar range, using protein bioconjugates as reference standards. The applicability of the method is demonstrated through the quantification of the c-myc-olfactory receptors (ORs, c-myc-OR1740 and c-myc-OR7D4) in plasma membrane nanovesicles (NVs). We also show by direct observation with Atomic Force Microscopy that nanovesicles deposit and flatten without rupturing on glass and gold substrates following approximately a diffusive law. We show that on glass surface coverages larger than 20-25% of the substrate can be reproducibly achieved under practical nanovesicle concentrations and reasonable time scales, while keeping to the minimum the presence of background residuals coming from the nanovesicles production process. On the other hand, on functionalized gold substrates surface coverages around 10-15% were achieved. Then, the role of surface chemistry was studied showing that modification of gold substrates indicates a higher affinity of natural nanovesicles for acid modified surfaces as compared to amino or alcohol modified surfaces. Nanovesicles deposition in acid modified gold surfaces and glass have been exploited for the generation of an array of multiple nanovesicles. Present results constitute an important step in the practical realization of biosensor devices based on natural nanovesicles integrating G-protein coupled membrane receptors. When olfactory receptors are genetically expressed in closed vesicles from natural yeast membrane fractions the verification of their capability for capturing specific odorant molecules are critical for the design of artificial noses. Thus, we demonstrated by Surface Plasmon Resonance (SPR) measurements on L1 Biacore chips that the receptors were functional. Despite the fact that the expression of olfactory receptors in nanovesicles is low, a fact that is coherent with the general expression level of GPCRs proteins in cells, the integration in nanovesicles together with a careful choice of the SPR experimental conditions and data analysis allowed us to obtain a concentration-dependent SPR response vs. odorant concentration with a sensitivity of 0.5-1.8RU/micromolar. The selectivity of OR carrying NV towards its specific odorant was proved in cross-check experiments with unspecific odorant molecules and control receptors. These results constitute a proof of concept that ORs embedded in nanovesicles properly respond to odorants and definitely open the perspective to use the surface plasmon resonance technique for the detection of small odorants at concentration in the micromolar range. / Vesícules naturals produïes a partir de cèl·lules modificades genèticament són prometedors components de sensat per utilitzar com a detectors en biodispositius. Això és particularment cert en el cas de receptors adjuntats a proteïna G (GPCRs) presents en molts processos cel·lulars, on la seva funcionalitat depèn estrictament del seu entorn lipídic. Els receptors de membrana estan involucrats en una gran varietat de vies bioquímiques i per tant són objectiu d’estudi per teràpia i desenvolupament de nous fàrmacs. Per tant, plataformes bioanalítiques i assajos d’unió receptor-lligand, utilitzant receptors transmembrana, requereixen la construcció de matrius de membranes lipídiques ben caracteritzades, actuant com a suport per evitar la desnaturalització de proteïnes durant el processament del bioxip. En aquesta tesi es presenta la producció i caracterització de nanovesícules de membrana (NV) provinents de cèl·lules de llevat Saccharomyces cerevisiae que contenen receptors olfactius (un membre de la família de GPCRs) heteròlogament expressats a la membrana. Hem demostrat que les fraccions de membrana, a partir de cèl·lules de llevat, en solució formen espontàniament nanovesícules esfèriques tancades. També s’ha demostrat, que després d’un procés de enginyeria genètica els receptors olfactius van ser expressats correctament a la membrana del llevat. També s’ha presentat un mètode simple per homogeneïtzar la mida de les nanovesícules. A més a més, es presenta per primer cop un nou mètode immunoquímic per la quantificació directa de les proteïnes transmembrana (GPCR) en el seu ambient lipídic natural. El mètode utilitza anticossos monoclonals en un assaig basat en ELISA amb alta detectabilitat. L’aplicació del mètode es demostra a través de la quantificació del receptors olfactius OR1740 i OR7D4 expressats en nanovesícules de membrana plasmàtica. També es presenta, mitjançant observació directa amb AFM, com les nanovesícules es depositen i s’aplanen sense trencar-se sobre substrats de vidre i or seguint la llei de difusió. Es demostra com en el cas del vidre els màxims recobriments superficials obtinguts són del 20-25% i en el cas del or funcionalitzat del 10-15%, controlant la concentració de nanovesícules, el temps de depòsit, la presència de residus procedents del procés de producció de les nanovesícules, la química de la superfície, la força iònica del medi, etc. Finalment, s’ha demostrat per SPR que els receptors expressats eren funcionals i que aquesta tècnica òptica permet la detecció de petites molècules, com són els odorants, a les concentracions en el rang micromolar. Els resultats presentats en aquesta tesis contribueixen donant un pas important a la realització de dispositius biosensors basats en nanovesícules naturals que integren receptors de membrana adjuntats a proteïna G.

This thesis presents the work carried out towards the implementation of RE-doped Si-based light emitting devices as integrated optoelectronic building blocks for Silicon Photonics. This work spans from the fundamentals such as the structure, the morphology of active layers containing Si-ncs and/or RE ions or the origin of the EL emission under different voltage excitations, to the development of advanced Si-based light emitting devices, providing insights on the device design, mask layout, device fabrication and the optoelectronic characterization. Also, novel layer architectures are proposed to overcome some of the inherent limitations of studied devices, paving the way towards efficient and reliable Si-based light emitting devices. This thesis is divided in two main blocks: one dedicated to the study of Er-doped Si-based light emitting devices emitting at 1.54 µm for on-chip optical data routing, and another one focussed on the structural and luminescence properties of Tb3+ and Ce3+ doped silicon oxide and oxynitride thin films with different layer compositions as enabling materials for sensing and RGB micro display applications. Also, different multilayer architectures containing alternated RE-doped single layers are explored. / Aquesta tesi presenta el treball dut a terme en la implementació de dispositius luminescents basats en silici i dopats amb terres rares pel desenvolupament de nous pilars optoelectrònics fonamentals compatibles amb la fotònica del silici. Aquest treball abasta des dels aspectes més fonamentals tals com l’estructura, la morfologia de les capes actives que contenen nano-cristalls de silici i/o ions de terres rares o l’estudi de l’origen de l’electroluminescència dels dispositius quan són polaritzats per diferents voltatges alterns, fins al desenvolupament de nous dispositius avançats basats en silici. En aquest cas, l’estudi portat a terme bé acompanyat d’una descripció detallada del disseny de cada dispositiu, el disseny de la màscara de procés elaborada especialment per a la fabricació dels dispositius en cas que sigui necessari, així com el procés de fabricació dels mateixos i una caracterització optoelectrònica detallada amb les pertinents conclusions al final de cada capítol. També, al final d’aquesta tesi es proposen nous dissenys i arquitectures de dispositiu que pretén establir un mapa de ruta per millorar els dispositius estudiats en aquest tesi, i que està basat en l’experiència adquirida durant els quatre anys d’investigació en dispositius electroluminescents basats en silici.

In this thesis I present a geophysical study that aims to define and characterize the crustal structure in the northern part of Morocco, especially beneath the Rif Cordillera. The geophysical data used in this thesis was acquired during the RIFSIS (2011) survey within the framework of the RIFSIS project, which was designated to overcome the lack of results on crustal structure in the northern part of Morocco. The RIFSIS project is mainly based on the acquisition of new wide-angle seismic profiles and on the integration of these data with the various seismic data sets currently available, such as the GASSIS, PICASSO, Topo-Iberia projects. The present work is based on the processing, modeling and interpretation of Wide-Angle seismic (WA) profiles, Receiver Functions (RFs) data projects, and has been focused on the crust below Morocco. The WA data was acquired during the RIFSIS survey in the Rif Cordillera from two profiles, the north-south profile (~430 km long) and the east-west profile (~330 km long). Both profiles extend across the Rif orogen, from the Middle Atlas to the Betic Range, and from the Gharb Basin to the Algerian border, respectively. The RFs data was acquired from the temporary deployment of seismological broadband stations from the Topo-Iberia and PICASSO projects. These stations were deployed along all the north Morocco territory. In this thesis I also have worked with Pn tomography of the uppermost mantle seismic velocity and anisotropy in the Euro-Mediterranean region. The results of the Pn tomography show significant features well correlated with the geological structures and evidence the heterogeneous character of the Euro-Mediterranean lithosphere. The processing of RIFSIS seismic data provides us the tectonic crustal structure model of the Rif Cordillera, whereas the modeling of WA data from travel-time forward modeling provides 2-D seismic velocity distribution of the crust and uppermost mantle. Furthermore, from the results that these projects provide, it becomes possible to infer the geometry of the crustal-mantle boundary, also presented in this thesis. These velocity-depth models hold major variations in the crustal thickness, especially the EW profile that shows a rapid change of 15-20 km in Moho depths within 30 km horizontal distances. Maximum depths around 50 km are found below the external Rif domain, while thinnest values of about 29 km are located eastwards, in the foreland and Atlasic terrances up to the Algerian border. The model along NS profile displays also marked differences in crustal thickness, ranging from 40 km beneath the Betics and internal Rif sampled domains, to 47-48 km beneath external Rif, and a progressive thinning southwards till Middle Atlas domain where the Moho is found at 32 km depth. In this thesis I also processed the WA data as low-fold WA seismic stack images. The deployment logistics during the RIFSIS project allowed that all the stations recorded all shots, not just aligned with the two profiles. From these profiles we can obtain a deep crustal image in­between the profiles and the shots. Although some uncertainties may be inherent to those approaches, a large crustal root, reaching more than 50 km, is well documented in the central part of the Rif Cordillera. At the same time I performed a RFs study in the Rif domain area, to determine the crustal thickness beneath the northern Morocco. The results obtained from the different methodologies used in this thesis reveal a crustal root well-documented in the central part of the Rif Cordillera, which is supported by the area large negative Bouguer anomaly values. / Aquesta tesi es centra en l’estudi de l’escorça cortical en el nord del Marroc, especialment sota de la Serralada del Rif, amb el principal objectiu de cobrir la falta de dades i resultats sobre l’estructura cortical de la zona. Per això, es realitza el processat, modelat i interpretació geològica de dos perfils geofísics de sísmica de gran angle adquirits durant la campanya sísmica RIFSIS (2011) en la Serralada del Rif, que s’han processat posteriorment com a com perfils low­fold. S’ha treballat amb dades de funcions receptores dels projectes Topo-Iberia i PICASSO. També s’ha treballat amb imatges tomografies del mantell superior sota la regió Euro-Mediterrània a partir d’ones Pn. A partir d’aquestes dades s’han pogut obtenir els resultats següents: 1. Dos models de velocitats de propagació d’ones P (Vp) i de la geometria de la Moho. 2. Obtenció d’imatges de l’escorça profunda al llarg dels perfils sísmics i entre els perfils i els dispars a partir dels perfils low-fold. 3. Obtenció d’un mapa de la topografia de la Moho del nord del Marroc a partir de les funcions receptores. 4. Els resultats obtinguts de les tomografies de Pn són altament coincidents amb les imatges tomogràfiques 3-D i mostren significants característiques ben correlacionades amb les estructures geològiques de l’Euro-Mediterrani. Amb aquestes metodologies s’ha pogut documentar una arrel cortical en la part central de la Serralada del Rif, resultats que coincideixen amb l’anomalia negativa de Bouguer observada en la regió. Finalment es proposa que la convergència de les plaques Ibèrica i Africana, juntament amb la possibilitat d’un slab sota el mar d’Alboran adjunt a la litosfera a la part central del Rif, podria ser l’explicació de la formació de l’arrel cortical sota el mateix Rif.

Tesi realitzada al Centre de Recerca en Agrigenòmica (CRAG) / La sobreexpresión del dominio de membrana de la Hidroximetil glutaril CoA reductasa (HMGR) en Arabidopsis thaliana desencadena la formación de agregados vesiculares de retículo endoplasmático (RE), esta capacidad morfogénica tiene precedentes claros en levaduras y mamíferos. En A. thaliana dos genes (HMG1 y HMG2) codifican tres isoformas de HMGR (HMGR1S, HMGR1L y HMGR2), las proteínas HMGR1S y HMGR1L derivan del mismo gen y tienen la misma secuencia, pero la isoforma 1L tiene una extensión en N-terminal de cincuenta residuos de aminoácidos, las tres isoformas de la HMGR se insertan co-traduccionalmente en la membrana del RE. En la presente tesis se determinó la localización subcelular de la HMGR1S de A. thaliana expresando el dominio de membrana de la HMGR1S unido a GFP, se observó que esta proteína quimérica se localiza en el RE formando agregados, con la ayuda de un marcador rojo T3RE, se observó en ensayos de colocalización en N. benthamiana que los agregados de HMGR son parte funcional del RE. Se determinó la organización estructural de las vesículas de HMGR formadas por el dominio de HMGR1S por primera vez a nivel de ultraestructura en hojas de N. benthamiana, así como también de una línea transgénica de A. thaliana, en este análisis se observaron distintas conformaciones lamelares, de círculos concéntricos y cristaloides presentes en los agregados. Se realizaron ensayos de inmunolocalización a nivel de la ultraestructura de los agregados de 1SGFP en A. thaliana, con anticuerpos contra GFP y anticuerpos contra el dominio catalítico de la HMGR, y se observó el marcaje de ambos anticuerpos, indicándonos la presencia en estos agregados, tanto de la 1SGFP como de la HMGR endógena, confirmando que el dominio de membrana de HMGR determina su localización subcelular. Los dominios de RE en el que se acumulan 1S:GFP y/o HMGR fueron denominados como Dominios ER-HMGR. Por otro lado, a pesar de las dificultades técnicas, un estudio anterior sugiere la coincidencia física entre vesículas de HMGR y ER bodies, estas últimas son estructuras derivadas de retículo endoplasmático que cumplen un rol importante en el mecanismo de defensa de las plantas en condiciones de estrés. La identidad estructural y funcional de los ER bodies no ha sido caracterizada aún. En este sentido en la presente tesis se profundizo en el estudio de la coincidencia física y funcional entre vesículas de HMGR y ER bodies, como su caracterización estructural en distintos tipos celulares de A. thaliana. Además, se determinó el papel biológico del dominio de membrana de HMGR en respuesta a estrés utilizando líneas mutantes knock out para el gen HMG1. En este análisis se observó que el gen de la HMGR1 es necesario para la derivación de Dominos ER-HMGR hacia vacuola en respuesta a estrés. / The overexpression of hydroxymetyl-glutaryl CoA reductase (HMGR) membrane domain in Arabidopsis thaliana triggers the endoplasmic reticulum (ER) vesicular aggregates formation, this morphogenetic capability has clear precedents in yeast and mammal cells. In the present thesis, the subcellular localization of A. thaliana HMGR1S isoform was determined through the expression of the HMGR1S membrane domain attached to GFP. This chimerical protein 1S:GFP is localized in the ER forming aggregates, the use of a red lumen ER marker T3RE allowed through colocalization assays in N. benthamiana, determine the 1S:GFP aggregates are a functional part of the ER. The ultra-structural organization of 1S:GFP aggregates induced by the HMGR membrane domain expression was determined for the first time at this level, in N. benthamiana agroinfiltrated and transgenic lines of A. thaliana. In this analysis, was observed that the 1S:GFP aggregates are conformed by smooth ER membranes distributed with different organization patterns, such as lamellar, whorls and mainly crystalloid. The immuno localization assays of 1S:GFP aggregates in A. thaliana at ultra-structural level, with anti-GFP and anti-CD1 (CD1:catalytic domain of HMGR1), revealed the presence in ER aggregates of 1S:GFP and HMGR distributed homogenously, therefore these aggregates are a specific domain characterized by the presence of HMGR and denominated as ER-HMGR domain. On the other hand, a previous study suggested the physical coincidence between HMGR vesicles and ER bodies, the last ones are ER derived structures which accumulates specific hydrolytic enzymes and has an important role in plant defense against stress conditions and pathogen attack. In the present work, the study of the physical coincidence between HMGR vesicles and ER bodies and the structural characterization in different tissues and cells was detailed. Additionally, the biologic role of the HMGR1S membrane domain in stress conditions was determined, using HMG1 gene knock out lines. In this analysis it was observed that the HMG1 gene is necessary to derive ER-HMGR domains to vacuole in response to stress conditions.

Tesi realitzada a l'Institut de Biologia Evolutiva (CSIC-UPF) / The arid areas of North Africa and Arabia cover a surface of more than 13 million square kilometers and are characterized by their extreme temperatures, diversity of desert habitats and well-adapted flora and fauna. The study of the evolution of their biota sheds light on the diversification processes in some of the world’s harshest environments and contributes to our knowledge on large-scale biogeographic patterns and processes. The general aim of this dissertation is to investigate the evolution of the biota of the arid areas of North Africa and Arabia, through the study of the systematics and biogeography of two representative reptile groups, Stenodactylus and Ptyodactylus. The geckos of these two genera are among the most common faunal elements of the arid North African and Arabian environments, yet they are very distinct with regard to their morphology, ecology and patterns of species divergence. On the one hand, the naked toe Stenodactylus species are relatively divergent morphologically and have conquered different arid and hyper-arid habitats across plains or dunes. On the other hand, Ptyodactylus geckos are fairly conserved morphologically across their entire range and are very well adapted to a common type of structural habitat characterized by the rocky substrate. The results presented herein provide a thorough insight into their phylogenetic relationships, review and update their taxonomy, explore their historical biogeography and, altogether, assemble a broader image of diversity patterns within a temporal framework for the arid North Africa and Arabia. Highlighting the most relevant results, in this dissertation an unprecedentedly large set of samples from North Africa, the Sahel and Arabia was compiled for the two selected groups of study: 249 samples of Stenodactylus from 150 localities and 382 samples of Ptyodactylus from 221 localities. Regarding molecular data, 770 DNA sequences of five different markers were newly produced for the former genus, and 1443 of six different markers for the latter. Stenodactylus originated in Arabia in the Late Oligocene. The Arabian Clade B, including five species, and the mainly African Clade C, with the same number of species, split approximately 22 Ma ago, coinciding with the opening of the Red Sea. Diversification within several clades in the phylogeny occurred during the Late Miocene, a time when a general increase in aridification initiated. Based on mitochondrial and nuclear molecular data, as well as morphological characters, a new species of Stenodactylus is described. Stenodactylus sharqiyahensis sp. nov. is shown to be endemic to the sand desert of Al Sharqiyah, in northeastern Oman. Nomenclatural actions undertaken for the stability of the nomina of North African species of Stenodactylus include the designation of a lectotype for the nomen Stenodactylus guttatus which places it under the synonymy of P. hasselquistii, ensuring continuity of the prevailing usage of S. petrii, and the application to the International Commission of Zoological Nomenclature to accept a new name-bearing type for S. sthenodactylus, in order to maintain its prevailing usage. Regarding Ptyodactylus, its broad sampling includes numerous new records for some of the species, and especially for the P. hasselquistii species complex, an important extension of its known distribution range both in northeastern Africa and Arabia. All the formerly known and newly-delimited species are shown to be mostly allopatric, except some known cases where activity patterns are adjusted to avoid competition. This is hypothesized to relate to their preference for the same type of structural habitat, conditioning their apparently conserved morphology. The onset of the diversification in the genus Ptyodactylus is estimated to have taken place in the Late Oligocene. In the northeastern African and Arabian clades A, B and C, diversification started during the Late Miocene, approximately 9.5-12 Ma ago, posterior to that in the western African P. togoensis and P. oudrii. The molecular data show that many species have high levels of genetic variability. Ptyodactylus ragazzii presents two clades in the phylogeny, from East and West Africa respectively, and given that topological tests reject their sister relationship, the available name P. togoensis is assigned to the West African populations. Multilocus coalescence-based analyses with the use of GMYC and BPP methodologies result in the delimitation of 17 putative species in the P. hasselquistii species complex. These species are grouped in two major clades: Clade A, with nine species, is distributed across northeastern Africa and a large part of north, central and eastern Arabian Peninsula, and Clade B, with eight species, is restricted to southern Arabia. / Les zones àrides del Nord d'Àfrica i d'Aràbia s'estenen per diversos milions de kilòmetres quadrats i l'estudi en aquesta àrea dels patrons de biodiversitat a gran escala, es veu condicionat de forma essencial per la dificultat d'obtenir un mostreig complet, per l'escassa informació de la seva biota i per la complexitat dels processos que han generat la biodiversitat actualment existent. En el marc limitat d'una tesi doctoral, i considerant les premisses anteriorment descrites, s'han seleccionat dos gèneres de rèptils àmpliament distribuïts en aquestes àrees, el gènere Stenodactylus i el Ptyodactylus. Ambdós gèneres formen part dels elements més comuns de la fauna del Nord d'Àfrica i d'Aràbia, malgrat presentar importants diferències pel que fa a la seva morfologia, ecologia i patrons de divergència. Per un costat, les espècies dels dragons del gènere Stenodactylus són relativament divergents entre elles respecte a la morfologia, i han conquistat diferents ambients àrids i hiperàrids, incloent-hi alguns dels entorns més hostils dels deserts. Per altra banda, els dragons del gènere Ptyodactylus són més aviat conservats pel que fa a la morfologia, sempre presentant els característics coixinets triangulars dividits en múltiples làmines en les seves potes, i estan ben adaptats als substrats rocosos, que constitueixen el seu hàbitat estructural. Els resultats aquí presentats aporten un coneixement profund en les seves relacions filogenètiques, es revisa i s'actualitza la seva taxonomia, s'explora la seva biogeografia històrica, i en conjunt, es dona una imatge general dels patrons de diversitat dins d'un marc temporal pels ambients àrids del Nord d'Àfrica i d'Aràbia.

In recent years, the LOC community has focused most of its research in the biomedical and biotechnology fields, due to the need of portable, low power consumption and low cost theranostics microdevices. Some developing countries do not have suitable medical diagnostics technologies and the supply and storage of the reagents is in many cases limited as well as the access to energy. Furthermore, developed countries are experimenting population aging needing novel low cost efficient disease-screening technologies. The introduction of LOC and microfluidics allow the integration of complex functions that could lead to the developing of more accurate, cheap and reliable theranostic tools. Current focus of application is focused mostly in drug delivery 1, cellular analysis 2, and disease diagnosis 3. Microfluidics is improving the developing of novel point-of-care devices, but there are some challenges that are slowing down the massive production of these LOC. These areas include new methods for sample collection, world-to-chip interfaces, sample pre-treatment, improvement of long-term stability of reagents, working with complex sample specimens, multiple detection of biomarkers and simplify the read-out 4. The main aim of this thesis work was to create novel, cheap and with a high degree of automatization miniaturized biosensing devices with the objective to facilitate Point-of-Care diagnostics in the near future. Our efforts have been focused into developing a LOC system with electrochemical sensing capabilities adjustable to any biomarker, depending only on sample volumes and required analysis times. The devices integrate low-cost label-free biosensors exploiting microfluidics-based self-functionalization, or specialization. The biosensor functionalization takes place in situ and selectively, just before the sensing, and their area keeps dry and inactive until the test starts. The reagents and the sensing parts are kept separated and brought into contact just before the test, avoiding the need of complex fabrication and storage methods to guarantee functionalization integrity. The novel design reduces the cost of the final instrumentation, by simplifying the measurements, while keeping sensitivities and LODs relevant for the application. Furthermore, since the interaction of antibody and protein is time and concentration dependent, our device has the capability to adjust its sensitivity. We have tuned and characterized our system sensitivity using different biomarkers. The development of our novel devices was possible by exploiting synergies in disciplines previously studied in our group. Particularly, in fields such as microfluidics 5-8, surface functionalization 9-14 and electrochemical biosensors 15-19. Summarizing, we are proposing novel microfluidic devices with integrated biosensors. The systems are based on the principle of laminar co-flow in order to perform an on-chip selective surface bio-functionalization of LOC integrated biosensors. This method has the advantage of performing the surface modification protocols “in situ” before the detection. The system can be easily scaled to incorporate several sensors with different biosensing targets in a single chip. We are proposing a novel voltage and impedance differential measurements; that allow us to simplify the read-out. As biomedical application we focus our attention on the detection of prostate cancer biomarkers. Bibliography 1. I. U. Khan, C. A. Serra, N. Anton and T. Vandamme, Journal of Controlled Release, 2013, 172, 1065-1074. 2. H. Andersson and A. Van den Berg, Sensors and Actuators B: Chemical, 2003, 92, 315-325. 3. M. J. Cima, Annual Review of Chemical and Biomolecular Engineering, 2011, 2, 355-378. 4. C. D. Chin, V. Linder and S. K. Sia, Lab on a Chip, 2012, 12, 2118-2134. 5. R. Rodriguez-Trujillo, C. A. Mills, J. Samitier and G. Gomila, Microfluidics and Nanofluidics, 2007, 3, 171-176. 6. R. Rodriguez-Trujillo, O. Castillo-Fernandez, M. Garrido, M. Arundell, A. Valencia and G. Gomila, Biosensors and Bioelectronics, 2008, 24, 290-296. 7. O. Castillo-Fernandez, R. Rodriguez-Trujillo, G. Gomila and J. Samitier, Microfluidics and Nanofluidics, 2014, 16, 91-99. 8. J. Comelles, V. Hortigüela, J. Samitier and E. Martínez, Langmuir, 2012, 28, 13688-13697. 9. E. Prats-Alfonso, F. García-Martín, N. Bayo, L. J. Cruz, M. Pla-Roca, J. Samitier, A. Errachid and F. Albericio, Tetrahedron, 2006, 62, 6876-6881. 10. J. Vidic, M. Pla-Roca, J. Grosclaude, M.-A. Persuy, R. Monnerie, D. Caballero, A. Errachid, Y. Hou, N. Jaffrezic-Renault, R. Salesse, E. Pajot-Augy and J. Samitier, Analytical Chemistry, 2007, 79, 3280-3290. 11. Y. Hou, S. Helali, A. Zhang, N. Jaffrezic-Renault, C. Martelet, J. Minic, T. Gorojankina, M.-A. Persuy, E. Pajot-Augy, R. Salesse, F. Bessueille, J. Samitier, A. Errachid, V. Akimov, L. Reggiani, C. Pennetta and E. Alfinito, Biosensors and Bioelectronics, 2006, 21, 1393-1402. 12. S. Rodríguez Seguí, M. Pla, J. Minic, E. Pajot‐Augy, R. Salesse, Y. Hou, N. Jaffrezic‐Renault, C. A. Mills, J. Samitier and A. Errachid, Analytical Letters, 2006, 39, 1735-1745. 13. A. Lagunas, J. Comelles, E. Martínez and J. Samitier, Langmuir, 2010, 26, 14154-14161. 14. A. Lagunas, J. Comelles, S. Oberhansl, V. Hortigüela, E. Martínez and J. Samitier, Nanomedicine: Nanotechnology, Biology and Medicine, 2013, 9, 694-701. 15. M. Castellarnau, N. Zine, J. Bausells, C. Madrid, A. Juárez, J. Samitier and A. Errachid, Materials Science and Engineering: C, 2008, 28, 680-685. 16. M. Castellarnau, N. Zine, J. Bausells, C. Madrid, A. Juárez, J. Samitier and A. Errachid, Sensors and Actuators B: Chemical, 2007, 120, 615-620. 17. M. Kuphal, C. A. Mills, H. Korri-Youssoufi and J. Samitier, Sensors and Actuators B: Chemical, 2012, 161, 279-284. 18. D. Caballero, E. Martinez, J. Bausells, A. Errachid and J. Samitier, Analytica Chimica Acta, 2012, 720, 43-48. 19. M. Barreiros dos Santos, J. P. Agusil, B. Prieto-Simón, C. Sporer, V. Teixeira and J. Samitier, Biosensors and Bioelectronics, 2013, 45, 174-180. / En años recientes, la comunidad de LOC ha enfocado todos sus esfuerzos en la investigación de nuevas aplicaciones para la biomedicina y biotecnología. Algunos países en vías de desarrollados no tienen tecnologías de diagnóstico adecuadas, además el suministro y almacenamiento de los reactivos es en muchos casos limitado, y en ocasiones cuentan con un acceso limitado al consumo de energía. Por otra parte, los países desarrollados se han encontrado con una población envejecida, y por lo tanto se ha generado la necesidad de contar con nuevas tecnologías para el diagnóstico de enfermedades las cuales sean accesibles y orientadas a una terapia más personalizada. Tanto la microfluídica como los LOC han permitido la integración de funciones de análisis complejas capaces de desarrollar herramientas de diagnostico más precisas, de bajo coste y confiables. Actualmente toda la atención se ha centrado en el diseño de aplicaciones para administración de fármacos 1, análisis celular 2 y diagnostico de enfermedades 3. La introducción de la microfluídica ha servido para mejorar el desarrollo de nuevos dispositivos point-of-care, pero todavía existen algunos problemas que han evitado la producción masiva de estos LOC. Las áreas en las que se pretende conseguir una mejora son la recolección de la muestra, mejora de la interfaz entre el chip y el usuario, tratamiento previo de la muestra, mejorar la estabilidad de los reactivos, trabajo con muestras complejas, detección múltiple de biomarcadores y simplificación del sistema de medida 4. Nuestros esfuerzos se han dedicado en desarrollar un sistema LOC con capacidad de detección electroquímica ajustable a cualquier biomarcador, dependiendo únicamente en la cantidad de muestra y los tiempos de análisis. Nuestros dispositivos microfluídicos cuentan con biosensores integrados de bajo coste con capacidad de auto-funcionalización. La funcionalización de los biosensores se realiza in-situ y selectivamente, antes de la detección, manteniendo el área de detección inerte hasta el inicio de la prueba. Los reactivos y el área de detección se almacenan por separado y entran en contacto hasta el inicio del experimento, lo cual facilita el método de fabricación. Se ha podido desarrollar este trabajo gracias a los estudios previos realizados en nuestro grupo en distintas disciplinas, tales como: microfluídica 5-8, funcionalización de superficies 9-14 y biosensores electroquímicos 15-19. Bibliografía 1. I. U. Khan, C. A. Serra, N. Anton and T. Vandamme, Journal of Controlled Release, 2013, 172, 1065-1074. 2. H. Andersson and A. Van den Berg, Sensors and Actuators B: Chemical, 2003, 92, 315-325. 3. M. J. Cima, Annual Review of Chemical and Biomolecular Engineering, 2011, 2, 355-378. 4. C. D. Chin, V. Linder and S. K. Sia, Lab on a Chip, 2012, 12, 2118-2134. 5. R. Rodriguez-Trujillo, C. A. Mills, J. Samitier and G. Gomila, Microfluidics and Nanofluidics, 2007, 3, 171-176. 6. R. Rodriguez-Trujillo, O. Castillo-Fernandez, M. Garrido, M. Arundell, A. Valencia and G. Gomila, Biosensors and Bioelectronics, 2008, 24, 290-296. 7. O. Castillo-Fernandez, R. Rodriguez-Trujillo, G. Gomila and J. Samitier, Microfluidics and Nanofluidics, 2014, 16, 91-99. 8. J. Comelles, V. Hortigüela, J. Samitier and E. Martínez, Langmuir, 2012, 28, 13688-13697. 9. E. Prats-Alfonso, F. García-Martín, N. Bayo, L. J. Cruz, M. Pla-Roca, J. Samitier, A. Errachid and F. Albericio, Tetrahedron, 2006, 62, 6876-6881. 10. J. Vidic, M. Pla-Roca, J. Grosclaude, M.-A. Persuy, R. Monnerie, D. Caballero, A. Errachid, Y. Hou, N. Jaffrezic-Renault, R. Salesse, E. Pajot-Augy and J. Samitier, Analytical Chemistry, 2007, 79, 3280-3290. 11. Y. Hou, S. Helali, A. Zhang, N. Jaffrezic-Renault, C. Martelet, J. Minic, T. Gorojankina, M.-A. Persuy, E. Pajot-Augy, R. Salesse, F. Bessueille, J. Samitier, A. Errachid, V. Akimov, L. Reggiani, C. Pennetta and E. Alfinito, Biosensors and Bioelectronics, 2006, 21, 1393-1402. 12. S. Rodríguez Seguí, M. Pla, J. Minic, E. Pajot‐Augy, R. Salesse, Y. Hou, N. Jaffrezic‐Renault, C. A. Mills, J. Samitier and A. Errachid, Analytical Letters, 2006, 39, 1735-1745. 13. A. Lagunas, J. Comelles, E. Martínez and J. Samitier, Langmuir, 2010, 26, 14154-14161. 14. A. Lagunas, J. Comelles, S. Oberhansl, V. Hortigüela, E. Martínez and J. Samitier, Nanomedicine: Nanotechnology, Biology and Medicine, 2013, 9, 694-701. 15. M. Castellarnau, N. Zine, J. Bausells, C. Madrid, A. Juárez, J. Samitier and A. Errachid, Materials Science and Engineering: C, 2008, 28, 680-685. 16. M. Castellarnau, N. Zine, J. Bausells, C. Madrid, A. Juárez, J. Samitier and A. Errachid, Sensors and Actuators B: Chemical, 2007, 120, 615-620. 17. M. Kuphal, C. A. Mills, H. Korri-Youssoufi and J. Samitier, Sensors and Actuators B: Chemical, 2012, 161, 279-284. 18. D. Caballero, E. Martinez, J. Bausells, A. Errachid and J. Samitier, Analytica Chimica Acta, 2012, 720, 43-48. 19. M. Barreiros dos Santos, J. P. Agusil, B. Prieto-Simón, C. Sporer, V. Teixeira and J. Samitier, Biosensors and Bioelectronics, 2013, 45, 174-180.

For this study the emerging contaminant ß-blocker Metoprolol (MET) has been selected due that it is a highly prescribed pharmaceutical and it has been detected in waste water treatment plants influents, thus, in natural waters. Several studies, focused on the toxicological potential of Metoprolol, indicate its potential environmental relevance and its recalcitrant nature. To remove MET from water, different Advanced Oxidation Processes (AOPs) were used. MET removal was studied, in different reactors with natural and artificial light, by photolysis, UVC/H2O2, photocatalysis, Fenton, photo-Fenton, bicarbonate-activated hydrogen peroxide (with cobalt or iron) processes. The different set-ups and technologies tested have been compared in order to establish the efficiency of the processes. The experiments were normally carried out with 50 mg/L of initial MET in Milli-Q water, free pH, and 25 ± 5 ºC. Thus, photolysis experiments were done in (solarbox (SB), Compound Parabolic Collector (CPC), Black light blue lamps (BLB) and UVC254 nm (UVC) reactors). The best result obtained was 93.5% of MET removal in UVC reactor after 240 minutes. UVC/H2O2 experiments were carried out in UVC reactor. Different H2O2 concentrations and pH were tested and the maximum removals were MET (98%) and TOC (70.7%) for 125 mg H2O2/L. Photocatalysis was carried in SB and CPC with different TiO2 concentrations (0.05, 0.10 and 0.40 g /L). Experiments were also carried out varying the initial MET concentration (25, 50 and 100 mg/L), pH and the water matrix with 0.4 g TiO2/L in SB or adding 25 and 150 mg/L of H2O2. The best results obtained were a complete MET removal and 45.7% of mineralization in SB and 81.5% of MET degradation and 29.2% of mineralization in CPC. The dark-Fenton experiments were carried out at pH in a reactor of 2 L. Two different concentrations of Fe (II) (2.5 mg/L or 10 mg/L) and H2O2 (25 mg/L or 150 mg/L) were used. To improve the Fenton process, the total Iron (II) concentration was divided in equal parts (5) and added at constant periods of time (12 minutes) during 1 hour. With the highest concentrations of iron and H2O2 the maximum MET conversion was 87.0% and mineralization 15.6%. Photo-Fenton experiments were done at pH 3, and temperature of 14 or 25 ºC in four reactors (BLB, SB, CPC and UVC). Two different concentrations of Fe (II) (2.5 mg/L or 10 mg/L) and H2O2 (25 mg/L or 150 mg/L) were used. With the highest iron and H2O2 concentrations, the best results in MET degradation were observed (BLB: 100% in 7 min; SB: 97.3% in 7 min; CPC: 98.3% in 3 min). The dark- Bicarbonate/hydrogen peroxide experiments were carried out with 5 mg/L of MET in drinking water, pH 6.2, and room temperature in a reactor of 0.5 L. To improve the process, cobalt (II) or iron (II) as catalyzer were added in the batch reactor. A complete MET conversion in 40 minutes was achieved. The efficiency of different AOPs and reactors tested was compared from the ratio between accumulated energy and MET eliminated. The energy is better used in CPC (0.065 kJ/mg) than in SB (0.275 kJ/mg). For photo-Fenton process (0.04 kJ/mg) UVC and (0.05 kJ/mg) BLB reactors exhibit a much better performance than (0.30 kJ/mg) SB and (0.26 kJ/mg) CPC reactors. From the intermediates identified, a possible MET fragmentation was proposed for the different processes, where, mainly oxidative attacks were detected. On the other hand, the irradiation in the photocatalytic reactor (SB) was measured by o-NB actinometry, based on pH or o-NB concentration. In addition, this work has demonstrated that the o-NB actinometry, followed by o-NB concentration consumption, could be used in the presence of the catalyst TiO2. / En este trabajo se ha estudiado la eficacia de varios Procesos de Oxidación Avanzada (UVC/H2O2, fotocatálisis, Fenton, foto-Fenton, bicarbonato/H2O2 con y sin catalizador) para degradar el fármaco Metoprolol (MET). Además se ha comparado la eficiencia energética de los diferentes procesos y diferentes instalaciones en la eliminación de MET. Los experimentos se realizaron con 50 mg/L iniciales de MET en agua Mili-Q, pH libre y 25ºC. En el caso de la fotólisis, se usaron cuatro instalaciones (solarbox (SB), concentradores parabólicos compuestos (CPC), lámparas black light blue (BLB) y UVC254 nm (UVC)) y el mejor resultado obtenido fue (UVC: 93,5%). Los experimentos UVC se realizaron a diferentes pH y concentraciones de H2O2 con una eliminación de MET de 98%. Los experimentos de fotocatálisis se llevaron a cabo con luz natural y artificial, variando la concentración de TiO2 (0,05, 0,10 y 0,40 g/L). También se realizaron experimentos con 0,4 g/L TiO2 pero variando la concentración inicial del MET (25, 50 y 100 mg/L), el pH, la matriz acuosa y adicionando peróxido de hidrógeno. Los mejores resultados fueron (CPC: 81,5% y SB: 100%). Los experimentos de Fenton se realizaron a pH 3,0 en un reactor de 2 L. Para mejorar el proceso, la adición del Fe (II) se dividió en 5 adiciones realizadas a intervalos constantes de tiempo durante 60 minutos. La mejor degradación de MET fue 87%. Los experimentos de foto-Fenton se realizaron a pH 3,0 y temperaturas de 14ºC y 25ºC en cuatro instalaciones diferentes (BLB, SB, CPC y UVC). Los mejores resultados obtenidos para la eliminación de MET fueron (BLB: 100%; SB: 97,3% y CPC: 98,3%). Los experimentos con bicarbonato/H2O2 se realizaron con 5 mg/L iniciales de MET en agua potable, pH libre y temperatura ambiental en un reactor de 0,5 L. Adicionalmente se utilizó Co (II) y Fe (II) como catalizadores. Eliminación de MET de 100%. Se realizó la identificación de los diferentes intermedios y se han establecido los posibles caminos de degradación del MET. Finalmente se realizó un estudio de un método actinométrico para poder realizar mediciones de radiación en un reactor fotocatalítico en presencia de un catalizador en suspensión.

In this doctoral thesis the interaction of emergent organic pollutants, such as alkylphenolic and perfluorinated compounds (APCs and PFCs, respectively) with sewage sludge and soil samples was studied. These two families of organic compounds were selected because of their ubiquitous presence and persistence in environmental matrices and to know mechanisms responsible for their interaction. With respect to the behaviour of APCs in sewage sludges, concretely octylphenol (OP), nonylphenol (NP) and nonylphenol-mono-ethoxylate (NP1EO), the application of a leaching test to fresh sludge samples and to samples dried at 40 °C showed that leaching yields of APCs decreased after the drying treatment (from 1.3-35% for fresh sludge samples to 0.8-3.4% for dried samples). This was attributed to differences in the characteristics of sludge organic matter in fresh and dried sludge samples. Thus, in the fresh sludge samples, APCs were mainly associated with the particulate matter in solution, showing high leaching yields. In contrast, drying treatment provoked the transformation of organic compounds into simpler molecules, thus leading to a decrease in the particulate matter in solution, thus favouring the association of APCs with the organic matter of the solid phase, and decreasing the leaching yield. Therefore, sewage sludge drying is recommended before their potential addition to agricultural soils. For studying the sorption behavior of NP and NP1EO in soils, sorption and desorption tests were applied to several soils with different characteristics such as pH, organic carbon (OC) and dissolved organic carbon. Sorption and desorption isotherms of NP and NP1EO were linear within the concentration range studied, and the solid-liquid distribution coefficients (Kd) derived from the isotherms correlated with the OC of soils, suggesting that soil organic matter governed the sorption of NP and NP1EO through a hydrophobic interaction with the alkyl chain of APCs. Exception to this pattern was the sorption of NP1EO in those soils with the highest DOC/OC ratio, which were better described by the Freundlich equation (N>1), due to the competition between the organic matter in solution and that in the solid phase for NP1EO sorption at low initial concentrations. Moreover, Kd values were usually higher for NP than for NP1EO, according with the higher hydrophobicity of NP. Desorption yields of NP and NP1EO were lower than 5 %, thus indicating that the sorption of APCs in soils was considerably irreversible. Sorption behaviour of PFCs, concretely perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA), perfluorobutane sulfonate (PFBS), in sewage sludge and soil samples led to similar conclusions to those obtained for APCs. Sorption and desorption isotherms showed a linear pattern in the two environmental matrices, with satisfactory correlations between Kd and the organic matter content of the sample, suggesting a hydrophobic interaction between the pefluorinated carbon chain and the organic matter of sewage sludges and soils, predominantly Van der Waals forces. Potential additional role of Ca/Mg ion bridges to overcome the electrostatic repulsions of deprotonated PFCs with negatively charged surface sites of sludges at the sample pH was also suggested. Sorption affinity of the PFCs for the sludge and soil samples increased with the hydrophobicity of the compound, that is, in the sequence: PFBS < PFOA < PFOS. In contrast, PFOS showed the highest irreversibility in the two environmental matrices, whereas PFOA and PFBS were desorbed in higher rates. This suggested that the physicochemical properties of the compound, such as solubility and octanol-water partitioning coefficient, together with the organic matter content of sewage sludges and soils controlled the sorption behaviour of PFCs. / En esta tesis doctoral se ha estudiado la interacción de contaminantes orgánicos persistentes, tales como compuestos alquilfenólicos (APCs) y perfluorados (PFCs) con lodos de depuradora y suelos. Por lo que se refiere a la lixiviación de APCs en muestras de lodo, concretamente de octilfenol (OP), nonilfenol (NP) y nonilfenol-mono-etoxilado (NP1EO), ésta disminuyó después de un tratamiento de secado a 40 °C. Este comportamiento se atribuyó al hecho que, en el lodo fresco, los APCs se encontraban mayoritariamente asociados a la materia orgánica particulada en solución, mientras que el secado condujo a una transformación de la materia orgánica en moléculas más sencillas, provocando que los APCs quedaran mayoritariamente retenidos en la fase sólida del lodo, disminuyendo su lixiviación. En relación con la interacción de APCs en suelos, se evaluó la sorción y desorción de NP y NP1EO en suelos con distintos contenidos del carbono orgánico (OC). La sorción y desorción de estos compuestos fue lineal para el intervalo de concentraciones estudiado y los coeficientes de distribución sólido-líquido (Kd) obtenidos mostraron una buena correlación con el OC, lo que sugirió una interacción hidrófobica entre la cadena alquilo del APC y la materia orgánica de los suelos. La desorción de NP y NP1EO fue inferior a 5% para todas las combinaciones APC-suelo, demostrando una sorción significativamente irreversible de estos compuestos. Los estudios de sorción de PFCs, concretamente de sulfonato de perfluoroctano (PFOS), ácido perfluorooctanoico (PFOA) y sulfonato de perfluorobutano (PFBS), en lodos y suelos llevaron a conclusiones similares a las obtenidas con APCs. Las isotermas de sorción de PFCs también mostraron un comportamiento lineal en ambas matrices ambientales con una buena correlación entre Kd y OC, lo que sugirió una interacción hidrofóbica entre la cadena perfluorada y la materia orgánica. Se observó que la afinidad de sorción de PFCs en lodos y suelos aumentó con la hidrofobicidad del compuesto (PFBS < PFOA < PFOS), mientras que la desorción disminuyó, siendo el PFOS el más irreversiblemente sorbido, lo que permitió concluir que la hidrofobicidad del compuesto gobierna su sorción en lodos y suelos.

Force plays an essential role in all fields of biology. Measurement of these forces with high precision provides information about the structure, dynamics, intra and intermolecular interactions, and the mechanical properties of the biomolecules and, in general, about molecular basis of diverse biological phenomena. Different techniques have been developed to address this task, particularly at the single molecule level. The main objective accomplished in this work of thesis is the development of sensors technologies applied to force spectroscopy measurements and the demonstration of its possibilities in real molecular studies. Scanning probe microscopy (SPM) is a fast growing technology that has been the source for the development of an immense variety of applications to investigate materials and molecules at nanoscale. As another technology, SPM is constantly improving through different advances in instrumentation level and the emergence of new applications. From the analysis of the main limitations of quartz tuning fork (QTF) based nanosensors on one side and the conventional force spectroscopy with cantilever tips on the other side, the main considerations have been determined for the technological developments on force microscopy applications. One of the main limitations of tuning fork probes is that they are usually custom-made because no commercial probes suitable for a wide range of experiments are available. The custom-made devices show considerable variation in dynamic response, poor lateral resolution and the characterization of the sensors remains unclear for force quantification. A new controller is developed to ensure the same dynamic response of different sensors in order to maintain the conditions in which the measurements are conducted. Also, a new method to improve lateral resolution of the QTF probes when working in liquid is proposed in this thesis based on attaching a standard AFM tip to the end of the fiber probe which has been previously sharpened. A method to calculate the spring constant of the QTF based sensors from easily measurable parameters is presented in this thesis. The method is based on a finite element analysis (FEA) model which includes the electrical part and can be used to calculate the spring constant of a QTF accurately for quantitative measurements. Results obtained in real biological experiments are promising and show the possibilities of the shear force microscopy improvements developed in this work of thesis. In a first experiment, a self-assembled monolayer (SAM) of micropatterned antibodies was imaged with three different techniques and in a second experiment, a molecular interaction analysis was done between biotin- streptavidin complex and results are compared with those obtained with AFM tip. The main problem for comparing steered molecular dynamics (SMD) simulation results with experimental data is that SMD simulations were restricted to nanosecond timescales (due to the high computational demand of all-atomistic simulations). A high-speed force spectroscopy methodology has been developed to achieve rates comparable to SMD simulations. The validation of the technique is performed with titin unfolding measurements allowing the direct comparison of experimental and simulated forces. / La fuerza juega un papel esencial en todos los campos de la biología. Las fuerzas experimentadas y generadas por las biomoléculas son múltiples en la naturaleza y pueden ir desde los subpiconewtons hasta varios nanonewtons. La medida de estas fuerzas con alta precisión proporciona información acerca de la estructura, la dinámica, las interacciones intra e intermoleculares y las propiedades mecánicas de las biomoléculas. Se han desarrollado diferentes técnicas para hacer frente a esta tarea, en particular a nivel de moléculas individuales. El principal objetivo en este trabajo de tesis ha sido el desarrollo de tecnologías de sensores aplicadas a la espectroscopia de fuerzas y la demostración de sus posibilidades en estudios moleculares reales. La microscopía de sonda de barrido (SPM) es una tecnología de rápido crecimiento que ha sido la fuente para el desarrollo de una inmensa variedad de aplicaciones para investigar materiales y moléculas a la nanoescala. A partir del análisis de las principales limitaciones de los nanosensores basados en tuning fork (TF) por un lado y la espectroscopia de fuerzas convencional con puntas de AFM por otro lado, se han determinado las principales consideraciones para los desarrollos tecnológicos. Una vez que la tecnología se ha desarrollado, se han llevado a cabo diferentes experimentos biológicos con el objetivo de demostrar las posibilidades de las tecnologías desarrolladas en aplicaciones reales: (i) Diferentes técnicas de imagen de biomoléculas se han comparado con sensores TF. La muestra estudiada ha sido una monocapa auto-ensamblada (SAM) de anticuerpos microestructurada. (ii) Se han realizado medidas cuantitativas de interacción molecular entre el sistema biotina- estreptavidina midiendo las energías de adhesión a diferentes velocidades de tracción y los resultados se han comparado con los obtenidos con medidas de AFM. (iii) Medidas de desplegamiento de la proteína titina se han realizado con la nueva técnica desarrollada de espectroscopia de fuerzas a velocidades alcanzadas por simulación (~4 milímetros por segundo) pudiendo comparar los resultados experimentales con simulaciones.