When small non-coding RNAs meet the ribosome: Tuning the translational machinery

The functions of ribosomes in translation are complex and involve different types of activities critical for decoding the genetic code, linkage of amino acids via amide bonds to form polypeptide chains, as well as the release and proper targeting of the synthesized protein. Non-protein-coding RNAs (ncRNAs) have been recognized to be crucial in establishing regulatory networks.1 However all of the recently discovered ncRNAs involved in translation regulation target the mRNA rather than the ribosome. The main goal of this project is to identify potential novel ncRNAs that directly bind and possibly regulate the ribosome during protein biosynthesis. To address this question we applied various stress conditions to the archaeal model organism Haloferax volcanii and deep-sequenced the ribosome-associated small ncRNA interactome. In total we identified 6.250 ncRNA candidates. Significantly, we observed the emersed presence of tRNA-derived fragments (tRFs). These tRFs have been identified in all domains of life and represent a growing, yet functionally poorly understood, class of ncRNAs. Here we present evidence that tRFs from H. volcanii directly bind to ribosomes. In the presented genomic screen of the ribosome-associated RNome a 26 residue long fragment originating from the 5’ part of valine tRNA was by far the most abundant tRF. The Val-tRF is processed in a stress- dependent manner and was found to primarily target the small ribosomal subunit in vitro and in vivo. As a consequence of ribosome binding, Val-tRF reduces protein synthesis by interfering with peptidyl transferase activity. Therefore this tRF functions as ribosome-bound small ncRNA capable of regulating gene expression in H. volcanii under environmental stress conditions probably by fine-tuning the rate of protein production.2 Currently we are investigating the binding site of this tRF on the 30S subunit in more detail.

Evidence for a role of sterol 27-hydroxylase in glucocorticoid metabolism in vivo

The intracellular availability of glucocorticoids is regulated by the enzymes 11β-hydroxysteroid dehydrogenase 1 (HSD11B1) and 11β-hydroxysteroid dehydrogenase 2 (HSD11B2). The activity of HSD11B1 is measured in the urine based on the (tetrahydrocortisol+5α-tetrahydrocortisol)/tetrahydrocortisone ((THF+5α-THF)/THE) ratio in humans and the (tetrahydrocorticosterone+5α-tetrahydrocorticosterone)/tetrahydrodehydrocorticosterone ((THB+5α-THB)/THA) ratio in mice. The cortisol/cortisone (F/E) ratio in humans and the corticosterone/11-dehydrocorticosterone (B/A) ratio in mice are markers of the activity of HSD11B2. In vitro agonist treatment of liver X receptor (LXR) down-regulates the activity of HSD11B1. Sterol 27-hydroxylase (CYP27A1) catalyses the first step in the alternative pathway of bile acid synthesis by hydroxylating cholesterol to 27-hydroxycholesterol (27-OHC). Since 27-OHC is a natural ligand for LXR, we hypothesised that CYP27A1 deficiency may up-regulate the activity of HSD11B1. In a patient with cerebrotendinous xanthomatosis carrying a loss-of-function mutation in CYP27A1, the plasma concentrations of 27-OHC were dramatically reduced (3.8 vs 90-140 ng/ml in healthy controls) and the urinary ratios of (THF+5α-THF)/THE and F/E were increased, demonstrating enhanced HSD11B1 and diminished HSD11B2 activities. Similarly, in Cyp27a1 knockout (KO) mice, the plasma concentrations of 27-OHC were undetectable (<1 vs 25-120 ng/ml in Cyp27a1 WT mice). The urinary ratio of (THB+5α-THB)/THA was fourfold and that of B/A was twofold higher in KO mice than in their WT littermates. The (THB+5α-THB)/THA ratio was also significantly increased in the plasma, liver and kidney of KO mice. In the liver of these mice, the increase in the concentrations of active glucocorticoids was due to increased liver weight as a consequence of Cyp27a1 deficiency. In vitro, 27-OHC acts as an inhibitor of the activity of HSD11B1. Our studies suggest that the expression of CYP27A1 modulates the concentrations of active glucocorticoids in both humans and mice and in vitro.

Theoretical models of planetary system formation II. Post-formation evolution

Aims. We extend the results of planetary formation synthesis by computing the long-term evolution of synthetic systems from the clearing of the gas disk into the dynamical evolution phase. Methods. We use the symplectic integrator SyMBA to numerically integrate the orbits of planets for 100 Myr, using populations from previous studies as initial conditions. Results. We show that within the populations studied, mass and semimajor axis distributions experience only minor changes from post-formation evolution. We also show that, depending upon their initial distribution, planetary eccentricities can statistically increase or decrease as a result of gravitational interactions. We find that planetary masses and orbital spacings provided by planet formation models do not result in eccentricity distributions comparable to observed exoplanet eccentricities, requiring other phenomena, such as stellar fly-bys, to account for observed eccentricities.

Characterization of template switching and recombination during Moloney murine leukemia virus replication

Retroviruses uniquely co-package two copies of their genomic RNA within each virion. The two copies are used as templates for synthesis of the proviral DNA during the process of reverse transcription. Two template switches are required to complete retroviral DNA synthesis by the retroviral enzyme, reverse transcriptase. With two RNA genomes present in the virion, reverse transcriptase can make template switches utilizing only one of the RNA templates (intramolecular) or utilizing both RNA templates (intermolecular) during the process of reverse transcription. The results presented in this study show that during a single cycle of Moloney murine leukemia virus replication, both nonrecombinant and recombinant proviruses predominantly underwent intramolecular minus- and plus-strand transfers during the process of reverse transcription. This is the first study to examine the nature of the required template switches occurring during MLV replication and these results support the previous findings for SNV, and the hypothesis that the required template switches are ordered events. This study also determined rates for deletion and a rate of recombination for a single cycle of MLV replication. The rates reported here are comparable to the rates previously reported for both SNV and MLV. ^

Function and regulation of anionic phospholipids in mitochondria of Saccharomyces cerevisiae

As the major anionic phospholipids predominantly found in the mitochondrial inner membrane of eukaryotic cells, cardiolipin (CL) and its precursor phosphatidylglycerol (PG) are of great importance in many critical mitochondrial processes. Pgs1Δ cells of Saccharomyces cerevisiae lacking both PG and CL display severe mitochondrial defects. Translation of several proteins including products of four mitochondrial DNA (mtDNA) encoded genes (COX1, COX2, COX3, and COB ) and one nuclear-encoded gene (COX4) is inhibited. The molecular basis of this phenotype was analyzed using a combined biochemical, molecular and genetic approach. ^ Using a mitochondrial targeted green fluorescence protein (mtGFP) fused to the COX4 promoter and its 5′ and 3′ untranslated regions (UTRs), lack of mtGFP expression independent of carbon source and strain background was confirmed to be at the translational level. The translational defect was not due to deficiency of mitochondrial respiratory function but rather caused directly by the lack of PG/CL in the mitochondrial membrane. Re-introduction of a functional PGS1 gene restored PG synthesis and expression of the above mtGFP. Deletional analysis of the 5′ UTR of COX4 mRNA revealed the presence of a 50 nt sequence as a cis-acting element inhibiting COX4 translation. Using similar constructs with HIS3 and lacZ as reporter genes, extragenic spontaneous mutations that allowed expression of His3p and β-galactosidase were isolated, which appeared to be recessive and derived from loss-of-function mutations as determined by mating analysis. Using a tetracycline repressible plasmid-borne PGS1 expression system and an in vivo mitochondrial protein translation method, the translation of mtDNA encoded COX1 and COX3 mRNAs was shown to be significantly inhibited in parallel with reduced levels of PG/CL content. Therefore, the cytoplasmic translation machinery appears to be able to sense the level of PG/CL in mitochondria and regulate COX4 translation coordinately with the mtDNA encoded subunits. ^ The essential requirement of PG and CL in mitochondrial function was further demonstrated in the study of CL synthesis by factors affecting mitochondrial biogenesis such as carbon source, growth phase or mitochondrial mutations at the level of transcription. We have also demonstrated that CL synthesis is dependent on the level of PG and INO2/INO4 regulatory genes. ^

The molecular mechanisms of sensing nucleoside analogue-induced DNA damage

Nucleoside analogues are antimetabolites effective in the treatment of a wide variety of solid tumors and hematological malignancies. Upon being metabolized to their active triphosphate form, these agents are incorporated into DNA during replication or excision repair synthesis. Because DNA polymerases have a greatly decreased affinity for primers terminated by most nucleoside analogues, their incorporation causes stalling of replication forks. The molecular mechanisms that recognize blocked replication may contribute to drug resistance but have not yet been elucidated. Here, several molecules involved in sensing nucleoside analogue-induced stalled replication forks have been identified and examined for their contribution to drug resistance. ^ The phosphorylation of the DNA damage sensor, H2AX, was characterized in response to nucleoside analogues and found to be dependent on both time and drug concentration. This response was most evident in the S-phase fraction and was associated with an inhibition of DNA synthesis, S-phase accumulation, and activation of the S-phase checkpoint pathway (Chk1-Cdc25A-Cdk2). Exposure of the Chk1 inhibitor, 7-hydroxystaurosporine (UCN-01), to cultures previously treated with nucleoside analogues caused increased apoptosis, clonogenic death, and a further log-order increase in H2AX phosphorylation, suggesting enhanced DNA damage. Ataxia-telangiectasia mutated (ATM) has been identified as a key DNA damage signaling kinase for initiating cell cycle arrest, DNA repair, and apoptosis while the Mre11-Rad50-Nbs1 (MRN) complex is known for its functions in double-strand break repair. Activated ATM and the MRN complex formed distinct nuclear foci that colocalized with phosphorylated H2AX after inhibition of DNA synthesis by the nucleoside analogues, gemcitabine, ara-C, and troxacitabine. Since double-strand breaks were undetectable, this response was likely due to stalling of replication forks. A similar DNA damage response was observed in human lymphocytes after exposure to ionizing radiation and in acute myelogenous leukemia blasts during therapy with the ara-C prodrug, CP-4055. Deficiencies in ATM, Mre11, and Rad50 led to a two- to five-fold increase in gemcitabine sensitivity, suggesting that these molecules contribute to drug resistance. Based on these results, a model is proposed for the sensing of nucleoside analogue-induced stalled replication forks that includes H2AX, ATM, and the Mre11-Rad50-Nbs1 complex. ^

Mechanisms Underlying the Heterogeneous Sensitivities of Cancer Cells to Proteasome Inhibitors

The mechanisms underlying cellular response to proteasome inhibitors have not been clearly elucidated in solid tumor models. Evidence suggests that the ability of a cell to manage the amount of proteotoxic stress following proteasome inhibition dictates survival. In this study using the FDA-approved proteasome inhibitor bortezomib (Velcade®) in solid tumor cells, we demonstrated that perhaps the most critical response to proteasome inhibition is repression of global protein synthesis by phosphorylation of the eukaryotic initiation factor 2-α subunit (eIF2α). In a panel of 10 distinct human pancreatic cancer cells, we showed marked heterogeneity in the ability of cancer cells to induce eIF2α phosphorylation upon stress (eIF2α-P); lack of inducible eIF2α-P led to excessive accumulation of aggregated proteins, reactive oxygen species, and ultimately cell death. In addition, we examined complementary cytoprotective mechanisms involving the activation of the heat shock response (HSR), and found that induction of heat shock protein 70 kDa (Hsp72) protected against proteasome inhibitor-induced cell death in human bladder cancer cells. Finally, investigation of a novel histone deacetylase 6 (HDAC6)-selective inhibitor suggested that the cytoprotective role of the cytoplasmic histone deacetylase 6 (HDAC6) in response to proteasome inhibition may have been previously overestimated.

Chemical and Sr isotopic compositions of pore fluids and marine carbonates from ODP Hole 130-807A

The 87Sr/86Sr ratios and Sr concentrations in sediment and pore fluids are used to evaluate the rates of calcite recrystallization at ODP Site 807A on the Ontong Java Plateau, an 800-meter thick section of carbonate ooze and chalk. A numerical model is used to evaluate the pore fluid chemistry and Sr isotopes in an accumulating section. The deduced calcite recrystallization rate is 2% per million years (%/Myr) near the top of the section and decreases systematically in older parts of the section such that the rate is close to 0.1/age (in years). The deduced recrystallization rates have important implications for the interpretation of Ca and Mg concentration profiles in the pore fluids. The effect of calcite recrystallization on pore fluid chemistry is described by the reaction length, L, which varies by element, and depends on the concentration in pore fluid and solid. When L is small compared to the thickness of the sedimentary section, the pore fluid concentration is controlled by equilibrium or steady-state exchange with the solid phase, except within a distance L of the sediment-water interface. When L is large relative to the thickness of sediment, the pore fluid concentration is mostly controlled by the boundary conditions and diffusion. The values of L for Ca, Sr, and Mg are of order 15, 150, and 1500 meters, respectively. L_Sr is derived from isotopic data and modeling, and allows us to infer the values of L_Ca and L_Mg. The small value for L_Ca indicates that pore fluid Ca concentrations, which gradually increase down section, must be equilibrium values that are maintained by solution-precipitation exchange with calcite and do not reflect Ca sources within or below the sediment column. The pore fluid Ca measurements and measured alkalinity allow us to calculate the in situ pH in the pore fluids, which decreases from 7.6 near the sediment-water interface to 7.1+/-0.1 at 400-800 mbsf. While the calculated pH values are in agreement with some of the values measured during ODP Leg 130, most of the measurements are artifacts. The large value for L_Mg indicates that the pore fluid Mg concentrations at 807A are not controlled by calcite-fluid equilibrium but instead are determined by the changing Mg concentration of seawater during deposition, modified by aqueous diffusion in the pore fluids. We use the pore fluid Mg concentration profile at Site 807A to retrieve a global record for seawater Mg over the past 35 Myr, which shows that seawater Mg has increased rapidly over the past 10 Myr, rather than gradually over the past 60 Myr. This observation suggests that the Cenozoic rise in seawater Mg is controlled by continental weathering inputs rather than by exchange with oceanic crust. The relationship determined between reaction rate and age in silicates and carbonates is strikingly similar, which suggests that reaction affinity is not the primary determinant of silicate dissolution rates in nature.

Fabrication of high frequency SAW resonators using AlN/Diamond/Si technology

The synthesis of AlN on diamond is a great challenge, not only because of the between an AlN/diamond interface, but also because of the high surface roughness of the diamond layers [8, 9]. In the case of microcrystalline diamond, the last problem was solved by polishing. However, polishing nanocrystalline diamond is not straightforward. For the diamond synthesis by CVD, silicon was used as a substrate. The diamond/Si interface presents a smoother diamond than the diamond/air interface. This paper reports on the fabrication of high frequency SAW resonators using AlN/Diamond/Si technology.

Aspectos de la medición dinámica instantánea de emisiones de motores. Aplicación al desarrollo de un equipo portátil y una metodología para estudios de contaminación de vehículos en tráfico real.

Esta Tesis Doctoral se encuadra en el ámbito de la medida de emisiones contaminantes y de consumo de combustible en motores de combustión interna alternativos cuando se utilizan como plantas de potencia para propulsión de vehículos ligeros de carretera, y más concretamente en las medidas dinámicas con el vehículo circulando en tráfico real. En este ámbito, el objetivo principal de la Tesis es estudiar los problemas asociados a la medición en tiempo real con equipos embarcados de variables medioambientales, energéticas y de actividad, de vehículos ligeros propulsados por motores térmicos en tráfico real. Y como consecuencia, desarrollar un equipo y una metodología apropiada para este objetivo, con el fin de realizar consiguientemente un estudio sobre los diferentes factores que influyen sobre las emisiones y el consumo de combustible de vehículos turismo en tráfico real. La Tesis se comienza realizando un estudio prospectivo sobre los trabajos de otros autores relativos al desarrollo de equipos portátiles de medida de emisiones (Portable Emission Measurement Systems – PEMS), problemas asociados a la medición dinámica de emisiones y estudios de aplicación en tráfico real utilizando este tipo de equipos. Como resultado de este estudio se plantea la necesidad de disponer de un equipo específicamente diseñado para ser embarcado en un vehículo que sea capaz de medir en tiempo real las concentraciones de emisiones y el caudal de gases de escape, al mismo tiempo que se registran variables del motor, del vehículo y del entorno como son la pendiente y los datos meteorológicos. De esta forma se establecen las especificaciones y condiciones de diseño del equipo PEMS. Aunque al inicio de esta Tesis ya existían en el mercado algunos sistemas portátiles de medida de emisiones (PEMS: Portable Emissions Measurement Systems), en esta Tesis se investiga, diseña y construye un nuevo sistema propio, denominado MIVECO – PEMS. Se exponen, discuten y justifican todas las soluciones técnicas incorporadas en el sistema que incluyen los subsistema de análisis de gases, subsistemas de toma de muestra incluyendo caudalímetro de gases de escape, el subsistema de medida de variables del entorno y actividad del vehículo y el conjunto de sistemas auxiliares. El diseño final responde a las hipótesis y necesidades planteadas y se valida en uso real, en banco de rodillos y en comparación con otro equipos de medida de emisiones estacionarios y portátiles. En esta Tesis se presenta también toda la investigación que ha conducido a establecer la metodología de tratamiento de las señales registradas en tiempo real que incluye la sincronización, cálculos y propagación de errores. La metodología de selección y caracterización de los recorridos y circuitos y de las pautas de conducción, preparación del vehículo y calibración de los equipos forma también parte del legado de esta Tesis. Para demostrar la capacidad de medida del equipo y el tipo de resultados que pueden obtenerse y que son útiles para la comunidad científica, y las autoridades medioambientales en la parte final de esta Tesis se plantean y se presentan los resultados de varios estudios de variables endógenas y exógenas que afectan a las emisiones instantáneas y a los factores de emisión y consumo (g/km) como: el estilo de conducción, la infraestructura vial, el nivel de congestión del tráfico, tráfico urbano o extraurbano, el contenido de biocarburante, tipo de motor (diesel y encendido provocado), etc. Las principales conclusiones de esta Tesis son que es posible medir emisiones másicas y consumo de motores de vehículos en uso real y que los resultados permiten establecer políticas de reducción de impacto medio ambiental y de eficiencia energética, pero, se deben establecer unas metodologías precisas y se debe tener mucho cuidado en todo el proceso de calibración, medida y postratamientos de los datos. Abstract This doctoral thesis is in the field of emissions and fuel consumption measurement of reciprocating internal combustion engines when are used as power-trains for light-duty road vehicles, and especially in the real-time dynamic measurements procedures when the vehicle is being driven in real traffic. In this context, the main objective of this thesis is to study the problems associated with on-board real-time measuring systems of environmental, energy and activity variables of light vehicles powered by internal combustion engines in real traffic, and as a result, to develop an instrument and an appropriate methodology for this purpose, and consequently to make a study of the different factors which influence the emissions and the fuel consumption of passenger cars in real traffic. The thesis begins developing a prospective study on other authors’ works about development of Portable Emission Measurement Systems (PEMS), problems associated with dynamic emission measurements and application studies on actual traffic using PEMS. As a result of this study, it was shown that a measuring system specifically designed for being on-board on a vehicle, which can measure in real time emission concentrations and exhaust flow, and at the same time to record motor vehicle and environment variables as the slope and atmospheric data, is needed; and the specifications and design parameters of the equipment are proposed. Although at the beginning of this research work there were already on the market some PEMS, in this Thesis a new system is researched, designed and built, called MIVECO – PEMS, in order to meet such measurements needs. Following that, there are presented, discussed and justify all technical solutions incorporated in the system, including the gas analysis subsystem, sampling and exhaust gas flowmeter subsystem, the subsystem for measurement of environment variables and of the vehicle activity and the set of auxiliary subsystems. The final design meets the needs and hypotheses proposed, and is validated in real-life use and chassis dynamometer testing and is also compared with other stationary and on-board systems. This thesis also presents all the research that has led to the methodology of processing the set of signals recorded in real time including signal timing, calculations and error propagation. The methodology to select and characterize of the routes and circuits, the driving patterns, and the vehicle preparation and calibration of the instruments and sensors are part of the legacy of this thesis. To demonstrate the measurement capabilities of the system and the type of results that can be obtained and that are useful for the scientific community and the environmental authorities, at the end of this Thesis is presented the results of several studies of endogenous and exogenous variables that affect the instantaneous and averaged emissions and consumption factors (g/km), as: driving style, road infrastructure, the level of traffic congestion, urban and extra-urban traffic, biofuels content, type of engine (diesel or spark ignition) etc. The main conclusions of this thesis are that it is possible to measure mass emissions and consumption of vehicle engines in actual use and that the results allow us to establish policies to reduce environmental impact and improve energy efficiency, but, to establish precise methodologies and to be very careful in the entire process of calibration, measurement and data post-treatment is necessary.

Aplicación de un tratamiento biológico a las aguas residuales provenientes de una destilería de alcohol de caña, utilizando un reactor UASB

En este trabajo se llevó a cabo el tratamiento de vinazas mediante dos tecnologías anaerobias. Se dividió en cuatro estudios técnicos. El primero fue el arranque y estabilización del reactor UASB (Upflow Anaerobic Sludge Blanket), en dónde se evaluó la estabilización mediante la eficiencia de remoción de DQO y la granulación del lodo. El segundo estudio evaluó el rendimiento del reactor UASB frente a diferentes Cva. El tercer estudio evaluó el efecto del TRH sobre la eficiencia del reactor UASB, y el cuarto de ellos fue evaluar el rendimiento del RABF (Reactor Anaerobio de Biomasa Fija). El reactor UASB de 2,6 L de capacidad, fue arrancado por lotes, con seis ensayos utilizando vinaza como sustrato. Se obtuvieron eficiencias de remoción en DQO en un rango de 79-91%, en los seis lotes. Se obtuvo formación de gránulos con diámetro (Ø) de 0,85-1,15 mm y un coeficiente de esfericidad (Є) de 0,7-0,77. Se logró la granulación de lodos tras 2 meses de operación. Alcanzada la estabilización del reactor UASB, se siguió una operación en flujo continuo. Las Cva probadas de 1, 2, 4 y 6 gDQO/L.d para el reactor UASB dan una respuesta bastante favorable con respecto al rendimiento del reactor, ya que presento eficiencias de remoción de DQOs del 51 hasta el 76%, eficiencias similares a los reportados por la literatura. En el estudio de TRH se operó con Cva de 6 gDQO/L.d y los TRH fueron de 24, 12 ,5 ,3 y 1 día. El % de eliminación de DQO fue de 51, 60, 57, 60 y 63 % remoción en DQOsoluble, respectivamente. Se alcanzó una producción de biogás máximo de 5.283 ml/d, pero al reducir el TRH se observó una reducción proporcional del volumen total de biogás. El %CH4 contenido en el biogás aumento al disminuir el TRH, reflejando valores de 80 al 92 % de CH4. El RABF con un volumen de 8,2 L, utilizo tubos de plástico corrugado como medio de soporte para las bacterias. Se aplicaron las siguientes Cva; 0,5, 1, 3 y 6 gDQO/L.d. El reactor RABF presento una excelente remoción de la materia orgánica (80% DQOs), una producción de biogás estable, y un contenido en CH4 del biogás muy interesante. Sin embargo, para una Cva superior a 3 gDQO/L.d empezó un comportamiento inesperado de reducción de capacidad. Las condiciones hidrodinámicas del reactor UASB son decisivas para la formación de los gránulos, condición previa para iniciar el flujo continuo. Al operar el reactor UASB en modo continuo, se pudo evaluar las mejores condiciones de operación para este tipo de residuo (vinaza). La Cva de 6 gDQO/L.d para el reactor UASB alimentado con vinaza bruta representa el límite de su capacidad. Sin embargo, al aumentar la Cva se genera una mayor producción de biogás y metano. La eficiencia de remoción de la DQO soluble es independiente del TRH, para una Cva de 6 g DQO/L•d y las condiciones de TRH probadas (24, 12, 5, 3 y 1 días). Los valores de remoción de DQO alcanzados son un poco superior a los valores de biodegradabilidad anaerobia de la vinaza observados de 50 %. De manera general, la reducción del TRH o bien la dilución de la vinaza no presenta un efecto significativo sobre la remoción de la materia orgánica soluble, pero si lo presenta en la remoción de sulfatos reduciendo indirectamente su toxicidad. El soporte termoplástico inoculado en el RABF y alimentado con vinaza bruta, actuó como un filtro, además de obtener buenos resultados en eliminación de DQO, pero dada las dimensiones y la altura del relleno se frena la evacuación del metano. This work was carried out by treatment vinasses with two anaerobic technologies. It was divided into four technical studies. The first was the start up and stabilization Upflow Anaerobic Sludge Blanket (UASB) reactor, where the stability was evaluated by the removal efficiency of COD and sludge granulation. The second study evaluated the performance of the UASB reactor against different OLR. The third study evaluated the effect of HRT on the efficiency of the UASB reactor, and the fourth of which was evaluate the performance Fixed Biomass Anaerobic (FBA) reactor. The UASB reactor of 2,6 L capacity, was started in batch, with six assays using vinasse as substrate. Were obtained removal efficiencies of COD in the range of 79- 91% in the six batches. Forming granules were obtained with a diameter (Ø) of 0,85- 1,15 mm and sphericity coefficient (Є) of 0,7 to 0,77. Sludge granulation was achieved after 2 months of operation. Once stabilization is achieved of the UASB reactor, it was followed by a continuous flow operation. The OLR tested 1, 2, 4 and 6 gCOD/L.d for UASB reactor gives a very favorable response regarding the performance of the reactor, as presented COD5 removal efficiencies of 51 to 76%, similar efficiencies those reported in the literature The HRT study was operated with an OLR of 6 gCOD/L.d and HRT were 24, 12, 5, 3 and 1 day. The removal efficiency was 51, 60, 57, 60 and 63% in soluble COD, respectively. It reached a maximum biogas production of 5.283 ml / d, but by reducing the HRT showed a proportional reduction in the total volume of biogas. The %CH4 content in the biogas increased with decreasing TRH, reflecting values of 80 to 92% of CH4. The FBA reactor with a volume of 8,2 L, used corrugated plastic tubes as carrier for bacteria transportation. The following OLR was applied, 0,5, 1, 3 and 6 gCOD/L.d. The FBA reactor showed an excellent removal of organic matter (80% CODS), a stable biogas production, and CH4 content very interesting. However, for more than 3 gCOD/L.d OLR began with unexpected behavior of capacity reduction. The UASB reactor hydrodynamic conditions are decisive for the formation of the granules, precondition to start the continuous flow. By operating the UASB reactor in continuous mode, it was possible to evaluate the best operating conditions for this type of waste (vinasse). The OLR of 6 gCOD/L.d for the UASB reactor fed with raw vinasse represents the limit of its capacity. However, with increasing OLR creates increased biogas production and methane. The removal efficiency of soluble COD is independent of HRT for OLR of 6 gCOD/L.d and HRT conditions tested (24, 12, 5, 3 and 1 day). COD Removal values achieved are slightly higher than the values of the vinasse anaerobic biodegradability of observed at 50%. Generally, reduction of HRT or vinasse dilution does not present a significant effect on the removal of the soluble organic matter; however if it occurs in the removal of sulfate reducing indirectly its toxicity. The thermoplastic support inoculated in FBA reactor and fed with raw vinasse, acted as a filter, in addition to obtaining good results in COD removal, but given the size and height of the filling slows evacuation of methane.

Análisis energético de las centrales de ciclo combinado con gasificación integrada y combustión mediante lazo químico

La combustión mediante lazo químico permite una integración de captura de CO2 en una planta térmica de potencia sin penalización energética. Se alcanza una menor destrucción exergética, alcanzándose un rendimiento térmico idóneo. Este proyecto se centra en el estudio del rendimiento energético en una planta de potencia con gasificación integrada cuando la cámara de combustión de las turbinas de gas se sustituye por un sistema de combustión CLC. Después del diseño termodinámico y optimización de algunos parámetros del ciclo, el rendimiento de la planta de potencia es evaluado bajo distintas condiciones de trabajo y comparado con el que se obtendría en una planta de ciclo combinado convencional con gasificación integrada y captura pre-combustión. Abstract Chemical-looping combustion allows an integration of CO2 capture in a thermal power plant without energy penalty. A less exergy destruction in the combustion chemical transformation is achieved, leading to a greater overall thermal efficiency. This Project focus on the study of the energetic performance in a cycle power plant with integrated gasification when the chamber combustion is supplied by a combustion system CLC. After thermodynamic modeling and optimization of some cycle parameters, the power plant performance is evaluated under diverse working conditions and compared to a conventional integrated gasification combined cycle with pre-combustion capture.

Sistema autónomo para carga de baterías mediante una instalación fotovoltaica con aplicación directa al vehículo eléctrico y usos

La situación energética actual es insostenible y como consecuencia se plantea un escenario próximo orientado a conseguir un futuro energético sostenible que permita el desarrollo económico y el bienestar social. La situación ambiental actual está afectada directamente por la combustión de combustibles fósiles que en 2013 constituyeron el 81% de la energía primaria utilizada por el ser humano y son la principal fuente antropogénica de gases de efecto invernadero. Los informes del IPCC1, ponen de manifiesto que el cambio climático se ha consolidado durante los últimos años y en la conferencia de la ONU sobre cambio climático de París que se celebrará a finales de 2015, se pretende que los gobiernos suscriban un acuerdo universal para limitar las emisiones de gases de efecto invernadero y evitar que el incremento de la temperatura media global supere los 2°C. Por otra parte, en el interior de las ciudades es especialmente preocupante, por su efecto directo sobre la salud humana, el impacto ambiental producido por las emisiones de NOx que generan el transporte de personas y mercancías. El sector del transporte fue responsable en 2012 del 27,9% del consumo final de energía. Una vez expuesto el escenario energético y ambiental actual, en esta tesis, se analiza la eficiencia de un sistema autónomo fotovoltaico para la carga de baterías de vehículos eléctricos y el uso del mismo con otras cargas, con el objetivo de aprovechar al máximo la energía eléctrica generada y contribuir a la utilización de energía limpia que no produzca impacto ambiental. Como primer paso para el desarrollo de la tesis se hizo un estudio de trabajos previos comenzando por las primeras aplicaciones de la energía fotovoltaica en los vehículos solares para después pasar a trabajos más recientes enfocados al suministro de energía a los vehículos eléctricos. También se hizo este estudio sobre las metodologías de simulación en los sistemas fotovoltaicos y en el modelado de distintos componentes. Posteriormente se eligieron, dentro de la amplia oferta existente en el mercado, los componentes con características técnicas más adecuadas para este tipo de instalaciones y para las necesidades que se pretenden cubrir. A partir de los parámetros técnicos de los componentes elegidos para configurar la instalación autónoma y utilizando modelos contrastados de distintos componentes, se ha desarrollado un modelo de simulación en ordenador del sistema completo con el que se han hecho simulaciones con distintos modos de demanda de energía eléctrica, según los modos de carga disponibles en el vehículo eléctrico para corriente alterna monofásica de 230 V. También se han simulado distintos tamaños del generador fotovoltaico y del sistema de acumulación de energía eléctrica para poder determinar la influencia de estos parámetros en los balances energéticos del sistema. Utilizando recursos propios el doctorando ha realizado la instalación real de un sistema fotovoltaico que incluye sistema de acumulación e inversor en un edificio de su propiedad. Para la realización de la tesis, La Fundación de Fomento e Innovación Industrial (F2I2) ha facilitado al doctorando un dispositivo que permite realizar la alimentación del vehículo eléctrico en modo 2 (este modo emplea un adaptador que incorpora dispositivos de seguridad y se comunica con el vehículo permitiendo ajustar la velocidad de recarga) y que ha sido necesario para los trabajos desarrollados. Se ha utilizado la red eléctrica como sistema de apoyo de la instalación fotovoltaica para permitir la recarga en el modo 2 que requiere más potencia que la proporcionada por el sistema fotovoltaico instalado. Se han analizado mediante simulación distintos regímenes de carga que se han estudiado experimentalmente en la instalación realizada, a la vez que se han hecho ensayos que se han reproducido mediante simulación con los mismos valores de radiación solar y temperatura con objeto de contrastar el modelo. Se han comparado los resultados experimentales con los obtenidos mediante simulación con objeto de caracterizar el comportamiento del sistema de acumulación (energía eléctrica suministrada y tensión de salida en las baterías) y del generador fotovoltaico (energía eléctrica fotovoltaica suministrada). Por último, se ha realizado un estudio económico de la instalación autónoma fotovoltaica ejecutada y simulada. En el mismo se ha planteado la utilización de fondos propios (como realmente se ha llevado a cabo) y la utilización de financiación, para determinar dos posibles escenarios que pudieran ser de utilidad a un propietario de vehículo eléctrico. Se han comparado los resultados obtenidos en los dos escenarios propuestos del estudio económico del sistema, en cuanto a los parámetros de tiempo de retorno de la inversión, valor actual neto de la inversión y tasa interna de retorno de la misma. Las conclusiones técnicas obtenidas, permiten la utilización del sistema con los modos de carga ensayados y otro tipo de cargas que aprovechen la generación eléctrica del sistema. Las baterías ofrecen mejor comportamiento cuando el aporte fotovoltaico está presente, pero no considera adecuado la conexión de cargas elevadas a un sistema de acumulación de gel (plomo-acido) como el que se ha utilizado, debido al comportamiento de este tipo de baterías ante demandas de intensidad de corriente eléctrica elevadas. Por otra parte, el comportamiento de este tipo de baterías con valores de intensidad de corriente eléctrica inferiores a 10 A en ausencia de energía fotovoltaica, con el objetivo de utilizar la generación de energía eléctrica diaria acumulada en el sistema, sí resulta interesante y ofrece un buen comportamiento del sistema de acumulación. Las circunstancias actuales de mercado, que carece de sistemas de acumulación de litio con precios de compra interesantes, no han permitido poder experimentar este sistema de acumulación en la instalación autónoma fotovoltaica ejecutada, tampoco se ha podido obtener el favor de ningún fabricante para ello. Actualmente hay disponibles sistemas de acumulación en litio que no se comercializan en España y que serían adecuados para el sistema de acumulación de energía propuesto en este estudio, que deja abierta las puertas para futuros trabajos de investigación. Las conclusiones económicas obtenidas, rentabilizan el uso de una instalación autónoma fotovoltaica con consumo instantáneo, sin acumulación de energía eléctrica. El futuro de conexión a red por parte de estas instalaciones, cuando se regule, aportará un incentivo económico para rentabilizar con menos tiempo las instalaciones autónomas fotovoltaicas, esto también deja la puerta abierta a futuros trabajos de investigación. El sistema de acumulación de energía aporta el mayor peso económico de inversión en este tipo de instalaciones. La instalación estudiada aporta indicadores económicos que la hacen rentable, pero se necesitaría que los precios de acumulación de la energía en sistemas eficientes estén comprendidos entre 100-200 €/kWh para que el sistema propuesto en este trabajo resulte atractivo a un potencial propietario de un vehículo eléctrico. ABSTRACT The current energy situation is untenable; it poses a scenario next focused on reaching a sustainable energy future, to allow economic development and social welfare. The environmental current situation is affected directly by the combustion of fossil fuels that in 2013 constituted 81 % of the primary energy used by the human being and they are the principal source human of greenhouse gases. The reports of the IPCC2, they reveal that the climate change has consolidated during the last years and in the conference of the UNO on climate change of Paris that will be celebrated at the end of 2015, there is claimed that the governments sign a universal agreement to limit the emission of greenhouse gases and to prevent that the increase of the global average temperature overcomes them 2°C. On the other hand, inside the cities it is specially worrying, for his direct effect on the human health, the environmental impact produced by the NOx emissions that generate the persons' transport and goods. The sector of the transport was responsible in 2012 of 27,9 % of the final consumption of energy. Once exposed the scenario and present environmental energy, in this thesis, it has analyzed the efficiency of an autonomous photovoltaic system for charging electric vehicles, and the use of the same with other workloads, with the objective to maximize the electrical energy generated and contribute to the use of clean energy that does not produce environmental impact. Since the first step for the development of the thesis did to itself a study of previous works beginning for the first applications of the photovoltaic power in the solar vehicles later to go on to more recent works focused on the power supply to the electrical vehicles. Also this study was done on the methodologies of simulation in the photovoltaic systems and in the shaped one of different components. Later they were chosen, inside the wide existing offer on the market, the components with technical characteristics more adapted for this type of facilities and for the needs that try to cover. From the technical parameters of the components chosen to form the autonomous installation and using models confirmed of different components, a model of simulation has developed in computer of the complete system with which simulations have been done by different manners of demand of electric power, according to the available manners of load in the electrical vehicle for single-phase alternating current of 230 V. Also there have been simulated different sizes of the photovoltaic generator and of the system of accumulation of electric power to be able to determine the influence of these parameters in the energy balances of the system. Using own resources the PhD student has realized a real installation of a photovoltaic system that includes system of accumulation and investing in a building of his property. For the accomplishment of the thesis, The Foundation of Promotion and Industrial Innovation (F2I2) it has facilitated to the PhD student a device that allows to realize the supply of the electrical vehicle in way 2 (this way uses an adapter that incorporates safety devices and communicates with the vehicle allowing to fit the speed of recharges) and that has been necessary for the developed works. The electrical network has been in use as system of support of the photovoltaic installation for allowing it her recharges in the way 2 that more power needs that provided by the photovoltaic installed system. There have been analyzed by means of simulation different rate of load that have been studied experimentally in the realized installation, simultaneously that have done to themselves tests that have reproduced by means of simulation with the same values of solar radiation and temperature in order the model contrasted. The experimental results have been compared by the obtained ones by means of simulation in order to characterize the behavior of the system of accumulation (supplied electric power and tension of exit in the batteries) and of the photovoltaic generator (photovoltaic supplied electric power). Finally, there has been realized an economic study of the autonomous photovoltaic executed and simulated installation. In the same one there has appeared the utilization of own funds (since really it has been carried out) and the utilization of financing, to determine two possible scenes that could be of usefulness to an owner of electrical vehicle. There have been compared the results obtained in both scenes proposed of the economic study of the system, as for the parameters of time of return of the investment, current clear value of the investment and rate hospitalizes of return of the same one. The technical obtained conclusions, they make the utilization of the system viable with the manners of load tested and another type of loads of that they take advantage the electrical generation of the system. The batteries offer better behavior when the photovoltaic contribution is present, but he does not consider to be suitable the connection of loads risen up to a system of accumulation of gel (lead - acid) as the one that has been in use, due to the behavior of this type of batteries before demands of intensity of electrical current raised. On the other hand, the behavior of this type of batteries with low values of intensity of electrical current to 10 To in absence of photovoltaic power, with the aim to use the generation of daily electric power accumulated in the system, yes turns out to be interesting and offers a good behavior of the system of accumulation. The current circumstances of market, which lacks systems of accumulation of lithium with interesting purchase prices, have not allowed to be able to experience this system of accumulation in the autonomous photovoltaic executed installation, neither one could have obtained the favor of any manufacturer for it. Nowadays there are available systems of accumulation in lithium that is not commercialized in Spain and that they would be adapted for the system of accumulation of energy proposed in this study, which makes the doors opened for future works of investigation. The economic obtained conclusions; they make more profitable the use of an autonomous photovoltaic installation with instantaneous consumption, without accumulation of electric power. The future of connection to network on the part of these facilities, when it is regulated, will contribute an economic incentive to make profitable with less time the autonomous photovoltaic facilities, this also leaves the door opened for future works of investigation. The system of accumulation of energy contributes the major economic weight of investment in this type of facilities. The studied installation contributes economic indicators that make her profitable, but it would be necessary that the prices of accumulation of the energy in efficient systems are understood between 100-200 € in order that the system proposed in this work turns out to be attractive to a proprietary potential of an electrical vehicle.

13C NMR study of the effects of leptin treatment on kinetics of hepatic intermediary metabolism

The recent discovery of leptin receptors in peripheral tissue raises questions about which of leptin’s biological actions arise from direct effects of the hormone on extraneural tissues and what intracellular mechanisms are responsible for leptin’s effects on carbohydrate and lipid metabolism. The present study is focused on the action of leptin on hepatic metabolism. Nondestructive 13C NMR methodology was used to follow the kinetics of intermediary metabolism by monitoring flux of 13C-labeled substrate through several multistep pathways. In perfused liver from either ob/ob or lean mice, we found that acute treatment with leptin in vitro modulates pathways controlling carbohydrate flux into 13C-labeled glycogen, thereby rapidly enhancing synthesis by an insulin-independent mechanism. Acute treatment of ob/ob liver also caused a rapid stimulation of long-chain fatty acid synthesis from 13C-labeled acetyl-CoA by the de novo synthesis route. Chronic leptin treatment in vivo induced homeostatic changes that resulted in a tripling of the rate of glycogen synthesis via the gluconeogenic pathway from [2-13C]pyruvate in ob/ob mouse liver perfused in the absence of the hormone. Consistent with the 13C NMR results, leptin treatment of the ob/ob mouse in vivo resulted in significantly increased hepatic glycogen synthase activity. Chronic treatment with leptin in vivo exerted the opposite effect of acute treatment in vitro and markedly decreased hepatic de novo synthesis of fatty acids in ob/ob mouse liver. In agreement with the 13C NMR findings, activities of hepatic acetyl-CoA carboxylase and fatty acid synthase were significantly reduced by chronic treatment of the ob/ob mouse with leptin. Our data represent a demonstration of direct effects of leptin in the regulation of metabolism in the intact functioning liver.