Towards a new test of time dilation: Laser spectroscopy on lithium ions stored at a velocity of 33.8 % of the speed of light

The subject of the presented thesis is the accurate measurement of time dilation, aiming at a quantitative test of special relativity. By means of laser spectroscopy, the relativistic Doppler shifts of a clock transition in the metastable triplet spectrum of ^7Li^+ are simultaneously measured with and against the direction of motion of the ions. By employing saturation or optical double resonance spectroscopy, the Doppler broadening as caused by the ions' velocity distribution is eliminated. From these shifts both time dilation as well as the ion velocity can be extracted with high accuracy allowing for a test of the predictions of special relativity. A diode laser and a frequency-doubled titanium sapphire laser were set up for antiparallel and parallel excitation of the ions, respectively. To achieve a robust control of the laser frequencies required for the beam times, a redundant system of frequency standards consisting of a rubidium spectrometer, an iodine spectrometer, and a frequency comb was developed. At the experimental section of the ESR, an automated laser beam guiding system for exact control of polarisation, beam profile, and overlap with the ion beam, as well as a fluorescence detection system were built up. During the first experiments, the production, acceleration and lifetime of the metastable ions at the GSI heavy ion facility were investigated for the first time. The characterisation of the ion beam allowed for the first time to measure its velocity directly via the Doppler effect, which resulted in a new improved calibration of the electron cooler. In the following step the first sub-Doppler spectroscopy signals from an ion beam at 33.8 %c could be recorded. The unprecedented accuracy in such experiments allowed to derive a new upper bound for possible higher-order deviations from special relativity. Moreover future measurements with the experimental setup developed in this thesis have the potential to improve the sensitivity to low-order deviations by at least one order of magnitude compared to previous experiments; and will thus lead to a further contribution to the test of the standard model.

Transport of nanoparticles into polymersomes : a minimal model system of particles passage through biological membranes

This thesis focuses on the design and characterization of a novel, artificial minimal model membrane system with chosen physical parameters to mimic a nanoparticle uptake process driven exclusively by adhesion and softness of the bilayer. The realization is based on polymersomes composed of poly(dimethylsiloxane)-b-poly(2-methyloxazoline) (PMDS-b-PMOXA) and nanoscopic colloidal particles (polystyrene, silica), and the utilization of powerful characterization techniques. rnPDMS-b-PMOXA polymersomes with a radius, Rh ~100 nm, a size polydispersity, PD = 1.1 and a membrane thickness, h = 16 nm, were prepared using the film rehydratation method. Due to the suitable mechanical properties (Young’s modulus of ~17 MPa and a bending modulus of ~7⋅10-8 J) along with the long-term stability and the modifiability, these kind of polymersomes can be used as model membranes to study physical and physicochemical aspects of transmembrane transport of nanoparticles. A combination of photon (PCS) and fluorescence (FCS) correlation spectroscopies optimizes species selectivity, necessary for a unique internalization study encompassing two main efforts. rnFor the proof of concepts, the first effort focused on the interaction of nanoparticles (Rh NP SiO2 = 14 nm, Rh NP PS = 16 nm; cNP = 0.1 gL-1) and polymersomes (Rh P = 112 nm; cP = 0.045 gL-1) with fixed size and concentration. Identification of a modified form factor of the polymersome entities, selectively seen in the PCS experiment, enabled a precise monitor and quantitative description of the incorporation process. Combining PCS and FCS led to the estimation of the incorporated particles per polymersome (about 8 in the examined system) and the development of an appropriate methodology for the kinetics and dynamics of the internalization process. rnThe second effort aimed at the establishment of the necessary phenomenology to facilitate comparison with theories. The size and concentration of the nanoparticles were chosen as the most important system variables (Rh NP = 14 - 57 nm; cNP = 0.05 - 0.2 gL-1). It was revealed that the incorporation process could be controlled to a significant extent by changing the nanoparticles size and concentration. Average number of 7 up to 11 NPs with Rh NP = 14 nm and 3 up to 6 NPs with Rh NP = 25 nm can be internalized into the present polymersomes by changing initial nanoparticles concentration in the range 0.1- 0.2 gL-1. Rapid internalization of the particles by polymersomes is observed only above a critical threshold particles concentration, dependent on the nanoparticle size. rnWith regard possible pathways for the particle uptake, cryogenic transmission electron microscopy (cryo-TEM) has revealed two different incorporation mechanisms depending on the size of the involved nanoparticles: cooperative incorporation of nanoparticles groups or single nanoparticles incorporation. Conditions for nanoparticle uptake and controlled filling of polymersomes were presented. rnIn the framework of this thesis, the experimental observation of transmembrane transport of spherical PS and SiO2 NPs into polymersomes via an internalization process was reported and examined quantitatively for the first time. rnIn a summary the work performed in frames of this thesis might have significant impact on cell model systems’ development and thus improved understanding of transmembrane transport processes. The present experimental findings help create the missing phenomenology necessary for a detailed understanding of a phenomenon with great relevance in transmembrane transport. The fact that transmembrane transport of nanoparticles can be performed by artificial model system without any additional stimuli has a fundamental impact on the understanding, not only of the nanoparticle invagination process but also of the interaction of nanoparticles with biological as well as polymeric membranes. rn

Effects of doxorubicin cancer therapy on autophagy and the ubiquitin-proteasome system in long-term cultured adult rat cardiomyocytes

The clinical use of anthracyclines in cancer therapy is limited by dose-dependent cardiotoxicity that involves cardiomyocyte injury and death. We have tested the hypothesis that anthracyclines affect protein degradation pathways in adult cardiomyocytes. To this aim, we assessed the effects of doxorubicin (Doxo) on apoptosis, autophagy and the proteasome/ubiquitin system in long-term cultured adult rat cardiomyocytes. Accumulation of poly-ubiquitinated proteins, increase of cathepsin-D-positive lysosomes and myofibrillar degradation were observed in Doxo-treated cardiomyocytes. Chymotrypsin-like activity of the proteasome was initially increased and then inhibited by Doxo over a time-course of 48 h. Proteasome 20S proteins were down-regulated by higher doses of Doxo. The expression of MURF-1, an ubiquitin-ligase specifically targeting myofibrillar proteins, was suppressed by Doxo at all concentrations measured. Microtubule-associated protein 1 light chain 3B (LC3)-positive punctae and both LC3-I and -II proteins were induced by Doxo in a dose-dependent manner, as confirmed by using lentiviral expression of green fluorescence protein bound to LC3 and live imaging. The lysosomotropic drug chloroquine led to autophagosome accumulation, which increased with concomitant Doxo treatment indicating enhanced autophagic flux. We conclude that Doxo causes a downregulation of the protein degradation machinery of cardiomyocytes with a resulting accumulation of poly-ubiquitinated proteins and autophagosomes. Although autophagy is initially stimulated as a compensatory response to cytotoxic stress, it is followed by apoptosis and necrosis at higher doses and longer exposure times. This mechanism might contribute to the late cardiotoxicity of anthracyclines by accelerated aging of the postmitotic adult cardiomyocytes and to the susceptibility of the aging heart to anthracycline cancer therapy.

Light curing through glass ceramics with a second- and a third-generation LED curing unit: effect of curing mode on the degree of conversion of dual-curing resin cements

Objectives The aim of this study was to measure the degree of conversion (DC) of five dual-curing resin cements after different curing modes with a second- and a third-generation light-emitting diode (LED) curing unit. Additionally, irradiance of both light curing units was measured at increasing distances and through discs of two glass ceramics for computer-aided design/manufacturing (CAD/CAM). Materials and methods Irradiance and spectra of the Elipar FreeLight 2 (Standard Mode (SM)) and of the VALO light curing unit (High Power Mode (HPM) and Xtra Power Mode (XPM)) were measured with a MARC radiometer. Irradiance was measured at increasing distances (control) and through discs (1.5 to 6 mm thickness) of IPS Empress CAD and IPS e.max CAD. DC of Panavia F2.0, RelyX Unicem 2 Automix, SpeedCEM, BisCem, and BeautiCem SA was measured with an attenuated total reflectance–Fourier transform infrared spectrometer when self-cured (negative control) or light cured in SM for 40 s, HPM for 32 s, or XPM for 18 s. Light curing was performed directly (positive control) or through discs of either 1.5- or 3-mm thickness of IPS Empress CAD or IPS e.max CAD. DC was analysed with Kruskal–Wallis tests followed by pairwise Wilcoxon rank sum tests (α = 0.05). Results Maximum irradiances were 1,545 mW/cm2 (SM), 2,179 mW/cm2 (HPM), and 4,156 mW/cm2 (XPM), and all irradiances decreased by >80 % through discs of 1.5 mm, ≥95 % through 3 mm, and up to >99 % through 6 mm. Generally, self-curing resulted in the lowest DC. For some cements, direct light curing did not result in higher DC compared to when light cured through ceramic discs. For other cements, light curing through ceramic discs of 3 mm generally reduced DC. Conclusions Light curing was favourable for dual-curing cements. Some cements were more susceptible to variations in curing mode than others. Clinical relevance When light curing a given cement, the higher irradiances of the third-generation LED curing unit resulted in similar DC compared to the second-generation one, though at shorter light curing times.

Aeromonas salmonicida subsp. salmonicida in the light of its type-three secretion system

Aeromonas salmonicida subsp. salmonicida is an important pathogen in salmonid aquaculture and is responsible for the typical furunculosis. The type-three secretion system (T3SS) is a major virulence system. In this work, we review structure and function of this highly sophisticated nanosyringe in A. salmonicida. Based on the literature as well as personal experimental observations, we document the genetic (re)organization, expression regulation, anatomy, putative functional origin and roles in the infectious process of this T3SS. We propose a model of pathogenesis where A. salmonicida induces a temporary immunosuppression state in fish in order to acquire free access to host tissues. Finally, we highlight putative important therapeutic and vaccine strategies to prevent furunculosis of salmonid fish.

A steerable UV laser system for the calibration of liquid argon time projection chambers

A number of liquid argon time projection chambers (LAr TPCs) are being built or are proposed for neutrino experiments on long- and short baseline beams. For these detectors, a distortion in the drift field due to geometrical or physics reasons can affect the reconstruction of the events. Depending on the TPC geometry and electric drift field intensity, this distortion could be of the same magnitude as the drift field itself. Recently, we presented a method to calibrate the drift field and correct for these possible distortions. While straight cosmic ray muon tracks could be used for calibration, multiple coulomb scattering and momentum uncertainties allow only a limited resolution. A UV laser instead can create straight ionization tracks in liquid argon, and allows one to map the drift field along different paths in the TPC inner volume. Here we present a UV laser feed-through design with a steerable UV mirror immersed in liquid argon that can point the laser beam at many locations through the TPC. The straight ionization paths are sensitive to drift field distortions, a fit of these distortion to the linear optical path allows to extract the drift field, by using these laser tracks along the whole TPC volume one can obtain a 3D drift field map. The UV laser feed-through assembly is a prototype of the system that will be used for the MicroBooNE experiment at the Fermi National Accelerator Laboratory (FNAL).

Yellow laser light generation by frequency doubling of the output from a master oscillator fiber power amplifier system

We present a power-scalable approach for yellow laser-light generation based on standard Ytterbium (Yb) doped fibers. To force the cavity to lase at 1154 nm, far above the gain-maximum, measures must be taken to fulfill lasing condition and to suppress competing amplified spontaneous emission (ASE) in the high-gain region. To prove the principle we built a fiber-laser cavity and a fiber-amplifier both at 1154 nm. In between cavity and amplifier we suppressed the ASE by 70 dB using a fiber Bragg grating (FBG) based filter. Finally we demonstrated efficient single pass frequency doubling to 577 nm with a periodically poled lithium niobate crystal (PPLN). With our linearly polarized 1154 nm master oscillator power fiber amplifier (MOFA) system we achieved slope efficiencies of more than 15 % inside the cavity and 24 % with the fiber-amplifier. The frequency doubling followed the predicted optimal efficiency achievable with a PPLN crystal. So far we generated 1.5 W at 1154nm and 90 mW at 577 nm. Our MOFA approach for generation of 1154 nm laser radiation is power-scalable by using multi-stage amplifiers and large mode-area fibers and is therefore very promising for building a high power yellow laser-light source of several tens of Watt.

Design, Calibration, and Evaluation of Depth-of-Interaction-Capable PET Detector Modules

High-resolution, small-bore PET systems suffer from a tradeoff between system sensitivity, and image quality degradation. In these systems long crystals allow mispositioning of the line of response due to parallax error and this mispositioning causes resolution blurring, but long crystals are necessary for high system sensitivity. One means to allow long crystals without introducing parallax errors is to determine the depth of interaction (DOI) of the gamma ray interaction within the detector module. While DOI has been investigated previously, newly available solid state photomultipliers (SSPMs) well-suited to PET applications and allow new modules for investigation. Depth of interaction in full modules is a relatively new field, and so even if high performance DOI capable modules were available, the appropriate means to characterize and calibrate the modules are not. This work presents an investigation of DOI capable arrays and techniques for characterizing and calibrating those modules. The methods introduced here accurately and reliably characterize and calibrate energy, timing, and event interaction positioning. Additionally presented is a characterization of the spatial resolution of DOI capable modules and a measurement of DOI effects for different angles between detector modules. These arrays have been built into a prototype PET system that delivers better than 2.0 mm resolution with a single-sided-stopping-power in excess of 95% for 511 keV g's. The noise properties of SSPMs scale with the active area of the detector face, and so the best signal-to-noise ratio is possible with parallel readout of each SSPM photodetector pixel rather than multiplexing signals together. This work additionally investigates several algorithms for improving timing performance using timing information from multiple SSPM pixels when light is distributed among several photodetectors.

Pathways leading to apoptosis resistance in a murine B -cell lymphoma cell system

The aberrant activation of signal transduction pathways has long been linked to uncontrolled cell proliferation and the development of cancer. The activity of one such signaling module, the Mitogen-Activated Protein Kinase (MAPK) pathway, has been implicated in several cancer types including pancreatic, breast, colon, and lymphoid malignancies. Interestingly, the activation of MAP-Kinase-Kinase-Kinase proteins often leads to the additional activation of NF-κB, a transcription factor that acts as a cell survival signal through its control of antiapoptotic genes. We have investigated the role of a specific dimer form of the NF-κB transcription factor family, NF-κB1 (p50) homodimers, in its control of the proto-oncogene, Bcl-2, and we have identified the MEK/ERK (MAPK) signaling cascade as a mediator of NF-κB1 activity. ^ Two murine B cell lymphoma cell lines were used for these studies: LY-as, an apoptosis proficient line with low Bcl-2 protein expression and no nuclear NF-κB activity, and LY-ar, a nonapoptotic line with constitutive p50 homodimer activity and 30 times more Bcl-2 protein expression than LY-as. Experiments modulating p50 activity correlated the activation of p50 homodimers with Bcl-2 expression and additional gel shift experiments demonstrated that the Bcl-2 P1 promoter had NF-κB sites with which recombinant p50 was able to interact. In vitro transcription revealed that p50 enhanced the production of transcripts derived from the Bcl-2 P1 promoter. These data strongly suggest that Bcl-2 is a target gene for p50-mediated transcription and suggest that the activation of p50 homodimers contributes to the expression of Bcl-2 observed in LY-ar cells. ^ Studies of upstream MAPK pathways that could influence NF-κB activity demonstrated that LY-ar cells had phosphorylated ERK proteins while LY-as cells did not. Treatment of LY-ar cells with the MEK inhibitors PD 98059, U0126, and PD 184352 led to a loss of phosphorylated ERK, a reversal of nuclear p50 homodimer DNA binding, and a decrease in the amount of Bcl-2 protein expression. Similarly, the activation of the MEK/ERK pathway in LY-as cells by phorbol ester led to Bcl-2 expression that could be blocked by PD 98059. Furthermore, treatment of LY-ar cells with TNFα, an IKK activator, did not change the suppressive effect of PD 98059 on p50 homodimer activity, suggesting an IKK-independent pathway for p50 homodimer activation. Lastly, all three MEK inhibitors sensitized LY-ar cells to radiation-induced apoptosis. ^ These data indicate that the activation of the MEK/ERK MAP-Kinase signaling pathway acts upstream of p50 homodimer activation and Bcl-2 expression in this B cell lymphoma cell system and suggest that the activation of MEK/ERK may be a key step in the progression of lymphoma to advanced-staged disease. Other researchers have used MEK inhibitors to inhibit cell growth and sensitize a number of tumors to chemotherapies. In light of our data, MEK inhibitors may additionally be useful clinically to radiosensitize cancers of lymphoid origin. ^

Properties of seawater and particulate matter (fluorescence) from a WETLabs Eco-FL sensor mounted on the continuous surface water sampling system during the Tara Oceans expedition 2009-2013

The Tara Oceans Expedition (2009-2013) sampled the world oceans on board a 36 m long schooner, collecting environmental data and organisms from viruses to planktonic metazoans for later analyses using modern sequencing and state-of-the-art imaging technologies. Tara Oceans Data are particularly suited to study the genetic, morphological and functional diversity of plankton. The present data set provides continuous measurements made with a WETLabs Eco-FL sensor mounted on the flowthrough system between June 4th, 2011 and March 30th, 2012. Data was recorded approximately every 10s. Two issues affected the data: 1. Periods when the water 0.2µm filtered water were used as blanks and 2. Periods where fluorescence was affected by non-photochemical quenching (NPQ, chlorophyll fluorescence is reduced when cells are exposed to light, e.g. Falkowski and Raven, 1997). Median data and their standard deviation were binned to 5min bins with period of light/dark indicated by an added variable (so that NPQ affected data could be neglected if the user so chooses). Data was first calibrated using HPLC data collected on the Tara (there were 36 data within 30min of each other). Fewer were available when there was no evident NPQ and the resulting scale factor was 0.0106 mg Chl m-3/count. To increase the calibration match-ups we used the AC-S data which provided a robust estimate of Chlorophyll (e.g. Boss et al., 2013). Scale factor computed over a much larger range of values than HPLC was 0.0088 mg Chl m-3/count (compared to 0.0079 mg Chl m-3/count based on manufacturer). In the archived data the fluorometer data is merged with the TSG, raw data is provided as well as manufacturer calibration constants, blank computed from filtered measurements and chlorophyll calibrated using the AC-S. For a full description of the processing of the Eco-FL please see Taillandier, 2015.

Interactive effects of light, nitrogen source, and carbon dioxide on energy metabolism in the diatom Thalassiosira pseudonana

Due to the ongoing effects of climate change, phytoplankton are likely to experience enhanced irradiance, more reduced nitrogen, and increased water acidity in the future ocean. Here, we used Thalassiosira pseudonana as a model organism to examine how phytoplankton adjust energy production and expenditure to cope with these multiple, interrelated environmental factors. Following acclimation to a matrix of irradiance, nitrogen source, and CO2 levels, the diatom's energy production and expenditures were quantified and incorporated into an energetic budget to predict how photosynthesis was affected by growth conditions. Increased light intensity and a shift from inline image to inline image led to increased energy generation, through higher rates of light capture at high light and greater investment in photosynthetic proteins when grown on inline image. Secondary energetic expenditures were adjusted modestly at different culture conditions, except that inline image utilization was systematically reduced by increasing pCO2. The subsequent changes in element stoichiometry, biochemical composition, and release of dissolved organic compounds may have important implications for marine biogeochemical cycles. The predicted effects of changing environmental conditions on photosynthesis, made using an energetic budget, were in good agreement with observations at low light, when energy is clearly limiting, but the energetic budget over-predicts the response to inline image at high light, which might be due to relief of energetic limitations and/or increased percentage of inactive photosystem II at high light. Taken together, our study demonstrates that energetic budgets offered significant insight into the response of phytoplankton energy metabolism to the changing environment and did a reasonable job predicting them.

A Light-Weight On-Chip Monitoring Network for Dynamic Adaptation and Calibration

Current nanometer technologies suffer within-die parameter uncertainties, varying workload conditions, aging, and temperature effects that cause a serious reduction on yield and performance. In this scenario, monitoring, calibration, and dynamic adaptation become essential, demanding systems with a collection of multi purpose monitors and exposing the need for light-weight monitoring networks. This paper presents a new monitoring network paradigm able to perform an early prioritization of the information. This is achieved by the introduction of a new hierarchy level, the threshing level. Targeting it, we propose a time-domain signaling scheme over a single-wire that minimizes the network switching activity as well as the routing requirements. To validate our approach, we make a thorough analysis of the architectural trade-offs and expose two complete monitoring systems that suppose an area improvement of 40% and a power reduction of three orders of magnitude compared to previous works.

On-chip Monitoring: A Light-Weight Interconnection Network Approach

Current nanometer technologies are subjected to several adverse effects that seriously impact the yield and performance of integrated circuits. Such is the case of within-die parameters uncertainties, varying workload conditions, aging, temperature, etc. Monitoring, calibration and dynamic adaptation have appeared as promising solutions to these issues and many kinds of monitors have been presented recently. In this scenario, where systems with hundreds of monitors of different types have been proposed, the need for light-weight monitoring networks has become essential. In this work we present a light-weight network architecture based on digitization resource sharing of nodes that require a time-to-digital conversion. Our proposal employs a single wire interface, shared among all the nodes in the network, and quantizes the time domain to perform the access multiplexing and transmit the information. It supposes a 16% improvement in area and power consumption compared to traditional approaches.

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.