Molecular interaction of the human metalloprotease meprin beta with the amyloid precursor protein, ADAM10, and ADAM17 based on proteomics

Die Zinkendopeptidasen Meprin α und β sind Schlüsselkomponenten in patho(physiologischen) Prozessen wie Entzündung, Kollagenassemblierung und Angiogenese. Nach ihrer Entdeckung in murinen Bürstensaummembranen und humanen Darmepithelien, wurden weitere Expressionsorte identifiziert, z.B. Leukozyten, Krebszellen und die humane Haut. Tiermodelle, Zellkulturen und biochemische Analysen weisen auf Funktionen der Meprine in der Epithelialdifferenzierung, Zellmigration, Matrixmodellierung, Angiogenese, Bindegewebsausbildung und immunologische Prozesse hin. Dennoch sind ihre physiologischen Substrate weitgehend noch unbekannt. Massenspektrometrisch basierte Proteomics-Analysen enthüllten eine einzigartige Spaltspezifität für saure Aminosäurereste in der P1´ Position und identifizierten neue biologische Substratkandidaten. Unter den 269 extrazellulären Proteinen, die in einem Substratscreen identifiziert wurden, stellten sich das amyloid precursor protein (APP) and ADAM10 (a disintegrin and metalloprotease 10) als sehr vielversprechende Kandidaten heraus. Mehrere Schnittstellen innerhalb des APP Proteins, hervorgerufen durch verschiedenen Proteasen, haben unterschiedlichen Auswirkungen zur Folge. Die β-Sekretase BACE (β-site APP cleaving enzyme) prozessiert APP an einer Schnittstelle, welche als initialer Schritt in der Entwicklung der Alzheimer Erkrankung gilt. Toxische Aβ (Amyloid β)-Peptide werden in den extrazellulären Raum freigesetzt und aggregieren dort zu senilen Plaques. Membran verankertes Meprin β hat eine β-Sekretase Aktivität, die in einem Zellkultur-basierten System bestätigt werden konnte. Die proteolytische Effizienz von Meprin β wurde in FRET (Fluorescence Resonance Energy Transfer)-Analysen bestimmt und war um den Faktor 104 höher als die von BACE1. Weiterhin konnte gezeigt werden, dass Meprin β die ersten zwei Aminosäuren prozessiert und somit aminoterminal einen Glutamatrest freisetzt, welcher nachfolgend durch die Glutaminylzyklase in ein Pyroglutamat zykliert werden kann. Trunkierte Aβ-Peptide werden nur in Alzheimer Patienten generiert. Aufgrund einer erhöhten Hydrophobie weisen diese Peptide eine höhere Tendenz zur Aggregation auf und somit eine erhöhte Toxizität. Bis heute wurde keine Protease identifiziert, welche diese Schnittstelle prozessiert. Die Bildung der Meprin vermittelten N-terminalen APP Fragmenten wurde in vitro und in vivo detektiert. Diese N-APP Peptide hatten keine cytotoxischen Auswirkungen auf murine und humane Gehirnzellen, obwohl zuvor N-APP als Ligand für den death receptor (DR) 6 identifiziert wurde, der für axonale Degenerationsprozesse verantwortlich ist. rnIm nicht-amyloidogenen Weg prozessiert ADAM10 APP und entlässt die Ektodomäne von der Zellmembran. Wir konnten das ADAM10 Propeptid als Substrat von Meprin β identifizieren und in FRET Analysen, in vitro und in vivo zeigen, dass die Meprin vermittelte Prozessierung zu einer erhöhten ADAM10 Aktivität führt. Darüber hinaus wurde ADAM10 als Sheddase für Meprin β identifiziert. Shedding konnte durch Phorbol 12-myristate 13-acetate (PMA) oder durch das Ionophor A23187 hervorgerufen werden, sowie durch ADAM10 Inhibitoren blockiert werden. rnDiese Arbeit konnte somit ein komplexes proteolytisches Netwerk innerhalb der Neurophysiologie aufdecken, welches für die Entwicklung der Alzheimer Demenz wichtig sein kann.rn

Biologischer Lichtsammler (LHCII) für Halbleiternanokristalle (Quantum Dots)

Der light harvesting complex II (LHCII) ist ein pflanzliches Membranprotein, das in seiner trimeren Form über 40 Chlorophylle bindet. In der Pflanze kann er besonders effizient Licht sammeln und die Anregungsenergie anschließend fast verlustfrei über andere chlorophyll-bindende Proteine an die Reaktionszentren weiterleiten. Aufgrund dieser besonderen Eigenschaften war es ein Ziel dieser Arbeit, rekombinanten LHCII mit synthetischen Komponenten zu kombinieren, die zur Ladungstrennung befähigt sind. Zu diesem Zweck wurden unter anderem Halbleiternanokristalle (Quantum Dots, QDs) ausgewählt, die je nach Zusammensetzung sowohl als Energieakzeptoren als auch als Energiedonoren in Frage kamen. Durch Optimierung des Puffers gelang es, die Fluoreszenzquantenausbeute der QDs in wässriger Lösung zu erhöhen und zu stabilisieren, so dass die Grundvoraussetzungen für die spektroskopische Untersuchung verschiedener LHCII-QD-Hybridkomplexe erfüllt waren.rnUnter Verwendung bereits etablierter Affinitätssequenzen zur Bindung des LHCII an die QDs konnte gezeigt werden, dass die in dieser Arbeit verwendeten Typ-I QDs aus CdSe und ZnS sich kaum als Energie-Donoren für den LHCII eignen. Ein Hauptgrund lag im vergleichsweise kleinen Försterradius R0 von 4,1 nm. Im Gegensatz dazu wurde ein R0 von 6,4 nm für den LHCII als Donor und Typ-II QDs aus CdTe, CdSe und ZnS als Akzeptor errechnet, wodurch in diesem System eine höhere Effizienz des Energietransfers zu erwarten war. Fluoreszenzspektroskopische Untersuchungen von Hybridkomplexen aus LHCII und Typ-II QDs ergaben eine hohe Plausibilität für einen Fluoreszenz Resonanz Energietransfer (FRET) vom Lichtsammler auf die QDs. Weitere QD-Affinitätssequenzen für den LHCII wurden identifiziert und deren Bindekonstanten ermittelt. Versuche mit dem Elektronenakzeptor Methylviologen lieferten gute Hinweise auf eine LHCII-sensibilisierte Ladungstrennung der Typ-II QDs, auch wenn dies noch anhand alternativer Messmethoden wie z.B. durch transiente Absorptionsspektroskopie bestätigt werden muss. rnEin weiteres Ziel war die Verwendung von LHCII als Lichtsammler in dye-sensitized solar cells (DSSC). Geeignete dotierte TiO2-Platten wurden ermittelt, das Verfahren zur Belegung der Platten optimiert und daher mit wenig Aufwand eine hohe LHCII-Belegungsdichte erzielt. Erste Messungen von Aktionsspektren mit LHCII und einem zur Ladungstrennung fähigen Rylenfarbstoff zeigen eine, wenn auch geringe, LHCII sensibilisierte Ladungstrennung. rnDie Verwendung von Lanthanide-Binding-Tags (LBTs) ist ein potentielles Verfahren zur in vivo-Markierung von Proteinen mit Lanthanoiden wie Europium und Terbium. Diese Metalle besitzen eine überdurchschnittlich lange Lumineszenzlebensdauer, so dass sie leicht von anderen fluoreszierenden Molekülen unterschieden werden können. Im Rahmen der vorliegenden Arbeit gelang es, eine LBT in rekombinanten LHCII einzubauen und einen Lumineszenz Resonanz Energietransfer (LRET) vom Europium auf den LHCII nachzuweisen.rn

Lichtsammler hinter Glas : Stabilisierung und Funktionalisierung des pflanzlichen Hauptlichtsammelkomplexes (LHCII) durch biomineralische Einbettung in Nanopartikel

Der Haupt-Lichtsammelkomplex des Fotosystems II (LHCII) setzt sich aus einem Proteinanteil und nicht-kovalent gebundenen Pigmenten – 8 Chlorophyll a, 6 Chlorophyll b und 4 Carotinoide - zusammen. Er assembliert in vivo zu einem Trimer, in dem die Monomereinheiten ebenfalls nicht-kovalent miteinander wechselwirken. Die ausgesprochen hohe Farbstoffdichte und die Tatsache, dass der Komplex rekombinant hergestellt werden kann, machen den LHCII zu einem interessanten Kandidaten für technische Anwendungen wie einer Farbstoffsolarzelle. Allerdings muss hierzu seine thermische Stabilität drastisch erhöht werden.rnDer Einschluss von Proteinen/Enzymen in Silikat erhöht deren Stabilität gegenüber Hitze signifikant. LHCII sollte als erster rekombinanter Membranproteinkomplex mittels kovalent verbundener, polykationischen Sequenzen in Silikat eingeschlossen werden. Hierzu wurde der Komplex auf zwei Weisen polykationisch modifiziert: Auf Genebene wurde die Sequenz des R5-Peptids in den N-terminalen Bereich des LHCP-Gens eingeführt und ein Protokoll zur Überexpression, Rekonstitution und Trimerisierung etabliert. Außerdem wurde eine kovalente Modifikation des trimeren LHCII mit dem Arginin-reichen Protamin über heterobifunktionelle Crosslinker entwickelt. Beide resultierenden LHCII-Derivate waren in der Lage, Silikat autogen zu fällen. Die Stabilisierung der so in Silikat präzipitierten Komplexe war jedoch deutlich geringer als bei nicht-modifizierten Komplexen, die durch eine Spermin-induzierte Copräzipitation eingeschlossenen wurden. Dabei zeigte sich, dass für den Anteil der eingebauten Komplexe und das Ausmaß an Stabilisierung die Größe und klare partikuläre Struktur des Silikats entscheidend ist. Kleine Partikel mit einem Durchmesser von etwa 20 nm führten zu einem Einbau von rund 75 % der Komplexe, und mehr als 80 % des Energietransfers innerhalb des Komplexes blieben erhalten, wenn für 24 Stunden bei 50°C inkubiert wurde. Nicht in Silikat eingeschlossene Komplexe verloren bei 50°C ihren Komplex-internen Energietransfer binnen weniger Minuten. Es war dabei unerheblich, ob die Partikelgröße durch die Wahl des Puffers und des entsprechenden pH-Wertes, oder aber durch Variation des Spermin-zu-Kieselsäure-Verhältnisses erreicht wurde. Wurden die polykationisch veränderten Komplexe in solchen Copräzipitationen verwendet, so erhöhte sich der Anteil an eingebauten Komplexen auf über 90 %, jedoch wurde nur bei der R5-modifizierten Variante vergleichbare Ausmaße an Stabilisierung erreicht. Ein noch höherer Anteil an Komplexen wurde in das Silikatpellet eingebaut, wenn LHCII kovalent mit Silanolgruppen modifiziert wurde (95 %); jedoch war das Ausmaß der Stabilisierung wiederum geringer als bei einer Copräzipitation. Die analysierten Fällungssysteme waren außerdem in der Lage, Titandioxid zu fällen, wobei der Komplex in dieses eingebaut wurde. Allerdings muss das Stabilisierungspotential hier noch untersucht werden. Innerhalb eines Silikatpräzipitats aggregierten die Komplexe nicht, zeigten aber einen inter-trimeren Energietransfer, der sehr wahrscheinlich auf einem Förster Resonanz Mechanismus basiert. rnDies und das hohe Maß an Stabilisierung eröffnen neue Möglichkeiten, rekombinanten LHCII in technischen Applikationen als Lichtsammelkomponente zu verwenden.rn

Factors improving the vibration transfer of the floating mass transducer at the round window

With the placement of a floating mass transducer (FMT) at the round window, a new approach of coupling an implantable hearing system to the cochlea has been introduced. The aim of the present experimental study is to examine the influence of different ways of FMT placement at the round window on the vibration energy transfer to the cochlea.

Enhanced (1)G(4) emission in NaLaF4: Pr3+, Yb3+ and charge transfer in NaLaF4: Ce3+, Yb3+ studied by fourier transform luminescence spectroscopy

A high resolution luminescence study of NaLaF4: 1%Pr3+, 5%Yb3+ and NaLaF4: 1%Ce3+, 5%Yb3+ in the UV to NIR spectral range using a InGaAs detector and a fourier transform interferometer is reported. Although the Pr3+(P-3(0) -> (1)G(4), Yb3+(F-2(7/2) -> F-2(5/2)) energy transfer step takes place, significant Pr3+ (1)G(4) emission around 993, 1330 and 1850 nm is observed. No experimental proof for the second energy transfer step in the down-conversion process between Pr3+ and Yb3+ can be given. In the case of NaLaF4: Ce3+, Yb3+ it is concluded that the observed Yb3+ emission upon Ce3+ 5d excitation is the result of a charge transfer process instead of down-conversion. (C) 2010 Elsevier B.V. All rights reserved.

Highly Efficient IR to NIR Upconversion in Gd2O2S: Er3+ for Photovoltaic Applications

Upconversion (UC) is a promising option to enhance the efficiency of solar cells by conversion of sub-bandgap infrared photons to higher energy photons that can be utilized by the solar cell. The UC quantum yield is a key parameter for a successful application. Here the UC luminescence properties of Er3+-doped Gd2O2S are investigated by means of luminescence spectroscopy, quantum yield measurements, and excited state dynamics experiments. Excitation into the maximum of the 4I15/2 → 4I13/2 Er3+ absorption band around 1500 nm induces very efficient UC emission from different Er3+ excited states with energies above the silicon bandgap, in particular, the emission originating from the 4I11/2 state around 1000 nm. Concentration dependent studies reveal that the highest UC quantum yield is realized for a 10% Er3+-doping concentration. The UC luminescence is compared to the well-known Er3+-doped β-NaYF4 UC material for which the highest UC quantum yield has been reported for 25% Er3+. The UC internal quantum yields were measured in this work for Gd2O2S: 10%Er3+ and β-NaYF4: 25%Er3+ to be 12 ± 1% and 8.9 ± 0.7%, respectively, under monochromatic excitation around 1500 nm at a power of 700 W/m2. The UC quantum yield reported here for Gd2O2S: 10%Er3+ is the highest value achieved so far under monochromatic excitation into the 4I13/2 Er3+ level. Power dependence and lifetime measurements were performed to understand the mechanisms responsible for the efficient UC luminescence. We show that the main process yielding 4I11/2 UC emission is energy transfer UC.

THE CALIBRATION AND IMPLEMENTATION OF CR-39 NUCLEAR TRACK DETECTOR MATERIAL AS A FAST NEUTRON MICRODOSIMETER

A detailed microdosimetric characterization of the M. D. Anderson 42 MeV (p,Be) fast neutron beam was performed using the techniques of microdosimetry and a 1/2 inch diameter Rossi proportional counter. These measurements were performed at 5, 15, and 30 cm depths on the central axis, 3 cm inside, and 3 cm outside the field edge for 10 $\times$ 10 and 20 $\times$ 20 cm field sizes. Spectra were also measured at 5 and 15 cm depth on central axis for a 6 $\times$ 6 cm field size. Continuous slowing down approximation calculations were performed to model the nuclear processes that occur in the fast neutron beam. Irradiation of the CR-39 was performed using a tandem electrostatic accelerator for protons of 10, 6, and 3 MeV and alpha particles of 15, 10, and 7 MeV incident energy on target at angles of incidence from 0 to 85 degrees. The critical angle as well as track etch rate and normal incidence diameter versus linear energy transfer (LET) were obtained from these measurements. The bulk etch rate was also calculated from these measurements. Dose response of the material was studied, and the angular distribution of charged particles created by the fast neutron beam was measured with CR-39. The efficiency of CR-39 was calculated versus that of the Rossi chamber, and an algorithm was devised for derivation of LET spectra from the major and minor axis dimensions of the observed tracks. The CR-39 was irradiated in the same positions as the Rossi chamber, and the derived spectra were compared directly. ^

A versatile toolkit to produce sensitive FRET biosensors to visualize signaling in time and space

Genetically encoded, ratiometric biosensors based on fluorescence resonance energy transfer (FRET) are powerful tools to study the spatiotemporal dynamics of cell signaling. However, many biosensors lack sensitivity. We present a biosensor library that contains circularly permutated mutants for both the donor and acceptor fluorophores, which alter the orientation of the dipoles and thus better accommodate structural constraints imposed by different signaling molecules while maintaining FRET efficiency. Our strategy improved the brightness and dynamic range of preexisting RhoA and extracellular signal-regulated protein kinase (ERK) biosensors. Using the improved RhoA biosensor, we found micrometer-sized zones of RhoA activity at the tip of F-actin bundles in growth cone filopodia during neurite extension, whereas RhoA was globally activated throughout collapsing growth cones. RhoA was also activated in filopodia and protruding membranes at the leading edge of motile fibroblasts. Using the improved ERK biosensor, we simultaneously measured ERK activation dynamics in multiple cells using low-magnification microscopy and performed in vivo FRET imaging in zebrafish. Thus, we provide a construction toolkit consisting of a vector set, which enables facile generation of sensitive biosensors.

Enhancement of photosynthetic carbon assimilation efficiency by phytoplankton in the future coastal ocean

A mesocosm experiment was conducted to evaluate the effects of future climate conditions on photosynthesis and productivity of coastal phytoplankton. Natural phytoplankton assemblages were incubated in field mesocosms under the ambient condition (present condition: ca. 400 ppmv CO2 and ambient temp.), and two future climate conditions (acidification condition: ca. 900 ppmv CO2 and ambient temp.; greenhouse condition: ca. 900 ppmv CO2 and 3 °C warmer than ambient). Photosynthetic parameters of steady-state light responses curves (LCs; measured by PAM fluorometer) and photosynthesis-irradiance curves (P-I curves; estimated by in situ incorporation of 14C) were compared to three conditions during the experiment period. Under acidification, electron transport efficiency (alpha LC) and photosynthetic 14C assimilation efficiency (alpha) were 10% higher than those of the present condition, but maximum rates of relative electron transport (rETRm,LC) and photosynthetic 14C assimilation (PBmax) were lower than the present condition by about 19% and 7%, respectively. In addition, rETRm,LC and alpha LC were not significantly different between and greenhouse conditions, but PBmax and alpha of greenhouse conditions were higher than those of the present condition by about 9% and 30%, respectively. In particular, the greenhouse condition has drastically higher PBmax and alpha than the present condition more than 60% during the post-bloom period. According to these results, two future ocean conditions have major positive effects on the photosynthesis in terms of energy utilization efficiency for organic carbon fixation through the inorganic carbon assimilation. Despite phytoplankton taking an advantage on photosynthesis, primary production of phytoplankton was not stimulated by future conditions. In particular, biomass of phytoplankton was depressed under both acidification and greenhouse conditions after the the pre-bloom period, and more research is required to suggest that some factors such as grazing activity could be important for regulating phytoplankton bloom in the future ocean.

Combined effects of different CO2 levels and N sources on the diazotrophic cyanobacterium Trichodesmium

To predict effects of climate change and possible feedbacks, it is crucial to understand the mechanisms behind CO2 responses of biogeochemically relevant phytoplankton species. Previous experiments on the abundant N2 fixers Trichodesmium demonstrated strong CO2 responses, which were attributed to an energy reallocation between its carbon (C) and nitrogen (N) acquisition. Pursuing this hypothesis, we manipulated the cellular energy budget by growing Trichodesmium erythraeum IMS101 under different CO2 partial pressure (pCO2) levels (180, 380, 980 and 1400?µatm) and N sources (N2 and NO3-). Subsequently, biomass production and the main energy-generating processes (photosynthesis and respiration) and energy-consuming processes (N2 fixation and C acquisition) were measured. While oxygen fluxes and chlorophyll fluorescence indicated that energy generation and its diurnal cycle was neither affected by pCO2 nor N source, cells differed in production rates and composition. Elevated pCO2 increased N2 fixation and organic C and N contents. The degree of stimulation was higher for nitrogenase activity than for cell contents, indicating a pCO2 effect on the transfer efficiency from N2 to biomass. pCO2-dependent changes in the diurnal cycle of N2 fixation correlated well with C affinities, confirming the interactions between N and C acquisition. Regarding effects of the N source, production rates were enhanced in NO3-grown cells, which we attribute to the higher N retention and lower ATP demand compared with N2 fixation. pCO2 effects on C affinity were less pronounced in NO3- users than N2 fixers. Our study illustrates the necessity to understand energy budgets and fluxes under different environmental conditions for explaining indirect effects of rising pCO2.

Novel Inductor‐less Conversion Strategy Based on Multiphase Transformer‐Coupled Converters: Analysis, Design and Applications

There are many the requirements that modern power converters should fulfill. Most of the applications where these converters are used, demand smaller converters with high efficiency, improved power density and a fast dynamic response. For instance, loads like microprocessors demand aggressive current steps with very high slew rates (100A/mus and higher); besides, during these load steps, the supply voltage of the microprocessor should be kept within tight limits in order to ensure its correct performance. The accomplishment of these requirements is not an easy task; complex solutions like advanced topologies - such as multiphase converters- as well as advanced control strategies are often needed. Besides, it is also necessary to operate the converter at high switching frequencies and to use capacitors with high capacitance and low ESR. Improving the dynamic response of power converters does not rely only on the control strategy but also the power topology should be suited to enable a fast dynamic response. Moreover, in later years, a fast dynamic response does not only mean accomplishing fast load steps but output voltage steps are gaining importance as well. At least, two applications that require fast voltage changes can be named: Low power microprocessors. In these devices, the voltage supply is changed according to the workload and the operating frequency of the microprocessor is changed at the same time. An important reduction in voltage dependent losses can be achieved with such changes. This technique is known as Dynamic Voltage Scaling (DVS). Another application where important energy savings can be achieved by means of changing the supply voltage are Radio Frequency Power Amplifiers. For example, RF architectures based on ‘Envelope Tracking’ and ‘Envelope Elimination and Restoration’ techniques can take advantage of voltage supply modulation and accomplish important energy savings in the power amplifier. However, in order to achieve these efficiency improvements, a power converter with high efficiency and high enough bandwidth (hundreds of kHz or even tens of MHz) is necessary in order to ensure an adequate supply voltage. The main objective of this Thesis is to improve the dynamic response of DC-DC converters from the point of view of the power topology. And the term dynamic response refers both to the load steps and the voltage steps; it is also interesting to modulate the output voltage of the converter with a specific bandwidth. In order to accomplish this, the question of what is it that limits the dynamic response of power converters should be answered. Analyzing this question leads to the conclusion that the dynamic response is limited by the power topology and specifically, by the filter inductance of the converter which is found in series between the input and the output of the converter. The series inductance is the one that determines the gain of the converter and provides the regulation capability. Although the energy stored in the filter inductance enables the regulation and the capability of filtering the output voltage, it imposes a limitation which is the concern of this Thesis. The series inductance stores energy and prevents the current from changing in a fast way, limiting the slew rate of the current through this inductor. Different solutions are proposed in the literature in order to reduce the limit imposed by the filter inductor. Many publications proposing new topologies and improvements to known topologies can be found in the literature. Also, complex control strategies are proposed with the objective of improving the dynamic response in power converters. In the proposed topologies, the energy stored in the series inductor is reduced; examples of these topologies are Multiphase converters, Buck converter operating at very high frequency or adding a low impedance path in parallel with the series inductance. Control techniques proposed in the literature, focus on adjusting the output voltage as fast as allowed by the power stage; examples of these control techniques are: hysteresis control, V 2 control, and minimum time control. In some of the proposed topologies, a reduction in the value of the series inductance is achieved and with this, the energy stored in this magnetic element is reduced; less stored energy means a faster dynamic response. However, in some cases (as in the high frequency Buck converter), the dynamic response is improved at the cost of worsening the efficiency. In this Thesis, a drastic solution is proposed: to completely eliminate the series inductance of the converter. This is a more radical solution when compared to those proposed in the literature. If the series inductance is eliminated, the regulation capability of the converter is limited which can make it difficult to use the topology in one-converter solutions; however, this topology is suitable for power architectures where the energy conversion is done by more than one converter. When the series inductor is eliminated from the converter, the current slew rate is no longer limited and it can be said that the dynamic response of the converter is independent from the switching frequency. This is the main advantage of eliminating the series inductor. The main objective, is to propose an energy conversion strategy that is done without series inductance. Without series inductance, no energy is stored between the input and the output of the converter and the dynamic response would be instantaneous if all the devices were ideal. If the energy transfer from the input to the output of the converter is done instantaneously when a load step occurs, conceptually it would not be necessary to store energy at the output of the converter (no output capacitor COUT would be needed) and if the input source is ideal, the input capacitor CIN would not be necessary. This last feature (no CIN with ideal VIN) is common to all power converters. However, when the concept is actually implemented, parasitic inductances such as leakage inductance of the transformer and the parasitic inductance of the PCB, cannot be avoided because they are inherent to the implementation of the converter. These parasitic elements do not affect significantly to the proposed concept. In this Thesis, it is proposed to operate the converter without series inductance in order to improve the dynamic response of the converter; however, on the other side, the continuous regulation capability of the converter is lost. It is said continuous because, as it will be explained throughout the Thesis, it is indeed possible to achieve discrete regulation; a converter without filter inductance and without energy stored in the magnetic element, is capable to achieve a limited number of output voltages. The changes between these output voltage levels are achieved in a fast way. The proposed energy conversion strategy is implemented by means of a multiphase converter where the coupling of the phases is done by discrete two-winding transformers instead of coupledinductors since transformers are, ideally, no energy storing elements. This idea is the main contribution of this Thesis. The feasibility of this energy conversion strategy is first analyzed and then verified by simulation and by the implementation of experimental prototypes. Once the strategy is proved valid, different options to implement the magnetic structure are analyzed. Three different discrete transformer arrangements are studied and implemented. A converter based on this energy conversion strategy would be designed with a different approach than the one used to design classic converters since an additional design degree of freedom is available. The switching frequency can be chosen according to the design specifications without penalizing the dynamic response or the efficiency. Low operating frequencies can be chosen in order to favor the efficiency; on the other hand, high operating frequencies (MHz) can be chosen in order to favor the size of the converter. For this reason, a particular design procedure is proposed for the ‘inductorless’ conversion strategy. Finally, applications where the features of the proposed conversion strategy (high efficiency with fast dynamic response) are advantageus, are proposed. For example, in two-stage power architectures where a high efficiency converter is needed as the first stage and there is a second stage that provides the fine regulation. Another example are RF power amplifiers where the voltage is modulated following an envelope reference in order to save power; in this application, a high efficiency converter, capable of achieving fast voltage steps is required. The main contributions of this Thesis are the following: The proposal of a conversion strategy that is done, ideally, without storing energy in the magnetic element. The validation and the implementation of the proposed energy conversion strategy. The study of different magnetic structures based on discrete transformers for the implementation of the proposed energy conversion strategy. To elaborate and validate a design procedure. To identify and validate applications for the proposed energy conversion strategy. It is important to remark that this work is done in collaboration with Intel. The particular features of the proposed conversion strategy enable the possibility of solving the problems related to microprocessor powering in a different way. For example, the high efficiency achieved with the proposed conversion strategy enables it as a good candidate to be used for power conditioning, as a first stage in a two-stage power architecture for powering microprocessors.

Free-form optical systems for nonimaging applications : Sistemas ópticos anamórficos para aplicaciones anidólicas

La óptica anidólica es una rama de la óptica cuyo desarrollo comenzó a mediados de la década de 1960. Este relativamente nuevo campo de la óptica se centra en la transferencia eficiente de la luz, algo necesario en muchas aplicaciones, entre las que destacamos los concentradores solares y los sistemas de iluminación. Las soluciones de la óptica clásica a los problemas de la transferencia de energía de la luz sólo son adecuadas cuando los rayos de luz son paraxiales. La condición paraxial no se cumple en la mayoría de las aplicaciones para concentración e iluminación. Esta tesis contiene varios diseños free-form (aquellos que no presentan ninguna simetría, ni de rotación ni lineal) cuyas aplicaciones van destinadas a estos dos campos. El término nonimaging viene del hecho de que estos sistemas ópticos no necesitan formar una imagen del objeto, aunque no formar la imagen no es una condición necesaria. Otra palabra que se utiliza a veces en lugar de nonimaging es la palabra anidólico, viene del griego "an+eidolon" y tiene el mismo significado. La mayoría de los sistemas ópticos diseñados para aplicaciones anidólicas no presentan ninguna simetría, es decir, son free-form (anamórficos). Los sistemas ópticos free-form están siendo especialmente relevantes durante los últimos años gracias al desarrollo de las herramientas para su fabricación como máquinas de moldeo por inyección y el mecanizado multieje. Sin embargo, solo recientemente se han desarrollado técnicas de diseño anidólicas capaces de cumplir con estos grados de libertad. En aplicaciones de iluminación el método SMS3D permite diseñar dos superficies free-form para controlar las fuentes de luz extensas. En los casos en que se requiere una elevada asimetría de la fuente, el objeto o las restricciones volumétricos, las superficies free-form permiten obtener soluciones de mayor eficiencia, o disponer de menos elementos en comparación con las soluciones de simetría de rotación, dado que las superficies free-form tienen más grados de libertad y pueden realizar múltiples funciones debido a su naturaleza anamórfica. Los concentradores anidólicos son muy adecuados para la captación de energía solar, ya que el objetivo no es la reproducción de una imagen exacta del sol, sino sencillamente la captura de su energía. En este momento, el campo de la concentración fotovoltaica (CPV) tiende hacia sistemas de alta concentración con el fin de compensar el gasto de las células solares multi-unión (MJ) utilizadas como receptores, reduciendo su área. El interés en el uso de células MJ radica en su alta eficiencia de conversión. Para obtener sistemas competitivos en aplicaciones terrestres se recurre a sistemas fotovoltaicos de alta concentración (HCPV), con factores de concentración geométrica por encima de 500x. Estos sistemas se componen de dos (o más) elementos ópticos (espejos y/o lentes). En los sistemas presentados a lo largo de este trabajo se presentan ejemplos de concentradores HCPV con elementos reflexivos como etapa primaria, así como concentradores con elementos refractivos (lente de Fresnel). Con la necesidad de aumentar la eficiencia de los sistemas HCPV reales y con el fin de proporcionar la división más eficiente del espectro solar, células conteniendo cuatro o más uniones (con un potencial de alcanzar eficiencias de más del 45% a una concentración de cientos de soles) se exploran hoy en día. En esta tesis se presenta una de las posibles arquitecturas de división del espectro (spectrum-splitting en la literatura anglosajona) que utilizan células de concentración comercial. Otro campo de aplicación de la óptica nonimaging es la iluminación, donde es necesario proporcionar un patrón de distribución de la iluminación específico. La iluminación de estado sólido (SSL), basada en la electroluminiscencia de materiales semiconductores, está proporcionando fuentes de luz para aplicaciones de iluminación general. En la última década, los diodos emisores de luz (LED) de alto brillo han comenzado a reemplazar a las fuentes de luz convencionales debido a la superioridad en la calidad de la luz emitida, elevado tiempo de vida, compacidad y ahorro de energía. Los colimadores utilizados con LEDs deben cumplir con requisitos tales como tener una alta eficiencia, un alto control del haz de luz, una mezcla de color espacial y una gran compacidad. Presentamos un colimador de luz free-form con microestructuras capaz de conseguir buena colimación y buena mezcla de colores con una fuente de LED RGGB. Una buena mezcla de luz es importante no sólo para simplificar el diseño óptico de la luminaria sino también para evitar hacer binning de los chips. La mezcla de luz óptica puede reducir los costes al evitar la modulación por ancho de pulso y otras soluciones electrónicas patentadas para regulación y ajuste de color. Esta tesis consta de cuatro capítulos. Los capítulos que contienen la obra original de esta tesis son precedidos por un capítulo introductorio donde se presentan los conceptos y definiciones básicas de la óptica geométrica y en el cual se engloba la óptica nonimaging. Contiene principios de la óptica no formadora de imagen junto con la descripción de sus problemas y métodos de diseño. Asimismo se describe el método de Superficies Múltiples Simultáneas (SMS), que destaca por su versatilidad y capacidad de controlar varios haces de rayos. Adicionalmente también se describe la integración Köhler y sus aplicaciones en el campo de la energía fotovoltaica. La concentración fotovoltaica y la iluminación de estado sólido son introducidas junto con la revisión de su estado actual. El Segundo y Tercer Capítulo contienen diseños ópticos avanzados con aplicación en la concentración solar principalmente, mientras que el Cuarto Capítulo describe el colimador free-form con surcos que presenta buena mezcla de colores para aplicaciones de iluminación. El Segundo Capítulo describe dos concentradores ópticos HCPV diseñados con el método SMS en tres dimensiones (SMS3D) que llevan a cabo integración Köhler en dos direcciones con el fin de proporcionar una distribución de irradiancia uniforme libre de aberraciones cromáticas sobre la célula solar. Uno de los diseños es el concentrador XXR free-form diseñado con el método SMS3D, donde el espejo primario (X) y la lente secundaria (R) se dividen en cuatro sectores simétricos y llevan a cabo la integración Köhler (proporcionando cuatro unidades del array Köhler), mientras que el espejo intermedio (X) presenta simetría rotacional. Otro concentrador HCPV presentado es el Fresnel-RXI (FRXI) con una lente de Fresnel funcionando como elemento primario (POE) y una lente RXI como elemento óptico secundario (SOE), que presenta configuración 4-fold con el fin de realizar la integración Köhler. Las lentes RXI son dispositivos nonimaging conocidos, pero su aplicación como elemento secundario es novedosa. Los concentradores XXR y FRXI Köhler son ejemplos académicos de muy alta concentración (más de 2,000x, mientras que los sistemas convencionales hoy en día no suelen llegar a 1,000x) preparados para las células solares N-unión (con N>3), que probablemente requerirán una mayor concentración y alta uniformidad espectral de irradiancia con el fin de obtener sistemas CPV terrestres eficientes y rentables. Ambos concentradores están diseñados maximizando funciones de mérito como la eficiencia óptica, el producto concentración-aceptancia (CAP) y la uniformidad de irradiancia sobre la célula libre de la aberración cromática (integración Köhler). El Tercer Capítulo presenta una arquitectura para la división del espectro solar basada en un módulo HCPV con alta concentración (500x) y ángulo de aceptancia alto (>1º) que tiene por objeto reducir ambas fuentes de pérdidas de las células triple unión (3J) comerciales: el uso eficiente del espectro solar y la luz reflejada de los contactos metálicos y de la superficie de semiconductor. El módulo para la división del espectro utiliza el espectro solar más eficiente debido a la combinación de una alta eficiencia de una célula de concentración 3J (GaInP/GaInAs/Ge) y una de contacto posterior (BPC) de concentración de silicio (Si), así como la técnica de confinamiento externo para la recuperación de la luz reflejada por la célula 3J con el fin de ser reabsorbida por la célula. En la arquitectura propuesta, la célula 3J opera con su ganancia de corriente optimizada (concentración geométrica de 500x), mientras que la célula de silicio trabaja cerca de su óptimo también (135x). El módulo de spectrum-splitting consta de una lente de Fresnel plana como POE y un concentrador RXI free-form como SOE con un filtro paso-banda integrado en él. Tanto POE como SOE realizan la integración Köhler para producir homogeneización de luz sobre la célula. El filtro paso banda envía los fotones IR en la banda 900-1,150nm a la célula de silicio. Hay varios aspectos prácticos de la arquitectura del módulo presentado que ayudan a reducir la complejidad de los sistemas spectrum-splitting (el filtro y el secundario forman una sola pieza sólida, ambas células son coplanarias simplificándose el cableado y la disipación de calor, etc.). Prototipos prueba-de-concepto han sido ensamblados y probados a fin de demostrar la fabricabilidad del filtro y su rendimiento cuando se combina con la técnica de reciclaje de luz externa. Los resultados obtenidos se ajustan bastante bien a los modelos y a las simulaciones e invitan al desarrollo de una versión más compleja de este prototipo en el futuro. Dos colimadores sólidos con surcos free-form se presentan en el Cuarto Capítulo. Ambos diseños ópticos están diseñados originalmente usando el método SMS3D. La segunda superficie ópticamente activa está diseñada a posteriori como una superficie con surcos. El diseño inicial de dos espejos (XX) está diseñado como prueba de concepto. En segundo lugar, el diseño RXI free-form es comparable con los colimadores RXI existentes. Se trata de un diseño muy compacto y eficiente que proporciona una muy buena mezcla de colores cuando funciona con LEDs RGB fuera del eje óptico como en los RGB LEDs convencionales. Estos dos diseños son dispositivos free-form diseñados con la intención de mejorar las propiedades de mezcla de colores de los dispositivos no aplanáticos RXI con simetría de revolución y la eficiencia de los aplanáticos, logrando una buena colimación y una buena mezcla de colores. La capacidad de mezcla de colores del dispositivo no-aplanático mejora añadiendo características de un aplanático a su homólogo simétrico sin pérdida de eficiencia. En el caso del diseño basado en RXI, su gran ventaja consiste en su menor coste de fabricación ya que el proceso de metalización puede evitarse. Aunque algunos de los componentes presentan formas muy complejas, los costes de fabricación son relativamente insensibles a la complejidad del molde, especialmente en el caso de la producción en masa (tales como inyección de plástico), ya que el coste del molde se reparte entre todas las piezas fabricadas. Por último, las últimas dos secciones son las conclusiones y futuras líneas de investigación. ABSTRACT Nonimaging optics is a branch of optics whose development began in the mid-1960s. This rather new field of optics focuses on the efficient light transfer necessary in many applications, among which we highlight solar concentrators and illumination systems. The classical optics solutions to the problems of light energy transfer are only appropriate when the light rays are paraxial. The paraxial condition is not met in most applications for the concentration and illumination. This thesis explores several free-form designs (with neither rotational nor linear symmetry) whose applications are intended to cover the above mentioned areas and more. The term nonimaging comes from the fact that these optical systems do not need to form an image of the object, although it is not a necessary condition not to form an image. Another word sometimes used instead of nonimaging is anidolic, and it comes from the Greek “an+eidolon” and has the same meaning. Most of the optical systems designed for nonimaging applications are without any symmetry, i.e. free-form. Free-form optical systems become especially relevant lately with the evolution of free-form tooling (injection molding machines, multi-axis machining techniques, etc.). Nevertheless, only recently there are nonimaging design techniques that are able to meet these degrees of freedom. In illumination applications, the SMS3D method allows designing two free-form surfaces to control very well extended sources. In cases when source, target or volumetric constrains have very asymmetric requirements free-form surfaces are offering solutions with higher efficiency or with fewer elements in comparison with rotationally symmetric solutions, as free-forms have more degrees of freedom and they can perform multiple functions due to their free-form nature. Anidolic concentrators are well suited for the collection of solar energy, because the goal is not the reproduction of an exact image of the sun, but instead the collection of its energy. At this time, Concentration Photovoltaics (CPV) field is turning to high concentration systems in order to compensate the expense of multi-junction (MJ) solar cells used as receivers by reducing its area. Interest in the use of MJ cells lies in their very high conversion efficiency. High Concentration Photovoltaic systems (HCPV) with geometric concentration of more than 500x are required in order to have competitive systems in terrestrial applications. These systems comprise two (or more) optical elements, mirrors and/or lenses. Systems presented in this thesis encompass both main types of HCPV architectures: concentrators with primary reflective element and concentrators with primary refractive element (Fresnel lens). Demand for the efficiency increase of the actual HCPV systems as well as feasible more efficient partitioning of the solar spectrum, leads to exploration of four or more junction solar cells or submodules. They have a potential of reaching over 45% efficiency at concentration of hundreds of suns. One possible architectures of spectrum splitting module using commercial concentration cells is presented in this thesis. Another field of application of nonimaging optics is illumination, where a specific illuminance distribution pattern is required. The Solid State Lighting (SSL) based on semiconductor electroluminescence provides light sources for general illumination applications. In the last decade high-brightness Light Emitting Diodes (LEDs) started replacing conventional light sources due to their superior output light quality, unsurpassed lifetime, compactness and energy savings. Collimators used with LEDs have to meet requirements like high efficiency, high beam control, color and position mixing, as well as a high compactness. We present a free-form collimator with microstructures that performs good collimation and good color mixing with RGGB LED source. Good light mixing is important not only for simplifying luminaire optical design but also for avoiding die binning. Optical light mixing may reduce costs by avoiding pulse-width modulation and other patented electronic solutions for dimming and color tuning. This thesis comprises four chapters. Chapters containing the original work of this thesis are preceded by the introductory chapter that addresses basic concepts and definitions of geometrical optics on which nonimaging is developed. It contains fundamentals of nonimaging optics together with the description of its design problems, principles and methods, and with the Simultaneous Multiple Surface (SMS) method standing out for its versatility and ability to control several bundles of rays. Köhler integration and its applications in the field of photovoltaics are described as well. CPV and SSL fields are introduced together with the review on their background and their current status. Chapter 2 and Chapter 3 contain advanced optical designs with primarily application in solar concentration; meanwhile Chapter 4 portrays the free-form V-groove collimator with good color mixing property for illumination application. Chapter 2 describes two HCPV optical concentrators designed with the SMS method in three dimensions (SMS3D). Both concentrators represent Köhler integrator arrays that provide uniform irradiance distribution free from chromatic aberrations on the solar cell. One of the systems is the XXR free-form concentrator designed with the SMS3D method. The primary mirror (X) of this concentrator and secondary lens (R) are divided in four symmetric sectors (folds) that perform Köhler integration; meanwhile the intermediate mirror (X) is rotationally symmetric. Second HCPV concentrator is the Fresnel-RXI (FRXI) with flat Fresnel lens as the Primary Optical Element (POE) and an RXI lens as the Secondary Optical Element (SOE). This architecture manifests 4-fold configuration for performing Köhler integration (4 array units), as well. The RXI lenses are well-known nonimaging devices, but their application as SOE is novel. Both XXR and FRXI Köhler HCPV concentrators are academic examples of very high concentration (more than 2,000x meanwhile conventional systems nowadays have up to 1,000x) prepared for the near future N-junction (N>3) solar cells. In order to have efficient and cost-effective terrestrial CPV systems, those cells will probably require higher concentrations and high spectral irradiance uniformity. Both concentrators are designed by maximizing merit functions: the optical efficiency, concentration-acceptance angle (CAP) and cell-irradiance uniformity free from chromatic aberrations (Köhler integration). Chapter 3 presents the spectrum splitting architecture based on a HCPV module with high concentration (500x) and high acceptance angle (>1º). This module aims to reduce both sources of losses of the actual commercial triple-junction (3J) solar cells with more efficient use of the solar spectrum and with recovering the light reflected from the 3J cells’ grid lines and semiconductor surface. The solar spectrum is used more efficiently due to the combination of a high efficiency 3J concentration cell (GaInP/GaInAs/Ge) and external Back-Point-Contact (BPC) concentration silicon (Si) cell. By employing external confinement techniques, the 3J cell’s reflections are recovered in order to be re-absorbed by the cell. In the proposed concentrator architecture, the 3J cell operates at its optimized current gain (at geometrical concentration of 500x), while the Si cell works near its optimum, as well (135x). The spectrum splitting module consists of a flat Fresnel lens (as the POE), and a free-form RXI-type concentrator with a band-pass filter embedded in it (as the SOE), both POE and SOE performing Köhler integration to produce light homogenization. The band-pass filter sends the IR photons in the 900-1,150nm band to the Si cell. There are several practical aspects of presented module architecture that help reducing the added complexity of the beam splitting systems: the filter and secondary are forming a single solid piece, both cells are coplanar so the heat management and wiring is simplified, etc. Two proof-of-concept prototypes are assembled and tested in order to prove filter manufacturability and performance, as well as the potential of external light recycling technique. Obtained measurement results agree quite well with models and simulations, and show an opened path to manufacturing of the Fresnel RXI-type secondary concentrator with spectrum splitting strategy. Two free-form solid V-groove collimators are presented in Chapter 4. Both free-form collimators are originally designed with the SMS3D method. The second mirrored optically active surface is converted in a grooved surface a posteriori. Initial two mirror (XX) design is presented as a proof-of-concept. Second, RXI free-form design is comparable with existing RXI collimators as it is a highly compact and a highly efficient design. It performs very good color mixing of the RGGB LED sources placed off-axis like in conventional RGB LEDs. Collimators described here improve color mixing property of the prior art rotationally symmetric no-aplanatic RXI devices, and the efficiency of the aplanatic ones, accomplishing both good collimation and good color mixing. Free-form V-groove collimators enhance the no-aplanatic device's blending capabilities by adding aplanatic features to its symmetric counterpart with no loss in efficiency. Big advantage of the RXI design is its potentially lower manufacturing cost, since the process of metallization may be avoided. Although some components are very complicated for shaping, the manufacturing costs are relatively insensitive to the complexity of the mold especially in the case of mass production (such as plastic injection), as the cost of the mold is spread in many parts. Finally, last two sections are conclusions and future lines of investigation.

Vehicle to vehicle energy exchange in smart grid applications

This paper presents a novel vehicle to vehicle energy exchange market (V2VEE) between electric vehicles (EVs) for decreasing the energy cost to be paid by some users whose EVs must be recharged during the day to fulfil their daily scheduled trips and also reducing the impact of charging on the electric grid. EVs with excess of energy in their batteries can transfer this energy among other EVs which need charge during their daily trips. These second type of owners can buy the energy directly to the electric grid or they can buy the energy from other EV at lower price. An aggregator is responsible for collecting all information among vehicles located in the same area at the same time and make possible this energy transfer.

Energy yield determination of concentrator solar cells using laboratory measurements

The annual energy conversion efficiency is calculated for a four junction inverted metamorphic solar cell that has been completely characterized in the laboratory at room temperature using measurements fit to a comprehensive optoelectronic model of the multijunction solar cells. A simple model of the temperature dependence is used redict the performance of the solar cell under varying temperature and spectra characteristic of Golden, CO for an entire year. The annual energy conversion efficiency is calculated by integrating the predicted cell performance over the entire year. The effects of geometric concentration, CPV system thermal characteristics, and luminescent coupling are ighlighted.

Membrane-bound state of the colicin E1 channel domain as an extended two-dimensional helical array

Atomic level structures have been determined for the soluble forms of several colicins and toxins, but the structural changes that occur after membrane binding have not been well characterized. Changes occurring in the transition from the soluble to membrane-bound state of the C-terminal 190-residue channel polypeptide of colicin E1 (P190) bound to anionic membranes are described. In the membrane-bound state, the α-helical content increases from 60–64% to 80–90%, with a concomitant increase in the average length of the helical segments from 12 to 16 or 17 residues, close to the length required to span the membrane bilayer in the open channel state. The average distance between helical segments is increased and interhelix interactions are weakened, as shown by a major loss of tertiary structure interactions, decreased efficiency of fluorescence resonance energy transfer from an energy donor on helix V of P190 to an acceptor on helix IX, and decreased resonance energy transfer at higher temperatures, not observed in soluble P190, implying freedom of motion of helical segments. Weaker interactions are also shown by a calorimetric thermal transition of low cooperativity, and the extended nature of the helical array is shown by a 3- to 4-fold increase in the average area subtended per molecule to 4,200 Å2 on the membrane surface. The latter, with analysis of the heat capacity changes, implies the absence of a developed hydrophobic core in the membrane-bound P190. The membrane interfacial layer thus serves to promote formation of a highly helical extended two-dimensional flexible net. The properties of the membrane-bound state of the colicin channel domain (i.e., hydrophobic anchor, lengthened and loosely coupled α-helices, and close association with the membrane interfacial layer) are plausible structural features for the state that is a prerequisite for voltage gating, formation of transmembrane helices, and channel opening.