Kinetische Untersuchungen zur Synthese von Blockcopolymeren mit Methacrylatsegmenten durch neue anionische Polymerisationssysteme

Ein metallfreies Reaktionssystem mit Bis(triphenylphosphoranyliden)ammonium-Kationen als Gegenionen sowie das Additiv Lithium-2-methoxyethoxid wurden auf ihre Eignung zur anionischen Synthese von Blockcopolymeren mit (Meth)acrylatsegmenten bei moderaten Temperaturen im Strömungsrohr-Reaktor untersucht. Das metallfreie System ist zur lebenden Polymerisation von Methacrylaten in THF mit engen Molekulargewichtsverteilungen bei Reaktionszeiten < 1 s bis zu Temperaturen von 0 °C geeignet. In Gegenwart von Metallionen (Li+) findet eine Verlangsamung der Polymerisation unter Verlust der Reaktionskontrolle statt, Lithiumenolate verursachen nun breite, multimodale Molekulargewichtsverteilungen. Eine lebende Polymerisation von Acrylaten ist nicht möglich, massenspektroskopische Untersuchungen der Produkte weisen auf einen komplexen Reaktionsmechanismus mit Abbruch- und Übertragungsreaktionen hin. Die Synthese von Poly(styrol)-block-Poly(1,4-butadien)-block-Poly(methylmethacrylat)-Copolymeren in Toluol ist mit Lithium-2-methoxyethoxid als Additiv für die MMA-Polymerisation bei moderaten Mischtemperaturen (T < 0 °C) im Strömungsrohr-Reaktor möglich, die Effektivität des Wechselschritts von Polybutadien zu PMMA beträgt im Durchschnitt ca. 50 %. Untersuchungen verschiedener Reaktionsparameter, wie z.B. der Endfunktionalisierung des Polybutadiens mit 1,1-Diphenylethylen und der Temperatur während der Verkappung und der MMA-Polymerisation, geben keine eindeutigen Hinweise auf die Ursache dieses Phänomens. MALDI-TOF-Massenspektren des unreagierten Polybutadien Precursors zeigen die Anlagerung von 1-3 Molekülen Methylmethacrylat und keinen Abbruch durch Backbiting, was auf die Ausbildung stabiler Aggregate hindeutet.

Spectroscopic studies on Cyclodextrin and Metal Organic Framework based potential nanovectors for delivery of Anticancer and Antiviral drugs

The aim of this work is to contribute to the development of new multifunctional nanocarriers for improved encapsulation and delivery of anticancer and antiviral drugs. The work focused on water soluble and biocompatible oligosaccharides, the cyclodextrins (CyDs), and a new family of nanostructured, biodegradable carrier materials made of porous metal-organic frameworks (nanoMOFs). The drugs of choice were the anticancer doxorubicin (DOX), azidothymidine (AZT) and its phosphate derivatives and artemisinin (ART). DOX possesses a pharmacological drawback due to its self-aggregation tendency in water. The non covalent binding of DOX to a series of CyD derivatives, such as g-CyD, an epichlorohydrin crosslinked b-CyD polymer (pb-CyD) and a citric acid crosslinked g-CyD polymer (pg-CyD) was studied by UV visible absorption, circular dichroism and fluorescence. Multivariate global analysis of multiwavelength data from spectroscopic titrations allowed identification and characterization of the stable complexes. pg-CyD proved to be the best carrier showing both high association constants and ability to monomerize DOX. AZT is an important antiretroviral drug. The active form is AZT-triphosphate (AZT-TP), formed in metabolic paths of low efficiency. Direct administration of AZT-TP is limited by its poor stability in biological media. So the development of suitable carriers is highly important. In this context we studied the binding of some phosphorilated derivatives to nanoMOFs by spectroscopic methods. The results obtained with iron(III)-trimesate nanoMOFs allowed to prove that the binding of these drugs mainly occurs by strong iono-covalent bonds to iron(III) centers. On the basis of these and other results obtained in partner laboratories, it was possible to propose this highly versatile and “green” carrier system for delivery of phosphorylated nucleoside analogues. The interaction of DOX with nanoMOFs was also studied. Finally the binding of the antimalarial drug, artemisinin (ART) with two cyclodextrin-based carriers,the pb-CyD and a light responsive bis(b-CyD) host, was also studied.

Laser driven proton acceleration and beam shaping

In the race to obtain protons with higher energies, using more compact systems at the same time, laser-driven plasma accelerators are becoming an interesting possibility. But for now, only beams with extremely broad energy spectra and high divergence have been produced. The driving line of this PhD thesis was the study and design of a compact system to extract a high quality beam out of the initial bunch of protons produced by the interaction of a laser pulse with a thin solid target, using experimentally reliable technologies in order to be able to test such a system as soon as possible. In this thesis, different transport lines are analyzed. The first is based on a high field pulsed solenoid, some collimators and, for perfect filtering and post-acceleration, a high field high frequency compact linear accelerator, originally designed to accelerate a 30 MeV beam extracted from a cyclotron. The second one is based on a quadruplet of permanent magnetic quadrupoles: thanks to its greater simplicity and reliability, it has great interest for experiments, but the effectiveness is lower than the one based on the solenoid; in fact, the final beam intensity drops by an order of magnitude. An additional sensible decrease in intensity is verified in the third case, where the energy selection is achieved using a chicane, because of its very low efficiency for off-axis protons. The proposed schemes have all been analyzed with 3D simulations and all the significant results are presented. Future experimental work based on the outcome of this thesis can be planned and is being discussed now.

In vivo electroporation and ubiquitin promoter--a protocol for sustained gene expression in the lung

BACKGROUND: Gene therapy applications require safe and efficient methods for gene transfer. Present methods are restricted by low efficiency and short duration of transgene expression. In vivo electroporation, a physical method of gene transfer, has evolved as an efficient method in recent years. We present a protocol involving electroporation combined with a long-acting promoter system for gene transfer to the lung. METHODS: The study was designed to evaluate electroporation-mediated gene transfer to the lung and to analyze a promoter system that allows prolonged transgene expression. A volume of 250 microl of purified plasmid DNA suspended in water was instilled into the left lung of anesthetized rats, followed by left thoracotomy and electroporation of the exposed left lung. Plasmids pCiKlux and pUblux expressing luciferase under the control of the cytomegalovirus immediate-early promoter/enhancer (CMV-IEPE) or human polyubiquitin c (Ubc) promoter were used. Electroporation conditions were optimized with four pulses (200 V/cm, 20 ms at 1 Hz) using flat plate electrodes. The animals were sacrificed at different time points up to day 40, after gene transfer. Gene expression was detected and quantified by bioluminescent reporter imaging (BLI) and relative light units per milligram of protein (RLU/mg) was measured by luminometer for p.Pyralis luciferase and immunohistochemistry, using an anti-luciferase antibody. RESULTS: Gene expression with the CMV-IEPE promoter was highest 24 h after gene transfer (2932+/-249.4 relative light units (RLU)/mg of total lung protein) and returned to baseline by day 3 (382+/-318 RLU/mg of total lung protein); at day 5 no expression was detected, whereas gene expression under the Ubc promoter was detected up to day 40 (1989+/-710 RLU/mg of total lung protein) with a peak at day 20 (2821+/-2092 RLU/mg of total lung protein). Arterial blood gas (PaO2), histological assessment and cytokine measurements showed no significant toxicity neither at day 1 nor at day 40. CONCLUSIONS: These results provide evidence that in vivo electroporation is a safe and effective tool for non-viral gene delivery to the lungs. If this method is used in combination with a long-acting promoter system, sustained transgene expression can be achieved.

A STUDY ON REGIONAL CIRCULAR ECONOMY SYSTEM AND ITS CONSTRUCTION, OPERATION AND SUGGESTION FOR SHANGHAI

The characteristics of the traditional linear economic model are high consumption, high emission and low efficiency. Economic development is still largely at the expense of the environment and requires a natural resource investment. This can realize rapid economic development but resource depletion and environmental pollution become increasingly serious. In the 1990's a new economic model, circular economics, began to enter our vision. The circular economy maximizes production and minimizes the impact of economic activities on the ecological environment through organizing the activities through the closed-loop feedback cycle of "resources - production - renewable resource". Circular economy is a better way to solve the contradictions between the economic development and resource shortages. Developing circular economy has become the major strategic initiatives to achieving sustainable development in countries all over the world. The evaluation of the development of circular economics is a necessary step for regional circular economy development. Having a quantitative evaluation of circular economy can better monitor and reveal the contradictions and problems in the process of the development of recycling economy. This thesis will: 1) Create an evaluation model framework and new types of industries and 2) Make an evaluation of the Shanghai circular economy currently to analyze the situation of Shanghai in the development of circular economy. I will then propose suggestions about the structure and development of Shanghai circular economy.

Collective Voluntary Agreements and the Production of Less Polluting Products

Recently, some industries have collectively agreed not to produce models that do not meet an energy efficiency (and hence an environmental) standard. This paper presents a simple model that can be used to examine a voluntary collective agreement to limit or completely eliminate the low efficiency model of a given product (e.g., a low efficiency washing machine). We show that, when there is competition between firms, a collective agreement to limit or even eliminate production of the polluting model can actually increase profits for all firms in the industry. This suggests that a collective agreement of this type might actually be beneficial to firms, while at the same time improving environmental quality. However, the implicit enforcement that comes from the public nature of the commitment is necessary to ensure this outcome. This suggests that, by promoting such agreements, policymakers may be able to achieve substantial environmental gains with relatively little inducement. The impact on social welfare will then depend on whether these gains are sufficiently large to offset consumer losses from reductions in product variety and the associated price increases.

Network-based genome-wide association study of Crohn's disease

Genome-wide association studies (GWAS) have rapidly become a standard method for disease gene discovery. Many recent GWAS indicate that for most disorders, only a few common variants are implicated and the associated SNPs explain only a small fraction of the genetic risk. The current study incorporated gene network information into gene-based analysis of GWAS data for Crohn's disease (CD). The purpose was to develop statistical models to boost the power of identifying disease-associated genes and gene subnetworks by maximizing the use of existing biological knowledge from multiple sources. The results revealed that Markov random field (MRF) based mixture model incorporating direct neighborhood information from a single gene network is not efficient in identifying CD-related genes based on the GWAS data. The incorporation of solely direct neighborhood information might lead to the low efficiency of these models. Alternative MRF models looking beyond direct neighboring information are necessary to be developed in the future for the purpose of this study.^

The two -state model of receptor activation: The agonist and the efficacy

The Two State model describes how drugs activate receptors by inducing or supporting a conformational change in the receptor from “off” to “on”. The beta 2 adrenergic receptor system is the model system which was used to formalize the concept of two states, and the mechanism of hormone agonist stimulation of this receptor is similar to ligand activation of other seven transmembrane receptors. Hormone binding to beta 2 adrenergic receptors stimulates the intracellular production of cyclic adenosine monophosphate (cAMP), which is mediated through the stimulatory guanyl nucleotide binding protein (Gs) interacting with the membrane bound enzyme adenylylcyclase (AC). ^ The effects of cAMP include protein phosphorylation, metabolic regulation and transcriptional regulation. The beta 2 adrenergic receptor system is the most well known of its family of G protein coupled receptors. Ligands have been scrutinized extensively in search of more effective therapeutic agents at this receptor as well as for insight into the biochemical mechanism of receptor activation. Hormone binding to receptor is thought to induce a conformational change in the receptor that increases its affinity for inactive Gs, catalyzes the release of GDP and subsequent binding of GTP and activation of Gs. ^ However, some beta 2 ligands are more efficient at this transformation than others, and the underlying mechanism for this drug specificity is not fully understood. The central problem in pharmacology is the characterization of drugs in their effect on physiological systems, and consequently, the search for a rational scale of drug effectiveness has been the effort of many investigators, which continues to the present time as models are proposed, tested and modified. ^ The major results of this thesis show that for many b2 -adrenergic ligands, the Two State model is quite adequate to explain their activity, but dobutamine (+/−3,4-dihydroxy-N-[3-(4-hydroxyphenyl)-1-methylpropyl]- b -phenethylamine) fails to conform to the predictions of the Two State model. It is a weak partial agonist, but it forms a large amount of high affinity complexes, and these complexes are formed at low concentrations much better than at higher concentrations. Finally, dobutamine causes the beta 2 adrenergic receptor to form high affinity complexes at a much faster rate than can be accounted for by its low efficiency activating AC. Because the Two State model fails to predict the activity of dobutamine in three different ways, it has been disproven in its strictest form. ^

Determinación del coeficiente de nascencia en coberturas cespitosas : oasis del Norte de Mendoza (Argentina)

Uno de los principales problemas que se presenta para la investigación en césped acerca de la adaptabilidad de diferentes especies y sus mezclas o la introducción de otras nuevas, consiste en la dificultad de calcular la densidad de plantas, en forma lo suficientemente aproximada a los resultados reales como para permitir comparaciones estadísticamente válidas de la calidad o usos del cultivo resultante. Se propone una nueva metodología para reemplazar aquella tradicional de cálculo de necesidades de semilla por peso, con densidades determinadas por prueba y error según el resultado final por aspecto y corrección de cantidad de mezcla sólo por valor cultural. La propuesta considera cálculos matemáticos para determinar la cantidad exacta de semillas a emplear para obtener una densidad final de siembra controlada, teniendo en cuenta peso específico de la semilla y valor cultural, afectando el dato resultante por un factor de corrección (CN) que relaciona el valor teórico obtenido con la respuesta a campo. Es decir que la cantidad de semilla a emplear debe calcularse en función de la densidad de plantas deseada. En el presente trabajo el factor CN se determina para diferentes especies. Se parte de la hipótesis que es posible mejorar las formas de selección de especies y preparación de mezclas a través del cálculo de la necesidad de semillas por cm2 y que algunas de las especies utilizadas poseen baja eficiencia en la relación semilla-planta, por lo que se debería ajustar su proporción en las mezclas comerciales locales según este factor de cultivo. Como metodología se propone: a. análisis de poder germinativo, pureza y peso específico de semillas comerciales; b. con cuatro repeticiones, en bloques al azar, realizar siembras individuales de las especies en condiciones ideales de sustrato, iluminación y humedad, y siembra en las mismas condiciones de campo que un cultivo tradicional en condiciones de especie pura y consociadas (mezclas); c. conteo del número de plántulas obtenidas en cada caso por método propuesto por Lush y Franz; d. con los resultados, estimar los coeficientes (CN) promedio y sus respectivos intervalos de confianza. Esta nueva manera de calcular las cantidades exactas de cada componente de una mezcla para césped abre un campo muy importante a la investigación de especies más adecuadas a cada ambiente, dado que el método tradicional conduce a resultados muy ambiguos y de difícil comparación.

Riego con equipos presurizados

En un área de aprox. 2 000 000 ha del sur de Córdoba (Argentina) se evaluaron los equipos de riego con el fin de conocer su funcionamiento, el grado de uniformidad con que trabajan y la eficiencia de riego lograda por los productores. Se realizaron 21 evaluaciones sobre equipos operando de acuerdo con la programación establecida por sus usuarios; 14 sobre pivote, una sobre avance lateral, 4 sobre enrolladores (3 de cañón y uno de baja presión) y 2 sobre side roll. Los parámetros de calidad de riego brindaron coeficiente medio de uniformidad = 81,4 %, con uniformidad de distribución = 73,23 %. En el 80 % de los casos, la lámina aplicada fluctuó entre 10 y 20 mm siendo su promedio = 17 mm. De los resultados se puede inferir que -en general- la superficie asignada a cada equipo es siempre mayor que su capacidad para realizar oportunamente una óptima reposición del agua al suelo y que, si bien los coeficientes de uniformidad y distribución del agua pueden considerarse aceptables, la programación del riego es mala en todos los establecimientos evaluados poniéndose de manifiesto en la baja eficiencia de almacenamiento y repercutiendo directamente sobre la producción de los cultivos regados.

Multiphase Buck Converter with Minimum Time Control Strategy for RF Envelope Modulation

Power amplifier supplied with constant supply voltage has very low efficiency in the transmitter. A DC-DC converter in series with a linear regulator can be used to obtain voltage modulation. Since this converter should be able to change the output voltage very fast, a multiphase buck converter with a minimum time control strategy is proposed. To modulate supply voltage of the envelope amplifier, the multiphase converter works with some particular duty cycle (i/n, i=1, 2 ... n, n is the number of phase) to generate discrete output voltages, and in these duty cycles the output current ripple can be completely cancelled. The transition times for the minimum time are pre-calculated and inserted in a look-up table. The theoretical background, the system model that is necessary in order to calculate the transition times and the experimental results obtained with a 4-phase buck prototype are given

Series combination of a switched dc-dc converter and a linear regulator for high frequency RF envelope amplifier

Classical linear amplifiers such as A, AB and B offer very good linearity suitable for RF power amplifiers. However, its inherent low efficiency limits its use especially in base-stations that manage tens or hundreds of Watts. The use of linearization techniques such as Envelope Elimination and Restoration (EER) allow an increase of efficiency keeping good linearity. This technique requires a very fast dc-dc power converter to provide variable voltage supply to the power amplifier. In this paper, several alternatives are analyzed to implement the envelope amplifier based on a cascade association of a switched dc-dc converter and a linear regulator. A simplified version of this approach is also suitable to operate with Envelope Tracking technique.

Development and experimental evaluation of a complete solar thermophotovoltaic system

We present a practical implementation of a solar thermophotovoltaic (TPV) system. The system presented in this paper comprises a sunlight concentrator system, a cylindrical cup-shaped absorber/emitter (made of tungsten coated with HfO2), and an hexagonal-shaped water-cooled TPV generator comprising 24 germanium TPV cells, which is surrounding the cylindrical absorber/emitter. This paper focuses on the development of shingled TPV cell arrays, the characterization of the sunlight concentrator system, the estimation of the temperature achieved by the cylindrical emitters operated under concentrated sunlight, and the evaluation of the full system performance under real outdoor irradiance conditions. From the system characterization, we have measured short-circuit current densities up to 0.95 A/cm2, electric power densities of 67 mW/cm2, and a global conversion efficiency of about 0.8%. To our knowledge, this is the first overall solar-to-electricity efficiency reported for a complete solar thermophotovoltaic system. The very low efficiency is mainly due to the overheating of the cells (up to 120 °C) and to the high optical concentrator losses, which prevent the achievement of the optimum emitter temperature. The loss analysis shows that by improving both aspects, efficiencies above 5% could be achievable in the very short term and efficiencies above 10% could be achieved with further improvements.

Scar, a WASp-related protein, activates nucleation of actin filaments by the Arp2/3 complex

The Arp2/3 complex, a stable assembly of two actin-related proteins (Arp2 and Arp3) with five other subunits, caps the pointed end of actin filaments and nucleates actin polymerization with low efficiency. WASp and Scar are two similar proteins that bind the p21 subunit of the Arp2/3 complex, but their effect on the nucleation activity of the complex was not known. We report that full-length, recombinant human Scar protein, as well as N-terminally truncated Scar proteins, enhance nucleation by the Arp2/3 complex. By themselves, these proteins either have no effect or inhibit actin polymerization. The actin monomer-binding W domain and the p21-binding A domain from the C terminus of Scar are both required to activate Arp2/3 complex. A proline-rich domain in the middle of Scar enhances the activity of the W and A domains. Preincubating Scar and Arp2/3 complex with actin filaments overcomes the initial lag in polymerization, suggesting that efficient nucleation by the Arp2/3 complex requires assembly on the side of a preexisting filament—a dendritic nucleation mechanism. The Arp2/3 complex with full-length Scar, Scar containing P, W, and A domains, or Scar containing W and A domains overcomes inhibition of nucleation by the actin monomer-binding protein profilin, giving active nucleation over a low background of spontaneous nucleation. These results show that Scar and, likely, related proteins, such as the Cdc42 targets WASp and N-WASp, are endogenous activators of actin polymerization by the Arp2/3 complex.

Saccharomyces cerevisiae Cells with Defective Spindle Pole Body Outer Plaques Accomplish Nuclear Migration via Half-Bridge–organized Microtubules

Cnm67p, a novel yeast protein, localizes to the microtubule organizing center, the spindle pole body (SPB). Deletion of CNM67 (YNL225c) frequently results in spindle misorientation and impaired nuclear migration, leading to the generation of bi- and multinucleated cells (40%). Electron microscopy indicated that CNM67 is required for proper formation of the SPB outer plaque, a structure that nucleates cytoplasmic (astral) microtubules. Interestingly, cytoplasmic microtubules that are essential for spindle orientation and nuclear migration are still present in cnm67Δ1 cells that lack a detectable outer plaque. These microtubules are attached to the SPB half- bridge throughout the cell cycle. This interaction presumably allows for low-efficiency nuclear migration and thus provides a rescue mechanism in the absence of a functional outer plaque. Although CNM67 is not strictly required for mitosis, it is essential for sporulation. Time-lapse microscopy of cnm67Δ1 cells with green fluorescent protein (GFP)-labeled nuclei indicated that CNM67 is dispensable for nuclear migration (congression) and nuclear fusion during conjugation. This is in agreement with previous data, indicating that cytoplasmic microtubules are organized by the half-bridge during mating.