<p><p>Heterotrimeric G proteins play invaluable roles in cellular processes involving transmembrane signaling, particularly at sites of neuronal connectivity within the central nervous system (CNS). G&alpha;z is a member of the G&alpha;i subfamily of heterotrimeric G proteins that displays unique biochemical characteristics and is primarily expressed in neuronal and neuroendocrine cells. Studies in Gz&ndash;null mice over the past decade reveal that Gz significantly impacts responses to psychoactive drugs, and is capable of coupling to D2 dopamine, 5&ndash;HT1A serotonin, &mu;&ndash;opioid, and &alpha;2A&ndash;adrenergic receptors. These studies have suggested that Gz may play a critical role in diseases and disorders involving disruptions of monoamine neurotransmitter signaling in the brain such as depression, anxiety, drug abuse, ADHD, schizophrenia, drug addiction, and pain sensitivity. Much is still unknown about the roles and mechanisms of action of Gz in biology. </p></p><p><p>In this thesis, I have built on what is known regarding G&#945;z biochemistry by conducting a series of studies that provide further understanding of its role in the CNS, particularly in neuronal development and seizure susceptibility. Gz interacts with several proteins that act as regulators and effectors: RGSZ, adenylyl cyclase, EYA2, and Rap1GAP being the best characterized. A finding regarding its impact of Gz on neurotrophin signaling through RAP1GAP in particular has led to much of the work described here. The studies presented in this thesis indicate that G&alpha;z inhibits BDNF-stimulated axon growth in cortical neurons, establishing an endogenous role for G&alpha;z in regulation of neurotrophin signaling in the CNS that may have important implications for development and plasticity. Furthermore, G&alpha;z was shown to be uniquely distributed to synaptic vesicles suggesting that one mechanism underlying Gz biology may be the regulation of vesicle loading, docking, or release. Finally, I demonstrate that Gz-null mice are hypersusceptible to pilocarpine&ndash;induced seizures, and provide histology data indicating increased levels of zinc in the hippocampus. Taken together, these findings suggest that Gz plays a regulatory role at the intersection of neurotrophin and GPCR signaling in the CNS. </p></p> / Dissertation

No description available.

Methamphetamine abuse is a serious public health concern affecting millions of people worldwide, and there are currently no viable pharmacotherapies to treat methamphetamine abuse. Methamphetamine increases extracellular dopamine (DA) concentrations through an interaction with the DA transporter (DAT) and the vesicular monoamine transporter-2 (VMAT2), leading to reward and abuse. While numerous studies have focused on DAT as a target for the discovery of pharmacotherapies to treat psychostimulant abuse, these efforts have been met with limited success. Taking into account the fact that methamphetamine interacts with VMAT2 to increase DA extracellular concentrations; the focus of the current work was to develop novel compounds that interact with VMAT2 to inhibit the effects of methamphetamine. Lobeline, the principal alkaloid found in Lobelia inflata, inhibits VMAT2 binding and function. Inhibition of VMAT2 was hypothesized to be responsible for the observed lobeline-induced inhibition of methamphetamine-evoked DA release in striatal slices and decrease in methamphetamine self-administration in rats. Lobeline has recently completed Phase Ib clinical trials demonstrating safety in methamphetamine abusers. Lobeline is also a potent inhibitor of nicotinic acetylcholine receptors (nAChRs), limiting selectivity for VMAT2. Chemical defunctionalization of the lobeline molecule afforded analogs, meso-transdiene (MTD) and lobelane, which exhibited decreased affinity for nAChRs. MTD, an unsaturated analog of lobeline, exhibited similar affinity for VMAT2 and increased affinity for DAT compared to lobeline. Conformationally-restricted MTD analogs exhibited decreased affinity for DAT compared to MTD, while retaining affinity at VMAT2. One analog, UKMH-106 exhibited high affinity and selectivity for VMAT2 and inhibited METH-evoked DA release from striatal slices. Unfortunately, the MTD analogs exhibited poor water solubility which limited further investigation of these promising analogs. Importantly lobelane, a saturated analog of lobeline, exhibited increased affinity and selectivity for VMAT2 compared to lobeline. To improve water solubility, a N-1,2-dihydroxypropyl (diol) moiety was incorporated into the lobelane molecule. GZ-793A, an N-1,2-diol analog, potently and competitively inhibited VMAT2 function, exhibiting over 50-fold selectivity for VMAT2 over DAT, serotonin transporters and nAChRs. GZ-793A released DA from preloaded synaptic vesicles, fitting a two-site model with the high-affinity site inhibited by tetrabenazine and reserpine (classical VMAT2 inhibitors), suggesting a VMAT2-mediated mechanism of release. Further, low concentrations of GZ-793A that selectively interact with high-affinity sites on VMAT2 to evoke DA release, inhibit methamphetamine-evoked DA release from synaptic vesicles. Results showed that increasing concentrations of GZ-793A produced a rightward shift in the METH concentration response; however, the Schild regression revealed a slope different from unity, consistent with surmountable allosteric inhibition. In addition, GZ-793A specifically inhibited methamphetamine-evoked DA release in striatal slices and methamphetamine self-administration in rats. To examine the possibility that GZ-793A produced DA depletion, the effect of a behaviorally active dose of GZ-793A on DA content in striatal tissue and striatal vesicles was determined. GZ-793A administration did not alter DA content in striatal tissue or vesicles and pretreatment with GZ-793A prior to methamphetamine administration did not exacerbate the DA depleting effects of methamphetamine. Importantly, GZ-793A was shown to protect against methamphetamine-induced striatal DA depletions. Thus, GZ-793A represents an exciting new lead in the development of pharmacotherapies to treat methamphetamine abuse.

Methamphetamine interacts with vesicular monoamine transporter-2 (VMAT2) to inhibit dopamine (DA) uptake and promotes DA release from presynaptic vesicles, increasing cytosolic DA available for methamphetamine-induced reverse transport by DA transporters. By inhibiting VMAT2, lobelane, a defunctionalized, saturated lobeline analog, decreases methamphetamine-evoked DA release and methamphetamine self-administration in rats. In this dissertation structure-activity relationships around the lobelane structure were investigated on racemic lobelane analogs with varying methylene linker lengths at central piperidine ring. Affinity for dihydrotetrabenazine (DTBZ) sites on VMAT2 and for inhibition of VMAT2 function was determined to be 0.88-63 and 0.024-4.6 µM, respectively, and positively correlated. The most potent and selective analog, (±)-cis-2-benzyl-6-(3-phenylpropyl)piperidine [(±)-GZ-730B], for VMAT2 uptake was identified as the lead. The ability of (±)-GZ-730B to inhibit methamphetamine-evoked [3H]DA release from striatal synaptic vesicles and endogenous DA release from striatal slices was determined. The lead analog-induced inhibition of methamphetamine-evoked vesicular [3H]DA release did not translate to inhibition of methamphetamine-evoked DA release in the more intact striatal slices. Moreover, poor water solubility of these lobelane analogs prohibited further in vivo work. Subsequent work focused on analogs with the C-3 and C-4 carbons in the piperidine ring eliminated to afford racemic acyclic lobelane analogs. Generally, acyclic analogs exhibited greater water solubility and less lipophilicity compared to lobelane. Acyclic analogs exhibited affinities (Ki = 0.096-17 μM) for [3H]DTBZ sites that correlated positively with affinity (Ki = 3.3-300 nM) for inhibition of [3H]DA uptake. Pure enantiomers of potent racemic analogs were synthesized, and found to potently, selectively, and competitively inhibit [3H]DA uptake at VMAT2 and to release vesicular [3H]DA in a biphasic manner. Lead enantiomer (R)-N-(1-phenylpropan-2-yl)-3-phenylpropan-1-amine [(R)-GZ-924] inhibited methamphetamine-evoked [3H]DA release from striatal synaptic vesicles, but not from the more intact striatal slices. Surprisingly, (R)-GZ-924 inhibited nicotine-evoked [3H]DA overflow from striatal slices, revealing nonspecific effects. Importantly, (R)-GZ-924 inhibited methamphetamine self-administration in rats. However, the analog also inhibited food-maintained responding, revealing a lack of specificity. The lead analog will not be pursued further as a pharmacotherapy due to the lack of specificity. Further evaluation of the pharmacophore is needed to discover analogs which specifically inhibit the neurochemical and behavioral effect of methamphetamine.

The aim of the thesis is to determine the market value of the company GZ Media, a.s. to 1.1.2015. The first part provides the basic information about the company. This chapter is than followed by financial analysis which is assessing the overall financial health of the company for the years 2010 to 2014. Strategic analysis focuses on the market and competitive position of the company and provides the forecast of the sales. The next chapter analyzes and forecasts the value generators and defines the main indicators later used for financial plan. The value of the company is calculated based on the DCF Equity method.

No description available.

From many years YSZ is used as the top coat material for TBC's, as it has good phase stability up to 1200°C, higher fracture toughness, lower thermal conductivity, erosion resistance &amp; higher coefficient of thermal expansion. But, it has a drawbacks at high temperature such as sintering and transformation of phases. For this reason new ceramic materials with pyrochlores crystal structure such as Gd2Zr2O7 are being considered as it has high melting points, phase stability, lower thermal conductivity and CMAS resistance. But it has low fracture toughness when compared to YSZ. In order to take advantage of low thermal conductivity and high thermal stability of gadolinium zirconate and avoiding the drawbacks of low coefficient of thermal expansion and low toughness using YSZ, a double/multi-layer coatings approach is being used. Therefore, multi-layer TBCs are sprayed and compared with single layer coating in this work. These coatings are processed by suspension plasma spraying. For single layer coating YSZ is used, for double layer coating YSZ as the intermediate coating and Gd2Zr2O7 as the top coat is used. Additionally, a triple layer coating system comprising YSZ, Gd2Zr2O7 and dense Gd2Zr2O7 as top coat is also sprayed. The as sprayed coatings are characterized for microstructure analysis using optical microscope and scanning electron microscope (SEM), elemental analysis of TGO using Energy-Dispersive Spectrometer (EDS). XRD analysis was done to identify various phases in the coating. Porosity analysis using Archimedes principle was carried out. Thermal cyclic fatigue (TCF) test of the sprayed coatings was carried out at 1100°C. Failure analysis of the TCF specimens was carried out using SEM/EDS. TCF results showed that the triple layer coatings (dense Gd2Zr2O7/Gd2Zr2O7/YSZ) had higher thermal cyclic fatigue life and lower TGO thickness when compared to single layer (YSZ) and double layer (Gd2Zr2O7/YSZ) TBCs.

Le visage humain joue un rôle important dans la communication interpersonnelle. La dysfonction du visage ou le défigurement due aux traumatismes ou pathologies peuvent entraver les activités sociales normales. Le traitement chirurgical est souvent nécessaire. De nos jours, le résultat du traitement chirurgical et l’état d’établissement ne sont estimé qu’avec les méthodes qualitatives telles que l’observation visuelle et la palpation. Dans l’attente de fournir des critères quantitatifs, cette thèse a pour l’objectif de modéliser la mimique faciale utilisant MEF (Méthode d’Éléments Finis) sur la base des données d’IRM (Imagerie par Résonance Magnétique). Un modèle sujet-spécifique du visage a été construit sur la base de la segmentation des données IRM ; il contient des parties osseuses, muscles de la mimique (p.ex. le muscle grand zygomatique), les tissues mous sous-cutanées et la peau. L’identification des tissus mous biologiques a été réalisée via des essais de traction bi-axiale et la modélisation numérique. Ensuite, le modèle géométrique a été maillé pour effectuer des calculs EF simulant trois mouvements mimiques du visage (sourire, prononciation du son « Pou » et « O »). Les muscles ont été modélisés comme un matériau quasi-incompressible, transversalement isotrope et hyperélastique, avec la capacité d’activation. Des informations pertinentes (p.ex. l’amplitude de contraction du muscle) utilisées dans la simulation ont été extraites de la mesure des données d’IRM. Il est à noter que les mêmes données expérimentales d’IRM telles qu’ils ont utilisées dans la modélisation ont été prises comme une référence de validation pour les résultats de simulation. Cette étude peut être appliquée cliniquement dans l’évaluation du traitement faciale et le rétablissement postopérative. / Human face plays an important role interpersonal communication. Facial dysfunction or disfigurement due to trauma or pathologies may impede normal social activities. Surgical treatment is often necessary. Nowadays, treatment outcome and rehabilitation condition are estimated only by qualitative methods, such as visual observation and palpation. In expectation of providing quantitative criteria, this thesis proposes to model facial mimics using FEM (Finite Element Method) on the basis of MRI (Magnetic Resonance Imaging) data. A subject-specific face model was reconstructed based on segmentation of MRI data; it contains bony parts, mimic muscles (e.g. zygomaticus major muscle), subcutaneous soft tissues and skin. Identification of biological soft tissues was conducted through bi-axial tension tests and numerical modeling. Then the geometric model was meshed to conduct FE calculations simulating three facial mimic movements (smile, pronunciation of sound “Pou” and “O”). Muscle was modeled as quasi-incompressible, transversely-isotropic, hyperelastic material, with activation ability. Relevant information (e.g. contraction amplitude of muscle) used in simulation was extracted from measurement of MRI data. It is to be noted that the same experimental MRI data as used in modeling was taken as validation reference for simulation results. This study can be applied clinically in evaluation of facial treatment andpostoperative recovery.

In this thesis, patterning methods to fabricate various functional particle patterns on substrates were developed, with the main aim of modifying the properties and functions of the substrates. Two classes of model substrates were selected; topographically patterned and smooth substrates. For the first model system, i.e., topographically patterned substrates, replication molding was used to topographically pattern substrates of different materials. The topographically patterned substrates, including TiO2, block-copolymer substrates (PS-b-P2VP and PS-b-P4VP), and microrings (TiO2 and Au), were then used to assemble silica (SiO2) microparticles for functional applications. By the assembly of microparticles on topographically patterned substrates, the wettability of the former could be reversibly switched from hydrophobic to hydrophilic. Moreover, a platform for the preparation of Janus particles by orthogonal functionalization of the top and bottom sides of microparticles assembled on topographically patterned substrates was developed. Clusters of superparamagnetic nanoparticles were stamped on the second class of model substrates, i.e., smooth silanized silicon substrates. A capillary stamping approach combined with an external permanent magnetic field or electromagnets was realized to print magnetic nanoparticle-based inks. In this way, ordered arrays of clusters of magnetic nanoparticles were produced.