The Wingless (Wnt) signaling pathway is one of the critical developmental pathways for control of cell differentiation, proliferation, and cell growth. The DEP domain, located on the C-terminus of Dishevelled (Dvl), plays a role in cytoplasm-membrane association, which branches the canonical and non-canonical Wnt signaling pathway within the cell. It has been suggested that the DEP domain requires the recruitment of ionic lipids, such as phosphatidic acid, to regulate its localization to the plasma membrane and association to the frizzle receptor. However, the physical mechanism for DEP association to the plasma membrane is still unknown. We show that mDvl2-DEP interacts with phosphatidic acid at a distinct patch on the surface formed by a positively charged surface area by NMR spectroscopy. The binding of this interaction was also found at physiologically relevant concentration using fluorescence spectroscopy. We also determined that the interaction is pH-dependent and regulated through a 'histidine switch' mechanism at His464 and His465 where there is increased association of mDvl2-DEP to the plasma membrane at higher pH values (7.5). This association is based on tertiary structure conformational changes with rearrangement of the loop regions by a change in local pH, not its interaction with phosphatidic acid. Overall, our work will contribute to elucidate how cells regulate their developmental pathways through localized molecular interactions. / Master of Science

Submitted by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-06-13T13:02:39Z No. of bitstreams: 1 GilbertoHanssenAndrovandiDissertacao2017.pdf: 1917004 bytes, checksum: 702e3ab7456ce3dd676c377dd4288b5c (MD5) / Approved for entry into archive by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-06-13T13:02:53Z (GMT) No. of bitstreams: 1 GilbertoHanssenAndrovandiDissertacao2017.pdf: 1917004 bytes, checksum: 702e3ab7456ce3dd676c377dd4288b5c (MD5) / Made available in DSpace on 2017-06-13T13:02:53Z (GMT). No. of bitstreams: 1 GilbertoHanssenAndrovandiDissertacao2017.pdf: 1917004 bytes, checksum: 702e3ab7456ce3dd676c377dd4288b5c (MD5) Previous issue date: 2017-03-09 / This paper investigates evidences of moral hazard in small and medium-sized Brazilian banks (SMB) that issued Time Deposits with Special Deposit Guarantee (DPGE), this product was created in 2008 Crisis, addressed to wholesale or institutional investors, and coverage by deposit insurance from Credit Guarantee Fund (FGC). For this purpose, we will use a differences-in-differences estimator (DID), comparing the results between DPGE issuers (treatment group) and DPGE nonissuers (control group) considering two banking risk measures (NPL and Basel Index) Data Collection comprehends information between June 2007 and December 2015 grouped in a panel data with significant events in the Brazilian banking market such as impact of global crisis of 2007-2008, the domestic securitization crisis and idiosyncratic events with potential contagion. Our evidence does not suggest the occurrence of moral hazard associated with the set of banks that emit DPGE. The result is in line with recent literature findings suggesting that there is not necessarily increased risk-taking by banks with the introduction of deposit insurance guarantees, provided that it is supported by an adequate design of the deposit insurance scheme by policymakers. / Este estudo investiga a evid??ncia de risco moral nos bancos brasileiros de porte pequeno e m??dio (BPM) que captaram recursos em Dep??sitos a Prazo com Garantia Especial (DPGE). O produto foi criado durante a crise global de liquidez de 2007/2008, direcionado ao investidor de atacado ou institucional, com cobertura por seguro-dep??sito at?? R$ 20 Milh??es do Fundo Garantidor de Cr??dito (FGC). Para tal, utilizaremos uma estima????o pelo m??todo de diferen??as em diferen??as (DD), com a compara????o dos resultados entre as institui????es emissoras de DPGE (grupo de tratamento) e n??o emissoras de DPGE (grupo de controle) considerando duas medidas de risco banc??rias (NPL e ??ndice de Basil??ia). O per??odo compreende informa????es semestrais compreendidas entre junho de 2007 e dezembro de 2015 agrupadas em painel de dados, com captura de eventos significativos no mercado banc??rio brasileiro como o impacto da crise global de 2007/2008, crise do mercado de cess??o dom??stico e eventos idiossincr??ticos com potencial de cont??gio. Os resultados encontrados n??o permitiram validar a evidencia de risco moral associado ao grupo de bancos emissores de DPGE.O resultado est?? em linha com achados recentes descobertas da literatura sugerindo que n??o h?? necessariamente aumento da tomada de risco pelos bancos com a introdu????o de garantias de seguro- dep??sito.

Regulators of G protein signaling (RGS) are a diverse group of proteins, which play a fundamental role in modulation of G protein coupled receptor signal transduction. RGS proteins are primarily known as GTPase activating proteins (GAPs) for Gá subunits. In addition to the RGS domain, which is responsible for GAP activity, most RGS proteins also contain other structural motifs. The R7 family of RGS proteins for example, which consists of RGS-6, 7, 9 and 11 gene products, also contains DEP, DHEX and GGL domains. All R7 RGS proteins are obligatory binding partners with G protein beta subunit, G beta sub five, which binds to the GGL domain. In my dissertation work, I provide insights into significance of the multi-domain architecture of G beta sub five-RGS7. I have identified a novel intramolecular interaction within the G beta sub five-RGS7 complex; between the DEP domain of RGS7 and G beta sub five subunit. My experimental evidence supports the idea that G beta sub five-RGS7 can exist in at least two hypothetical conformations: "closed" where the DEP domain and G beta sub five subunit are bound to each other, and "open" where DEP and G beta sub five are not interacting, and as a result both these proteins can associate with other binding partners. My results indicate that in its "open" conformation, G beta sub five-RGS7 can selectively inhibit calcium mobilization elicited by stimulated muscarinic acetylcholine receptor type 3 (M3R). This inhibition is mediated by direct interaction between the third intracellular loop of M3R and the DEP domain of RGS7. In addition to the effect on M3R signaling, I observed that the G beta sub five-RGS7 complex redistributes from the cytosol to endocytic vesicles in an M3R-specific manner. These results identify a novel molecular mechanism that can impart receptor-subtype selectivity on signal transduction via G protein-coupled receptors. Lastly, I have identified a small group of compounds that inhibits the DEP-G beta sub five interaction. These compounds may serve as starting points for design of G beta sub five-RGS7 modulators in the future.

Filtration processes, whether on the microfluidic, clinical treatment systems, or industrial scale (e.g., point-of-care diagnostics, dialysis, and biopharmaceutical manufacturing, respectively), are often inseparable from membrane clogging (fouling). As a consequence, most, if not all, filtration systems require frequent maintenance to maintain functionality and efficiency. The thesis of this project hypothesizes that Dielectrophoresis can be combined with standard filtration to reduce filter fouling, extending membrane life, and enabling continuous operation. This project investigates a method to reduce fouling, add specificity and efficiency, and decrease the cost and challenge of filtration based biofluid separations. To substantiate this thesis, we designed, fabricated, and tested a filtration system to filter micron diameter particles in suspension using Millipore™ membranes together with fabricated electrodes in a cross-flow filtration system. This prototype device elicits a repulsive dielectrophoretic (DEP) force via the application of a direct current (5-20 volts) sourced from a computer controlled voltage sequencer, designed to levitate and remove larger particles (> 6 µm) before particulate-membrane interaction. Analysis of the results shows a sufficient decrease in particles adhered to the filtration membrane, as compared to control, suggesting DC DEP may be a valid effector in this device. We are convinced that further research will augment the results validating the proof-of-concept thesis presented herein.

No description available.

La perméabilité vasculaire est une caractéristique cruciale de l’angiogenèse. Les acteurs principaux sont les cellules endothéliales qui la régulent en réponse à divers facteurs perméabilisant, tels que le « Vascular Endothelial Growth Factor » (VEGF). Dans le contexte pathologique du cancer, les cellules tumorales produisent de grandes quantités de VEGF qui stimulent la perméabilité, ce qui leur permet d’infiltrer le réseau vasculaire. Il est connu que la tyrosine kinase Src contrôle cette modulation de la perméabilité. Puisque notre laboratoire a préalablement démontré que la phosphatase de type récepteur (PTP) DEP-1 est impliquée dans l’activation de Src en réponse au VEGF, nous avons émis l'hypothèse que DEP-1 pourrait aussi jouer un rôle dans la perméabilité des cellules endothéliales. Grâce à des expériences de transfections d’ARN interférant, nous démontrons que DEP-1 est important pour la régulation de la phosphorylation de la VE-Cadhérine, un médiateur critique de la perméabilité. L’impact de DEP-1 sur la dissociation de jonctions intercellulaires est également démontré par microscopie à immunofluorescence de cellules endothéliales. DEP-1 est également nécessaire à l’augmentation de la perméabilité induite par VEGF in vitro. Deux résidus tyrosine retrouvés dans la queue carboxy-terminale de DEP-1 sont essentiels à l’activation de Src en réponse au VEGF. Suite à la transfection d’un plasmide encodant DEP-1 muté pour ces deux résidus, nous démontrons aussi leur implication dans la régulation de la perméabilité in vitro par DEP-1. Ces travaux permettent ainsi d’approfondir nos connaissances sur un nouveau régulateur potentiel de la perméabilité vasculaire. / Endothelial cell permeability is a crucial step of angiogenesis. The main actors behind permeability are endothelial cells who accomplish this in response to permeabilizing factors, most notably Vascular Endothelial Growth Factor (VEGF). In a pathological context, migrating tumor cells produce great quantities of VEGF that stimulate an increase of vascular permeability, which allows them to intravasate into the vasculature. Src has been shown to mediate this process. Our laboratory has previously shown that the protein tyrosine phosphatase DEP-1 is involved in the regulation of VEGF-dependant activation of Src. These data thus suggested that DEP-1 might play a role in endothelial cell permeability. Here, we show through siRNA experiments that DEP-1 is important for the regulatory phosphorylation of VE-Cadherin which is critical for the induction of permeability. The impact of DEP-1 on intercellular junction dissociation is also demonstrated through immunofluorescence microscopy of endothelial cells. We further show that DEP-1 is absolutely required for the VEGF-dependent increase of permeability as illustrated by in vitro permeability assay on siRNA-transfected endothelial cells. Finally, we show that tyrosine residues in DEP-1’s carboxy-terminal tail, which are crucial for mediating Src activity in response to VEGF, are implicated in VEGF-dependant increase in permeability by transfecting plasmids coding for DEP-1 mutants of these tyrosine residues. These findings shed light on a novel potential key regulator of in vivo permeability.

A tendência atual, imposta pela competitividade do mercado, exige bovinos mais eficientes e adequados ao sistema de produção, visando atender a demanda pela qualidade de carne. Em função disso, o melhoramento genético vem sendo utilizado, de forma a incrementar a eficiência dos rebanhos, proporcionando assim, maior rentabilidade ao sistema. Assim, o presente estudo foi conduzido com o propósito de estudar a variabilidade da progênie de touros submetidos à avaliação genética, através da análise das Diferenças Esperadas na Progênie (DEP) dos filhos de 359 reprodutores. As DEP das progênies foram estimadas por procedimentos REML e as avaliações de touros por metodologias alternativas, foram realizadas com o auxílio do software SAS. Foram utilizados 45.014 dados de peso à desmama (PD), 45.014 de peso ao sobreano (PS) e 45.014 de ganho de peso da desmama ao sobreano (GP) de bovinos de corte da raça Nelore, criados em uma fazenda em região tropical do Brasil. As DEPs dos touros foram estimadas de acordo com três métodos: uso da média aritmética, média harmônica acrescida da média da característica estudada e média harmônica simples. Os resultados sugerem que o uso da média harmônica na estimação de DEPs com o objetivo de ajustar o valor genético aditivo de um touro para a dispersão dos valores genéticos de suas progênies não trouxe diferenças importantes em relação à estimativa pela média aritmética, na população estudada. No entanto, dadas as implicações e importância de novas metodologias, são necessários maiores estudos em busca de se encontrar uma metodologia que permita maior uniformidade das progênies de touros. / The strong competition in beef market imposed an actual trend of searching for quality in beef production systems, trying to attend the consumers\' demand. Due to the necessity of increasing productivity, animal breeding programs have been utilized to increase cattle efficiency, offering more profitability to the production system. The present study was carried out to contribute to knowledge about sires\' progeny variability, using Expected Progeny Difference (EPD) from calves of 359 sires. EPD\'s were estimated by REML procedures. The prediction od bull\'s EPD\'s were performed by software SAS, using 45,014 data of weaning weight (PD), 45,014 of post-weaning weight (PS) and 45,014 of weight gain post-weaning (GP) from Nellore beef cattle, raised in a tropical area of Brazil. Those data were analyzed according with three methods: application of arithmetic mean, harmonic mean of EPD added by trait mean and only harmonic mean. Results suggested that estimation of EPDs using harmonic mean on condition that adjust sire\'s additive genetic value to dispersion form its calves\' genetic values did not make difference to the prediction trough arithmetic mean in the population studied. Due to the implications and importance of alternative methods to adjust bull\'s EPDs to variability of progeny, further studies are necessary to search for better progeny\'s uniformity.

Submitted by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-08-17T20:39:30Z No. of bitstreams: 1 LucianoRodriguesLaraDissertacao2017.pdf: 1892610 bytes, checksum: 98e4dc2c74fcef79b192fd26eac8010e (MD5) / Approved for entry into archive by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-08-17T20:39:43Z (GMT) No. of bitstreams: 1 LucianoRodriguesLaraDissertacao2017.pdf: 1892610 bytes, checksum: 98e4dc2c74fcef79b192fd26eac8010e (MD5) / Made available in DSpace on 2017-08-17T20:39:43Z (GMT). No. of bitstreams: 1 LucianoRodriguesLaraDissertacao2017.pdf: 1892610 bytes, checksum: 98e4dc2c74fcef79b192fd26eac8010e (MD5) Previous issue date: 2017-03-29 / The 2008 crisis has confirmed the fact that financial crisis can have damaging effects on the real side of the economy. One of the regulators responses to this crisis was the proposition of new capital requirements, known as Basel III. This paper aims to study the effects of two macroprudential policies: capital requirements with countercyclical capital buffers and reserve requirements. This is done using a dynamic stochastic general equilibrium model (DSGE) developed by Gertler and Karadi (2011). Changes in monetary policy and an increase in the total factor productivity are evaluated. The two policies works together to stabilize the economy. / A crise de 2008 confirmou o fato de que crises financeiras podem gerar efeitos danosos no lado real da economia. Uma das respostas dos reguladores a essa crise foi a proposi????o de novos requerimentos de capital, conhecidos como Basil??ia III. Este trabalho tem por objetivo estudar os efeitos de duas pol??ticas macroprudenciais: requerimentos de capital com buffer de capital contrac??clico e dep??sitos compuls??rios. Isto ?? feito utilizando um modelo din??mico estoc??stico de equil??brio geral (DSGE) desenvolvido por Gertler e Karadi (2011). S??o avaliadas altera????es na pol??tica monet??ria e eleva????o da produtividade total dos fatores. ?? verificada a complementariedade entre as duas pol??ticas no objetivo de estabilizar a economia.

The Pennsylvania Department of Environmental Protection has set designated uses for all of the 83,000 miles of waterways based on their quality. Only the highest quality streams are afforded the highest protection with a High Quality (HQ) or Exceptional Value (EV). Little Sewickley Creek is designated as a High Quality-Trout Stocking Fishery and may warrant redesignation as an Exceptional Value waterway. The aim of the study was to provide technical data on the stream to support an EV reclassification. The report has compiled previous data and collected new data in order to file a formal petition to the PA DEP. The petition process involves consideration of physical, chemical, biological data on the stream, along with the land use of the watershed. The report gives an example of a stream located just outside of Pittsburgh that has extremely high biodiversity and deserves the highest protection to preserve it for future generations. / Bayer School of Natural and Environmental Sciences; / Environmental Science and Management (ESM) / MS; / Thesis;

Texte remanié de: Thèse de doctorat--Histoire médiévale--Nancy 2, 1997. Titre de soutenance : Les peuplements castraux de la montagne du Doubs. / Bibliogr. p. 195-197. Notes bibliogr. Glossaire.