Phosphorylation Mechanism of Phosphomevalonate Kinase: Implications for Rational Engineering of Isoprenoid Biosynthetic Pathway Enzymes

Mevalonate pathway is of important clinical, pharmaceutical and biotechnological relevance. However, lack of the understanding of the phosphorylation mechanism of the kinases in this pathway has limited rationally engineering the kinases in industry. Here the phosphorylation reaction mechanism of a representative kinase in the mevalonate pathway, phosphomevalonate kinase, was studied by using molecular dynamics and hybrid QM/MM methods. We find that a conserved residue (Ser106) is reorientated to anchor ATP via a stable H-bond interaction. In addition, Ser213 located on the α-helix at the catalytic site is repositioned to further approach the substrate, facilitating the proton transfer during the phosphorylation. Furthermore, we elucidate that Lys101 functions to neutralize the negative charge developed at the β-, γ-bridging oxygen atom of ATP during phosphoryl transfer. We demonstrate that the dissociative catalytic reaction occurs via a direct phosphorylation pathway. This is the first study on the phosphorylation mechanism of a mevalonate pathway kinase. The elucidation of the catalytic mechanism not only sheds light on the common catalytic mechanism of GHMP kinase superfamily, but also provides the structural basis for engineering the mevalonate pathway kinases to further exploit their applications in the production of a wide range of fine chemicals such as biofuels or pharmaceuticals. 

Use of Remote Sensing for Collection of Data Elements for Linear Referencing Systems, 2002

This report evaluates the use of remotely sensed images in implementing the Iowa DOT LRS that is currently in the stages of system architecture. The Iowa Department of Transportation is investing a significant amount of time and resources into creation of a linear referencing system (LRS). A significant portion of the effort in implementing the system will be creation of a datum, which includes geographically locating anchor points and then measuring anchor section distances between those anchor points. Currently, system architecture and evaluation of different data collection methods to establish the LRS datum is being performed for the DOT by an outside consulting team.

A biochemical analysis into the co-translational folding of a G protein-coupled receptor; GPR35

The folding and targeting of membrane proteins poses a major challenge to the cell, as they must remain insertion competent while their highly hydrophobic transmembrane (TM) domains are transferred from the ribosome, through the aqueous cytosol and into the lipid bilayer. The biogenesis of a mature membrane protein takes place through the insertion and integration into the lipid bilayer. A number of TM proteins have been shown to gain some degree of secondary structure within the ribosome tunnel and to retain this conformation throughout maturation. Although studies into the folding and targeting of a number of membrane proteins have been carried out to date, there is little information on one of the largest class of eukaryotic membrane proteins; the G-protein-coupled receptors (GPCRs). This project studies the early folding events of the human ortholog of GPR35. To analyse the structure of the 1st TM domain, intermediates were generated and assessed by the biochemical method of pegylation (PEG-MAL). A structurally-similar microbial opsin (Bacterioopsin) was also used to investigate the differences in the early protein folding within eukaryotic and prokaryotic translation systems. Results showed that neither the 1st TM domain of GPR35 nor Bacterioopsin were capable of compacting in the ribosome tunnel before their N-terminus reached the ribosome exit point. The results for this assay remained consistent whether the proteins were translated in a eukaryotic or prokaryotic translation system. To examine the communication mechanism between the ribosome, the nascent chain and the protein targeting pathway, crosslinking experiments were carried out using the homobifunctional lysine cross-linker BS3. Specifically, the data generated here show that the nascent chain of GPR35 reaches the ribosomal protein uL23 in an extended conformation and interacts with the SRP protein as it exits the ribosome tunnel. This confirms the role of SRP in the co-translational targeting of GPR35. Using these methods insights into the early folding of GPCRs has been obtained. Further experiments using site-directed mutagenesis to reduce hydrophobicity in the 1st TM domain of GPR35, highlighted the mechanisms by which GPCRs are targeted to the endoplasmic reticulum. Confirming that hydrophobicity within the signal anchor sequence is essential of SRP-dependent targeting. Following the successful interaction of the nascent GPR35 and SRP, GPR35 is successfully targeted to ER membranes, shown here as dog pancreas microsomes (DPMs). Glycosylation of the GPR35 N-terminus was used to determine nascent chain structure as it is inserted into the ER membrane. These glycosylation experiments confirm that TM1 has obtained its compacted state whilst residing in the translocon. Finally, a site-specific cross-linking approach using the homobifunctional cysteine cross-linker, BMH, was used to study the lateral integration of GPR35 into the ER. Cross-linking of GPR35 TM1 and TM2 could be detected adjacent to a protein of ~45kDa, believed to be Sec61α. The loss of this adduct, as the nascent chain extends, showed the lateral movement of GPR35 TM1 from the translocon was dependent on the subsequent synthesis of TM2.

Adesão a recomendações de ingestão de água, em idosos: preditores psicossociais e efeitos da comunicação de informação com uso de âncoras de referência

Sendo os idosos um grupo de risco face a desidratação, torna-se particularmente relevante desenvolver estratégias eficazes de comunicação em saúde para o incremento da adesão a recomendações médicas para ingestão de água. Com este objetivo procurámos avaliar os efeitos da comunicação de uma recomendação, com recurso a uma referência familiar (e.g. garrafa de 1.5 Litros) na qual os idosos pudessem “ancorar” o seu julgamento e decisão de adesão à recomendação, com base numa mais eficaz compreensão de informação de quantidade. Dos 70 idosos que participaram no estudo, apenas 56 foram considerados na análise, por cumprirem os critérios de inclusão na amostra. Esta amostra incluiu idosos autónomos, não institucionalizados, responsáveis pelas suas escolhas alimentares e com indicadores de hidratação aceitáveis. Os resultados mostraram que numa condição com âncora familiar (mas não na condição sem), a adesão associada ao próprio nível de consumo existente, do idoso aumentou após a recomendação. Análises adicionais identificaram as crenças face à água e as suas várias funções, como um preditor psicossocial positivo da adesão, o mesmo não acontecendo com atitudes face à água, evitamento de informação alimentar negativa e numeracia; Abstract: Being the elderly a risk group with regard to dehydration it is necessary to develop effective health communication strategies to increase adherence to health recommendations concerning water intake. Accordingly, we aimed to assess the effects of communicating a recommendation with the addition of a familiar reference (e.g. bottle of 1.5 Liters) in which the elderly could anchor their judgment and decision making to adhere to the recommendation, based on a better comprehension of the quantified information presented. From the 70 older adults that have participated in the study, only 56 were considered in the analysis, for fulfilling the sample inclusion criteria. This sample included older adults considered to be autonomous, non-institutionalized, responsible for their good choices and with acceptable hydration levels. Results showed that in a condition with a familiar anchor (but not in a condition without) the adherence in line with the elderly own current consumption level, increased after the recommendation. Additional analysis identified beliefs concerning water and its various functions as a positive psychosocial predictor of adherence, the same not happening for attitudes towards water, avoidance of negative food information and numeracy.

More Than A Movement: Unpacking Contemporary Anti-Human Trafficking Efforts in Washington, D.C.

Trafficking in persons has attracted seemingly boundless attention over the last two decades and the work aimed at fighting it is best understood when this cause is contextualized against the backdrop of other social forces—economic, social, and cultural—shaping contemporary nonprofit activities. This project argues that the paid and volunteer labor that takes place in metro Washington, D.C., to combat trafficking in persons can be understood as both a movement and an industry. In addition to arguing that anti-trafficking work is part of a nonprofit industrial complex that situates activist and advocacy work firmly inside state and economic institutions, this project is concerned with the ways in which trafficking work and workers conduct their business collectively. As an organizational study, it identifies the key players in the D.C. region focused on this issue and traces their interactions, collaborations, and cooperation. Significantly, this project suggests that despite variations in objectives, methods, priorities, and characterizations of trafficking, thirty organizations in metro D.C. working on this issue “get along” because they are bound by the benign common goal of raising awareness. Awareness, in this context, is best understood as both a cultural anchor facilitating cohesion and as a social currency allowing groups to opt into joint efforts. The dissertation concludes that organizations centralize awareness in their collective activities over more drastic priorities around which consensus would need to be gained. This is a lost opportunity for making sense of the ways that individual bodies—men, women, and children—experience not just trafficking, but the world around them.

A interseccionalidade de gênero, raça e classe no Programa Ciência sem Fronteiras : um estudo sobre estudantes brasileiros com destino aos EUA

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Educação, Departamento de Pós-Graduação, Mestrado Profissional em Educação, 2015.

TAKING THE PERSPECTIVE OF A SELLER AND A BUYER: IMPLICATIONS FOR PRICE ELICITATION AND PRICE FRAMING

This dissertation consists of two essays which investigate how assuming the role of a seller or a buyer affects valuations in a price elicitation task (essay I) and how different presentations of an equivalent price affect evaluations when a consumer plays the dual roles of a buyer and a seller in transactions involving trade-ins (essay II). Sellers’ willingness to accept (WTA) to give up a good is typically higher than buyers' willingness to pay (WTP) to obtain the good. Essay I proposes that valuation processes of sellers and buyers are guided by a motivational orientation of “getting the best.” For a seller (buyer) indicating WTA (WTP), getting the best implies receiving as much as possible to give up a specific good (giving up as little as possible to get the specific good). Results of six studies suggest that the WTA-WTP elicitation task activates different directional goals, leading to the WTA-WTP disparity. The different directional goals lead sellers and buyers to focus on different aspects and bias their cognitive reasoning and interpretation of information. By connecting the valuation process to the general motivation of getting the best, this research provides a unifying framework to explain the disparate interpretations of the WTA-WTP disparity. Many new purchases and replacement decisions involve consumers’ trading in their old products. In such transactions, the overall exchange may be priced either as separate transactions (partitioned) with price tags for the payment and the receipt or as a single net price (consolidated) which takes into account the value of the trade-in. Essay II examines whether consumers prefer a partitioned price versus a consolidated price presentation. The findings suggest that when consumers are trading in a product which has a low value relative to the price of a new product, they prefer a consolidated price. In contrast, when trading in a product which has high value, they prefer a partitioned price. The results suggest that consumers use the price of the new product as an anchor to evaluate the trade-in value, and the perception of the trade-in value influences the overall evaluation especially when the transaction is partitioned.

Objeção da consciência e aborto previsto em lei : vivências em um serviço público de saúde

Tese (doutorado)—Universidade de Brasília, Faculdade de Ciências da Saúde, Departamento de Enfermagem, Programa de Pós-Graduação em Enfermagem, 2015.

O interface do Campo Grande

Dissertação para obtenção do grau de Mestre em Arquitectura com Especialização em Urbanismo, apresentada na Universidade de Lisboa - Faculdade de Arquitectura.

A relação diagnóstica em contexto de creche e jardim-de-infância

O presente estudo enquadra-se no domínio das Ciências da Educação e na área específica da Análise da Acão Educativa. Não se afastando das preocupações inscritas nos objetivos de um mestrado de autor "A Criança em Diferentes Contextos Educativos”, também se compagina com as nossas determinações pessoais e profissionais, empreendidas na convicção da investigação como um projeto de vida. Para o efeito, arquitetámos o nosso universo de estudo a partir da observação naturalista em sete salas de creche e de jardim-de-infância do distrito de Évora. Da população estudada fizeram parte diferentes acores educativos, entre eles, as crianças dos 3 meses aos 6 anos de idade, as educadoras e as auxiliares de Acão educativa. Pretendemos, a partir dos sinais emitidos pelas crianças, retirar indicadores de necessidades educativas, para compreender as atitudes e comportamentos dos agentes educativos (educadores e auxiliares) na sua prática pedagógica, inferindo das suas próprias necessidades formativas. Procurámos saber como eram feitos os diagnósticos de necessidades (educativas das crianças e formativas dos educadores) e como organizavam a sua relação com outros acores educativos, no sentido de um desenvolvimento holístico da criança. Dada a natureza e o objeto de estudo, optámos por uma metodologia qualitativa de carácter interpretativo, configurada num estudo de caso. Utilizámos a linha de Investigação-Acão/Formação, preconizada por Luís Barbosa, integrada na sua "Escola Sensível e Transformacionista", que radica os seus fundamentos na ideia de que o jardim-de-infância, tal como outra qualquer organização educativa, deve ser um "Observatório de Caracterização de necessidades educativas das crianças e formativas dos sues agentes educativos”, gerido na perspetiva da mudança. Neste enquadramento de ideias, ao qual aderimos por convicção, utilizámos, para caracterizar o real, a Observação como técnica privilegiada, pese embora as limitações que lhe são subjacentes. Minimizámos este facto, triangulando os dados com entrevistas semiestruturadas e notas de campo. Suportámo-nos da Técnica do Espelhamento, da Pedagogia de Ajuda, da Renomeação de Experiências e da Extensibilidade de Si, às quais, pela sua importância demos ênfase no enquadramento teórico. O nosso estudo permitiu-nos inferir que, a relação diagnostica nos processos de ensino/aprendizagem, naqueles contextos, parece estar correlacionada com o facto de que os agentes educativos ainda necessitam de adquirir um leque variado de competências, relacionadas com o Saber-Ser e o Saber-Estar, que lhes permitam atitudes relacionais pró-activas e maior sensibilidade para captar as emoções e as sensações das crianças. Este facto vem corroborar o facto de termos verificado que os agentes educativos não usam o diagnóstico de necessidades como âncora de organização das suas práticas educativas o que não contribui de forma sustentável para a promoção do bem-estar e do desenvolvimento integral da criança. ABSTRACT: The present study lies within the field of Educational Sciences in the specific area of analysis of education. Not moving away from the concerns listed in the goals of an author master degree "The Child in Different Educational Contexts”, it also fits our personal and professional decisions, undertaken in the belief of research as a project of life. We chose the universe of our study from naturalistic observation in seven rooms of Nurseries and Infant Schools in the district of Évora. The study population consisted of children from three months to six years old, educators and assistants of education. From signals emitted by children, we intended to take indicators of educational needs to understand the attitudes and behavior of the educators and their assistants in their pedagogical practice, inferring their own educational needs. We tried to know how the diagnoses of needs were made (educational needs of the children and training needs of the educators) and how they organized their relationship with other educational agents towards a holistic development of children. Considering the nature and purpose of the study we opted for a qualitative methodology of interpretative nature, in a case study. We used the action research training line, advocated by Luís Barbosa in his work "Escola Sensível e Transformacionista" (Sensitive and Transformational School), whose reason lies in the idea that Infant Schools, like other educational organizations, should be centers for the characterization of children’s educational needs and educational agents' formative needs, managed in terms of change. ln this framework of ideas, that we adopted by conviction, we used the observation as the main technique, despite its limitations. To minimize them we triangulated the data with semi structured interviews and field notes. We used the techniques of Mirroring, Pedagogy of Help, renaming of Experiences and Extensibility of Yourself, and, because of their importance, we focused on them in the theoretical framework. This study allowed us to infer that the diagnostic relationship in the teaching­learning processes, in those contexts, seem to be correlated with the fact that the educational agents still need to acquire a wide range of skills related to the Know How To Be, to provide them with proactive relational attitudes and a bigger sensibility to get the children's emotions and sensations. We also found that the educational agents don't use a diagnostic of needs as anchor of the organization of their educational practices, what does not contribute in a sustainable way for the promotion of the well being and integral development of the child.

Serviços educativos e turismo cultural no museu da tapeçaria de Portalegre Guy Fino

Este projecto faz uma abordagem à nova museologia e ao seu papel no mercado do turismo, com especial enfoque nos serviços educativos dos museus portugueses e na sua relação com o turismo cultural. O estudo do património artístico e cultural do Museu da Tapeçaria 1e Portalegre Guy Fino e da Manufactura de Tapeçarias de Portalegre, bem como a análise crítica dos serviços educativos de museus importantes a nível nacional e internacional, serviram de suporte à realização do projecto de criação de serviços educativos no referido Museu. O projecto tem como principal objectivo qualificar a oferta turístico-cultural que as Tapeçarias de Portalegre representam e está estruturado de forma a poderem ser desenvolvidas actividades em rede com os outros dois museus da cidade, o Museu Municipal e a Casa Museu José Régio. Foram escolhidas, como áreas temáticas, a História, a História da Arte e a Literatura. As actividades propostas para as áreas temáticas, são de três tipos: visitas dinamizadas, visitas-oficina e vistas-jogo, podendo algumas delas ser desenvolvidas on-line, de forma interactiva. Todas as actividades propostas exploram conteúdos programáticos que integram os planos curriculares do 1° Ciclo do Ensino Básico, podendo algumas delas ser desenvolvidas também por alunos do 2° Ciclo e por famílias de diferentes nacionalidades. O projecto constitui um primeiro passo para uma progressiva transformação das Tapeçarias de Portalegre numa atracção âncora do turismo cultural regional, sem perder de vista as suas inquestionáveis potencialidades de se tornar numa atracção de âmbito nacional e internacional. ABSTRACT: This project makes an approach to the new museology and its role on tourist market, with special focus on the educational services in portuguese museums and its relationship with cultural tourism. The investigation about the artistic and cultural patrimony of Guy Fino Museum and the Manufacture, as well as the critic analysis of educational services in important national and international museums, supported the present project of creation of educational services for Guy Fino Museum. The defined main objective of the project is qualifying the touristic and cultural offer represented by the Tapestries of Portalegre. lt is structured in a way that allows activities to be developed in network with other museums in the city: The Municipal Museum and the Museum House of Jose Regio. ln this way, the following areas were chosen: History, History of Arte and Literature. The activities proposed for the themes are three types: dynamic visits, visits - hands on and visits - games, some being developed on-line, in an interactive way. Twelve activities were conceptualized. Ali the proposed activities explore programmed contents that integrate curricular plans of primary school. Some of the activities can also be developed by students of secondary school and families. The translation of the material used should be in a way that they can be used and developed by foreign visitors. The project constitutes a first step for a progressive transformation of the Tapestries of Portalegre in an anchor attraction of the regional cultural tourism, being sure of their unquestionable potentialities to become a national and international attraction.

Teste para existência de dominância fiscal no Brasil entre 2011 e 2015

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Economia, Administração e Contabilidade, Departamento de Economia, Programa de Pós-Graduação em Economia, 2016.

Économie comportementale : retrouve-t-on un effet d’ancrage dans la LNH ?

De par leur nature scientifique, les sciences économiques visent, entre autre, à observer, qualifier, ainsi que quantifier des phénomènes économiques afin de pouvoir en dégager diverses prévisions. Ce mémoire se penche sur ces prévisions et, plus particulièrement, sur les facteurs pouvant biaiser les prévisionnistes au niveau comportemental en référant à l’effet d’ancrage, un biais propre à l’économie comportementale – une sous-discipline des sciences économiques. Il sera donc question de comprendre, par une analyse selon la discipline que représente l’économie comportementale, ce qui peut les affecter, avec un accent mis sur l’effet d’ancrage plus précisément. L’idée générale de ce dernier est qu’un agent peut être biaisé inconsciemment par la simple connaissance d’une valeur précédente lorsqu’il est demandé de faire une estimation ultérieure. De cette façon, une analyse des salaires des joueurs de la Ligne Nationale de Hockey (NHL) selon leurs performances passées et leurs caractéristiques personnelles, de 2007 à 2016, a été réalisée dans ce travail afin d’en dégager de possibles effets d’ancrage. Il est alors possible de constater que les directeurs généraux des équipes de la ligue agissent généralement de façon sensible et rationnelle lorsque vient le temps d’octroyer des contrats à des joueurs mais, néanmoins, une anomalie persiste lorsqu’on porte attention au rang auquel un joueur a été repêché. Dans un tel contexte, il semble pertinent de se référer à l’économie comportementale afin d’expliquer pourquoi le rang au repêchage reste une variable significative huit ans après l’entrée d’un joueur dans la NHL et qu’elle se comporte à l’inverse de ce que prévoit la théorie à ce sujet.

The State of our Toolbox: A Meta-analysis of Reliability Measurement Precision

My study investigated internal consistency estimates of psychometric surveys as an operationalization of the state of measurement precision of constructs in industrial and organizational (I/O) psychology. Analyses were conducted of samples used in research articles published in the Journal of Applied Psychology between 1975 and 2010 in five year intervals (K = 934) from 480 articles yielding 1427 coefficients. Articles and their respective samples were coded for test-taker characteristics (e.g., age, gender, and ethnicity), research settings (e.g., lab and field studies), and actual tests (e.g., number of items and scale anchor points). A reliability and inter-item correlations depository was developed for I/O variables and construct groups. Personality measures had significantly lower inter-item correlations than other construct groups. Also, internal consistency estimates and reporting practices were evaluated over time, demonstrating an improvement in measurement precision and missing data.

Économie comportementale : retrouve-t-on un effet d’ancrage dans la LNH ?

De par leur nature scientifique, les sciences économiques visent, entre autre, à observer, qualifier, ainsi que quantifier des phénomènes économiques afin de pouvoir en dégager diverses prévisions. Ce mémoire se penche sur ces prévisions et, plus particulièrement, sur les facteurs pouvant biaiser les prévisionnistes au niveau comportemental en référant à l’effet d’ancrage, un biais propre à l’économie comportementale – une sous-discipline des sciences économiques. Il sera donc question de comprendre, par une analyse selon la discipline que représente l’économie comportementale, ce qui peut les affecter, avec un accent mis sur l’effet d’ancrage plus précisément. L’idée générale de ce dernier est qu’un agent peut être biaisé inconsciemment par la simple connaissance d’une valeur précédente lorsqu’il est demandé de faire une estimation ultérieure. De cette façon, une analyse des salaires des joueurs de la Ligne Nationale de Hockey (NHL) selon leurs performances passées et leurs caractéristiques personnelles, de 2007 à 2016, a été réalisée dans ce travail afin d’en dégager de possibles effets d’ancrage. Il est alors possible de constater que les directeurs généraux des équipes de la ligue agissent généralement de façon sensible et rationnelle lorsque vient le temps d’octroyer des contrats à des joueurs mais, néanmoins, une anomalie persiste lorsqu’on porte attention au rang auquel un joueur a été repêché. Dans un tel contexte, il semble pertinent de se référer à l’économie comportementale afin d’expliquer pourquoi le rang au repêchage reste une variable significative huit ans après l’entrée d’un joueur dans la NHL et qu’elle se comporte à l’inverse de ce que prévoit la théorie à ce sujet.