Molecular fMRI

Comprehensive analysis of brain function depends on understanding the dynamics of diverse neural signaling processes over large tissue volumes in intact animals and humans. Most existing approaches to measuring brain signaling suffer from limited tissue penetration, poor resolution, or lack of specificity for well-defined neural events. Here we discuss a new brain activity mapping method that overcomes some of these problems by combining MRI with contrast agents sensitive to neural signaling. The goal of this “molecular fMRI” approach is to permit noninvasive whole-brain neuroimaging with specificity and resolution approaching current optical neuroimaging methods. In this article, we describe the context and need for molecular fMRI as well as the state of the technology today. We explain how major types of MRI probes work and how they can be sensitized to neurobiological processes, such as neurotransmitter release, calcium signaling, and gene expression changes. We comment both on past work in the field and on challenges and promising avenues for future development.National Institutes of Health (U.S.) (Grants R21-MH102470 and U01-NS09045)Massachusetts Institute of Technology. Simons Center for the Social Brain (Seed Grant

Enacting the untranslatable: the sociolinguistic situatedness of Shakespearean emotions

This thesis explores the phenomenon of emotion in different cultural contexts, through the lens of Shakespearean performance. In particular, it investigates how differences in language may reflect, or even shape, how feelings in the playhouse are experienced, expressed, and emphasised. This involves the comparison of emotion concepts in early modern English to twenty-first-century English, as well as to untranslatable concepts in Russian, French, and German. The INTRODUCTION serves as an overview of the importance of both translation scholarship and emotion studies as they relate to Shakespeare and early modern England. CHAPTER ONE introduces the philosophical, scientific, and linguistic groundwork that will permit a confident exploration of the situated nature of emotions on the Shakespearean stage. In particular, this chapter presents a theoretical paradigm known as enaction, as well as the methodological tools of the Natural Semantic Metalanguage. CHAPTER TWO focuses on the experience of lovesickness as it pertains to the character of Mariana in Measure for Measure. The chapter begins with a study of the idea of love melancholy and the many ways it was considered a serious illness in Renaissance England, and then examines how twenty-first-century productions of the play navigate the dramaturgical implications of Mariana’s emotive improvisation. CHAPTER TWO then compares current English-language notions of lovesickness with the Russian untranslatable concept тоска (toska), and looks at how Russian productions of the play have embodied this emotion. CHAPTER THREE looks at Othello, through the lens of an emotion we tend to think of as positive: joy. However, in exploring the early modern English attitudes toward this particular feeling, this chapter uncovers the interrelatedness of Renaissance England’s concepts of joy and death. CHAPTER THREE then turns to an examination of joy’s French-language analogue: joie. An analysis of French translations and productions of Othello will show how joie carries surprisingly different connotations from the “joy” that native English speakers know. CHAPTER FOUR explores the concept of fear in Hamlet. The first half of the chapter examines the neuroscience behind the claim that fear is a universal human emotion, before demonstrating how fear in Renaissance England was bound up with concepts of the afterlife. The second half explores the German-language concept of Angst, and analyses how this emotion has coloured German theatre-makers’ relationship with Hamlet. The CODA looks more closely at the practical, theatrical implications of the disparity between the emotion concepts that emerged from Shakespeare’s specific time and those of today. In particular, the CODA outlines a means by which actors can use the Natural Semantic Metalanguage to deconstruct some of Shakespeare’s emotions, which may be considered “untranslatable” today, and synthesise them into a culturally relevant mode of expression

Inibição da quimiotaxia de neutrófilos pela alfa-1-glicoproteína ácida: relevância e mecanismos envolvidos

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Biológicas, Programa de Pós-Graduação em Farmacologia, Florianópolis, 2015.A chegada dos neutrófilos até o foco da inflamação desenvolve papelessencial na resposta inflamatória. Além de serem uma populaçãocelular fundamental para conter e eliminar o patógeno, seu acúmulo levaao dano tecidual uma vez que essas células não são capazes dediscriminar entre as células do hospedeiro e o agente causador dainflamação Neutrófilos se acumulam no local da inflamação através deuma série de eventos sequenciais, entre eles a quimiotaxia. Aquimiotaxia é responsável por guiar os neutrófilos até o foco exato dainflamação. Estudos experimentais demonstram que a proteína de faseaguda Alfa-1-glicoproteína ácida (AGP) é capaz de interferir nesseprocesso, pois inibe o rolamento, a adesão e a migração dos neutrófilosin vivo, assim como a quimiotaxia in vitro. Altamente glicosilada,aproximadamente 12% dos carboidratos da AGP são de ácido siálico.Dentre os receptores que reconhecem os ácidos siálicos estão as lectinasdo tipo Siglecs (Sialic acid binding Ig-like lectins). Siglecs apresentamem sua estrutura um domínio inibitório capaz de interferir na respostaintracelular ativada por outros receptores. Assim, a hipótese deste estudoé de que os resíduos de ácido siálico presentes na estrutura da AGPseriam responsáveis pela ativação de Siglec-5 e/ou 9, inibindo aquimiotaxia dos neutrófilos. Para investigar essa hipótese, foi geradauma AGP desialilada sem a presença desse carboidrato. Neutrófilostratados com essa proteína foram submetidos a um ensaio dequimiotaxia em câmara de Boyden. Esse ensaio revelou que a retiradado ácido siálico reverteu o perfil inibitório induzido pela proteínasialilada. Ainda mais, os resultados demonstram que o ácido siálico emsua forma livre também foi capaz de inibir a quimiotaxia de maneiraequivalente à AGP. Sabendo que a AGP é capaz de ligar-se à superfíciedos neutrófilos, procurou-se investigar se essa interação ocorre atravésda ligação do ácido siálico à Siglec-5 ou 9. Para isso foi feita umacitometria de fluxo e avaliada a intensidade de fluorescência emitidapela AGP ou por anticorpos fluorescentes anti-hSiglec-5 e 9, quando napresença de anticorpos neutralizantes hSiglec-5 e 9 ou da AGP nãomarcada, respectivamente. No entanto nenhuma diferença na média deintensidade de fluorescência foi observada. Em conjunto nossoresultados sugerem que o reconhecimento do ácido siálico é necessáriona inibição da quimiotaxia induzida pela AGP.Abstract : Neutrophil recruitment has a central role in host response and inresolution of inflammation, but if uncontrolled can lead to severe tissuedamage. Overwhelming activation of neutrophils result in aninappropriate infiltration of neutrophils known to elicit cell and tissuedamage that can lead to an organ dysfunction. The acute phase proteinAlpha-1-acid glycoprotein (AGP) was shown to inhibit neutrophilmigration, rolling and adhesion in vivo. AGP also inhibits neutrophilchemotaxis in vitro, and evidence suggests that AGP terminal sialic acidresidues interact with an inhibitory receptor known as Siglec (Sialicacid-binding immunoglobulin-like). However, how AGP affectsneutrophil-response is not completely understood. In this study, weexamine mechanisms related to the effect of AGP on neutrophilchemotaxis responses, and we hypothesized that Alpha-1-acidglycoprotein inhibits neutrophil chemotaxis in vitro by Siglec-5 e/or 9activation. AGP 6µM inhibits neutrophil migration in vitro, as does6µM of sialic acid, when compared with control neutrophils.Neuraminidase treatment of AGP reversed the suppressive effect of theprotein. To investigate how AGP interacts with neutrophils, flowcytometry analyses using AGP labeled with Alexa Fluor 488 andhSiglec-5 and 9 neutralizing antibodies, or AGP and hSiglec-5 and 9fluorescent antibodies were used. Although no difference in the MFIwas observed. Taken together these results suggest that sialic acidrecognition is required for its inhibitory function, and that the interactionof AGP with neutrophils depend on this carbohydrate. However, thisstudy was not able to determine if this interaction was due to the ligationof AGP sialic acid to the receptor Siglec-5 and/or -9

Mecanismos envolvidos na inibição da quimiotaxia de neutrófilos humanos pela alfa-1-glicoproteína ácida

Tese (doutorado) - Universidade Federal de Santa Catarina, Centro de Ciências Biológicas, Programa de Pós-Graduação em Farmacologia, Florianópolis, 2019.A migração dos neutrófilos é fundamental para uma resposta inflamatória eficaz, no entanto quando desregulada pode levar a uma infiltração excessiva dessas células nos tecidos resultando em dano. A proteína de fase aguda alfa-1 glicoproteína ácida (AGP) atua como um regulador da migração dos neutrófilos inibindo diferentes etapas nesse processo como o rolamento e adesão in vivo e quimiotaxia in vitro. No entanto, os mecanismos celulares pelo qual a AGP inibe a migração dos neutrófilos para os sítios inflamatórios ainda não foram completamente elucidados. Uma vez que muitas das atividades da AGP estão relacionadas a sua porção de carboidratos e a perda dos resíduos de ácido siálico compromete sua capacidade de modulação do sistema imune, nós levantamos a hipótese de que a AGP inibe a quimiotaxia de neutrófilos por meio da ativação de receptores Siglec (sialic-acid-binding immunoglobulin-like lectins)-5 e/ou de Siglec-9. Siglecs são uma família de receptores inibitórios expressos em neutrófilos que se ligam a ácido siálico, e quando ativados regulam negativamente a resposta inflamatória. Nesse contexto, o impacto da glicosilação da AGP em neutrófilos humanos ativados pelo fMLP foi avaliado após 40 minutos de incubação das células com diferentes concentrações da proteína, variando de 0,1 a 1 mg/ml. Para avaliar se os resíduos de ácido siálico da AGP influenciam no efeito inibitório, a AGP desialilada (tratada com neuraminidase, enzima que cliva o ácido siálico na sua conformação a-2,3 - a-2,6- ou a-2,8) também foi utilizada. Nossos resultados demonstraram que, in vitro, a AGP inibe a quimiotaxia e a polimerização de actina de neutrófilos induzida por fMLP, por um mecanismo dependente de seus resíduos de ácido siálico. Além disso, a inibição da polimerização de actina em resposta ao fMLP foi suprimida quando os neutrófilos foram pré-incubados com anticorpos neutralizantes anti-Siglec-5, mas não com anti-Siglec-9. Entretanto, a AGP não inibe o processo de fagocitose induzido por Escherichia coli. Os efeitos da AGP sobre a produção de espécies reativas de oxigênio (ROS) e ativação de integrinas b2 também foram investigadas. A produção de ROS pelos neutrófilos induzida pelo fMLP ou PMA foi inibida pela AGP sialilada e em concentrações mais elevadas também pela sua variante sem o ácido siálico. Ainda, o tratamento dessas células com AGP, previamente ao estímulo com fMLP, inibiu a mudança conformacional de ativação das integrinas b2 pela via inside-out de ativação. Juntos, esses resultados demonstram que a AGP inibe a quimiotaxia de neutrófilos mediada por fMLP, ao prejudicar a polimerização de actina e ativação das integrinas. Além disso, o efeito supressor da AGP na polimerização de actina ocorre por meio da interação entre os resíduos de ácido siálico da proteína e o receptor Siglec- 5 dos neutrófilos.Abstract: Neutrophil recruitment has a central role in host response and resolution of inflammation, but if uncontrolled it can lead to severe tissue damage. Due to its harmful effects, neutrophils migration must be tightly regulated to prevent an exacerbated infiltration. The acute phase protein Alpha-1-acid glycoprotein (AGP) has been shown to inhibit some steps of the neutrophil migration process, such as rolling and adhesion in vivo, and chemotaxis in vitro. However, the specific mechanism by which AGP inhibits the neutrophil migration to the focus of infection has not been fully elucidated yet. Since AGP activity is related to its carbohydrate portion, and the loss of its sialic acid residues implicates its capacity to modulate the immune system, we hypothesized that Siglec (sialic-acid-binding immunoglobulin-like lectins)-5 or ?9 activation by AGP inhibits the mechanisms related to human neutrophil chemotaxis. Siglecs are a family of inhibitory receptors expressed on neutrophils that bind to sialic acid to down regulate the inflammatory response. In this context, the role of AGP sialylation on human neutrophils activated by fMLP was evaluated after 40 minutes of incubation with protein ranging from 0.1 to 1 mg/ml. In order to evaluate whether AGP sialic acid residues influence on the inhibitory effect, a desialilated AGP (dAGP) (AGP treated with neuraminidase that cleaves sialic acid in a-2,3 - a-2,6- or a-2,8 conformation) was also used. Our results showed that AGP inhibits in vitro neutrophil chemotaxis and actin polymerization by a sialic acid-dependent mechanism. In addition, the inhibition of actin polymerization by AGP in response to fMLP was abrogated when neutrophils were incubated with an anti-Siglec-5, but not by an anti-Siglec-9, neutralizing antibody prior to AGP treatment. On the other hand, the neutrophil E.coli phagocytosis was not altered by the suppressive effect of AGP in actin polymerization. Siglec receptors are also involved in ROS production and integrin activation, therefore the effects of AGP on these neutrophil responses were also investigated. ROS production was shown to be inhibited by AGP and by its variant without sialic acid. Neutrophils treatment with AGP prior to fMLP stimulation also impaired the integrins activation conformational change and inhibited their inside-out pathway of activation. Taken together, our results demonstrated that AGP inhibits fMLP-mediated neutrophil chemotaxis by impairing actin polymerization and integrin activation. Moreover, the suppressive effect of AGP on actin polymerization relays on the interaction between the protein sialic acid residues and the neutrophil receptor Siglec-5

Screen-Based Analysis of Magnetic Nanoparticle Libraries Formed Using Peptidic Iron Oxide Ligands

The identification of effective polypeptide ligands for magnetic iron oxide nanoparticles (IONPs) could considerably accelerate the high-throughput analysis of IONP-based reagents for imaging and cell labeling. We developed a procedure for screening IONP ligands and applied it to compare candidate peptides that incorporated carboxylic acid side chains, catechols, and sequences derived from phage display selection. We found that only l-3,4-dihydroxyphenylalanine (DOPA)-containing peptides were sufficient to maintain particles in solution. We used a DOPA-containing sequence motif as the starting point for generation of a further library of over 30 peptides, each of which was complexed with IONPs and evaluated for colloidal stability and magnetic resonance imaging (MRI) contrast properties. Optimal properties were conferred by sequences within a narrow range of biophysical parameters, suggesting that these sequences could serve as generalizable anchors for formation of polypeptide–IONP complexes. Differences in the amino acid sequence affected T[subscript 1]- and T[subscript 2]-weighted MRI contrast without substantially altering particle size, indicating that the microstructure of peptide-based IONP coatings exerts a substantial influence and could be manipulated to tune properties of targeted or responsive contrast agents. A representative peptide–IONP complex displayed stability in biological buffer and induced persistent MRI contrast in mice, indicating suitability of these species for in vivo molecular imaging applications.National Institutes of Health (U.S.) (Grant R01-DA28299)National Institutes of Health (U.S.) (Grant R01-NS76462)National Institutes of Health (U.S.) (Grant R21-MH102470)Japan Society for the Promotion of Science (Postdoctoral Fellowship for Research Abroad

A Súmula de efeito vinculante face à Reforma do Poder Judiciário

TCC(graduação) - Universidade Federal de Santa Catarina. Centro de Ciências Jurídicas. Direito.O presente trabalho monográfico analisa o instituto jurídico da Súmula de efeito vinculante sob o enfoque da Reforma do Poder Judiciário (Emenda Constitucional nº 45, de 8 de dezembro de 2004). Nesse contexto, trazem-se à baila inúmeros temas que por si só poderiam ser alvos de estudos mais aprofundados, tais como os principais entraves ao livre acesso à Justiça, a utilização do stare decisis no sistema jurídico da common law e do direito jurisprudencial na civil law. De qualquer sorte, procurar-se-á demonstrar que a Súmula de efeito vinculante é uma das tentativas de solução de uma parte da crise do Poder Judiciário, tendo por base o princípio da segurança jurídica e o da isonomia, refutando, por conseguinte, a tese de violação ao princípio da separação dos Poderes, eis que se apresenta como ato típico da função jurisdicional

Levantamento das condições de saúde bucal no DF : planejamento de execução do levantamento e adaptação de instrumento de mensuração de qualidade de vida relacionada a DTM

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Ciências da Saúde, Departamento de Odontologia, Programa de Pós-Graduação em em Odontologia, 2019.As doenças bucais e suas sequelas são comprovadamente um dos maiores problemas de saúde pública do mundo, e devem ser objeto constante de estudo e direcionamento de recursos públicos e privados, no intuito de se evitar, deter ou reabilitar suas consequências. Os inquéritos de saúde bucal são recursos fundamentais para que se conheça a realidade epidemiológica de um determinado cenário. O presente estudo tem por objetivo realizar pesquisas metodológicas e desenvolver ferramentas que servirão de base para o 1o levantamento epidemiológico em saúde bucal do DF. Para tanto, promoveu-se extensa análise comparativa com inquéritos da mesma natureza realizados no Brasil e no mundo, em face da das recomendações da Organização Mundial da Saúde (OMS). Frente ao lapso da literatura, executou-se processo de validação transcultural para adaptação de ferramenta que mensura impacto da DTM na qualidade de vida das pessoas, propondo que esta ferramenta faça parte do levantamento em questão. Finalmente, sugere-se uma matriz que relaciona as principais barreiras à implantação dos projetos com desfechos dos estudos de implementação, elencando-se possíveis soluções para que se contornem os problemas.Oral diseases and their consequences are proven to be one of the biggest public health problems in the world, and should be a constant object of study and direction of public and private resources, in order to avoid, deter or rehabilitate their consequences. Oral health surveys are fundamental resources to know the epidemiological reality of a given scenario. This study aims to carry out methodological research and develop tools that will serve as the basis for the first oral health epidemiological survey of the DF. To this end, an extensive comparative analysis was carried out within similar surveys conducted in Brazil and worldwide, in light of the recommendations of the World Health Organization (WHO). In the void of the literature, a cross-cultural validation process was performed to adapt a tool that measures the impact of TMD on people's quality of life, proposing that this tool be part of the survey in question. Finally, we suggest a matrix that relates the main barriers to project implementation with outcomes of the implementation studies, listing possible solutions to overcome the problems

Oral health effects of botulinum toxin treatment for drooling:a systematic review

Drooling is a major morbidity in several neurological diseases. Intraglandular botulinum neurotoxin (BoNT) injections have been used to manage this condition. However, by decreasing salivary flow, BoNT injections may result in an increased risk of caries and other oral adverse effects. In this study, we aimed to assess whether, in patients with drooling, intraglandular BoNT injections are associated with increased dental caries development, modifications on salivary composition (oral pH, buffering capacity and osmolality) and cariogenic bacterial load. We performed a systematic review, searching PubMed, CENTRAL, Web of Science, and Scopus for all experimental and observational studies reporting on adverse effects of intraglandular BoNT injections in patients with drooling. Primary study selection, quality assessment, and data extraction were independently performed by two researchers. No studies were excluded based on their language, publication status or date of publication. Studies? quality was based on revised Cochrane Risk of Bias tools. Meta-analysis was not performed. We retrieved 1025 studies, of which 5 were included. Two studies were two randomized controlled trials and three quasi-experimental studies. None of the included studies found BoNT injections to be associated with dental caries development or with significant reductions in oral pH. One of the included primary studies even observed an increase in salivary buffer capacity. One study found an increase in Lactobacilli counts. As for the risk of bias, two studies were classified as having a critical risk, two as high risk and one as having some concerns. Currently, there is no evidence that, in patients with drooling, BoNT injections associate with increased risk of dental caries or disturbances in oral pH or salivary buffering capacity. However, the included primary studies had important limitations and differences in their methodologies