CLARITY and PACT-based imaging of adult zebrafish and mouse for whole-animal analysis of infections.

Visualization of infection and the associated host response has been challenging in adult vertebrates. Owing to their transparency, zebrafish larvae have been used to directly observe infection in vivo; however, such larvae have not yet developed a functional adaptive immune system. Cells involved in adaptive immunity mature later and have therefore been difficult to access optically in intact animals. Thus, the study of many aspects of vertebrate infection requires dissection of adult organs or ex vivo isolation of immune cells. Recently, CLARITY and PACT (passive clarity technique) methodologies have enabled clearing and direct visualization of dissected organs. Here, we show that these techniques can be applied to image host-pathogen interactions directly in whole animals. CLARITY and PACT-based clearing of whole adult zebrafish and Mycobacterium tuberculosis-infected mouse lungs enables imaging of mycobacterial granulomas deep within tissue to a depth of more than 1 mm. Using established transgenic lines, we were able to image normal and pathogenic structures and their surrounding host context at high resolution. We identified the three-dimensional organization of granuloma-associated angiogenesis, an important feature of mycobacterial infection, and characterized the induction of the cytokine tumor necrosis factor (TNF) within the granuloma using an established fluorescent reporter line. We observed heterogeneity in TNF induction within granuloma macrophages, consistent with an evolving view of the tuberculous granuloma as a non-uniform, heterogeneous structure. Broad application of this technique will enable new understanding of host-pathogen interactions in situ.

Dealing with the devil: a critical and creative look at the diabolical pact

This thesis is comprised of three parts: a critical dissertation, a creative work of fiction and a bridge piece that connects the two. The critical work is an examination of the Devil as a satirist in Faustian bargains. Through the usage of the Devil as a literary figure, his character has become a more secular being: a trickster rather than evil incarnate—a facilitator of sin rather than its originator. In the tragicomedy of pacts with the Devil, he acts as a mirror, reflecting mankind’s foibles and vanity, while elevating the reader in the process. The thesis considers the language, tone, purpose and conceits of several versions of the story. While the focus is primarily on American Literature, the influence of English, Scottish, French and German folklore and fiction are recognized as an essential component of the theme’s evolution. In the bridge piece, the pact with the Devil is literalized in a modern context; a corporate business of reaping souls is theorized in which techniques of persuasion are streamlined into an effective formula. Whether immersive or expository in approach, the portrayal of the supernatural depends on the literary principles of science fiction and fantasy in order to manipulate the reader and allow irrational concepts to obey rational laws. Such theories are cited to support how the Devil functions as a believable character. The novel, Could Be Much Worse, relates the story of an egocentric boss and his dependable employee, a scout who disguises himself as a taxi driver and seeks candidates who may succumb to temptation. Passengers’ monologues of desperation and pathos are interspersed throughout the protagonist’s day-to-day narrative. At times, the work is experimental, utilizing irregular storytelling techniques, alternative forms and conceits. Light-hearted, but nonetheless poignant, the story serves as a cautionary tale, illustrating the tedium of a bureaucratic job in a transmundane existence.

Desafios para um novo pacto sanitário: biotecnologia e risco

Este artigo discute a incorporação e o uso da biotecnologia na Saúde Pública, no contexto da sociedade de risco. Tendo por referência autores da teoria social contemporânea, analisam-se as implicações das práticas biotecnológicas. O artigo está dividido em três partes. Na primeira, são apresentados alguns exemplos de manipulação biológica desenvolvidos no âmbito da saúde e as consequências da utilização dessas técnicas na dinâmica ecológica das populações envolvidas. A partir desses exemplos, discute-se o que vem a ser esses seres biologicamente modificados, híbridos, e como ocorre sua incorporação nas práticas sociais, especialmente as de Saúde Pública. A segunda parte apresenta o referencial teórico utilizado para análise, que situa a sociedade contemporânea na etapa reflexiva da modernização e que tem na sociedade de risco uma de suas configurações. A última parte do artigo problematiza os usos da biotecnologia em saúde, mais especificamente em Saúde Pública, abordando os aspectos de risco dessa aplicação, propondo o necessário debate sobre um outro pacto sanitário.

Antimicrobial photodynamic action on dentin using a light-emitting diode light source

Objective: The aim of this study was the evaluation of two different photosensitizers activated by red light emitted by light-emitting diodes (LEDs) in the decontamination of carious bovine dentin. Materials and Methods: Fifteen bovine incisors were used to obtain dentin samples which were immersed in brain-heart infusion culture medium supplemented with 1% glucose, 2% sucrose, and 1% young primary culture of Lactobacillus acidophilus 108 CFU/mL and Streptococcus mutans 108 CFU/mL for caries induction. Three different concentrations of the Photogem solution, a hematoporphyrin derivative (1, 2, and 3 mg/mL) and two different concentrations of toluidine blue O (TBO), a basic dye (0.025 and 0.1 mg/mL) were used. To activate the photosensitizers two different light exposure times were used: 60 sec and 120 sec, corresponding respectively to the doses of 24 J/cm(2) and 48 J/cm(2). Results: After counting the numbers of CFU per milligram of carious dentin, we observed that the use of LED energy in association with Photogem or TBO was effective for bacterial reduction in carious dentin, and that the greatest effect on S. mutans and L. acidophilus was obtained with TBO at 0.1 mg/mL and a dose of 48 J/cm(2). It was also observed that the overall toxicity of TBO was higher than that of Photogem, and that the phototoxicity of TBO was higher than that of Photogem. Conclusion: Based on our data we propose a mathematical model for the photodynamic effect when different photosensitizer concentrations and light doses are used.

Antimicrobial mechanisms behind photodynamic effect in the presence of hydrogen peroxide

This study describes the use of methylene blue (MB) plus light (photodynamic inactivation, PDI) in the presence of hydrogen peroxide (H(2)O(2)) to kill Staphylococcus aureus, Escherichia coli, and Candida albicans. When H(2)O(2) was added to MB plus light there was an increased antimicrobial effect, which could be due to a change in the type of ROS generated or increased microbial uptake of MB. To clarify the mechanism, the production of ROS was investigated in the presence and absence of H(2)O(2). It was observed that ROS production was almost inhibited by the presence of H(2)O(2) when cells were not present. In addition, experiments using different sequence combinations of MB and H(2)O(2) were performed and MB optical properties inside the cell were analyzed. Spectroscopy experiments suggested that the amount of MB was higher inside the cells when H(2)O(2) was used before or simultaneously with PDI, and ROS formation inside C. albicans cells confirmed that ROS production is higher in the presence of H(2)O(2). Moreover enzymatic reduction of MB by E. coli during photosensitizer uptake to the photochemically inactive leucoMB could be reversed by the oxidative effects of hydrogen peroxide, increasing ROS formation inside the microorganism. Therefore, the combination of a photosensitizer such as MB and H(2)O(2) is an interesting approach to improve PDI efficiency.

Treatment of Osteomyelitis in the Feet of Diabetic Patients by Photodynamic Antimicrobial Chemotherapy

Objective: This article describes two inexpensive photodynamic antimicrobial chemotherapy (PACT) protocols to provide intensive local care on ulcerated feet of diabetic patients with osteomyelitis. Background Data: Patients with this condition generally have poor quality of life. The usual treatment consists of the administration of a cocktail of drugs including anti-inflammatories, promoters of blood circulation, and systemic antibiotics. However, depending on the conditions of the tissues, amputation may be required. Consequently, it is important to develop PACT protocols that can help avoid amputation. Materials and Methods: Two PACT protocols were applied to two diabetic patients with osteomyelitis. These protocols were based on several PACT sessions that consisted of: (1) local injection of mixtures of phenothiazines (2% in water) and Hypericum perforatum extract (10% in propylene glycol), and (2) illumination, lasting 10 min, applied to the lesion's interior and exterior using, respectively, an optical fiber and a non-coherent light source. The frequency of PACT was daily or every other day in the beginning, and weekly after tissue recovery begun. The patients were followed clinically and by radiographic testing. Results: Both PACT protocols helped cure these patients who were about to have amputation of their feet. Radiograms showed that bone had healed and that the bone's texture had improved. Conclusion: Here we have described efficient and affordable PACT protocols to treat osteomyelitis in the feet of diabetic patients. This treatment modality should be considered by vascular surgeons and by orthopedists to treat osteomyelitis that is resistant to conventional treatments.

Emerging Threats and Opportunities for Large-Scale Ecological Restoration in the Atlantic Forest of Brazil

Over the past 150 years, Brazil has played a pioneering role in developing environmental policies and pursuing forest conservation and ecological restoration of degraded ecosystems. In particular, the Brazilian Forest Act, first drafted in 1934, has been fundamental in reducing deforestation and engaging private land owners in forest restoration initiatives. At the time of writing (December 2010), however, a proposal for major revision of the Brazilian Forest Act is under intense debate in the National Assembly, and we are deeply concerned about the outcome. On the basis of the analysis of detailed vegetation and hydrographic maps, we estimate that the proposed changes may reduce the total amount of potential areas for restoration in the Atlantic Forest by approximately 6 million hectares. As a radically different policy model, we present the Atlantic Forest Restoration Pact (AFRP), which is a group of more than 160 members that represents one of the most important and ambitious ecological restoration programs in the world. The AFRP aims to restore 15 million hectares of degraded lands in the Brazilian Atlantic Forest biome by 2050 and increase the current forest cover of the biome from 17% to at least 30%. We argue that not only should Brazilian lawmakers refrain from revising the existing Forest Law, but also greatly step up investments in the science, business, and practice of ecological restoration throughout the country, including the Atlantic Forest. The AFRP provides a template that could be adapted to other forest biomes in Brazil and to other megadiversity countries around the world.

C-13-NMR, H-1-NMR, and FT-Raman study of radiation-induced modifications in radiation dosimetry polymer gels

H-1- and C-13-NMR spectroscopy and FT-Raman spectroscopy are used to investigate the properties of a polymer gel dosimeter post-irradiation. The polymer gel (PACT) is composed of acrylamide, N,N'-methylene-bisacrylamide, gelatin, and water. The formation of a polyacrylamide network within the gelatin matrix follows a dose dependence nonlinearly correlated to the disappearance of the double bonds from the dissolved monomers within the absorbed dose range of 0-50 Gy. The signal from the gelatin remains constant with irradiation. We show that the NMR spin-spin relaxation times (T-2) of PAGs irradiated to up to 50 Gy measured in a NMR spectrometer and a clinical magnetic resonance imaging scanner can be modeled using the spectroscopic intensity of the growing polymer network. More specifically, we show that the nonlinear T-2 dependence against dose can be understood in terms of the fraction of protons in three different proton pools. (C) 2000 John Wiley & Sons, Inc.

A reforma do poder judici??rio sob a ??tica do governo federal

Este artigo discute os principais aspectos do processo de reforma do Poder Judici??rio, tendo como ponto de partida os objetivos a que ela se prop??e: a amplia????o do acesso ?? Justi??a e a melhoria da qualidade dos servi??os prestados. Inicia-se destacando os problemas do sistema judicial brasileiro. Ressalta, a partir da??, o conjunto de medidas constante no ???Pacto em favor de um Judici??rio mais r??pido e republicano???, firmados pelos representantes m??ximos dos tr??s poderes. A primeira delas constitui a realiza????o de um diagn??stico do Judici??rio. A segunda a????o diz respeito ?? moderniza????o de sua gest??o, por meio, entre outros, da capacita????o de servidores no exerc??cio das atividades administrativas e da incorpora????o de novas tecnologias. Por fim, integram-se a esse conjunto iniciativas de altera????o legislativa.

A perspectiva brasileira sobre a pobreza: um estudo de caso do Programa Bolsa Fam??lia

Tendo como pano de fundo a dificuldade de definir a pobreza, este artigo visa retomar parte da discuss??o sobre o tema, utilizando como estudo de caso o Programa Bolsa Fam??lia. Inicia-se com a defini????o institucional de pobreza dada pelo soci??logo alem??o Georg Simmel passando-se, na seq????ncia, a uma abordagem mais substancial da pobreza e da rela????o entre trabalho e precariedade, desenvolvida por Robert Castel. Apresenta uma breve descri????o do funcionamento do Programa Bolsa Fam??lia e alguns aspectos relacionados aos conceitos e modos de tratamento da quest??o da renda e da educa????o, bem como da fam??lia e dos modelos familiares, baseando-se tamb??m nos resultados parciais de uma pesquisa de campo realizada nos estados da Bahia, Pernambuco, Alagoas e Goi??s. O texto trata ainda de alguns desafios administrativos na gest??o do Programa e os preconceitos morais e sociais recorrentemente veiculados em rela????o a ele. Por fim, na conclus??o, busca retomar a an??lise do Programa Bolsa Fam??lia, seus limites e desafios, defendendo a necessidade de participa????o de todos em um pacto social contra a pobreza e a desigualdade no Brasil.

Programa Bolsa Fam??lia: descentraliza????o, centraliza????o ou gest??o em redes?

O artigo analisa o processo de implementa????o e o modelo de gest??o do Programa Bolsa Fam??lia (PBF), no cerne do debate de centraliza????o-descentraliza????o. A hip??tese do estudo ?? que o PBF n??o apenas caminhou para o fortalecimento da descentraliza????o, superando alguns desafios da consolida????o do ???pacto federativo???, como tamb??m construiu uma configura????o mais complexa, pr??xima ?? gest??o em redes. Para tanto, ?? apresentada uma discuss??o te??rica acerca do federalismo, da descentraliza????o das pol??ticas sociais e do conceito de redes de pol??ticas p??blicas. Em seguida, s??o discernidos os princ??pios que fundamentaram a concep????o do programa, bem como as principais estrat??gias de implementa????o. Como conclus??o, argumenta-se que apesar de ainda persistirem desafios ?? gest??o compartilhada, a implementa????o do PBF se pautou n??o apenas no princ??pio da descentraliza????o, como tamb??m na transpar??ncia, controle e participa????o popular.

Perfil das condi????es de sa??de das capitais brasileiras na perspectiva dos Objetivos de Desenvolvimento do Mil??nio (ODM)

No contexto atual, o uso de indicadores sociais na avalia????o das pol??ticas p??blicas tem se tornado cada vez maior. Os indicadores sociais tamb??m t??m sido implementados no acompanhamento dos Objetivos de Desenvolvimento do Mil??nio (ODM), projeto da Organiza????o das Na????es Unidas (ONU) em que v??rios pa??ses, inclusive o Brasil, firmaram um pacto para tornar o mundo menos desigual. Sendo assim, para avaliar o cumprimento das metas estabelecidas nos ODM, este artigo pretende delinear perfis das capitais brasileiras quanto ??s condi????es de sa??de, sob a perspectiva dos ODM, a fim de contribuir para a formula????o e avalia????o de pol??ticas p??blicas e indicar um caminho para a escolha de indicadores importantes de serem monitorados. Foram delineados perfis, com a ado????o do m??todo Grade of Membership, quanto ?? sa??de nas capitais brasileiras. Perante os resultados obtidos, pode-se dizer que as condi????es de sa??de nas capitais brasileiras apresentam forte vi??s regional.

O jornalismo de proximidade como promotor da cultura e identidade de uma região: o caso do Porto Canal

Relatório de estágio apresentado à Escola Superior de Comunicação Social como parte dos requisitos para obtenção de grau de mestre em Jornalismo.

A transparência fiscal

Mestrado em Fiscalidade

Boas práticas de igualdade de género e sua relação nos domínios da satisfação laboral e clima organizacional

A Igualdade entre Homens e Mulheres e a não discriminação constituem princípios fundamentais da Constituição da Republica Portuguesa e do Tratado que institui a União Europeia – Tratado de Lisboa. A desigualdade entre Homens e Mulheres constitui uma violação dos direitos fundamentais, e impõe um pesado custo à economia na medida em que se desaproveitam talentos em função do género. A promoção de uma efetiva igualdade entre Homens e Mulheres constitui um dever fundamental do Estado. A promoção da participação ativa de Homens e Mulheres na vida política, ao nível da administração central, regional e local, é também um forte indicador da qualidade da democracia de um estado. Tomando por base a temática da Igualdade de Género, o Roteiro para a Igualdade entre Homens e Mulheres (2006-2010), o pacto Europeu para a Igualdade entre os Sexos (2006), a Estratégia para a Igualdade entre Homens e Mulheres 2010-2015 que elencam ações consideradas prioritárias – igualdade na independência económica; igualdade na remuneração por trabalho igual e por trabalho de igual valor; igualdade na tomada de decisões; promoção da dignidade e a integridade, pôr fim à violência de género; e questões horizontais – papéis desempenhados por Homens e Mulheres, legislação, governação e instrumentos no domínio da igualdade entre Homens e Mulheres, o objeto deste estudo centra-se na atividade do Estado Português, mais concretamente ao nível local. Procurou-se enquadrar esta temática na Gestão dos Recursos Humanos, na busca de um conhecimento mais aprofundado sobre a implementação de Boas Práticas de Igualdade de Género e a sua relação nos domínios da Satisfação Laboral assim como no Clima Organizacional. O presente estudo expõe uma abordagem quantitativa, de carácter descritivo, exploratório, correlacional e preditivo. O tratamento estatístico realizou-se com recurso ao programa IBM SPSS Statistics, versão 21. Os resultados encontrados apontam para uma associação positiva entre a existência de boas práticas de igualdade de género e a satisfação laboral dos trabalhadores, assim como do clima organizacional. São apresentadas pistas para a intervenção no domínio da função da gestão e desenvolvimento dos recursos humanos.