Interactive media on Chagas Disease: development and content

An interactive media on Chagas disease was developed as an educational tool, on the context of the scientific research and dissemination actions of the National Institute of Structural Biotechnology and Medicinal Chemistry in Infectious Diseases (INBEQMeDI). Different computational resources were used either in terms of hardware and software. The media contains 13 videos that range from 30 seconds to 4 minutes, all with information about Chagas disease, showing the social and economic aspects; the research made by the INBEQMeDI group; different aspects of the disease illustrated by slides arranged in a mobile carousel, and radio programs, with funny skits. The target audience for use of this feature is students aged 10 to 17 years. Teachers of areas of science and biology, through a partnership with the Agency of Education of the State of São Paulo, will be invited to plan a strategy for media use with their students.FAPESPCNP

Inhibition of human transthyretin aggregation by non-steroidal anti-inflammatory compounds: a structural and thermodynamic analysis

Transthyretin (TTR) is a homotetrameric protein that circulates in plasma and cerebral spinal fluid (CSF) whose aggregation into amyloid fibrils has been associated with at least two different amyloid diseases: senile systemic amyloidosis (SSA) and familial amyloid polyneuropathy (FAP). In SSA aggregates are composed of WT-TTR, while in FAP more than 100 already-described variants have been found in deposits. Until now, TTR-related diseases have been untreatable, although a new drug called Tafamidis has been approved only in Europe to specifically treat V30M patients. Thus, new strategies are still necessary to treat FAP caused by other variants of TTR. TTR has two channels in the dimer interface that bind to the hormone thyroxin and that have been used to accommodate anti-amyloidogenic compounds. These compounds stabilize the tetramers, rendering TTR less amyloidogenic. Here, we investigated the effects of three non-steroidal anti-inflammatory compounds-sulindac (SUL), indomethacin (IND) and lumiracoxib (LUM)-as tetramer stabilizers and aggregation inhibitors. WT-TTR and the very aggressive TTR variant L55P were used as models. These compounds were able to stabilize TTR against high hydrostatic pressure (HHP), increasing the ΔGf by several kcal. They were also effective in inhibiting WT-TTR and L55P acid- or HHP-induced aggregation; in particular, LUM and IND were very effective, inhibiting almost 100% of the aggregation of both proteins under certain conditions. The species formed when aggregation was performed in the presence of these compounds were much less toxic to cells in culture. The crystal structures of WT-TTR bound to the three compounds were solved at high resolution, allowing the identification of the relevant protein:drug interactions. We discuss here the ligand-binding features of LUM, IND and SUL to TTR, emphasizing the critical interactions that render the protein more stable and less amyloidogenic.CAPESCNPqInstituto Nacional de Ciência e Tecnologia de Biologia Estrutural e Bioimagem (INBEB)FAPERJFAPES

Abordagens da dermatite atópica no âmbito dermatológico atual

A dermatite atópica seria definida como doença inflamatória cutânea crônica, de caráter genético, caracterizada pela presença de episódios recorrentes de eczema associado a prurido, muita vezes intenso, apresentando como substrato alterações imunológicas cutâneas que produzem inflamação, podendo estar eventualmente associada a doenças respiratórias, como a asma e a rinite alérgica. Trata-se de doença multifatorial, com enfoque nas alterações sistêmicas e alérgicas ou nas manifestações cutâneas, de acordo com diferentes visões da doença. A conceituação da dermatite atópica é importante, porque a conduta terapêutica pode variar segundo as diferentes formas de analisá-la. O presente estudo tem como objetivo avaliar a dermatite atópica no contexto atual da dermatologia. Trata-se de uma revisão integrativa, com uso dos descritores DeCs (Descritores em Saúde) e o Medical Subject Headings (MeSH), nos idiomas português e inglês, com os seguintes termos: “Dermatite”(dermatitis), “Pele” (skin), “tratamento” (treatment).  Desta busca, foram encontrados 150 artigos, posteriormente submetidos aos critérios de seleção. A seleção resultou em 8 artigos que foram submetidos à análise seletiva, exploratória e interpretativa no contexto do presente estudo. A partir da revisão bibliográfica dos estudos analisados, conclui-se que, de fato, a dermatite atópica é complexa e precisa ser mais estudada

Avaliação da habilidade quimiotaxica e da adesão de eosinofilos de pacientes com rinite alergica : efeito da eotaxina e interleucina-5

Orientador: Edson AntunesDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias MedicasResumo: Neste estudo, investigamos se a IL-5, a eotaxina e a dexametasona modulam a migração e adesão de eosinófilos obtidos de indivíduos sadios e de pacientes com rinite alérgica. O ensaio de quimiotaxia e de adesão foi realizado com eosinófilos de sangue periférico usando-se sistema imunomagnético de separação celular. Para os ensaios de quimiotaxia, foram utilizadas placas ChemoTx-5, e para os ensaios de adesão, placas de adesão recobertas com fibronectina. Em ambos os ensaios, a medida da absorbância (490 nM) da peroxidase eosinofílica foi utilizada como índice de eosinófilos migrados ou aderidos. O PAF (10~ M) induziu quimiotaxia de mesma magnitude em eosinófilos de indivíduos sadios e de pacientes com rinite. A IL-5 (0,25 nglml) aumentou significativamente a quimiotaxia de eosinófilos induzida pelo PAF; porém, o aumento observado em eosinófilos de pacientes com rinite alérgica foi 65% maior (p<0,001) do que de indivíduos sadios. A eotaxina (100 nglml) não alterou a resposta quimiotáxica do PAF em nenhum dos grupos estudados. A dexametasona (100 J.1g/ml)reduziu significativamente a quimiotaxia, tanto no grupo de indivíduos sadios, como no de pacientes com rinite alérgica. A IL-5 (0,25 ng/ml) induziu resposta quimiotáxica significativamente maior em eosinófilos de pacientes com rinite, em comparação com eosinófilos de indivíduos sadios. A resposta quimiotáxica à IL-5 não foi modificada pela eotaxina (100 ng/ml) em eosinófilos de indivíduos sadios, mas foi marcantemente reduzida em eosinófilos de pacientes com rinite alérgica. A eotaxina (100 nglml) induziu resposta quimiotáxica significativamente maior em eosinófilos de pacientes com rinite, em comparação com eosinófilos de indivíduos sadios. A resposta quimiotáxica à eotaxina foi potencializada pela IL-5 (100 ng/ml)em eosinófilos de indivíduos sadios e de pacientes com rinite alérgica. Entretanto, o aumento observado nos eosinófilos de pacientes com nnite alérgica foi significativamente maior do que de indivíduos sadios. Em placas recobertas com fibronectina (10 J.1g1m,l) ligante oposto da molécula de adesão VLA-4, estudamos a adesão dos eosinófilos. A adesão espontânea foi maior com eosinófilos de pacientes com rinitealérgica do que de indivíduos sadios. A incubação dos eosinófilos com eotaxina (100 nglml) ou IL-5 (0,25 ng/ml) não modificou a adesão dos eosinófilos em nenhum dos grupos estudados. A incubação com dexametasona (100 J.1g1ml) reduziu significativamente a adesão celular em ambos os grupos experimentais. Nossos resultados mostram que eosinófilos de pacientes com rinite alérgica estão pré-ativados. Isto pode ser devido à exposição na circulação à IL-5, que leva ao aumento da expressão elou função da molécula de adesão VLA-4 e, conseqüentemente,de sua migraçãoAbstract: In this study, we tested whether IL-5, eotaxin and dexamethasone modulate the adhesion and migration of eosinophils obtained trom allergic rhinitis subjects, in comparison with healty individuais. The chemotaxis and adhesion assays were performed with eosinophils isolated from peripheral blood using an imunomagnetic cell separator. ChemoTx-5 plates were used for the chemotaxis assays, whereas fibronectin-coated plates were used for the adhesion assays. For both assays, measurement of eosinophil peroxidase activity was used as marker for migrated or adhered eosinophils. Platelet-activated factor (PAF; 10-8 M) induced a significant eosinophil chemotaxis in both studied groups. Interleukin-5 (IL-5, 0.25 nglml) significantly increased the PAF-induced chemotaxis in eosinophils trom both healthy individuais and rhinitis subjects. However, the increase in rhinitis subjects eosinophils was 65% higher (p<0.001) compared with healthy individuais eosinophils. Eotaxin (100 ng/ml) did not alter the PAF-induced eosinophil chemotaxis in either groups studied. Dexamethasone (100 Ilg/ml) significantly reduced PAF-induced chemotaxis in cells trom both groups. Interleukin-5 (0.25 nglm!) induced a higher chemotactic response in eosinophils trom rhinitis subjects compared with healthy individuais. Eotaxin (100 ng/ml) had no effect on IL-5-induced eosinophil chemotaxis in healthy individuais, but prevented the increase in eosinophil chemotaxis induced by IL-5 in rhinitis subjects. Eotaxin (100 ng/ml) induced a higher chemotactic response in eosinophils trom rhinitis subjects compared with healthy individuais. Eotaxin-induced eosinophil chemotaxis was significantly enhanced by IL-5 (0.25 nglml) in both group studied, but this enhancement was higher in eosinophils trom rhinitis subjects. In plated coated with fibronectin (an opposite ligand for VLA-4) we studied the eosinophil adhesion. The basal (spontaneous) cell adhesion was siginificantly higher (p<0.05) using eosinophils from rhinitis subjects compared with those trom healthy individuais. However, the pattem of cell adhesion in both groups was not significantlyaffectedby eotaxin (100 ng/ml) or IL-5 (0.25 nglm!). Dexamethasone (100 Ilg/ml) significantly reduced the eosinophil adhesion in cells from both studied groups. Ours results indicate that eosinophils trom rhinitissubjects are found primed, and that can be due to previous exposition to IL-5 while in peripheral blood. It is likelythat IL-5 increases the expression andlor function01adhesion molecule VLA- 4 in eosinophils leading consequently to an enhanced cell chemotaxisMestradoFarmacologiaMestre em Farmacologi

Structural Insights into Plasticity and Discovery of Flavonoid Allosteric Inhibitors of Flavivirus NS2B&ndash;NS3 Protease

Flaviviruses are among the most critical pathogens in tropical regions; they cause various severe diseases in developing countries but are not restricted to these countries. The development of antiviral therapeutics is crucial for managing flavivirus outbreaks. Ten proteins are encoded in the flavivirus RNA. The N2B&ndash;NS3pro protein complex plays a fundamental role in flavivirus replication and is a promising drug target; however, no flavivirus protease inhibitors have progressed to the preclinical stage. This study analyzed the structural models and plasticity of the NS2B&ndash;NS3pro protein complex of five medically important non-dengue flaviviruses (West Nile, Rocio, Ilh&eacute;us, yellow fever, and Saint Louis encephalitis). The flavonoids amentoflavone, tetrahydrorobustaflavone, and quercetin were selected for their exceptional binding energies as potential inhibitors of the NS2B&ndash;NS3pro protein complex. AutoDock Vina results ranged from &minus;7.0 kcal/mol to &minus;11.5 kcal/mol and the compounds preferentially acted non-competitively. Additionally, the first structural model for the NS2B&ndash;NS3pro protein complex was proposed for Ilh&eacute;us and Rocio viruses. The NS2B&ndash;NS3pro protease is an attractive molecular target for drug development. The three identified natural flavonoids showed great inhibitory potential against the viral species. Nevertheless, further in silico and in vitro studies are required to obtain more information regarding NS2B&ndash;NS3pro inhibition by these flavonoids and their therapeutic potential

Rocio Virus: An Updated View on an Elusive Flavivirus

Rocio virus (ROCV) is a mosquito-borne flavivirus and human pathogen. The virus is indigenous to Brazil and was first detected in 1975 in the Sao Paulo State, and over a period of two years was responsible for several epidemics of meningoencephalitis in coastal communities leading to over 100 deaths. The vast majority of ROCV infections are believed to be subclinical and clinical manifestations can range from uncomplicated fever to fatal meningoencephalitis. Birds are the natural reservoir and amplification hosts and ROCV is maintained in nature in a mosquito-bird-mosquito transmission cycle, primarily involving Psorophora ferox mosquitoes. While ROCV has remained mostly undetected since 1976, in 2011 it re-emerged in Goiás State causing a limited outbreak. Control of ROCV outbreaks depends on sustainable vector control measures and public education. To date there is no specific treatment or licensed vaccine available. Here we provide an overview of the ecology, transmission cycles, epidemiology, pathogenesis, and treatment options, aiming to improve our ability to understand, predict, and ideally avert further ROCV emergence

Caffeic Acid Has Antiviral Activity against Ilhéus Virus In Vitro

Ilhéus virus (ILHV) is a neglected mosquito-borne flavivirus. ILHV infection may lead to Ilhéus fever, an emerging febrile disease like dengue fever with the potential to evolve into a severe neurological disease characterized by meningoencephalitis; no specific treatments are available for this disease. This study assessed the antiviral properties of caffeic acid, an abundant component of plant-based food products that is also compatible with the socioeconomic limitations associated with this neglected infectious disease. The in vitro activity of caffeic acid on ILHV replication was investigated in Vero and A549 cell lines using plaque assays, quantitative RT-PCR, and immunofluorescence assays. We observed that 500 µM caffeic acid was virucidal against ILHV. Molecular docking indicated that caffeic acid might interact with an allosteric binding site on the envelope protein

Caffeic Acid Has Antiviral Activity against Ilh&eacute;us Virus In Vitro

Ilh&eacute;us virus (ILHV) is a neglected mosquito-borne flavivirus. ILHV infection may lead to Ilh&eacute;us fever, an emerging febrile disease like dengue fever with the potential to evolve into a severe neurological disease characterized by meningoencephalitis; no specific treatments are available for this disease. This study assessed the antiviral properties of caffeic acid, an abundant component of plant-based food products that is also compatible with the socioeconomic limitations associated with this neglected infectious disease. The in vitro activity of caffeic acid on ILHV replication was investigated in Vero and A549 cell lines using plaque assays, quantitative RT-PCR, and immunofluorescence assays. We observed that 500 &micro;M caffeic acid was virucidal against ILHV. Molecular docking indicated that caffeic acid might interact with an allosteric binding site on the envelope protein