A successful strategy for the recovering of active P21, an insoluble recombinant protein of Trypanosoma cruzi

Structural studies of proteins normally require large quantities of pure material that can only be obtained through heterologous expression systems and recombinant technique. in these procedures, large amounts of expressed protein are often found in the insoluble fraction, making protein purification from the soluble fraction inefficient, laborious, and costly. Usually, protein refolding is avoided due to a lack of experimental assays that can validate correct folding and that can compare the conformational population to that of the soluble fraction. Herein, we propose a validation method using simple and rapid 1D H-1 nuclear magnetic resonance (NMR) spectra that can efficiently compare protein samples, including individual information of the environment of each proton in the structure.Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)INBEQMeDIUniv Fed Uberlandia, Inst Ciencias Biomed, BR-38400 Uberlandia, MG, BrazilUniv São Paulo, Inst Fis Sao Carlos, Sao Carlos, SP, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, Vila Mariana, SP, BrazilUniv Fed Minas Gerais, Inst Ciencias Biol, Dept Biol Geral, Belo Horizonte, MG, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, Vila Mariana, SP, BrazilFAPESP: 2010/51867-6FAPESP: 2012/21153-7FAPEMIG: APQ-00621-11FAPEMIG: APQ-00305-12CAPES: 23038.005295/2011-40Web of Scienc

Mechanistic Insights into the Anti-angiogenic Activity of Trypanosoma cruzi Protein 21 and its Potential Impact on the Onset of Chagasic Cardiomyopathy

Chronic chagasic cardiomyopathy (CCC) is arguably the most important form of the Chagas Disease, caused by the intracellular protozoan Trypanosoma cruziit is estimated that 10-30% of chronic patients develop this clinical manifestation. The most common and severe form of CCC can be related to ventricular abnormalities, such as heart failure, arrhythmias, heart blocks, thromboembolic events and sudden death. Therefore, in this study, we proposed to evaluate the anti-angiogenic activity of a recombinant protein from T. cruzi named P21 (rP21) and the potential impact of the native protein on CCC. Our data suggest that the anti-angiogenic activity of rP21 depends on the protein's direct interaction with the CXCR4 receptor. This capacity is likely related to the modulation of the expression of actin and angiogenesis-associated genes. Thus, our results indicate that T. cruzi P21 is an attractive target for the development of innovative therapeutic agents against CCC.Univ Fed Sao Paulo, Escola Paulista Med, Departamento Microbiol Imunol Parasitol, BR-05508 Sao Paulo, SP, BrazilUniv Fed Uberlandia, Inst Ciencias Biomed, Dept Imunol, Lab Tripanosomatideos, Uberlandia, MG, BrazilUniv Fed Uberlandia, Inst Genet & Bioquim, Lab Bioquim & Toxinas Animais, Uberlandia, MG, BrazilCeTICS, Inst Butantan, Sao Paulo, BrazilUniv Fed Uberlandia, Fac Med, Centro Referencia Nacl Dermatol Sanitaria Hanseni, Lab Patol Mol & Biotecnol, Uberlandia, MG, BrazilUniv Fed Uberlandia, Inst Ciencias Biomed, Dept Immunol, Lab Osteoimunol & Imunol Tumores, Uberlandia, MG, BrazilUniv Fed Sao Paulo, Escola Paulista Med, Departamento Microbiol Imunol Parasitol, BR-05508 Sao Paulo, SP, BrazilUniv Fed Sao Paulo, Escola Paulista Med, Departamento Microbiol Imunol Parasitol, BR-05508 Sao Paulo, SP, BrazilWeb of Scienc

Serum Metabolite Profiling of Leprosy Patients Revealed a Biomarkers Pattern

Hansen's disease still represents a health problem in Brazil and worldwide. This disease, caused by Mycobacterium leprae, mainly affects the skin and the peripheral nervous system. Currently, clinical symptoms are used for the diagnosis of this disease; however, there is still no fast and reliable laboratory diagnosis. The early diagnosis of Hansen’s disease and monitoring strategies for populations at risk of becoming ill is a challenge for its eradication. This study sought to identify changes in the metabolic profile induced in the serum of Paucibacillary and Multibacillary patients. In this study, plasma was used from individuals with different forms of Hansen’s disease (PB = 42, Mb = 39), as well as from healthy individuals (control group, n = 37). The analysis of the metabolomic profile was carried out in the High-Performance Liquid Chromatography (HPLC) system coupled to a quadrupole-flight-time mass spectrometer. The metabolic profile allowed the identification of 14 metabolites differentially expressed in the serum; 14 were responsible for the differences between the PB and control groups. Likewise, 14 metabolites were expressed differently between the MB and control groups; 4 differentially expressed metabolites were responsible for the difference between the PB and MB groups. Moreover, using the Random Forest classification algorithm, it was possible to differentiate the groups of infected and uninfected patients with an accuracy of 98.62%. These metabolites must be validated so that these molecules can be used as biomarkers not only to classify clinical forms but also to assess disease progression or to be used as therapeutic targets.CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorTese (Doutorado)A hanseníase ainda representa um problema de saúde pública no Brasil e no mundo. Essa doença, causada pelo Mycobacterium leprae, afeta principalmente a pele e o sistema nervoso periférico. Atualmente sintomas clínicos são utilizados para o diagnóstico desta doença, no entanto, ainda não existe um diagnóstico laboratorial rápido e confiável. O diagnóstico precoce da hanseníase e estratégias de monitoramento de populações sob risco de adoecer é um desafio para eliminação. Esse estudo buscou identificar as mudanças no perfil metabólico induzido no soro de pacientes PB e MB. Nesse estudo foram utilizados amostra de soro de indivíduos portadores das diferentes formas de hanseníase (PB = 42, MB = 39) bem como de indivíduos sadios (controles, n=37). A análise do perfil metabolômico foi analisado no sistema de HPLC acoplado a um espectrômetro de massas do tipo quadrupolo-tempo-de-vôo. O perfil metabólico permitiu identificar 14 metabólitos diferencialmente expressos entre o grupo PB e controle. Da mesma forma, 14 metabólitos foram expressos de forma distinta entre o grupo MB e controle; 4 metabólitos diferencialmente expressos foram responsáveis pela diferença entre o grupo PB e MB. Ainda, utilizando o algoritmo de classificação Random Forest, foi possível diferenciar os grupos de doentes e não doentes com acurácia de 98,62 %. Esses metabólitos devem ser validados para que essas moléculas possam ser utilizadas como biomarcadores não só para classificar as formas clínicas, mas também para avaliar a progressão da doença ou serem utilizados como alvos terapêuticos

Molecular Characterisation of Newly Identified HIV-1 Infections in Curitiba, Brazil: Preponderance of Clade C Among Males With Recent Infections

As in many areas of Brazil, the AIDS epidemic in Curitiba is relatively stable, but surveillance is important to support public policy. The molecular characteristics of HIV may be instrumental for monitoring epidemic trends. We evaluated plasma HIV-1 RNA (n = 37) from 38 cases presenting with positive serology, who were among 820 consenting volunteers visiting the downtown counselling and serology testing centre. Seroprevalence was 4.6% (CI 95% 3.2-6.3) and the estimated HIV incidence, as defined by the BED assay, was 2.86 persons/years (CI 95% 1.04-4.68). An additional set of contemporaneous, anonymous samples from a local laboratory was also analysed (n = 20). Regions of the HIV-1 polymerase (n = 57) and envelope (n = 34) were evaluated for subtyping, determination of mosaic structure, primary drug resistance mutations (pDRM), envelope V3 loop motifs and amino acid signatures related to viral tropism. HIV-1 clade B was observed in 53% of cases; HIV-1C in 30% and BC mosaics in 14%, with one F genome and one CF mosaic. Clade C infection was associated with recent infections among males (p < 0.03). Stanford surveillance pDRM was observed in 8.8% of sequences, with 7% showing high level resistance to at least one antiretroviral drug. Tropism for CXCR4 co-receptor was predicted in 18% of envelope sequences, which were exclusively among clade B genomes and cases with serological reactivity to chronic infection

Resumos em andamento - Educação

Resumos em andamento - Educaçã

Resumos em andamento - Educação

Resumos em andamento - Educaçã