A Phosphoproteomic Approach towards the Understanding of the Role of TGF-β in Trypanosoma cruzi Biology

Transforming growth factor beta (TGF-β) plays a pivotal role in Chagas disease, not only in the development of chagasic cardiomyopathy, but also in many stages of the T. cruzi life cycle and survival in the host cell environment. The intracellular signaling pathways utilized by T. cruzi to regulate these mechanisms remain unknown. To identify parasite proteins involved in the TGF-β response, we utilized a combined approach of two-dimensional gel electrophoresis (2DE) analysis and mass spectrometry (MS) protein identification. Signaling via TGF-β is dependent on events of phosphorylation, which is one of the most relevant and ubiquitous post-translational modifications for the regulation of gene expression, and especially in trypanosomatids, since they lack several transcriptional control mechanisms. Here we show a kinetic view of T. cruzi epimastigotes (Y strain) incubated with TGF-β for 1, 5, 30 and 60 minutes, which promoted a remodeling of the parasite phosphorylation network and protein expression pattern. The altered molecules are involved in a variety of cellular processes, such as proteolysis, metabolism, heat shock response, cytoskeleton arrangement, oxidative stress regulation, translation and signal transduction. A total of 75 protein spots were up- or down-regulated more than twofold after TGF-β treatment, and from these, 42 were identified by mass spectrometry, including cruzipain–the major T. cruzi papain-like cysteine proteinase that plays an important role in invasion and participates in the escape mechanisms used by the parasite to evade the host immune system. In our study, we observed that TGF-β addition favored epimastigote proliferation, corroborating 2DE data in which proteins previously described to be involved in this process were positively stimulated by TGF-β

Terapia cognitivo-comportamental com intervenção familiar para crianças e adolescentes com transtorno obsessivo-compulsivo: uma revisão sistemática Cognitive behavioral therapy with family intervention for children and adolescents with obsessive-compulsive disorder: a systematic review

O transtorno obsessivo-compulsivo (TOC) é uma doença mental grave, com graves consequências para a dinâmica familiar. Desta forma, o envolvimento dos pais parece ser determinante na resolução dos sintomas desse transtorno. O objetivo deste estudo foi avaliar a qualidade da evidência para a recomendação de terapia cognitivo-comportamental (TCC) com intervenção familiar para crianças e adolescentes com TOC. A busca sistemática foi realizada nas bases de dados MEDLINE/PubMed, seguida da análise de resumos e artigos na íntegra por dois avaliadores independentes. Posteriormente, foi realizada a análise de evidência através do sistema Grading of Recommendations Assessment, Development and Evaluation (GRADE). O tamanho de efeito da intervenção foi calculado através do d de Cohen. Foram localizados 77 artigos no PubMed e mais 12 artigos após busca cruzada de referências. Destes, sete artigos foram incluídos na revisão, segundo os seguintes critérios: ser estudo de intervenção, envolver apenas crianças e/ou adolescentes e possuir diagnóstico clínico ou estruturado de TOC. A escala Children’s Yale-Brown Obsessive Compulsive Scale (CY-BOCS) foi utilizada por todos os artigos para a avaliação de desfecho, permitindo avaliar o tamanho de efeito das intervenções não controladas (d = 1,43), que resultou em uma diferença de médias de cerca 13 pontos (IC95% 11,84-14,39; p < 0,001). Por outro lado, uma alta heterogeneidade foi detectada entre os estudos (I² = 67%). A TCC com intervenção familiar parece ter um efeito importante na redução dos sintomas de TOC na infância e adolescência. No entanto, os poucos estudos disponíveis não nos permitem estabelecer um grau de evidência maior do que C para essa recomendação. Novos ensaios clínicos randomizados são necessários para confirmar essa recomendação.<br>Obsessive-compulsive disorder (OCD) is a severe mental disorder with serious consequences to family dynamics. Therefore, parental involvement seems to be a key factor for the successful treatment of this psychiatric disorder. The aim of this study was to evaluate the level of evidence available to allow recommendation of cognitive behavioral therapy (CBT) with family intervention for the treatment of children and adolescents with OCD. The systematic search was performed on MEDLINE/PubMed, followed by analysis of abstracts and full-length articles by two independent evaluators. Subsequently, an analysis of the evidence available was conducted using the Grading of Recommendations Assessment, Development and Evaluation (GRADE) system. The effect size of the intervention was calculated using Cohen’s d. We found 77 articles on PubMed, plus 12 articles via cross-reference search. Of these, seven articles were included in this review, according to the following criteria: intervention study, involving only children and/or adolescents, and a having a structured or clinical diagnosis of OCD. The Children’s Yale-Brown Obsessive Compulsive Scale (CY-BOCS) was used for outcome evaluation in all articles, thus enabling assessment of the effect size of non-controlled interventions (d = 1.43), resulting in a mean difference of 13.11 points (95%CI 11.84-14.39; p < 0.001). Conversely, a high heterogeneity was detected among the studies (I² = 67%). Family-based CBT seems to have an important effect on the reduction of OCD symptoms in children and adolescents. However, the small number of studies available do not allow us to establish an evidence level higher than C for this recommendation. New randomized clinical trials are needed to confirm this recommendation

Trypanosoma cruzi High Mobility Group B (TcHMGB) can act as an inflammatory mediator on mammalian cells

When an infection occurs, the innate immune cells recognize Pathogen Associated Molec ular Patterns (PAMPs) through their Pattern Recognition Receptors. This triggers an inflammatory response intended to kill the foreign microbe. But inflammation can also be triggered by the recognition of endogenous molecules called “Danger (or Damage) Asso ciated Molecular Patterns” (DAMPs) that are released by damaged or necrotic cells to “ring the alarm” of the immune system that repair is needed, so some of them are also known as “alarmins”. High Mobility group box 1 protein (HMGB1) is a prototypical alar min molecule released by injured cells and it is also actively secreted by cells of the innate immune system in response to invasion as well as to sterile damage. Trypanosoma cruzi, the causal agent of Chagas Disease, has its own HMGB protein that we called TcHMGB. Using in vitro and in vivo experimental systems, we demonstrated for the first time that TcHMGB is able to mediate inflammation on mammalian cells, inducing the expression of both pro-inflammatory and anti-inflammatory cytokines. Our results suggest that the parasite´s protein could have a role in the immune response and the pathogenesis of Cha gas disease, probably overlapping to some extent with the host cell DAMP molecules´ functions.Para citar este articulo: Cribb P, Perdomo V, Alonso VL, Manarin R, Barrios-Paya´n J, Marquina-Castillo B, et al. (2017) Trypanosoma cruzi High Mobility Group B (TcHMGB) can act as an inflammatory mediator on mammalian cells. PLoS Negl Trop Dis 11(2): e0005350. doi:10.1371/journal.pntd.0005350Fil: Cribb, Pamela. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario (IBR -CONICET); Argentina.Fil: Cribb, Pamela. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina.Fil: Perdomo, Virginia Gabriela. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario (IBR -CONICET); Argentina.Fil: Perdomo, Virginia Gabriela. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina.Fil: Alonso, Victoria Lucía. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina.Fil: Manarin, Romina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina.Fil: Barrios-Payán, Jorge. Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán. Sección de Patología Experimental; México.Fil: Marquina-Castillo, Brenda. Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán. Sección de Patología Experimental; México.Fil: Tavernelli, Luis. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario (IBR -CONICET); Argentina.Fil: Hernández-Pando, Rogelio. Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán. Sección de Patología Experimental; México