Our results suggest that central sympathetic inhibition exerts beneficial effects by increasing perfusion and reducing inflammatory marker expression and oxidative stress in the brains of rats with metabolic syndrome. Centrally acting antihypertensive drugs may be helpful in regulating cerebral microcirculatory function and vascular inflammation in metabolic syndrome.

BackgroundCardiac fibrosis is a consequence of chronic chagasic cardiomyopathy (CCC). In other cardiovascular diseases, the protagonist role of fibroblasts in cardiac fibrosis is well established. However, the role of cardiac fibroblasts (CFs) in fibrosis during the CCC is not clear. Here, our aim was to investigate the effect of Trypanosoma cruzi, the etiological agent of Chagas disease on CFs activation.MethodsCardiac fibroblasts were purified from primary cultures of mouse embryo cardiac cells. After two passages, cells were infected with T. cruzi (Y strain) and analyzed at different times for determination of infectivity, activation and production of extracellular matrix components (fibronectin, laminin and collagen IV) by immunofluorescence and western blot.ResultsAt second passage, cultures were enriched in CFs (95% of fibroblasts and 5% of cardiomyocytes), as revealed by presence of alpha-smooth muscle actin (α-SMA) and discoidin domain receptor 2 (DDR2) and absence of sarcomeric tropomyosin (ST) protein expression. Trypanosoma cruzi infection induced fibroblast-myofibroblast transition, with increased expression of α-SMA after 6 and 24 h post-infection (hpi). Fibronectin was increased at 6, 24 and 48 hpi, laminin was increased at 6 and 24 hpi and collagen IV was increased at 6 hpi.ConclusionsOur results showed that T. cruzi activates CFs, inducing activation and exacerbates ECM production. Furthermore, our data raise the possibility of the involvement of CFs in heart fibrosis during Chagas disease.

Chagas disease is responsible for more than 10,000 deaths per year and about 6 to 7 million infected people worldwide. In its chronic stage, patients can develop mega-colon, mega-esophagus, and cardiomyopathy. Differences in clinical outcomes may be determined, in part, by the genetic background of the parasite that causes Chagas disease. Trypanosoma cruzi has a high genetic diversity, and each group of strains may elicit specific pathological responses in the host. Conflicting results have been reported in studies using various combinations of mammalian host-T. cruzi strains. We previously profiled the transcriptomic signatures resulting from infection of L6E9 rat myoblasts with four reference strains of T. cruzi (Brazil, CL, Y, and Tulahuen). The four strains induced similar overall gene expression alterations in the myoblasts, although only 21 genes were equally affected by all strains. Cardiotrophin-like cytokine factor 1 (Clcf1) was one of the genes found to be consistently upregulated by the infection with all four strains of T. cruzi. This cytokine is a member of the interleukin-6 family that binds to glycoprotein 130 receptor and activates the JAK/STAT signaling pathway, which may lead to muscle cell hypertrophy. Another commonly upregulated gene was tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein theta (Ywhaq, 14-3-3 protein), present in the Cell Cycle Pathway. In the present work, we reanalyzed our previous microarray dataset, aiming at understanding in more details the transcriptomic impact that each strain has on JAK/STAT signaling and Cell Cycle pathways. Using Pearson correlation analysis between the expression levels of gene pairs in biological replicas from each pathway, we determined the coordination between such pairs in each experimental condition and the predicted protein interactions between the significantly altered genes by each strain. We found that although these highlighted genes were similarly affected by all four strains, the downstream genes or their interaction partners Nisimura et al. Transcriptomic Profiling of T. cruzi Infection were not necessarily equally affected, thus reinforcing the idea of the role of parasite background on host cell transcriptome. These new analyses provide further evidence to the mechanistic understanding of how distinct T. cruzi strains lead to diverse remodeling of host cell transcriptome.

Most investigations identified a control breach as the source of infections.

BACKGROUND Angiogenesis has been implicated in tissue injury in several noninfectious diseases, but its role in Chagas disease (CD) physiopathology is unclear.OBJECTIVES The present study aimed to investigate the effect of Trypanosoma cruzi infection on cardiac angiogenesis during the acute phase of experimental CD. METHODSThe signalling pathway involved in blood vessel formation and cardiac remodelling was evaluated in Swiss Webster mice infected with the Y strain of T. cruzi. The levels of molecules involved in the regulation of angiogenesis, such as vascular endothelial growth factor-A (VEGF-A), Flk-1, phosphorylated extracellular-signal-regulated protein kinase (pERK), hypoxiainducible factor-1α (HIF-1α), CD31, α-smooth muscle actin (α-SMA) and also the blood vessel growth were analysed during T. cruzi infection. Hearts were analysed using conventional histopathology, immunohistochemistry and western blotting.FINDINGS In this study, our data demonstrate that T. cruzi acute infection in mice induces exacerbated angiogenesis in the heart and parallels cardiac remodelling. In comparison with noninfected controls, the cardiac tissue of T. cruzi-infected mice presented higher levels of (i) HIF-1α, VEGF-A, Flk-1 and pERK; (ii) angiogenesis; (iii) α-SMA + cells in the tissue; and (iv) collagen -1 deposition around blood vessels and infiltrating throughout the myocardium. MAIN CONCLUSIONSWe observed cardiac angiogenesis during acute experimental T. cruzi infection parallels cardiac inflammation and remodelling.

Introduction: Breast cancer is the most prevalent cancer among women worldwide and in Brazil. Although, great scientific advances have provided therapeutic innovations, it is still a challenge therapeutic agents that act against metastasis. The three-dimensional (3D) systems of cell culture better mimic the molecular, morphological and functional features of in vivo tumor than traditional monolayer cultures (2D), working as an ideal platform for understanding cancer biology and to perform therapeutic response analysis.Objective: The aim of this study was to establish a scaffold-free 3D spheroid model to investigate in vitro metastatic potential and therapeutic response of tumor cells during doxorubicin treatment.Methodology: First, we established and characterized the 3D model by analyzing PI, Ki-67, E-cadherin and laminin staining in spheroids of MCF-7. Then, we produced spheroid from cells isolated from mouse mammary tumors produced from 4T1 and 67NR cell lines. We also characterized the migration/metastasis assay by evaluating E-cadherin and Vimentin expression (epithelial-mesenchymal transition markers) and cellular dispersion of MCF-7 cells. Even, we validated the model for in vitro therapeutic response studies using doxorubicin (dox), evaluating spheroid diameter, cell death, cell viability and migration/metastatic potential of cells.Results: Mammary tumor spheroids (MTS) produced from the MCF-7 cell line mimicked avascular tumor characteristics, such as necrotic center and peripheral proliferative cells. In addition, MTS exhibited adherent junction proteins and were able to produce their own extracellular matrix. We also demonstrate that the spheroid model supports the 3D culturing of cells isolated from mouse mammary tumors produced from 4T1 and 67NR cell lines. Through the cell migration assay, we demonstrated for the first time that this model reproduces the epithelial-mesenchymal transition process (EMT), characteristic as one of the steps of metastasis formation and verified by the loss of E-cadherin protein and increased vimentin expression, as cells move away from "tumor" in vitro. We also observed collective cell migration, another feature of the metastatic process. Dox induced cell death, reduced spheroid diameter and inhibited the metastatic potential of tumor cells in vitro, by reducing collective migration and inhibiting the EMT process, suggesting a new application for this drug in anti-cancer treatment. Conclusion:Our results demonstrate that 3D cultivation reproduced characteristics of breast tumors observed in vivo, including the metastatic process sensitive to dox treatment, suggesting that mammary tumor spheroids may be considered a powerful in vitro tool for the study of metastatic potential of tumor cells and new therapeutic approaches against breast cancer.

metastasis. β-lapachone (β-lap) is a natural naphthoquinone obtained from the inner bark of the lapacho trees, native of South America. This natural compound has several pharmacological effects, such as antibacterial, antifungal, antiviral, analgesic, anti-inflammatory activities, as well as, antitumor effects. The 3D systems of cell culture better recapitulate cell-cell and cell-matrix interactions, mimetizing the tumor morphology and behavior, responding in vitro to treatments in a more similar way to in vivo tumors than traditional 2D culture systems.Objective: Given the lack of studies using 3D culture using β-lap, the aim of this study was to evaluate the effect of β-lap treatment in breast tumor spheroids.Methodology: First, we produced our scaffold-free 3D model with MCF-7 human breast tumor cell line. Thereafter, we evaluated the cytotoxic and antimetastatic effect of β-lap in spheroids, evaluating spheroid diameter, cell death, migration/metastatic potential of cells and epithelial-mesenchymal markers (E-cadherin and vimentin).Results: Our results revealed that β-lap reduced spheroid diameter, induced cell death and inhibited the metastatic potential of tumor cells in vitro by reducing collective migration and inhibiting the EMT process. Conclusion:Our results revealed that β-lap reduced spheroid diameter, induced cell death and inhibited the metastatic potential of tumor cells in vitro by reducing collective migration and inhibiting the EMT process.

Introduction: Breast cancer is the second most common type of cancer in the world. Personalized therapy is an option in the fight for the cure of cancer, since tumor variability is a great challenge in the elaboration of therapeutic protocols. Only 5% of the compounds tested in vitro in 2D systems present in vivo antitumor activity. Otherwise, the three-dimensional (3D) cell culture systems, which better mimic the architecture and tumor behavior observed in vivo, respond to in vitro treatment in a similar way to tumors in patients when treated with the same chemotherapeutic agents, showing great potential for evaluation of specific tumor therapy. Objective: The aim of this study is to evaluate the similarity of tumor spheroids produced in vitro with their original tumors, as regards the morphological, molecular and functional characteristics for therapeutic response studies, aiming at personalized anticancer therapy. Methodology: First, we established the 3D culture with the MCF7 human breast cancer cell line and analyzed spheroid growth, death (7-AAD) and migration during doxorubicin or betalapachone treatment. At this point, we started to standardize the 3D culture with breast cancer cells from patients. Then we will perform molecular and functional analyzes. The molecular characterization will be done by analyzing the gene expression of mammary tumor biomarkers. Functional analyzes of the spheroids will be done through evaluation of proliferation, death and cell viability to compare with the pathological response after neoadjuvant therapy. Results: Our preclinical results demonstrate that it is possible to produce tumor spheroids both from cell lines and tumors isolated from mouse. The treatment of spheroids with doxorubicin or beta-lapachone were able to inhibit spheroid growth, induce apoptosis and inhibit metastasis in vitro. Conclusion: We established our 3D cell culture using cell lines and tumors isolated from mouse. Our next step will be perform the assays with tumor spheroids produced from tumor cells of patients and, once we keep the tumor identity in vitro, we will begin the therapeutic tests with different drugs already used in the clinic, aiming the production of a prototype of a kit for personalized therapy ("tumor antibiogram" or "chemogram").