Rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART): Study protocol for a randomized controlled trial

Background: Acute respiratory distress syndrome (ARDS) is associated with high in-hospital mortality. Alveolar recruitment followed by ventilation at optimal titrated PEEP may reduce ventilator-induced lung injury and improve oxygenation in patients with ARDS, but the effects on mortality and other clinical outcomes remain unknown. This article reports the rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART). Methods/Design: ART is a pragmatic, multicenter, randomized (concealed), controlled trial, which aims to determine if maximum stepwise alveolar recruitment associated with PEEP titration is able to increase 28-day survival in patients with ARDS compared to conventional treatment (ARDSNet strategy). We will enroll adult patients with ARDS of less than 72 h duration. The intervention group will receive an alveolar recruitment maneuver, with stepwise increases of PEEP achieving 45 cmH(2)O and peak pressure of 60 cmH2O, followed by ventilation with optimal PEEP titrated according to the static compliance of the respiratory system. In the control group, mechanical ventilation will follow a conventional protocol (ARDSNet). In both groups, we will use controlled volume mode with low tidal volumes (4 to 6 mL/kg of predicted body weight) and targeting plateau pressure <= 30 cmH2O. The primary outcome is 28-day survival, and the secondary outcomes are: length of ICU stay; length of hospital stay; pneumothorax requiring chest tube during first 7 days; barotrauma during first 7 days; mechanical ventilation-free days from days 1 to 28; ICU, in-hospital, and 6-month survival. ART is an event-guided trial planned to last until 520 events (deaths within 28 days) are observed. These events allow detection of a hazard ratio of 0.75, with 90% power and two-tailed type I error of 5%. All analysis will follow the intention-to-treat principle. Discussion: If the ART strategy with maximum recruitment and PEEP titration improves 28-day survival, this will represent a notable advance to the care of ARDS patients. Conversely, if the ART strategy is similar or inferior to the current evidence-based strategy (ARDSNet), this should also change current practice as many institutions routinely employ recruitment maneuvers and set PEEP levels according to some titration method.Hospital do Coracao (HCor) as part of the Program 'Hospitais de Excelencia a Servico do SUS (PROADI-SUS)'Brazilian Ministry of Healt

Estabelecimento de ensaio para triagem e identificação de drogas contra a neurotoxicidade causada pelo zika vírus

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2018-06-28T13:19:58Z No. of bitstreams: 1 Bruno Raphael Ribeiro Cavalcante Estabelecimento de ensaio...2018.pdf: 2621503 bytes, checksum: 989c5b4182bc08670dc62c69ef126308 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2018-06-28T13:29:00Z (GMT) No. of bitstreams: 1 Bruno Raphael Ribeiro Cavalcante Estabelecimento de ensaio...2018.pdf: 2621503 bytes, checksum: 989c5b4182bc08670dc62c69ef126308 (MD5)Made available in DSpace on 2018-06-28T13:29:00Z (GMT). No. of bitstreams: 1 Bruno Raphael Ribeiro Cavalcante Estabelecimento de ensaio...2018.pdf: 2621503 bytes, checksum: 989c5b4182bc08670dc62c69ef126308 (MD5) Previous issue date: 2018Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, BrasilINTRODUÇÃO: O vírus da Zika (ZIKV), um membro pertencente ao gênero Flavivirus e à família Flaviviridae, chamou a atenção da população com sua rápida expansão geográfica e o aumento da patogenicidade, incluindo Síndrome de Guillain- Barré e microcefalia. Até o momento, não existe nenhuma vacina aprovada ou terapia específica para prevenir ou tratar a infecção por ZIKV. Dadas as complicações da infecção por ZIKV e os potenciais danos à saúde pública, opções de tratamento efetivas, incluindo vacinas e intervenções farmacológicas, têm se tornado o foco de Universidades e indústrias em todo o mundo. OBJETIVO: Realizar ensaio para triagem e identificação de drogas contra a neurotoxicidade causada pelo ZIKV em células progenitoras neurais (NPCs) humanas. METODOLOGIA: A triagem de drogas consistiu na avaliação do potencial antiviral de cloroquina, fingolimod (FTY720), mefloquina, tenofovir, N,N-dimetilesfingosina (DMS), azidotimidina (AZT) e ácido betulínico, em 3 linhagens celulares, que incluíram células HepG2, fibroblastos dermais e NPCs humanos em uma plataforma de triagem de alto conteúdo. RESULTADOS: NPCs derivadas de células-tronco humanas de pluripotência induzida (iPSCs) foram geradas e caracterizadas com sucesso para testes de drogas in vitro. Todas as drogas avaliadas não apresentaram efeito antiviral satisfatório para a infecção com ZIKV, nem em HepG2 nem em fibroblastos, com exceção do ácido betulínico, que, embora tenha indicado modesto efeito antiviral em NPCs, acarretou menos morte celular após a infecção viral nesta linhagem. CONCLUSÃO: Este ensaio baseado na utilização de NPCs derivadas de iPSCs produziu resultados consistentes para a triagem de drogas quanto à avaliação de neurotoxicidade. Além disso, o ácido betulínico evidenciou um efeito citoprotetor em NPCs infectadas por ZIKV, as quais são muito suscetíveis aos efeitos da infecção viral.INTRODUCTION: Zika virus (ZIKV), a member of the genus Flavivirus and the Flaviviridae family, has drawn the population's attention with its rapid geographic expansion and increased pathogenicity, including Guillain-Barré syndrome and microcephaly. To date, there is no approved vaccine or therapy to prevent or treat ZIKV infection. Due to ZIKV infection complications and potential threat to public health, effective treatment options, including vaccines and pharmacological interventions, became the aims of Universities and industries around the world. OBJECTIVE: To perform an assay for drug screening to identify agents against ZIKV neurotoxicity in human neural progenitor cells. METHODOLOGY: The drug screening was performed in order to evaluate antiviral potential of chloroquine, fingolimod (FTY720), mefloquine, tenofovir, N,N-dimethylsphingosine (DMS), azidothymidine (AZT) and betulinic acid (BA) in 3 cell lines, including HepG2 cell line, human dermal fibroblasts and human NPCs on a high-content screening platform. RESULTS: human induced pluripotent stem cells (iPSCs)-derived NPCs were successfully generated and characterized for in vitro screening assays. All the screened drugs did not display a satisfactory antiviral effect after ZIKV infection, neither on HepG2 nor on fibroblasts, except for betulinic acid, which, although indicated a mild antiviral effect on NPCs, led to less cell death after viral infection in this cell line. CONCLUSIONS: The assay based on the use of iPSCs-derived NPCs presented in this work produced consistent results for drug screening for the evaluation of neurotoxicity. In addition, betulinic acid displayed a cytoprotective effect on NPCs infected by ZIKV, as they are very susceptible to the effects of viral infection

Generation of integration-free iPS cell lines from three sickle cell disease patients from the state of Bahia, Brazil

Sickle cell disease (SCD) is one of the most prevalent and severe monogenetic disorders, affecting several million people around the world. Clinical manifestations and complications of the disease include sickle cell pain crisis, silent cerebral infarct, stroke, nephropathy and early death. In this study, we generated induced pluripotent stem cell (iPSC) lines from three homozygous SCD patients from the state of Bahia, Brazil, where SCD is highly prevalent. Peripheral blood mononuclear cells were collected and erythroblasts were expanded for cell reprogramming with the use of non-integrative episomal vectors. The generated iPSC lines expressed high levels of pluripotency markers, presented a normal karyotype and were able to differentiate into the three germ layers in embryoid body spontaneous differentiation assays. Moreover, the expression of the episomal vectors was lost in all iPSC lines after 15 passages. These iPSC lines may help increasing the knowledge about SCD pathogenesis and can be a useful tool for drug testing and gene editing studies

Factors Associated to Arterial Stiffness in Patients With Symptomatic Peripheral Artery Disease

BACKGROUND: The aim of this study was to identify the clinical factors associated with arterial stiffness in patients with symptomatic peripheral artery disease. METHODS: In this cross-sectional study, 181 patients (67% men; mean aged 66 ± 9 years) were recruited and had their central arterial stiffness assessed by carotid-femoral pulse wave velocity (cf-PWV). Clinical characteristics are sociodemographic data, body mass index, comorbid conditions, and walking capacity. RESULTS: Multiple linear regression analysis showed that age (b = 0.182, P = 0.032), body mass index (b = 0.254, P = 0.002), and mean blood pressure (b = 0.249, P = 0.021) were positively associated with cf-PWV. CONCLUSIONS: Our results showed that the aging, elevated body mass index, and higher blood pressure are clinical factors associated with increased arterial stiffness in patients with peripheral artery disease

IGF-1-Overexpressing Mesenchymal Stem/Stromal Cells Promote Immunomodulatory and Proregenerative Effects in Chronic Experimental Chagas Disease

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2018-12-14T12:54:42Z No. of bitstreams: 1 Silva DN IGF-1- Overexpressing Mesenchymal....2018.pdf: 19423799 bytes, checksum: c3702b567dcf37a96ef282cd8a66c8a2 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2018-12-14T13:17:36Z (GMT) No. of bitstreams: 1 Silva DN IGF-1- Overexpressing Mesenchymal....2018.pdf: 19423799 bytes, checksum: c3702b567dcf37a96ef282cd8a66c8a2 (MD5)Made available in DSpace on 2018-12-14T13:17:36Z (GMT). No. of bitstreams: 1 Silva DN IGF-1- Overexpressing Mesenchymal....2018.pdf: 19423799 bytes, checksum: c3702b567dcf37a96ef282cd8a66c8a2 (MD5) Previous issue date: 2018FINEP, CNPq, and FAPESB for research funding.Hospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.Hospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / National Institute of Science and Technology for Regenerative Medicine. Rio de Janeiro, RJ, Brazil.Hospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.Hospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.Hospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, BrasilHospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, BrasilHospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, BrasilHospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, BrasilHospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, BrasilHospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / National Institute of Science and Technology for Regenerative Medicine. Rio de Janeiro, RJ, Brazil.Hospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / National Institute of Science and Technology for Regenerative Medicine. Rio de Janeiro, RJ, Brazil.Mesenchymal stem/stromal cells (MSCs) have been investigated for the treatment of diseases that affect the cardiovascular system, including Chagas disease. MSCs are able to promote their beneficial actions through the secretion of proregenerative and immunomodulatory factors, including insulin-like growth factor-1 (IGF-1), which has proregenerative actions in the heart and skeletal muscle. Here, we evaluated the therapeutic potential of IGF-1-overexpressing MSCs (MSC_IGF-1) in a mouse model of chronic Chagas disease. C57BL/6 mice were infected with Colombian strain Trypanosoma cruzi and treated with MSCs, MSC_IGF-1, or vehicle (saline) six months after infection. RT-qPCR analysis confirmed the presence of transplanted cells in both the heart and skeletal muscle tissues. Transplantation of either MSCs or MSC_IGF-1 reduced the number of inflammatory cells in the heart when compared to saline controls. Moreover, treatment with MSCs or MSC_IGF-1 significantly reduced TNF-α, but only MSC treatment reduced IFN-γ production compared to the saline group. Skeletal muscle sections of both MSC- and MSC_IGF-1-treated mice showed a reduction in fibrosis compared to saline controls. Importantly, the myofiber area was reduced in T. cruzi-infected mice, and this was recovered after treatment with MSC_IGF-1. Gene expression analysis in the skeletal muscle showed a higher expression of pro- and anti-inflammatory molecules in MSC_IGF-1-treated mice compared to MSCs alone, which significantly reduced the expression of TNF-α and IL-1β. In conclusion, our results indicate the therapeutic potential of MSC_IGF-1, with combined immunomodulatory and proregenerative actions to the cardiac and skeletal muscles

Arsenic Trioxide Triggers Apoptosis of Metastatic Oral Squamous Cells Carcinoma with Concomitant Downregulation of GLI1 in Hedgehog Signaling

Given the lack of advances in Oral Squamous Cell Carcinoma (OSCC) therapy in recent years, pharmacological strategies to block OSCC-related signaling pathways have gained prominence. The present study aimed to evaluate the therapeutic potential of Arsenic Trioxide (ATO) concerning its antitumoral effects and the inhibition of the Hedgehog (HH) pathway in OSCC. Initially, ATO cytotoxicity was assessed in a panel of cell lines. Cell viability, cell cycle, death patterns, and cell morphology were analyzed, as well as the effect of ATO on the expression of HH pathway components. After the cytotoxic assay, HSC3 cells were chosen for all in vitro assays. ATO increased apoptotic cell death and nuclear fragmentation in the sub-G1 cell cycle phase and promoted changes in cell morphology. In addition, the reduced expression of GLI1 indicated that ATO inhibits HH activity. The present study provides evidence of ATO as an effective cytotoxic drug for oral cancer treatment in vitro

Circulating miRNAs as Potential Biomarkers Associated with Cardiac Remodeling and Fibrosis in Chagas Disease Cardiomyopathy

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2019-09-27T16:01:25Z No. of bitstreams: 1 Nonaka Vasques K.C Circulating.pdf: 2693091 bytes, checksum: ede286cd2f7ad2cf644ed62421a5a459 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2019-09-27T16:22:08Z (GMT) No. of bitstreams: 1 Nonaka Vasques K.C Circulating.pdf: 2693091 bytes, checksum: ede286cd2f7ad2cf644ed62421a5a459 (MD5)Made available in DSpace on 2019-09-27T16:22:09Z (GMT). No. of bitstreams: 1 Nonaka Vasques K.C Circulating.pdf: 2693091 bytes, checksum: ede286cd2f7ad2cf644ed62421a5a459 (MD5) Previous issue date: 2019-01-20Bahia State Foundation for Research (FAPESB) and Institutos Nacionais de Ciência e Tecnologia (INCT; 465656/2014-5). Milena B. P. Soares is a recipient of CNPq fellowship.Hospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / D’Or Institute for Research and Education. Rio de Janeiro, RJ, Brazil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / São Rafael Hospital. Department of Cardiology. Salvador, BA, Brasil.Hospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / D’Or Institute for Research and Education. Rio de Janeiro, RJ, Brazil.Hospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil.Hospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / D’Or Institute for Research and Education. Rio de Janeiro, RJ, Brazil.Hospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil.Hospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / Universidade Federal da Bahia. Salvador, BA, Brasil.Messejana Hospital. Fortaleza, CE, Brasil.Messejana Hospital. Fortaleza, CE, Brasil.São Rafael Hospital. Department of Cardiology. Salvador, BA, Brasil.University of Southern Denmark. Institute of Regional Health Research. Vejle Hospital. Department of Clinical Genetics. Vejle, Denmark.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / National Institute of Science and Technology for Regenerative Medicine. Rio de Janeiro, RJ, Brazil.Salvador, BA, Brasil / D’Or Institute for Research and Education. Rio de Janeiro, RJ, Brazil / National Institute of Science and Technology for Regenerative Medicine. Rio de Janeiro, RJ, Brazil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / National Institute of Science and Technology for Regenerative Medicine. Rio de Janeiro, RJ, Brazil.Chagas disease (CD) affects approximately 6-7 million people worldwide, from which 30% develop chronic Chagas cardiomyopathy (CCC), usually after being asymptomatic for years. Currently available diagnostic methods are capable of adequately identifying infected patients, but do not provide information regarding the individual risk of developing the most severe form of the disease. The identification of biomarkers that predict the progression from asymptomatic or indeterminate form to CCC, may guide early implementation of pharmacological therapy. Here, six circulating microRNAs (miR-19a-3p, miR-21-5p, miR-29b-3p, miR-30a-5p, miR-199b-5p and miR-208a-3p) were evaluated and compared among patients with CCC (n = 28), CD indeterminate form (n = 10) and healthy controls (n = 10). MiR-19a-3p, miR-21-5p, and miR-29b-3p were differentially expressed in CCC patients when compared to indeterminate form, showing a positive correlation with cardiac dysfunction, functional class, and fibrosis, and a negative correlation with ejection fraction and left ventricular strain. Cardiac tissue analysis confirmed increased expression of microRNAs in CCC patients. In vitro studies using human cells indicated the involvement of these microRNAs in the processes of cardiac hypertrophy and fibrosis. Our study suggests that miRNAs are involved in the process of cardiac fibrosis and remodeling presented in CD and indicate a group of miRNAs as potential biomarkers of disease progression in CCC

Therapeutic miR-21 Silencing Reduces Cardiac Fibrosis and Modulates Inflammatory Response in Chronic Chagas Disease

Chagas disease, caused by the parasite Trypanosoma cruzi (T. cruzi), remains a serious public health problem for which there is no effective treatment in the chronic stage. Intense cardiac fibrosis and inflammation are hallmarks of chronic Chagas disease cardiomyopathy (CCC). Previously, we identified upregulation of circulating and cardiac miR-21, a pro-fibrotic microRNA (miRNA), in subjects with CCC. Here, we explored the potential role of miR-21 as a therapeutic target in a model of chronic Chagas disease. PCR array-based 88 microRNA screening was performed in heart samples obtained from C57Bl/6 mice chronically infected with T. cruzi and serum samples collected from CCC patients. MiR-21 was found upregulated in both human and mouse samples, which was corroborated by an in silico analysis of miRNA-mRNA target prediction. In vitro miR-21 functional assays (gain-and loss-of-function) were performed in cardiac fibroblasts, showing upregulation of miR-21 and collagen expression upon transforming growth factor beta 1 (TGFβ1) and T. cruzi stimulation, while miR-21 blockage reduced collagen expression. Finally, treatment of T. cruzi-infected mice with locked nucleic acid (LNA)-anti-miR-21 inhibitor promoted a significant reduction in cardiac fibrosis. Our data suggest that miR-21 is a mediator involved in the pathogenesis of cardiac fibrosis and indicates the pharmacological silencing of miR-21 as a potential therapeutic approach for CCC