Itoring patients after initial diagnosis/surgery. Even though each biomarker investigated in the present work is not exclusively associated with melanoma, their combination reveals a high specificity for melanoma detection.Supporting InformationFigure S1 95 CI of the AUC according to the stage ofdisease. Bonferroni adjusted confidence intervals of the AUC of total cfDNA (Panel A), integrity index 180/67 (Panel B), methylated purchase 4 IBP RASSF1A (Panel C), and BRAFV600E (Panel D) according to the stage of disease. The horizontal dashed line in each Panel represent the AUC value obtained for each biomarker by comparing all cases and controls. (TIF)Table S1 Descriptive Statistics according to the stage ofdisease. (DOC)Author ContributionsConceived and designed the experiments: CO PP. Performed the experiments: FS. Analyzed the data: PV CMC. Contributed reagents/ materials/analysis tools: DM MP. Wrote the paper: PP. Patients enrollment: VDG MG.
The Role of Reactive Oxygen Species in Anopheles aquasalis Response to Plasmodium vivax Infection???Ana C. Bahia1, Jose Henrique M. Oliveira2, Marina S. Kubota1, Helena R. C. Araujo3, Jose B. P. Lima4, ???Claudia Maria Rios-Velasquez5, Marcus Vinicius G. Lacerda6, Pedro L. Oliveira2,7, Yara M. Traub?Cseko1*., Paulo F. P. Pimenta3*.????1 Laboratorio de Biologia Molecular de Parasitas e Vetores, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil, 2 Laboratorio de Bioquimica de Artropodes ? ica, Programa 
de Biologia Molecular e Biotecnologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil, ?Hematofagos, Instituto de Bioquimica Me ? ica, Instituto Rene Rachou, Belo Horizonte, Brazil, 4 Laboratorio de Fisiologia e Controle de Artropodes Vetores, Instituto Oswaldo Cruz, ???3 Laboratorio de Entomologia Me ?^ ?Fiocruz, Rio de Janeiro, Brazil, 5 Laboratorio de Biodiversidade em Saude, Centro de Pesquisa Leonidas Maria Deane, Fiocruz, Manaus, Brazil, 6 Fundacao de Medicina ^ncia e Tecnologia em Entomologia Molecular, Rio de Janeiro, Brazil Tropical Dr. Heitor Vieira Dourado, Manaus, Brazil, 7 Instituto Nacional de CieAbstractMalaria affects millions of people worldwide and hundreds of thousands of people each year in Brazil. The mosquito Anopheles aquasalis is an important vector of Plasmodium vivax, the main human malaria parasite in the Americas. Reactive oxygen species (ROS) have been shown to have a role in insect innate immune responses as a potent pathogen-killing agent. We investigated the mechanisms of free radicals modulation after A. aquasalis ML 281 infection with P. vivax. ROS metabolism was evaluated in the vector by studying expression and activity of three key detoxification enzymes, one catalase and two superoxide dismutases (SOD3A and SOD3B). Also, the involvement of free radicals in the mosquito immunity was measured by silencing the catalase gene followed by infection of A. aquasalis with P. vivax. Catalase, SOD3A and SOD3B expression in whole A. aquasalis were at the same levels of controls at 24 h and upregulated 36 h after ingestion of blood containing P. vivax. However, in the insect isolated midgut, the mRNA for these enzymes was not regulated by P. vivax infection, while catalase activity was reduced 24 h after the infectious meal. RNAi-mediated silencing of catalase 1527786 reduced enzyme activity in the midgut, resulted in increased P. vivax infection and prevalence, and decreased bacterial load in the mosquito midgut. Our findings suggest that the interactions between A. aquasalis and.Itoring patients after initial diagnosis/surgery. Even though each biomarker investigated in the present work is not exclusively associated with melanoma, their combination reveals a high specificity for melanoma detection.Supporting InformationFigure S1 95 CI of the AUC according to the stage ofdisease. Bonferroni adjusted confidence intervals of the AUC of total cfDNA (Panel A), integrity index 180/67 (Panel B), methylated RASSF1A (Panel C), and BRAFV600E (Panel D) according to the stage of disease. The horizontal dashed line in each Panel represent the AUC value obtained for each biomarker by comparing all cases and controls. (TIF)Table S1 Descriptive Statistics according to the stage ofdisease. (DOC)Author ContributionsConceived and designed the experiments: CO PP. Performed the experiments: FS. Analyzed the data: PV CMC. Contributed reagents/ materials/analysis tools: DM MP. Wrote the paper: PP. Patients enrollment: VDG MG.
The Role of Reactive Oxygen Species in Anopheles aquasalis Response to Plasmodium vivax Infection???Ana C. Bahia1, Jose Henrique M. Oliveira2, Marina S. Kubota1, Helena R. C. Araujo3, Jose B. P. Lima4, ???Claudia Maria Rios-Velasquez5, Marcus Vinicius G. Lacerda6, Pedro L. Oliveira2,7, Yara M. Traub?Cseko1*., Paulo F. P. Pimenta3*.????1 Laboratorio de Biologia Molecular de Parasitas e Vetores, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil, 2 Laboratorio de Bioquimica de Artropodes ? ica, Programa de Biologia Molecular e Biotecnologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil, ?Hematofagos, Instituto de Bioquimica Me ? ica, Instituto Rene Rachou, Belo Horizonte, Brazil, 4 Laboratorio de Fisiologia e Controle de Artropodes Vetores, Instituto Oswaldo Cruz, ???3 Laboratorio de Entomologia Me ?^ ?Fiocruz, Rio de Janeiro, Brazil, 5 Laboratorio de Biodiversidade em Saude, Centro de Pesquisa Leonidas Maria Deane, Fiocruz, Manaus, Brazil, 6 Fundacao de Medicina ^ncia e Tecnologia em Entomologia Molecular, Rio de Janeiro, Brazil Tropical Dr. Heitor Vieira Dourado, Manaus, Brazil, 7 Instituto Nacional de CieAbstractMalaria affects millions of people worldwide and hundreds of thousands of people each year in Brazil. The mosquito Anopheles aquasalis is an important vector of Plasmodium vivax, the main human malaria parasite in the Americas. Reactive oxygen species (ROS) have been shown to have a role in insect innate immune responses as a potent pathogen-killing agent. We investigated the mechanisms of free radicals modulation after A. aquasalis infection with P. vivax. ROS metabolism was evaluated in the vector by studying expression and activity of three key detoxification enzymes, one catalase and two superoxide dismutases (SOD3A and SOD3B). Also, the involvement of free radicals in the mosquito immunity was measured by silencing the catalase gene followed by infection of A. aquasalis with P. vivax. Catalase, SOD3A and SOD3B expression in whole A. aquasalis were at the same levels of controls at 24 h and 
upregulated 36 h after ingestion of blood containing P. vivax. However, in the insect isolated midgut, the mRNA for these enzymes was not regulated by P. vivax infection, while catalase activity was reduced 24 h after the infectious meal. RNAi-mediated silencing of catalase 1527786 reduced enzyme activity in the midgut, resulted in increased P. vivax infection and prevalence, and decreased bacterial load in the mosquito midgut. Our findings suggest that the interactions between A. aquasalis and.