Professional Education

  • PhD, Federal University of Rio de Janeiro, Biochemistry (2011)

Stanford Advisors

Journal Articles

  • An Insight into the Transcriptome of the Digestive Tract of the Bloodsucking Bug, Rhodnius prolixus. PLoS neglected tropical diseases Ribeiro, J. M., Genta, F. A., Sorgine, M. H., Logullo, R., Mesquita, R. D., Paiva-Silva, G. O., Majerowicz, D., Medeiros, M., Koerich, L., Terra, W. R., Ferreira, C., Pimentel, A. C., Bisch, P. M., Leite, D. C., Diniz, M. M., Junior, J. L., Da Silva, M. L., Araujo, R. N., Gandara, A. C., Brosson, S., Salmon, D., Bousbata, S., González-Caballero, N., Silber, A. M., Alves-Bezerra, M., Gondim, K. C., Silva-Neto, M. A., Atella, G. C., Araujo, H., Dias, F. A., Polycarpo, C., Vionette-Amaral, R. J., Fampa, P., Melo, A. C., Tanaka, A. S., Balczun, C., Oliveira, J. H., Gonçalves, R. L., Lazoski, C., Rivera-Pomar, R., Diambra, L., Schaub, G. A., Garcia, E. S., Azambuja, P., Braz, G. R., Oliveira, P. L. 2014; 8 (1): e2594


    The bloodsucking hemipteran Rhodnius prolixus is a vector of Chagas' disease, which affects 7-8 million people today in Latin America. In contrast to other hematophagous insects, the triatomine gut is compartmentalized into three segments that perform different functions during blood digestion. Here we report analysis of transcriptomes for each of the segments using pyrosequencing technology. Comparison of transcript frequency in digestive libraries with a whole-body library was used to evaluate expression levels. All classes of digestive enzymes were highly expressed, with a predominance of cysteine and aspartic proteinases, the latter showing a significant expansion through gene duplication. Although no protein digestion is known to occur in the anterior midgut (AM), protease transcripts were found, suggesting secretion as pro-enzymes, being possibly activated in the posterior midgut (PM). As expected, genes related to cytoskeleton, protein synthesis apparatus, protein traffic, and secretion were abundantly transcribed. Despite the absence of a chitinous peritrophic membrane in hemipterans - which have instead a lipidic perimicrovillar membrane lining over midgut epithelia - several gut-specific peritrophin transcripts were found, suggesting that these proteins perform functions other than being a structural component of the peritrophic membrane. Among immunity-related transcripts, while lysozymes and lectins were the most highly expressed, several genes belonging to the Toll pathway - found at low levels in the gut of most insects - were identified, contrasting with a low abundance of transcripts from IMD and STAT pathways. Analysis of transcripts related to lipid metabolism indicates that lipids play multiple roles, being a major energy source, a substrate for perimicrovillar membrane formation, and a source for hydrocarbons possibly to produce the wax layer of the hindgut. Transcripts related to amino acid metabolism showed an unanticipated priority for degradation of tyrosine, phenylalanine, and tryptophan. Analysis of transcripts related to signaling pathways suggested a role for MAP kinases, GTPases, and LKBP1/AMP kinases related to control of cell shape and polarity, possibly in connection with regulation of cell survival, response of pathogens and nutrients. Together, our findings present a new view of the triatomine digestive apparatus and will help us understand trypanosome interaction and allow insights into hemipteran metabolic adaptations to a blood-based diet.

    View details for DOI 10.1371/journal.pntd.0002594

    View details for PubMedID 24416461

  • The Role of Reactive Oxygen Species in Anopheles aquasalis Response to Plasmodium vivax Infection PLOS ONE Bahia, A. C., Oliveira, J. H., Kubota, M. S., Araujo, H. R., Lima, J. B., Rios-Velasquez, C. M., Lacerda, M. V., Oliveira, P. L., Traub-Csekoe, Y. M., Pimenta, P. F. 2013; 8 (2)


    Malaria 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 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 P. vivax do not follow the model of ROS-induced parasite killing. It appears that P. vivax manipulates the mosquito detoxification system in order to allow its own development. This can be an indirect effect of fewer competitive bacteria present in the mosquito midgut caused by the increase of ROS after catalase silencing. These findings provide novel information on unique aspects of the main malaria parasite in the Americas interaction with one of its natural vectors.

    View details for DOI 10.1371/journal.pone.0057014

    View details for Web of Science ID 000315159200088

    View details for PubMedID 23441231

  • Mitochondrial Reactive Oxygen Species Modulate Mosquito Susceptibility to Plasmodium Infection PLOS ONE Goncalves, R. L., Oliveira, J. H., Oliveira, G. A., Andersen, J. F., Oliveira, M. F., Oliveira, P. L., Barillas-Mury, C. 2012; 7 (7)


    Mitochondria perform multiple roles in cell biology, acting as the site of aerobic energy-transducing pathways and as an important source of reactive oxygen species (ROS) that modulate redox metabolism.We demonstrate that a novel member of the mitochondrial transporter protein family, Anopheles gambiae mitochondrial carrier 1 (AgMC1), is required to maintain mitochondrial membrane potential in mosquito midgut cells and modulates epithelial responses to Plasmodium infection. AgMC1 silencing reduces mitochondrial membrane potential, resulting in increased proton-leak and uncoupling of oxidative phosphorylation. These metabolic changes reduce midgut ROS generation and increase A. gambiae susceptibility to Plasmodium infection.We provide direct experimental evidence indicating that ROS derived from mitochondria can modulate mosquito epithelial responses to Plasmodium infection.

    View details for DOI 10.1371/journal.pone.0041083

    View details for Web of Science ID 000306548900074

    View details for PubMedID 22815925

  • Energy metabolism affects susceptibility of Anopheles gambiae mosquitoes to Plasmodium infection INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY Oliveira, J. H., Goncalves, R. L., Oliveira, G. A., Oliveira, P. L., Oliveira, M. F., Barillas-Mury, C. 2011; 41 (6): 349-355


    Previous studies showed that Anopheles gambiae L3-5 females, which are refractory (R) to Plasmodium infection, express higher levels of genes involved in redox-metabolism and mitochondrial respiration than susceptible (S) G3 females. Our studies revealed that R females have reduced longevity, faster utilization of lipid reserves, impaired mitochondrial state-3 respiration, increased rate of mitochondrial electron leak and higher expression levels of several glycolytic enzyme genes. Furthermore, when state-3 respiration was reduced in S females by silencing expression of the adenine nucleotide translocator (ANT), hydrogen peroxide generation was higher and the mRNA levels of lactate dehydrogenase increased in the midgut, while the prevalence and intensity of Plasmodium berghei infection were significantly reduced. We conclude that there are broad metabolic differences between R and S An. gambiae mosquitoes that influence their susceptibility to Plasmodium infection.

    View details for DOI 10.1016/j.ibmb.2011.02.001

    View details for Web of Science ID 000290355300001

    View details for PubMedID 21320598

  • Blood Meal-Derived Heme Decreases ROS Levels in the Midgut of Aedes aegypti and Allows Proliferation of Intestinal Microbiota PLOS PATHOGENS Oliveira, J. H., Goncalves, R. L., Lara, F. A., Dias, F. A., Gandara, A. C., Menna-Barreto, R. F., Edwards, M. C., Laurindo, F. R., Silva-Neto, M. A., Sorgine, M. H., Oliveira, P. L. 2011; 7 (3)


    The presence of bacteria in the midgut of mosquitoes antagonizes infectious agents, such as Dengue and Plasmodium, acting as a negative factor in the vectorial competence of the mosquito. Therefore, knowledge of the molecular mechanisms involved in the control of midgut microbiota could help in the development of new tools to reduce transmission. We hypothesized that toxic reactive oxygen species (ROS) generated by epithelial cells control bacterial growth in the midgut of Aedes aegypti, the vector of Yellow fever and Dengue viruses. We show that ROS are continuously present in the midgut of sugar-fed (SF) mosquitoes and a blood-meal immediately decreased ROS through a mechanism involving heme-mediated activation of PKC. This event occurred in parallel with an expansion of gut bacteria. Treatment of sugar-fed mosquitoes with increased concentrations of heme led to a dose dependent decrease in ROS levels and a consequent increase in midgut endogenous bacteria. In addition, gene silencing of dual oxidase (Duox) reduced ROS levels and also increased gut flora. Using a model of bacterial oral infection in the gut, we show that the absence of ROS resulted in decreased mosquito resistance to infection, increased midgut epithelial damage, transcriptional modulation of immune-related genes and mortality. As heme is a pro-oxidant molecule released in large amounts upon hemoglobin degradation, oxidative killing of bacteria in the gut would represent a burden to the insect, thereby creating an extra oxidative challenge to the mosquito. We propose that a controlled decrease in ROS levels in the midgut of Aedes aegypti is an adaptation to compensate for the ingestion of heme.

    View details for DOI 10.1371/journal.ppat.1001320

    View details for Web of Science ID 000288994900014

    View details for PubMedID 21445237

  • Antioxidant pathways are up-regulated during biological nitrogen fixation to prevent ROS-induced nitrogenase inhibition in Gluconacetobacter diazotrophicus ARCHIVES OF MICROBIOLOGY Alqueres, S. M., Oliveira, J. H., Nogueira, E. M., Guedes, H. V., Oliveira, P. L., Camara, F., Baldani, J. I., Martins, O. B. 2010; 192 (10): 835-841


    Gluconacetobacter diazotrophicus, an endophyte isolated from sugarcane, is a strict aerobe that fixates N(2). This process is catalyzed by nitrogenase and requires copious amounts of ATP. Nitrogenase activity is extremely sensitive to inhibition by oxygen and reactive oxygen species (ROS). However, the elevated oxidative metabolic rates required to sustain biological nitrogen fixation (BNF) may favor an increased production of ROS. Here, we explored this paradox and observed that ROS levels are, in fact, decreased in nitrogen-fixing cells due to the up-regulation of transcript levels of six ROS-detoxifying genes. A cluster analyses based on common expression patterns revealed the existence of a stable cluster with 99.8% similarity made up of the genes encoding the ?-subunit of nitrogenase Mo-Fe protein (nifD), superoxide dismutase (sodA) and catalase type E (katE). Finally, nitrogenase activity was inhibited in a dose-dependent manner by paraquat, a redox cycler that increases cellular ROS levels. Our data revealed that ROS can strongly inhibit nitrogenase activity, and G. diazotrophicus alters its redox metabolism during BNF by increasing antioxidant transcript levels resulting in a lower ROS generation. We suggest that careful controlled ROS production during this critical phase is an adaptive mechanism to allow nitrogen fixation.

    View details for DOI 10.1007/s00203-010-0609-1

    View details for Web of Science ID 000281842600006

    View details for PubMedID 20697694

  • Blood-Feeding Induces Reversible Functional Changes in Flight Muscle Mitochondria of Aedes aegypti Mosquito PLOS ONE Goncalves, R. L., Machado, A. C., Paiva-Silva, G. O., Sorgine, M. H., Momoli, M. M., Oliveira, J. H., Vannier-Santos, M. A., Galina, A., Oliveira, P. L., Oliveira, M. F. 2009; 4 (11)


    Hematophagy poses a challenge to blood-feeding organisms since products of blood digestion can exert cellular deleterious effects. Mitochondria perform multiple roles in cell biology acting as the site of aerobic energy-transducing pathways, and also an important source of reactive oxygen species (ROS), modulating redox metabolism. Therefore, regulation of mitochondrial function should be relevant for hematophagous arthropods. Here, we investigated the effects of blood-feeding on flight muscle (FM) mitochondria from the mosquito Aedes aegypti, a vector of dengue and yellow fever.Blood-feeding caused a reversible reduction in mitochondrial oxygen consumption, an event that was parallel to blood digestion. These changes were most intense at 24 h after blood meal (ABM), the peak of blood digestion, when oxygen consumption was inhibited by 68%. Cytochromes c and a+a(3) levels and cytochrome c oxidase activity of the electron transport chain were all reduced at 24 h ABM. Ultrastructural and molecular analyses of FM revealed that mitochondria fuse upon blood meal, a condition related to reduced ROS generation. Consistently, BF induced a reversible decrease in mitochondrial H(2)O(2) formation during blood digestion, reaching their lowest values at 24 h ABM where a reduction of 51% was observed.Blood-feeding triggers functional and structural changes in hematophagous insect mitochondria, which may represent an important adaptation to blood feeding.

    View details for DOI 10.1371/journal.pone.0007854

    View details for Web of Science ID 000271851000011

    View details for PubMedID 19924237

  • Trypanosoma brucei brucei: Effects of ferrous iron and heme on ecto-nucleoside triphosphate diphosphohydrolase activity EXPERIMENTAL PARASITOLOGY Leite, M. S., Thomaz, R., Oliveira, J. H., Oliveira, P. L., Meyer-Femandes, J. R. 2009; 121 (2): 137-143


    Trypanosoma brucei brucei is the causative agent of animal African trypanosomiasis, also called nagana. Procyclic vector form resides in the midgut of the tsetse fly, which feeds exclusively on blood. Hemoglobin digestion occurs in the midgut resulting in an intense release of free heme. In the present study we show that the magnesium-dependent ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase) activity of procyclic T. brucei brucei is inhibited by ferrous iron and heme. The inhibition of E-NTPDase activity by ferrous iron, but not by heme, was prevented by pre-incubation of cells with catalase. However, antioxidants that permeate cells, such as PEG-catalase and N-acetyl-cysteine prevented the inhibition of E-NTPDase by heme. Ferrous iron was able to induce an increase in lipid peroxidation, while heme did not. Therefore, both ferrous iron and heme can inhibit E-NTPDase activity of T. brucei brucei by means of formation of reactive oxygen species, but apparently acting through distinct mechanisms.

    View details for DOI 10.1016/j.exppara.2008.10.018

    View details for Web of Science ID 000262658900004

    View details for PubMedID 19027737