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Made available in DSpace on 2021-06-25T15:03:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-04-16 Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Leprosy is a disease with a clinical spectrum of presentations that is also manifested in diverse histological features. At one pole, lepromatous lesions (L-pole) have phagocytic foamy macrophages heavily parasitized with freely multiplying intracellular Mycobacterium leprae. At the other pole, the presence of epithelioid giant cells and granulomatous formation in tuberculoid lesions (T-pole) lead to the control of M. leprae replication and the containment of its spread. The mechanism that triggers this polarization is unknown, but macrophages are central in this process. Over the past few years, leprosy has been studied using large scale techniques to shed light on the basic pathways that, upon infection, rewire the host cellular metabolism and gene expression. M. leprae is particularly peculiar as it invades Schwann cells in the nerves, reprogramming their gene expression leading to a stem-like cell phenotype. This modulatory behavior exerted by M. leprae is also observed in skin macrophages. Here, we used live M. leprae to infect (10:1 multiplicity of infection) monocyte-derived macrophages (MDMs) for 48 h and analyzed the whole gene expression profile using microarrays. In this model, we observe an intense upregulation of genes consistent with a cellular immune response, with enriched pathways including peptide and protein secretion, leukocyte activation, inflammation, and cellular divalent inorganic cation homeostasis. Among the most differentially expressed genes (DEGs) are CCL5/RANTES and CYP27B1, and several members of the metallothionein and metalloproteinase families. This is consistent with a proinflammatory state that would resemble macrophage rewiring toward granulomatous formation observed at the T-pole. Furthermore, a comparison with a dataset retrieved from the Gene Expression Omnibus of M. leprae-infected Schwann cells (MOI 100:1) showed that the patterns among the DEGs are highly distinct, as the Schwann cells under these conditions had a scavenging and phagocytic gene profile similar to M2-like macrophages, with enriched pathways rearrangements in the cytoskeleton, lipid and cholesterol metabolism and upregulated genes including MVK, MSMO1, and LACC1/FAMIN. In summary, macrophages may have a central role in defining the paradigmatic cellular (T-pole) vs. humoral (L-pole) responses and it is likely that the multiplicity of infection and genetic polymorphisms in key genes are gearing this polarization. Fiocruz MS, Lab Hanseniase, Inst Oswaldo Cruz, Rio De Janeiro, Brazil Inst Lauro de Souza Lima, Div Pesquisa & Ensino, Bauru, SP, Brazil Univ Estadual Paulista, Fac Med Botucatu, Dept Doencas Trop, Botucatu, SP, Brazil Univ Estadual Paulista, Fac Med Botucatu, Dept Doencas Trop, Botucatu, SP, Brazil FAPERJ: E_09/2019 FAPERJ: E_34/2014 -PENSA RIO CNPq: 313657/2018-1 CNPq: 4000170/20172 FAPESP: 2015/01744-9

Made available in DSpace on 2018-11-26T17:06:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-11-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Alexander von Humboldt Foundation This study reports three interaction events between the whale shark (Rhincodon typus) and fisheries in Brazilian waters. Two events were described as incidental captures in a gillnet fishery on the northern coast of Rio de Janeiro State, in southeastern Brazil. The last event took place at the Royal Charlotte Oceanic Bank (RCOB), in the southern region of Bahia State, where five fishermen filmed their interaction with a juvenile specimen. For one of the incidental captures, the fish was landed and consumed by locals, and for the other, the shark was released. In the RCOB, the fishermen touched the whale shark with their hands, wooden sticks and even rode on it. In addition, 74 interviews were conducted with fishermen from Bahia about sightings and possible interactions with R. typus. According to statements made by the fishermen, although not frequent, the conduct reported for the RCOB occasionally takes place in the region. Evaluating information of this nature is important to support government plans that regulate fishing activities in order to reduce incidental captures and the harassment of whale sharks. Encouraging the participation of fishermen in a collaborative monitoring program for R. typus may be a good way to better understand the threats to the species at a reduced cost, particularly for developing countries, such as Brazil. (C) 2016 Elsevier Ltd. All rights reserved. Univ Estadual Santa Cruz, Dept Ciencias Biol, Programa Posgrad Zool, Rodovia Jorge Amado,Km 16, BR-45662900 Ilheus, BA, Brazil Univ Estadual Norte Fluminense Darcy Ribeiro UENF, Av Alberto Lamego 2000,Parque Calif, BR-28013602 Campos Dos Goytacazes, RJ, Brazil Fiocruz MS, Inst Oswaldo Cruz, Pavilhao Mourisco,Sala 122,Av Brasil 4-365, BR-21040360 Rio De Janeiro, RJ, Brazil Leibniz Ctr Trop Marine Ecol ZMT, Syst Ecol Grp, Fahrenheitstr 6, D-28359 Bremen, Germany Univ Estadual Feira de Santana, Dept Ciencias Biol, Ave Transnordestina S-N, BR-44036900 Feira De Santana, BA, Brazil Univ Fed Sao Paulo, Inst Mar, Lab Ecol & Conservacao Marinha, Santos, Brazil Univ Estadual Paulista, Lab Pesquisa Elasmobranquios, Sao Vicente, Brazil Univ Estadual Paulista, Lab Pesquisa Elasmobranquios, Sao Vicente, Brazil Alexander von Humboldt Foundation: BEX 0128/14-7

Made available in DSpace on 2020-12-11T17:20:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-12-12 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Background Arboviruses and protozoans can cause neurologic disorders in horses. In Brazilian Amazon, several horses presenting signs compatible with disorders caused by these infectious agents have been observed. Objective To contribute to the knowledge of this epidemiological picture, we sought to construct a serological diagnostic panel for neurotrophic infectious agents in local horses. Material and methods A total of 213 blood samples from horses were collected from 29 farms in three municipalities. Samples were evaluated and considered positive when they met the following criteria: titers >= 1:80 with the indirect fluorescent antibody test (IFAT) for apicomplexan protozoans; positive recombinant enzyme-linked immunosorbent assay (ELISA) with subsequent titers >= 1:10 by the PRNt for viruses; and detection under direct microscopic examination for Trypanosoma evansi. Results No horses were found to be infected by T. evansi, and only two were infected Toxoplasma gondii and/or Neospora spp. The highest protozoan infection rate was observed for Sarcocystis neurona (40.3%; n = 86/213). Among the positive ELISA samples tested by the plaque reduction neutralization test (PRNT90), 92% (n = 76/83) were positive for St Louis Encephalitis virus, 43% (n = 6/14) were positive for West Nile virus and 33% (n = 16/48) were positive for Mayaro virus. Eighteen percent (n = 39/213) of horses were co-infected by S. neurona and at least one arbovirus, particularly SLEV and/or MAYV. Conclusion Samples positive for SLEV associated with S. neurona, including samples from horses that had recovered from neurological signs were frequent, and must be considered when investigating the possible causes of neurological diseases in South Roraima horses. Fiocruz MS, Oswaldo Cruz Inst, Lab Trypanosomatid Biol, Rio De Janeiro, RJ, Brazil Fed Inst Educ Sci & Technol Roraima, Caracarai, Roraima, Brazil Sao Paulo State Univ, Sch Agr & Vet Studies Jaboticabal, Jaboticabal, SP, Brazil Univ Sao Paulo, Sch Vet & Anim Sci, Dept Prevent Vet & Anim Hlth, Sao Paulo, Brazil Univ Sao Paulo, Ribeirao Preto Med Sch, Virol Res Ctr, Ribeirao Preto, Brazil Sao Paulo State Univ, Sch Agr & Vet Studies Jaboticabal, Jaboticabal, SP, Brazil CAPES: 001

Made available in DSpace on 2018-11-28T00:18:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-12-06 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundacao Oswaldo Cruz - Programa de Apoio a Pesquisas Estrategicas (FIOCRUZ-PAPES IV) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) The treatment of leishmaniasis still relies on drugs with potentially serious adverse effects. Herein, we tested a topical formulation of bacterial cellulose (BC) membranes containing Diethyldithiocarbamate (DETC), a superoxide dismutase 1 inhibitor. Leishmania-infected macrophages exposed to BC-DETC resulted in parasite killing, without pronounced toxic effects to host cells. This outcome was associated with lower SOD1 activity and higher production of superoxide and cytokine mediators. Topical application of BC-DETC significantly decreased lesion size, parasite load and the inflammatory response at the infection site, as well as the production of both IFN-gamma and TNF. Combination of topical BC-DETC plus intraperitoneal Sb-v also significantly reduced disease development and parasite load. The leishmanicidal effect of BC-DETC was extended to human macrophages infected with L. braziliensis, highlighting the feasibility of BC-DETC as a topical formulation for chemotherapy of cutaneous leishmaniasis caused by L. braziliensis. Fiocruz MS, Inst Goncalo Moniz, Salvador, BA, Brazil Univ Estadual Paulista, Inst Quim, Araraquara, SP, Brazil Univ Araraquara UNIARA, Araraquara, SP, Brazil Katholieke Univ Leuven, Rega Inst Med Res, Dept Microbiol & Immunol, Leuven, Belgium INCT, Inst Invest Imunol 3, Sao Paulo, Brazil Univ Estadual Paulista, Inst Quim, Araraquara, SP, Brazil CNPq: 482193/2012-3 Fundacao Oswaldo Cruz - Programa de Apoio a Pesquisas Estrategicas (FIOCRUZ-PAPES IV): 407437/2012-6 CNPq: 161535/2011-9

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) NIH Center for AIDS Research at Einstein In prior studies, we demonstrated that glucuronoxylomannan (GXM), the major capsular polysaccharide of the fungal pathogen Cryptococcus neoformans, interacts with chitin oligomers at the cell wall-capsule interface. the structural determinants regulating these carbohydrate-carbohydrate interactions, as well as the functions of these structures, have remained unknown. in this study, we demonstrate that glycan complexes composed of chitooligomers and GXM are formed during fungal growth and macrophage infection by C. neoformans. To investigate the required determinants for the assembly of chitin-GXM complexes, we developed a quantitative scanning electron microscopy-based method using different polysaccharide samples as inhibitors of the interaction of chitin with GXM. This assay revealed that chitin-GXM association involves noncovalent bonds and large GXM fibers and depends on the N-acetyl amino group of chitin. Carboxyl and O-acetyl groups of GXM are not required for polysaccharide-polysaccharide interactions. Glycan complex structures composed of cryptococcal GXM and chitin-derived oligomers were tested for their ability to induce pulmonary cytokines in mice. They were significantly more efficient than either GXM or chitin oligomers alone in inducing the production of lung interleukin 10 (IL-10), IL-17, and tumor necrosis factor alpha (TNF-alpha). These results indicate that association of chitin-derived structures with GXM through their N-acetyl amino groups generates glycan complexes with previously unknown properties. Univ Fed Rio de Janeiro, Inst Microbiol Prof Paulo de Goes, Rio de Janeiro, Brazil Univ Fed Rio de Janeiro, Inst Biofis Carlos Chagas Filho, Lab Ultraestrutura Celular Hertha Meyer, BR-21941 Rio de Janeiro, Brazil Albert Einstein Coll Med, Dept Microbiol & Immunol, Bronx, NY 10467 USA Albert Einstein Coll Med, Div Infect Dis, Dept Med, Bronx, NY 10467 USA Universidade Federal de São Paulo, Disciplina Biol Celular, São Paulo, Brazil Fiocruz MS, Fundacao Oswaldo Cruz, Ctr Desenvolvimento Tecnol, BR-21045900 Rio de Janeiro, Brazil Universidade Federal de São Paulo, Disciplina Biol Celular, São Paulo, Brazil NIH: AI033142 NIH: AI033774 NIH: AI052733 NIH: HL059842 Web of Science

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Millennium Institute for Gene Therapy (Brazil) Recently, we described a heterologous prime-boost strategy using plasmid DNA followed by replication-defective human recombinant adenovirus type 5 as a powerful strategy to elicit long-lived CD8(+) T-cell-mediated protective immunity against experimental systemic infection of mice with a human intracellular protozoan parasite, Trypanosoma cruzi. in the present study, we further characterized the protective long-lived CD8(+) T cells. We compared several functional and phenotypic aspects of specific CD8(+) T cells present 14 or 98 days after the last immunizing dose and found the following: (i) the numbers of specific cells were similar, as determined by multimer staining or by determining the number of gamma interferon (IFN-gamma)-secreting cells by enzyme-linked immunospot (ELISPOT) assay; (ii) these cells were equally cytotoxic in vivo; (iii) following in vitro stimulation, a slight decline in the frequency of multifunctional cells (CD107a(+) IFN-gamma(+) or CD107a(+) IFN-gamma(+) tumor necrosis factor alpha positive [TNF-alpha(+)]) was paralleled by a significant increase of CD107a singly positive cells after 98 days; (iv) the expression of several surface markers was identical, except for the reexpression of CD127 after 98 days; (v) the use of genetically deficient mice revealed a role for interleukin-12 (IL-12)/IL-23, but not IFN-gamma, in the maintenance of these memory cells; and (vi) subsequent immunizations with an unrelated virus or a plasmid vaccine or the depletion of CD4(+) T cells did not significantly erode the number or function of these CD8(+) T cells during the 15-week period. From these results, we concluded that heterologous plasmid DNA prime-adenovirus boost vaccination generated a stable pool of functional protective long-lived CD8(+) T cells with an effector memory phenotype. Universidade Federal de São Paulo, Escola Paulista Med, UNIFESP, CTCMol, BR-04044010 São Paulo, Brazil Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, BR-04044010 São Paulo, Brazil Fiocruz MS, Ctr Pesquisas Rene Rachou, BR-30190002 Belo Horizonte, MG, Brazil Univ Fed Minas Gerais, Inst Ciencias Biol, Dept Bioquim & Imunol, BR-31270901 Belo Horizonte, MG, Brazil Univ Massachusetts, Sch Med, Dept Med, Div Infect Dis & Immunol, Worcester, MA 01655 USA Univ Fed Minas Gerais, Dept Microbiol, Inst Ciencias Biol, BR-31270901 Belo Horizonte, MG, Brazil Universidade Federal de São Paulo, Escola Paulista Med, UNIFESP, CTCMol, BR-04044010 São Paulo, Brazil Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, BR-04044010 São Paulo, Brazil FAPESP: 2006/1983-4 FAPESP: 2009/06820-4 CNPq: 420067/2005-1 Web of Science

The dynamics of dengue virus (DENV) circulation depends on serotype, genotype and lineage replacement and turnover. In São José do Rio Preto, Brazil, we observed that the L6 lineage of DENV-1 (genotype V) remained the dominant circulating lineage even after the introduction of the L1 lineage. We investigated viral fitness and immunogenicity of the L1 and L6 lineages and which factors interfered with the dynamics of DENV epidemics. The results showed a more efficient replicative fitness of L1 over L6 in mosquitoes and in human and non-human primate cell lines. Infections by the L6 lineage were associated with reduced antigenicity, weak B and T cell stimulation and weak host immune system interactions, which were associated with higher viremia. Our data, therefore, demonstrate that reduced viral immunogenicity and consequent greater viremia determined the increased epidemiological fitness of DENV-1 L6 lineage in São José do Rio Preto., Author summary Since 2008 L6 is the endemic lineage that has circulated in SJRP. In 2010, the L1 lineage was first identified in the city. For a period, both lineages co-circulated, and then in 2013, L1 began to diminish until it was no longer detected in the population. Differences in replicative fitness are usually the main factor for clade replacement (CR) in DENV epidemics. However, despite the better viral fitness of the emerging lineage, the absence of CR could not be explained by these differences alone. Here, we combine epidemiological, phylogenetic, molecular and immunological analyses to provide a more precise understanding of the role of fitness in lineage dynamics with the persistence of L6 even after the introduction of L1 without CR. Differences in immune responses elicited by DENV-1 L1 and L6 lineages (genotype V), but not viral fitness in mosquito or human cells, explain the dynamics of circulating DENV in a city of Brazil.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2014-02-26T17:16:52Z No. of bitstreams: 1 WOS000185425500016.pdf: 436693 bytes, checksum: 6e53e418baab576ea898d4d9383e0c12 (MD5) Made available in DSpace on 2014-02-26T17:16:52Z (GMT). No. of bitstreams: 1 WOS000185425500016.pdf: 436693 bytes, checksum: 6e53e418baab576ea898d4d9383e0c12 (MD5) Previous issue date: 2003-10-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T14:02:23Z No. of bitstreams: 1 WOS000185425500016.pdf: 436693 bytes, checksum: 6e53e418baab576ea898d4d9383e0c12 (MD5) Made available in DSpace on 2014-05-20T14:02:24Z (GMT). No. of bitstreams: 1 WOS000185425500016.pdf: 436693 bytes, checksum: 6e53e418baab576ea898d4d9383e0c12 (MD5) Previous issue date: 2003-10-01 BaP1 is a 22.7-kD P-I-type zinc-dependent metalloproteinase isolated from the venom of the snake Bothrops asper, a medically relevant species in Central America. This enzyme exerts multiple tissue-damaging activities, including hemorrhage, myonecrosis, dermonecrosis, blistering, and edema. BaP1 is a single chain of 202 amino acids that shows highest sequence identity with metalloproteinases isolated front the venoms of snakes of the subfamily Crotalinae. It has six Cys residues involved in three disulfide bridges (Cys 117-Cys 197, Cys 159-Cys 181, Cys 157-Cys 164). It has the consensus sequence H(142)E(143)XXH(146)XXGXXH(152), as well as the sequence C164I165M166, which characterize the metzincin superfamily of metalloproteinases. The active-site cleft separates a major subdomain (residues 1-152), comprising four a-helices and a five-stranded beta-sheet, from the minor subdomain, which is formed by a single a-helix and several loops. The catalytic zinc ion is coordinated by the N-epsilon2 nitrogen atoms of His 142, His 146, and His 152, in addition to a solvent water molecule, which in turn is bound to Glu 143. Several conserved residues contribute to the formation of the hydrophobic pocket, and Met 166 serves as a hydrophobic base for the active-site groups. Sequence and structural comparisons of hemorrhagic and nonhemorrhagic P-I metalloproteinases from snake venoms revealed differences in several regions. In particular, the loop comprising residues 153 to 176 has marked structural differences between metalloproteinases with very different hemorrhagic activities. Because this region lies in close proximity to the active-site microenvironment, it may influence the interaction of these enzymes with physiologically relevant substrates in the extracellular matrix. Univ Costa Rica, Fac Microbiol, Inst Clodomiro Picado, San Jose, Costa Rica Univ Costa Rica, Escuela Med, Dept Bioquim, San Jose, Costa Rica Univ Liverpool, Royal Liverpool Univ Hosp, Dept Haematol, Liverpool, Merseyside, England Univ Liverpool, Liverpool Sch Trop Med, Venom Res Unit, Liverpool L3 5QA, Merseyside, England Fiocruz MS, Dept Fisiol & Farmacodinam, BR-21045900 Rio de Janeiro, Brazil Univ Virginia, Hlth Sci Ctr, Dept Microbiol, Charlottesville, VA 22908 USA UNESP, IBILCE, Dept Phys, BR-15054000 Sao Jose do Rio Preto, Brazil UNESP, IBILCE, Dept Phys, BR-15054000 Sao Jose do Rio Preto, Brazil

Made available in DSpace on 2014-12-03T13:08:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-11-01Bitstream added on 2014-12-03T13:24:42Z : No. of bitstreams: 1 WOS000328079200016.pdf: 265403 bytes, checksum: 8db1180d1a3f1a388e04e74b78254a54 (MD5) Since the emergence of West Nile virus (WNV) in North America in 1999, there have been several reports of WNV activity in Central and South American countries. To detect WNV in Brazil, we performed a serological survey of horses from different regions of Brazil using recombinant peptides from domain III of WNV. Positive samples were validated with the neutralisation test. Our results showed that of 79 ELISA-positive horses, nine expressed WNV-specific neutralising antibodies. Eight of the infected horses were from the state of Mato Grosso do Sul and one was from the state of Paraiba. Our results provide additional evidence for the emergence of WNV in Brazil and for its circulation in multiple regions of the country. Univ Sao Paulo, Fac Med Ribeirao Preto, Ctr Pesquisa Virol, BR-14049 Ribeirao Preto, SP, Brazil Fiocruz MS, Inst Oswaldo Cruz, Dept Virol, BR-21045900 Rio De Janeiro, RJ, Brazil Univ Julio de Mesquita Filho, Fac Ciencias Vet, Jaboticabal, SP, Brazil Univ Fed Mato Grosso do Sul, Escola Ciencias Vet, Campo Grande, MS, Brazil Univ Fed Campina Grande, Campina Grande, PB, Brazil Univ Uberaba, Uberaba, MG, Brazil Acad Mil Agulhas Negras, Resende, RJ, Brazil Univ Julio de Mesquita Filho, Fac Ciencias Vet, Jaboticabal, SP, Brazil FAPESP: 08/50167-6

Made available in DSpace on 2015-10-21T20:56:59Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-03-24. Added 1 bitstream(s) on 2015-10-22T09:52:42Z : No. of bitstreams: 1 WOS000353889600144.pdf: 503172 bytes, checksum: c34e46d02eedd63a2e54d831b00aa1d2 (MD5) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Pro-Reitoria de Pesquisa/UNESP (ProPe) Several differences concerning bacterial species, opportunistic pathogens, elements of the resistome as well as variations concerning the CFU/mL counts were identified in some of the five most marketed bottled mineral water from Araraquara city, Sao Paulo, Brazil. Two out of five brands tested were confirmed as potential source of opportunistic pathogens, including Mycobacterium gordonae, Ralstonia picketti and Burkholderia cepacia complex (Bcc). A total of one hundred and six isolates were recovered from four of these bottled mineral water brands. Betaproteobacteria was predominant followed by Alphaproteobacteria, Gammaproteobacteria and Firmicutes. Ninety percent of the bacteria isolated demonstrated resistance to seventeen of the nineteen antimicrobials tested. These antimicrobials included eight different classes, including 3rd and 4th generation cephalosporins, carbapenems and fluoroquinolones. Multidrug resistant bacteria were detected for fifty-nine percent of isolates in three water brands at counts up to 103 CFU/ml. Of major concern, the two bottled mineral water harboring opportunistic pathogens were also source of elements of the resistome that could be directly transferred to humans. All these differences found among brands highlight the need for continuous bacteriological surveillance of bottled mineral water. Univ Estadual Paulista, Sch Pharmaceut Sci, Araraquara, SP, Brazil Univ Buenos Aires, Fac Med, Dept Microbiol, Buenos Aires, DF, Argentina Fiocruz MS, ILMD, Manaus, Amazonas, Brazil Univ Estadual Paulista, Faculdade de Ciências Farmacêuticas de Araraquara, Departamento de Ciências Biológicas, Araraquara, SP, Brazil CAPES: 18207/12-0 ProPe: 05/2012

Background Understanding the mechanisms that influence the population dynamics and spatial genetic structure of the vectors of pathogens infecting humans is a central issue in tropical epidemiology. In view of the rapid changes in the features of landscape pathogen vectors live in, this issue requires new methods that consider both natural and human systems and their interactions. In this context, individual-based model (IBM) simulations represent powerful yet poorly developed approaches to explore the response of pathogen vectors in heterogeneous social-ecological systems, especially when field experiments cannot be performed. Methodology/Principal Findings We first present guidelines for the use of a spatially explicit IBM, to simulate population genetics of pathogen vectors in changing landscapes. We then applied our model with Triatoma brasiliensis, originally restricted to sylvatic habitats and now found in peridomestic and domestic habitats, posing as the most important Trypanosoma cruzi vector in Northeastern Brazil. We focused on the effects of vector migration rate, maximum dispersal distance and attraction by domestic habitat on T. brasiliensis population dynamics and spatial genetic structure. Optimized for T. brasiliensis using field data pairwise fixation index (FST) from microsatellite loci, our simulations confirmed the importance of these three variables to understand vector genetic structure at the landscape level. We then ran prospective scenarios accounting for land-use change (deforestation and urbanization), which revealed that human-induced land-use change favored higher genetic diversity among sampling points. Conclusions/Significance Our work shows that mechanistic models may be useful tools to link observed patterns with processes involved in the population genetics of tropical pathogen vectors in heterogeneous social-ecological landscapes. Our hope is that our study may provide a testable and applicable modeling framework to a broad community of epidemiologists for formulating scenarios of landscape change consequences on vector dynamics, with potential implications for their surveillance and control., Author Summary Worldwide, humans are modifying landscapes at an unprecedented rate. These modifications have an influence on the ecology of pathogen vectors, yet this issue has received relatively little input from modeling research. The current study presents guidelines for the use of a modeling framework for the representation of the dynamics and spatial genetic structure of pathogen vectors. It allows considering spatiotemporal landscape modifications explicitly, to represent human-altered modifications and consequences. We applied this modeling framework to Triatoma brasiliensis, vector of the pathogen Trypanosoma cruzi responsible for the Chagas disease, in the semi-arid Northeastern Brazil. Using field data of pairwise fixation index (FST) from microsatellite loci, we found that migration rate, maximum dispersal distance and attraction by domestic habitat were all key parameters to understand vector spatial genetic structure at the landscape level. At the interface across disciplines, this study provides to the community of epidemiologists a testable and applicable framework to foresee landscape modification consequences on vector dynamics and genetic structure, with potential implications for their surveillance and control.

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) In earlier studies, we reported that a heterologous prime-boost regimen using recombinant plasmid DNA followed by replication-defective adenovirus vector, both containing Trypanosoma cruzi genes encoding trans-sialidase (TS) and amastigote surface protein (ASP) 2, provided protective immunity against experimental infection with a reticulotropic strain of this human protozoan parasite. Herein, we tested the outcome of genetic vaccination of F1 (CB10XBALB/c) mice challenged with myotropic parasite strains (Brazil and Colombian). Initially, we determined that the coadministration during priming of a DNA plasmid containing the murine IL-12 gene improved the immune response and was essential for protective immunity elicited by the heterologous prime-boost regimen in susceptible male mice against acute lethal infections with these parasites. the prophylactic or therapeutic vaccination of resistant female mice led to a drastic reduction in the number of inflammatory infiltrates in cardiac and skeletal muscles during the chronic phase of infection with either strain. Analysis of the electrocardiographic parameters showed that prophylactic vaccination reduced the frequencies of sinus arrhythmia and atrioventricular block. Our results confirmed that prophylactic vaccination using the TS and ASP-2 genes benefits the host against acute and chronic pathologies caused by T. cruzi and should be further evaluated for the development of a veterinary or human vaccine against Chagas disease. Universidade Federal de São Paulo, Escola Paulista Med, Ctr Terapia Celular & Mol CTCMol, BR-04044010 São Paulo, Brazil Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Immunol & Parasitol, BR-04044010 São Paulo, Brazil Fiocruz MS, Inst Oswaldo Cruz, Lab Biol Celular, BR-21040360 Rio de Janeiro, RJ, Brazil Fiocruz MS, Ctr Pesquisas Goncalo Moniz, BR-40296710 Salvador, BA, Brazil Hosp Sao Rafael, BR-41253190 Salvador, BA, Brazil UNIFESP, Inst Saude Soc, Dept Biociencias, BR-11015020 Santos, SP, Brazil Fiocruz MS, Ctr Pesquisas Rene Rachou, BR-30190002 Belo Horizonte, MG, Brazil Univ Fed Minas Gerais, Inst Ciencias Biol, Dept Bioquim & Imunol, BR-31270901 Belo Horizonte, MG, Brazil Univ Massachusetts, Sch Med, Dept Med, Div Infect Dis & Immunol, Worcester, MA 01655 USA Univ Fed Santa Catarina, Dept Microbiol Immunol & Parasitol, BR-88040900 Florianopolis, SC, Brazil Universidade Federal de São Paulo, Escola Paulista Med, Ctr Terapia Celular & Mol CTCMol, BR-04044010 São Paulo, Brazil Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Immunol & Parasitol, BR-04044010 São Paulo, Brazil UNIFESP, Inst Saude Soc, Dept Biociencias, BR-11015020 Santos, SP, Brazil FAPESP: 2009/06820-4 FAPESP: 2013/13668/0 FAPESP: 2012/22514-3 Web of Science

Made available in DSpace on 2014-12-03T13:08:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-08-07Bitstream added on 2014-12-03T13:24:38Z : No. of bitstreams: 1 WOS000323109700082.pdf: 1393704 bytes, checksum: 25bc393488fc34ae69a2e38b45a346c2 (MD5) NPRP grant from the Qatar National Research Fund (a member of Qatar foundation) NCF (Netherlands Computer Facility Foundation) NWO (Nederlandse Organisatie voor Wetenschappelijk Onderzoek) Odo van Vloten Foundation Over the past two decades, several fungal outbreaks have occurred, including the high-profile 'Vancouver Island' and 'Pacific Northwest' outbreaks, caused by Cryptococcus gattii, which has affected hundreds of otherwise healthy humans and animals. Over the same time period, C. gattii was the cause of several additional case clusters at localities outside of the tropical and subtropical climate zones where the species normally occurs. In every case, the causative agent belongs to a previously rare genotype of C. gattii called AFLP6/VGII, but the origin of the outbreak clades remains enigmatic. Here we used phylogenetic and recombination analyses, based on AFLP and multiple MLST datasets, and coalescence gene genealogy to demonstrate that these outbreaks have arisen from a highly-recombining C. gattii population in the native rainforest of Northern Brazil. Thus the modern virulent C. gattii AFLP6/VGII outbreak lineages derived from mating events in South America and then dispersed to temperate regions where they cause serious infections in humans and animals. CBS KNAW Fungal Biodivers Ctr, Dept Yeast & Basidiomycete Res, Utrecht, Netherlands Canisius Wilhelmina Hosp, Dept Med Microbiol & Infect Dis, Nijmegen, Netherlands Univ Sao Paulo State, Dept Phytopathol, UNESP, Ilha Solteira, Brazil Hadassah Hebrew Univ Med Ctr, Dept Clin Microbiol & Infect Dis, Jerusalem, Israel Univ Birmingham, Sch Biosci, Birmingham, W Midlands, England Univ Ghent, Fac Pharmaceut Sci, Lab Pharmaceut Biotechnol, B-9000 Ghent, Belgium Ctr Genom Regulat, Barcelona, Spain UPF Doctor Aiguader, Barcelona, Spain Netherlands Inst Ecol NIOO KNAW, Ctr Limnol, Wageningen, Netherlands Ctr Wiskunde & Informat, Amsterdam, Netherlands Vrije Univ Amsterdam, Dept Operat Res, Amsterdam, Netherlands Univ British Columbia, Sch Environm Hlth, Vancouver, BC V5Z 1M9, Canada Inst Nacl Salud, Microbiol Grp, Bogota, Colombia Fiocruz MS, Fundacao Oswaldo Cruz, Lab Micol, Inst Pesquisa Clin Evandro Chagas, BR-21045900 Rio De Janeiro, Brazil Univ Sydney, Westmead Hosp, Ctr Infect Dis & Microbiol,Sydney Emerging Dis &, Mol Mycol Res Lab,Westmead Millennium Inst,Sydney, Westmead, NSW 2145, Australia Radboud Univ Nijmegen, Med Ctr, Dept Med Microbiol, NL-6525 ED Nijmegen, Netherlands Second Mil Med Univ, Changzheng Hosp, Shanghai Key Lab Mol Med Mycol, Shanghai, Peoples R China Univ Med Ctr, Dept Internal Med & Infect Dis, Utrecht, Netherlands Univ Sao Paulo State, Dept Phytopathol, UNESP, Ilha Solteira, Brazil NPRP grant from the Qatar National Research Fund (a member of Qatar foundation)5-298-3-06

Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) PROQUALI/UFJF Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Nucleoside triphosphate diphosphohydrolase (NTPDase) activity was recently characterized in Leishmania (Viannia) braziliensis promastigotes (Lb), and an antigenic conserved domain (r82-121) from the specific NTPDase 1 isoform was identified. in this work, mouse polyclonal antibodies produced against two synthetic peptides derived from this domain (LbB1LJ, r82-103; LbB2LJ, r102-121) were used. the anti-LbB1LJ or anti-LbB2LJ antibodies were immobilized on protein A-sepharose and immunoprecipitated the NTPDase 1 of 48 kDa and depleted approximately 40% of the phosphohydrolytic activity from detergent-homogenized Lb preparation. Ultrastructural immunocytochemical microscopy identified the NTPDase 1 on the parasite surface and in its subcellular cytoplasmic vesicles, mitochondria, kinetoplast and nucleus. the ATPase and ADPase activities of detergent-homogenized Lb preparation were partially inhibited by anti-LbB1LJ antibody (43-79%), which was more effective than that inhibition (18-47%) by anti-LbB2LJ antibody. in addition, the immune serum anti-LbB1LJ (67%) or anti-LbB2LJ (33%) was cytotoxic, significantly reducing the promastigotes growth in vitro. the results appoint the conserved domain from the L braziliensis NTPDase as an important target for inhibitor design and the potential application of these biomolecules in experimental protocols of disease control. (c) 2012 Elsevier Inc. All rights reserved. Univ Fed Juiz de Fora, Inst Ciencias Biol, Dept Bioquim, BR-36036900 Juiz de Fora, MG, Brazil Univ Fed Alfenas, Inst Ciencias Biomed, Dept Ciencias Biol, Alfenas, MG, Brazil Fiocruz MS, Inst Oswaldo Cruz, Lab Biol Estrut, Rio de Janeiro, RJ, Brazil Universidade Federal de São Paulo, Escola Paulista Med, Dept Biofis, São Paulo, Brazil Universidade Federal de São Paulo, Escola Paulista Med, Dept Biofis, São Paulo, Brazil FAPEMIG: CBB-APQ-01384-09 FAPEMIG: CBB-APQ 00754-09 Web of Science

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) National Institutes of Health Biomolecule Analysis Core Facility at the Border Biomedical Research Center/Biology/UTEP University of Texas at El Paso (National Institutes of Health) The glycosylphosphatidylinositol (GPI)-anchored mucins of Trypanosoma cruzi trypomastigotes play an important immunomodulatory role during the course of Chagas disease. Here, some biological activities of tGPI-mucins from four T cruzi isolates, including benznidazole-susceptible (BZS-Y), benznidazole-resistant (BZR-Y), CL, and Colombiana, were evaluated. GPI-mucins were able to differentially trigger the production of interleukin-12 and nitric oxide in BALB/c macrophages and modulate LLC-MK2 cell invasion. the significance of these variations was assessed after analysis of the terminal alpha-galactosyl residues. Enzymatic treatment with alpha-galactosidase indicated a differential expression of O-linked alpha-galactosyl residues among the strains, with higher expression of this sugar in BZS-Y and BZR-Y T cruzi populations followed by Colombiana and CL. Unweighted pair group method analysis of the carbohydrate anchor profile and biological parameters allowed the clustering of two groups. One group includes Y and CL strains (T cruzi II and VI), and the other group is represented by Colombiana strain (T. cruzi I). Fiocruz MS, Ctr Pesquisas Rene Rachou, BR-30190002 Belo Horizonte, MG, Brazil Universidade Federal de São Paulo, UNIFESP, Dept Ciencias Biol, Diadema, SP, Brazil Univ Texas El Paso, Dept Biol Sci, Border Biomed Res Ctr, El Paso, TX 79968 USA Universidade Federal de São Paulo, UNIFESP, Dept Ciencias Biol, Diadema, SP, Brazil CNPq: 305042/2010-6 CNPq: 552072/2009-5 CNPq: PAPES-IV-400138/2006-9 CNPq: PDJ-150880/2005-7 National Institutes of Health: 1R01AI070655-04 National Institutes of Health: 3R01AI070655-04S1 National Institutes of Health: 2G12RR008124-16A1 National Institutes of Health: 2G12RR008124-16A1S1 Biomolecule Analysis Core Facility at the Border Biomedical Research Center/Biology/UTEP University of Texas at El Paso (National Institutes of Health): 2G12RR008124-16A1 Biomolecule Analysis Core Facility at the Border Biomedical Research Center/Biology/UTEP University of Texas at El Paso (National Institutes of Health): 2G12RR008124-16A1S1 Biomolecule Analysis Core Facility at the Border Biomedical Research Center/Biology/UTEP University of Texas at El Paso (National Institutes of Health): G12MD007592 Web of Science

MHC class Ia-restricted CD8+ T cells are important mediators of the adaptive immune response against infections caused by intracellular microorganisms. Whereas antigen-specific effector CD8+ T cells can clear infection caused by intracellular pathogens, in some circumstances, the immune response is suboptimal and the microorganisms survive, causing host death or chronic infection. Here, we explored the cellular and molecular mechanisms that could explain why CD8+ T cell-mediated immunity during infection with the human protozoan parasite Trypanosoma cruzi is not optimal. For that purpose, we compared the CD8+ T-cell mediated immune responses in mice infected with T. cruzi or vaccinated with a recombinant adenovirus expressing an immunodominant parasite antigen. Several functional and phenotypic characteristics of specific CD8+ T cells overlapped. Among few exceptions was an accelerated expansion of the immune response in adenoviral vaccinated mice when compared to infected ones. Also, there was an upregulated expression of the apoptotic-signaling receptor CD95 on the surface of specific T cells from infected mice, which was not observed in the case of adenoviral-vaccinated mice. Most importantly, adenoviral vaccine provided at the time of infection significantly reduced the upregulation of CD95 expression and the proapoptotic phenotype of pathogen-specific CD8+ cells expanded during infection. In parallel, infected adenovirus-vaccinated mice had a stronger CD8 T-cell mediated immune response and survived an otherwise lethal infection. We concluded that a suboptimal CD8+ T-cell response is associated with an upregulation of CD95 expression and a proapoptotic phenotype. Both can be blocked by adenoviral vaccination., Author Summary Killer lymphocytes are important mediators of the immunological resistance against infections caused by virus, bacteria and parasites. In some circumstances, however, these lymphocytes are unable to properly eliminate the microorganisms which survive, causing death or establishing chronic infections. The purpose of our study was to understand why these killer cells do not succeed during infection with a human protozoan parasite. For that purpose, we compared the immune responses in animals infected or vaccinated. Many characteristics of these killer cells were similar. Among few exceptions was an accelerated immune response in vaccinated animals when compared to infected ones. Also, we observed on the surface of the killer lymphocytes from infected, but not from vaccinated animals, an increased expression of a protein involved in signaling cell death. Most importantly, vaccine significantly reduced the higher expression of this cell-death receptor. In parallel, these animals had a stronger immune response and cured infection. We concluded that a deficient killer cell response observed during infection was associated with an upregulation of this cell-death receptor and it was changed by vaccination.

Made available in DSpace on 2013-09-27T14:53:25Z (GMT). No. of bitstreams: 1 WOS000294833800002.pdf: 697909 bytes, checksum: 709430a3961b3abb5933dbf3f3753e5f (MD5) Previous issue date: 2011-10-01 Made available in DSpace on 2013-09-30T17:55:19Z (GMT). No. of bitstreams: 1 WOS000294833800002.pdf: 697909 bytes, checksum: 709430a3961b3abb5933dbf3f3753e5f (MD5) Previous issue date: 2011-10-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:15:29Z No. of bitstreams: 1 WOS000294833800002.pdf: 697909 bytes, checksum: 709430a3961b3abb5933dbf3f3753e5f (MD5) Made available in DSpace on 2014-05-20T13:15:29Z (GMT). No. of bitstreams: 1 WOS000294833800002.pdf: 697909 bytes, checksum: 709430a3961b3abb5933dbf3f3753e5f (MD5) Previous issue date: 2011-10-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Although the number of genes known to be associated with bovine spermatogenesis has increased in the past few years, regulation of this biological process remains poorly understood. Therefore, discovery of new male fertility genetic markers is of great value for assisted selection in commercially important cattle breeds, e.g., Nelore, that have delayed reproductive maturation and low fertility rates. The objective of the present study was to identify sequences associated with spermatogenesis that could be used as fertility markers. With RT-PCR, the following five transcripts preferentially expressed in adult testis were detected: TET(656) detected only in adult testis; TET(868) and TET(515) expressed preferentially in adult testis but also detected in fetal gonads of both sexes; and TET(456) and TET(262). expressed primarily in the testis, but also present in very low amounts in somatic tissues. Based on their homologies and expression profiles, we inferred that they had putative roles in spermatogenesis. Detection of sequences differentially expressed in testis, ovary, or both, was a useful approach for identifying new genes related to bovine spermatogenesis. The data reported here contributed to discovery of gene pathways involved in bovine spermatogenesis, with potential for prediction of fertility. (C) 2011 Elsevier B.V. All rights reserved. Univ São Paulo, Fac Med, Dept Pediat, BR-05403900 São Paulo, Brazil Univ São Paulo, Ctr Pesquisas Biotecnol, BR-05508900 São Paulo, Brazil Fiocruz MS, CPqRR, Ctr Pesquisas Rene Rachou, BR-30190002 Belo Horizonte, MG, Brazil UNESP, Fac Ciencias Agr & Vet, Dept Anim Reprod, BR-14884900 Jaboticabal, SP, Brazil UNESP, Fac Ciencias Agr & Vet, Dept Anim Reprod, BR-14884900 Jaboticabal, SP, Brazil FAPESP: 04/14738-2 CNPq: 305635/2009-3 CNPq: 300996/2006-7

Made available in DSpace on 2013-08-28T14:07:28Z (GMT). No. of bitstreams: 1 WOS000273911100001.pdf: 2830627 bytes, checksum: 0c7455b8252aa62fc1be0f88aa8c7d95 (MD5) Made available in DSpace on 2013-09-30T19:23:02Z (GMT). No. of bitstreams: 2 WOS000273911100001.pdf: 2830627 bytes, checksum: 0c7455b8252aa62fc1be0f88aa8c7d95 (MD5) WOS000273911100001.pdf.txt: 53135 bytes, checksum: e328cf4448342ee34ae938f9286c1ac4 (MD5) Previous issue date: 2009-12-24 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:23:50Z No. of bitstreams: 2 WOS000273911100001.pdf: 2830627 bytes, checksum: 0c7455b8252aa62fc1be0f88aa8c7d95 (MD5) WOS000273911100001.pdf.txt: 53135 bytes, checksum: e328cf4448342ee34ae938f9286c1ac4 (MD5) Made available in DSpace on 2014-05-20T13:23:50Z (GMT). No. of bitstreams: 2 WOS000273911100001.pdf: 2830627 bytes, checksum: 0c7455b8252aa62fc1be0f88aa8c7d95 (MD5) WOS000273911100001.pdf.txt: 53135 bytes, checksum: e328cf4448342ee34ae938f9286c1ac4 (MD5) Previous issue date: 2009-12-24 Ministerio de Ciência e Tecnologia (MCT) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Financiadora de Estudos e Projetos (FINEP) International Foundation for Science (IFS), Stockholm, Sweden Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Background: The pathogenic fungus Paracoccidioides brasiliensis is the agent of paracoccidioidomycosis (PCM). This is a pulmonary mycosis acquired by inhalation of fungal airborne propagules that can disseminate to several organs and tissues leading to a severe form of the disease. Adhesion and invasion to host cells are essential steps involved in the internalization and dissemination of pathogens. Inside the host, P. brasiliensis may use the glyoxylate cycle for intracellular survival.Results: Here, we provide evidence that the malate synthase of P. brasiliensis (PbMLS) is located on the fungal cell surface, and is secreted. PbMLS was overexpressed in Escherichia coli, and polyclonal antibody was obtained against this protein. By using Confocal Laser Scanning Microscopy, PbMLS was detected in the cytoplasm and in the cell wall of the mother, but mainly of budding cells of the P. brasiliensis yeast phase. PbMLSr and its respective polyclonal antibody produced against this protein inhibited the interaction of P. brasiliensis with in vitro cultured epithelial cells A549.Conclusion: These observations indicated that cell wall-associated PbMLS could be mediating the binding of fungal cells to the host, thus contributing to the adhesion of fungus to host tissues and to the dissemination of infection, behaving as an anchorless adhesin. Univ Fed Goias, Inst Ciencias Biol, Dept Bioquim & Biol Mol, Mol Biol Lab, BR-74001970 Goiania, Go, Brazil UNESP, Univ Estadual Paulista, Lab Micol Clin, Araraquara, SP, Brazil Fiocruz MS, Inst Oswaldo Cruz, Lab Patol, BR-21045900 Rio de Janeiro, Brazil UNESP, Univ Estadual Paulista, Lab Micol Clin, Araraquara, SP, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Millennium Institute for Vaccine Development and Technology Millennium Institute for Gene Therapy Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) A heterologous prime-boost strategy using plasmid DNA, followed by replication-defective recombinant adenovirus 5, is being proposed as a powerful way to elicit CD4(+) and CD8(+) T-cell-mediated protective immunity against intracellular pathogens. We confirmed this concept and furthered existing research by providing evidence that the heterologous prime-boost regimen using the gene encoding amastigote surface protein 2 elicited CD4(+) and CD8(+) T-cell-mediated protective immunity (reduction of acute parasitemia and prolonged survival) against experimental infection with Trypanosoma cruzi. Protective immunity correlated with the presence of in vivo antigen-specific cytotoxic activity prior to challenge. Based on this, our second goal was to determine the outcome of infection after heterologous prime-boost immunization of perforin-deficient mice. These mice were highly susceptible to infection. A detailed analysis of the cell-mediated immune responses in immunized perforin-deficient mice showed an impaired gamma interferon (IFN-gamma) secretion by immune spleen cells upon restimulation in vitro with soluble recombinant antigen. in spite of a normal numeric expansion, specific CD8(+) T cells presented several functional defects detected in vivo (cytotoxicity) and in vitro (simultaneous expression of CD107a/IFN-gamma or IFN-gamma/tumor necrosis factor alpha) paralleled by a decreased expression of CD44 and KLRG-1. Our final goal was to determine the importance of IFN-gamma in the presence of highly cytotoxic T cells. Vaccinated IFN-gamma-deficient mice developed highly cytotoxic cells but failed to develop any protective immunity. Our study thus demonstrated a role for perforin and IFN-gamma in a number of T-cell-mediated effector functions and in the antiparasitic immunity generated by a heterologous plasmid DNA prime-adenovirus boost vaccination strategy. Universidade Federal de São Paulo, Escola Paulista Med, CINTERGEN, BR-04044010 São Paulo, Brazil Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, BR-04044010 São Paulo, Brazil Univ Fed Rio de Janeiro, Inst Biofis Carlos Chagas Filho, Ctr Ciencias Saude, BR-21941 Rio de Janeiro, Brazil Fiocruz MS, Inst Oswaldo Cruz, Lab Biol Celular & Biol Interacoes, BR-21045900 Rio de Janeiro, Brazil Univ Fed Minas Gerais, Dept Bioquim & Imunol, Inst Ciencias Biol, BR-31270901 Belo Horizonte, MG, Brazil Fiocruz MS, Ctr Pesquisas Rene Rachou, BR-30190002 Belo Horizonte, MG, Brazil Univ Massachusetts, Sch Med, Dept Med, Div Infect Dis & Immunol, Worcester, MA 01605 USA Univ Fed Minas Gerais, Inst Ciencias Biol, Dept Microbiol, BR-31270901 Belo Horizonte, MG, Brazil Universidade Federal de São Paulo, Escola Paulista Med, CINTERGEN, BR-04044010 São Paulo, Brazil Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, BR-04044010 São Paulo, Brazil FAPESP: 2006/1983-4 Millennium Institute for Vaccine Development and Technology: CNPq-420067/2005-1 FAPEMIG: EDT 24.000 Web of Science

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The role of natural killer (NK) T cells in the development of lupus-like disease in mice is still controversial. We treated NZB/W mice with anti-NK1.1 monoclonal antibodies (mAbs) and our results revealed that administration of either an irrelevant immunoglobulin G2a (IgG2a) mAb or an IgG2a anti-NK1.1 mAb increased the production of anti-dsDNA antibodies in young NZB/W mice. However, the continuous administration of an anti-NK1.1 mAb protected aged NZB/W mice from glomerular injury, leading to prolonged survival and stabilization of the proteinuria. Conversely, the administration of the control IgG2a mAb led to an aggravation of the lupus-like disease. Augmented titres of anti-dsDNA in NZB/W mice, upon IgG2a administration, correlated with the production of BAFF/BLyS by dendritic, B and T cells. Treatment with an anti-NK1.1 mAb reduced the levels of interleukin-16, produced by T cells, in spleen cell culture supernatants from aged NZB/W. Adoptive transfer of NK T cells from aged to young NZB/W accelerated the production of anti-dsDNA in recipient NZB/W mice, suggesting that NK T cells from aged NZB/W are endowed with a B-cell helper activity. in vitro studies, using purified NK T cells from aged NZB/W, showed that these cells provided helper B-cell activity for the production of anti-dsDNA. We concluded that NK T cells are involved in the progression of lupus-like disease in mature NZB/W mice and that immunoglobulin of the IgG2a isotype has an enhancing effect on antibody synthesis due to the induction of BAFF/BLyS, and therefore have a deleterious effect in the NZB/W mouse physiology. Fiocruz MS, Oswaldo Cruz Fdn, IGM, BR-40296710 Salvador, BA, Brazil Univ São Paulo, Inst Heart INCOR, Immunol Lab, São Paulo, Brazil Universidade Federal de São Paulo, Discipline Immunol, Dept Microbiol Immunol & Parasitol, São Paulo, Brazil Universidade Federal de São Paulo, Discipline Immunol, Dept Microbiol Immunol & Parasitol, São Paulo, Brazil CNPq: 305835/2003-3 FAPESP: 95/9379-2 FAPESP: 97/06225-0 Web of Science

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2014-02-26T17:01:59Z No. of bitstreams: 1 WOS000252220200024.pdf: 152729 bytes, checksum: 04aa1cc1b09b8a4c91e2a052a03bf3ed (MD5) Made available in DSpace on 2014-02-26T17:01:59Z (GMT). No. of bitstreams: 1 WOS000252220200024.pdf: 152729 bytes, checksum: 04aa1cc1b09b8a4c91e2a052a03bf3ed (MD5) Previous issue date: 2007-11-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:24:56Z No. of bitstreams: 1 WOS000252220200024.pdf: 152729 bytes, checksum: 04aa1cc1b09b8a4c91e2a052a03bf3ed (MD5) Made available in DSpace on 2014-05-20T13:24:56Z (GMT). No. of bitstreams: 1 WOS000252220200024.pdf: 152729 bytes, checksum: 04aa1cc1b09b8a4c91e2a052a03bf3ed (MD5) Previous issue date: 2007-11-01 During April and May 2006, experiments were carried out in Brejo do Mutambal, Varzelandia Town, Minas Gerais State, to evaluate the attractiveness of phlebotomine sandflies to CDC light traps, baited with kairomones. of the 19 species of Lutzomyia already registered for the region, L. lutziana (Costa Lima), L. longipennis (Barreto), L. goiana (Martins, Falcao & Silva) and L. brasiliensis (Costa Lima) were recorded for the first time, thus increasing the diversity of phlebotomine sandflies fauna in this area to 23 species. The new registered species and distribution are shown and discussed herein. Univ Fed Minas Gerais, Inst Ciências Biol, Dept Parasitol, BR-31270901 Belo Horizonte, MG, Brazil Fiocruz MS, Ctr Pesquisas Rene Rachou, BR-30196002 Belo Horizonte, MG, Brazil Univ Estadual Paulista, Fac Farm, BR-14801902 Araraquara, SP, Brazil Univ Estadual Paulista, Fac Farm, BR-14801902 Araraquara, SP, Brazil

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2014-02-26T17:00:17Z No. of bitstreams: 0 Made available in DSpace on 2014-02-26T17:00:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-09-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T15:23:23Z No. of bitstreams: 0 Made available in DSpace on 2014-05-20T15:23:23Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-09-01 In the present work, the antitumor effect of fastuosain, a cysteine proteinase from Bromelia fastuosa, was investigated. In the intravenous model of lung colonization in C57Bl/6 mice, fastuosain and bromelain injected intraperitoneally were protective, and very few nodules of B16F10-Nex2 melanoma cells were detected. Tumor cells treated with fastuosain showed reduced expression of CD44 and decreased invasion through Matrigel, lost their cytoplasmic extensions and substrate adherence, and became round and detached, forming strongly bound cell clusters in suspension. Peritoneal cells recruited and activated by fastuosain treatment ( mainly monocytic cells and lymphocytes) migrated to the lung, where pulmonary melanoma metastases grew. Adoptive transference of peritoneal cells recruited by fastuosain had no protective effect against lung metastases in recipient mice. Treatment of green fluorescent protein - chimeric animals with fastuosain did not change the number of cells that migrated to the lung, compared to PBS-injected control mice, but the number of positive major histocompatibility complex class II cells increased with fastuosain treatment. Murine antibodies against fastuosain, bromelain, and cathepsins B and L cross-reacted in ELISA and recognized surface and cytoplasmic components expressed on B16F10-Nex2 cells. Anti-fastuosain antibodies were cytotoxic/lytic to B16F10-Nex2 cells. Antitumor effects of fastuosain involve mainly the direct effect of the enzyme and elicitation of protective antibodies. Univ Fed São Paulo, UNONEX, Unidade Oncol Expt, Dept Microbiol Immunol & Parasitol, BR-04023062 São Paulo, Brazil Univ Fed São Paulo, Discipline Parasitol, Dept Microbiol Immunol & Parasitol, BR-04023062 São Paulo, Brazil Univ Fed São Paulo, IBILCE, Dept Quim & Ciências Ambientais, BR-04023062 São Paulo, Brazil Fiocruz MS, Ctr Pesquisas Goncalo Moniz, Salvador, BA, Brazil Univ Fed São Paulo, IBILCE, Dept Quim & Ciências Ambientais, BR-04023062 São Paulo, Brazil

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2014-02-26T17:29:56Z No. of bitstreams: 1 WOS000247665800008.pdf: 187726 bytes, checksum: 0359e8c0ca21180daff770c76c42b3e4 (MD5) Made available in DSpace on 2014-02-26T17:29:56Z (GMT). No. of bitstreams: 1 WOS000247665800008.pdf: 187726 bytes, checksum: 0359e8c0ca21180daff770c76c42b3e4 (MD5) Previous issue date: 2007-07-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T15:24:42Z No. of bitstreams: 1 WOS000247665800008.pdf: 187726 bytes, checksum: 0359e8c0ca21180daff770c76c42b3e4 (MD5) Made available in DSpace on 2014-05-20T15:24:42Z (GMT). No. of bitstreams: 1 WOS000247665800008.pdf: 187726 bytes, checksum: 0359e8c0ca21180daff770c76c42b3e4 (MD5) Previous issue date: 2007-07-01 Extracts from Holostylis reniformis were tested in vivo against Plasmodium berghei and in vitro against a chloroquine-resistant strain of Plasmodium falciparum. The hexane extract of the roots was the most active, causing 67% reduction of parasitemia in vivo. From this extract, six lignans, including a new (7 ' R,8S,8 ' S)-3 ',4 '-methylenedioxy-4,5-dimethoxy-2,7 '-cyclolignan-7-one, were isolated and tested in vitro against P. falciparum. The three most active lignans showed 50% inhibitor concentrations of

Made available in DSpace on 2015-03-18T15:56:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-08-18Bitstream added on 2015-03-18T16:29:28Z : No. of bitstreams: 1 WOS000341302700089.pdf: 799858 bytes, checksum: d1be8c07fb4f88f06503b9b92110110a (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) National Institute of Science and Technology - Pharmaceutical Innovation (INCT-if) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) Plasmodium vivax is the most prevalent of the five species causing malaria in humans. The current available treatment for P. vivax malaria is limited and unsatisfactory due to at least two drawbacks: the undesirable side effects of primaquine (PQ) and drug resistance to chloroquine. Phenylalanine-alanine-PQ (Phe-Ala-PQ) is a PQ prodrug with a more favorable pharmacokinetic profile compared to PQ. The toxicity of this prodrug was evaluated in in vitro assays using a human hepatoma cell line (HepG2), a monkey kidney cell line (BGM), and human red blood cells deficient in the enzyme glucose-6-phosphate-dehydrogenase (G6PD). In addition, in vivo toxicity assays were performed with rats that received multiple doses of Phe-Ala-PQ to evaluate biochemical, hematological, and histopathological parameters. The activity was assessed by the inhibition of the sporogonic cycle using a chicken malaria parasite. Phe-Ala-PQ blocked malaria transmission in Aedes mosquitoes. When compared with PQ, it was less cytotoxic to BGM and HepG2 cells and caused less hemolysis of G6PD-deficient red blood cells at similar concentrations. The prodrug caused less alteration in the biochemical parameters than did PQ. Histopathological analysis of the liver and kidney did show differences between the control and Phe-Ala-PQ-treated groups, but they were not statistically significant. Taken together, the results highlight the prodrug as a novel lead compound candidate for the treatment of P. vivax malaria and as a blocker of malaria transmission. Univ Estadual Paulista UNESP, Fac Ciencias Farm, Dept Principios Ativos Nat & Toxicol, Sao Paulo, Brazil Fiocruz MS, Ctr Pesquisas Rene Rachou, Belo Horizonte, MG, Brazil Univ Estadual Paulista UNESP, Fac Odontol Araraquara, Dept Fisiol & Patol, Sao Paulo, Brazil Univ Estadual Paulista UNESP, Fac Ciencias Farm, Dept Anal Clin, Sao Paulo, Brazil Univ Estadual Paulista UNESP, Fac Ciencias Farm, Lab Pesquisa & Desenvolvimento Farmacos Lapdesf, Sao Paulo, Brazil Univ Estadual Paulista UNESP, Fac Ciencias Farm, Dept Principios Ativos Nat & Toxicol, Sao Paulo, Brazil Univ Estadual Paulista UNESP, Fac Odontol Araraquara, Dept Fisiol & Patol, Sao Paulo, Brazil Univ Estadual Paulista UNESP, Fac Ciencias Farm, Dept Anal Clin, Sao Paulo, Brazil Univ Estadual Paulista UNESP, Fac Ciencias Farm, Lab Pesquisa & Desenvolvimento Farmacos Lapdesf, Sao Paulo, Brazil FAPESP: 09/51075-5 FAPESP: 11/11239-9

Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Instituto Oswaldo Cruz (FIOCRUZ, Brazil) Background: Plasmodium vivax merozoite surface protein-1 (MSP-1) is an antigen considered to be one of the leading malaria vaccine candidates. PvMSP-1 is highly immunogenic and evidences suggest that it is target for protective immunity against asexual blood stages of malaria parasites. Thus, this study aims to evaluate the acquired cellular and antibody immune responses against PvMSP-1 in individuals naturally exposed to malaria infections in a malaria-endemic area in the north-eastern Amazon region of Brazil.Methods: the study was carried out in Paragominas, Para State, in the Brazilian Amazon. Blood samples were collected from 35 individuals with uncomplicated malaria. Peripheral blood mononuclear cells were isolated and the cellular proliferation and activation was analysed in presence of 19 kDa fragment of MSP-1 (PvMSP-1(19)) and Plasmodium falciparum PSS1 crude antigen. Antibodies IgE, IgM, IgG and IgG subclass and the levels of TNF, IFN-gamma and IL-10 were measured by enzyme-linked immunosorbent assay.Results: the prevalence of activated CD4(+) was greater than CD8(+) T cells, in both ex-vivo and in 96 h culture in presence of PvMSP-1(19) and PSS1 antigen. A low proliferative response against PvMSP-1(19) and PSS1 crude antigen after 96 h culture was observed. High plasmatic levels of IFN-gamma and IL-10 as well as lower TNF levels were also detected in malaria patients. However, in the 96 h supernatant culture, the dynamics of cytokine responses differed from those depicted on plasma assays; in presence of PvMSP-1(19) stimulus, higher levels of TNF were noted in supernatant 96 h culture of malaria patient's cells while low levels of IFN-gamma and IL-10 were verified. High frequency of malaria patients presenting antibodies against PvMSP-1(19) was evidenced, regardless class or IgG subclass. PvMSP-1(19)-induced antibodies were predominantly on non-cytophilic subclasses.Conclusions: the results presented here shows that PvMSP-1(19) was able to induce a high cellular activation, leading to production of TNF and emphasizes the high immunogenicity of PvMSP-1(19) in naturally exposed individuals and, therefore, its potential as a malaria vaccine candidate. Fiocruz MS, Inst Oswaldo Cruz, Lab Pesquisas Malaria, BR-21040900 Rio de Janeiro, RJ, Brazil Fiocruz MS, Ctr Pesquisa Diagnost & Treinamento Malaria CPD M, Reference Ctr Malaria Extra Amazonian Reg Secreta, BR-21040900 Rio de Janeiro, RJ, Brazil Fiocruz MS, Inst Oswaldo Cruz, Lab Biol Mol & Doencas Endem, BR-21040900 Rio de Janeiro, RJ, Brazil Univ São Paulo, Dept Analises Clin & Toxicol, São Paulo, Brazil Universidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, São Paulo, Brazil SVS, Inst Evandro Chagas, Programa Ensaios Clin Malaria, Belem, Para, Brazil Universidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, São Paulo, Brazil Web of Science