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... Lutz. 1961; 21: 99 194. 6 Borges MJ, Azevedo MO, Bonatelli R, Felipe MSS, Astolfi-Filho S. A practical method for the preparation of total DNA from filamentous fungi. Fungal. Genet. ... Br. J. Exp. Pathol. 1985; 66: 585 594. PubMed.... more
... Lutz. 1961; 21: 99 194. 6 Borges MJ, Azevedo MO, Bonatelli R, Felipe MSS, Astolfi-Filho S. A practical method for the preparation of total DNA from filamentous fungi. Fungal. Genet. ... Br. J. Exp. Pathol. 1985; 66: 585 594. PubMed. 10 Goihman-Yahr M, Pinel L, Albornoz MC. ...
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Decades of studies on antibody structure led to the tenet that the V region binds antigens while the C region interacts with immune effectors. In some antibodies, however, the C region affects affinity and/or specificity for the antigen.... more
Decades of studies on antibody structure led to the tenet that the V region binds antigens while the C region interacts with immune effectors. In some antibodies, however, the C region affects affinity and/or specificity for the antigen. One example is the 3E5 monoclonal murine IgG family, in which the mIgG3 isotype has different fine specificity to the Cryptococcus neoformans capsule polysaccharide than the other mIgG isotypes despite their identical variable sequences. Our group serendipitously found another pair of mIgG1/mIgG3 antibodies based on the 2H1 hybridoma to the C. neoformans capsule that recapitulated the differences observed with 3E5. In this work, we report the molecular basis of the constant domain effects on antigen binding using recombinant antibodies. As with 3E5, immunofluorescence experiments show a punctate pattern for 2H1‐mIgG3 and an annular pattern for 2H1‐mIgG1; these binding patterns have been associated with protective efficacy in murine cryptococcosis. A...
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Paracoccidioidomycosis (PCM) is one of the most frequent systemic mycoses in Latin America. It affects mainly male rural workers in impoverished regions, and the therapy can last up to two years or use drugs that are very toxic. Given the... more
Paracoccidioidomycosis (PCM) is one of the most frequent systemic mycoses in Latin America. It affects mainly male rural workers in impoverished regions, and the therapy can last up to two years or use drugs that are very toxic. Given the need for novel safe and effective approaches to treat PCM, we have been developing monoclonal antibodies (mAbs) that could be used not only to block specific fungal targets, but also modulate the host’s antifungal immunity. In this work we show the generation of and promising results with a mAb against HSP90, a molecular chaperone that is an important virulence factor in fungi. Using recombinant Paracoccidioides lutzii (Pb01) and P. brasiliensis (Pb18) HSP90 proteins produced in E. coli, we immunized mice and generated polyclonal antibodies and an IgG1 hybridoma mAb. The proteins were very immunogenic and both the polyclonal serum and mAb were used in immunofluorescence experiments, which showed binding of antibodies to the yeast cell surface. The ...
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Research Interests: Genetics, Microbiology, Population Genetics, Plant Biology, Biology, and 12 moreMedicine, Phylogenomics, Genetic Diversity, Ciencias Biológicas, Micología, Genotype, Fungal Genetics and Biology, Gran Chaco, Enseñanza - Aprendizaje Ciencias Naturales Y Exactas, Academic, Paracoccidioides, and paracoccidioidomycosis
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The order Chaetothyriales (Pezizomycotina, Ascomycetes) harbours obligatorily melanised fungi and includes numerous etiologic agents of chromoblastomycosis, phaeohyphomycosis and other diseases of vertebrate hosts. Diseases range from... more
The order Chaetothyriales (Pezizomycotina, Ascomycetes) harbours obligatorily melanised fungi and includes numerous etiologic agents of chromoblastomycosis, phaeohyphomycosis and other diseases of vertebrate hosts. Diseases range from mild cutaneous to fatal cerebral or disseminated infections and affect humans and cold-blooded animals globally. In addition, Chaetothyriales comprise species with aquatic, rock-inhabiting, ant-associated, and mycoparasitic life-styles, as well as species that tolerate toxic compounds, suggesting a high degree of versatile extremotolerance. To understand their biology and divergent niche occupation, we sequenced and annotated a set of 23 genomes of main the human opportunists within the Chaetothyriales as well as related environmental species. Our analyses included fungi with diverse life-styles, namely opportunistic pathogens and closely related saprobes, to identify genomic adaptations related to pathogenesis. Furthermore, ecological preferences of C...
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On the basis of multilocus phylogenetic data, Fonsecaea nubica was described in 2010 as a molecular sibling of F. monophora, an established agent of the human skin disease chomoblastomycosis in tropical zones. Genome analysis of these... more
On the basis of multilocus phylogenetic data, Fonsecaea nubica was described in 2010 as a molecular sibling of F. monophora, an established agent of the human skin disease chomoblastomycosis in tropical zones. Genome analysis of these pathogens is mandatory to identify genes involved in the interaction with host and virulence.
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Histoplasma capsulatum comprises a worldwide complex of saprobiotic fungi mainly found in nitrogen/phosphate (often bird guano) enriched soils. The microconidia of Histoplasma species may be inhaled by mammalian hosts, and is followed by... more
Histoplasma capsulatum comprises a worldwide complex of saprobiotic fungi mainly found in nitrogen/phosphate (often bird guano) enriched soils. The microconidia of Histoplasma species may be inhaled by mammalian hosts, and is followed by a rapid conversion to yeast that can persist in host tissues causing histoplasmosis, a deep pulmonary/systemic mycosis. Histoplasma capsulatum sensu lato is a complex of at least eight clades geographically distributed as follows: Australia, Netherlands, Eurasia, North American classes 1 and 2 (NAm 1 and NAm 2), Latin American groups A and B (LAm A and LAm B) and Africa. With the exception of the Eurasian cluster, those clades are considered phylogenetic species. Increased Histoplasma sampling (n = 234) resulted in the revision of the phylogenetic distribution and population structure using 1,563 aligned nucleotides from four protein-coding regions. The LAm B clade appears to be divided into at least two highly supported clades, which are geographic...
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The prevalence of invasive fungal infections worldwide has increased in the last decades. The development of specific drugs targeting pathogenic fungi without producing collateral damage to mammalian cells is a daunting pharmacological... more
The prevalence of invasive fungal infections worldwide has increased in the last decades. The development of specific drugs targeting pathogenic fungi without producing collateral damage to mammalian cells is a daunting pharmacological challenge. Indeed, many of the toxicities and drug interactions observed with contemporary antifungal therapies can be attributed to "nonselective" interactions with enzymes or cell membrane systems found in mammalian host cells. A computer-aided screening strategy against the TRR1 protein of Paracoccidioides lutzii is presented here. Initially, a bank of commercially available compounds from Life Chemicals provider was docked to model by virtual screening simulations. The small molecules that interact with the model were ranked and, among the best hits, twelve compounds out of 3,000 commercially-available candidates were selected. These molecules were synthesized for validation and in vitro antifungal activity assays for Paracoccidioides lu...
Research Interests: Molecular Dynamics Simulation, Biology, Enzyme Inhibitors, Medicine, Multidisciplinary, and 15 moreCell line, Virtual screening, Mice, Animals, Cell Death, Antifungal, PLoS one, Thioredoxin, Antifungal Agents, Microbial Sensitivity Tests, Recombinant Proteins, Biological Assay, Ligands, Paracoccidioides, and Thioredoxin reductase
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Research Interests: Biology and Bioscience
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Research Interests: Pharmacology, Chemistry, Organic Chemistry, Nanoparticles, Medicine, and 15 moreSafety, Mice, Animals, Biodistribution, In Vivo, Antifungal Agents, PLGA, Materials Testing, Drug Carriers, Lactic Acid, Paracoccidioides, paracoccidioidomycosis, Pharmacology and pharmaceutical sciences, succimer, and Drug combinations
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Research Interests: Microbiology, Melanin, Biology, Medicine, Cryptococcus Neoformans, and 6 moreYeast, Laccase, Exocytosis, Mbio, MACRÓFAGOS, and Leveduras
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Research Interests: Genetics, Mycology, Biology, Medicine, Clinical Microbiology, and 15 moreBiological Sciences, DNA, Virulence, Humans, Clinical, Polymerase Chain Reaction, Genome, Genomic DNA, Fungal genome size, Base Sequence, Paracoccidioides, Molecular Sequence Data, paracoccidioidomycosis, gene amplification, and Medical and Health Sciences
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Fifteen Paracoccidioides brasiliensis isolates were discriminated by the RAPD analysis into two groups with only 17% of genomic identity. The ability of P. brasiliensis isolates to invade tissues was studied in an experimental model using... more
Fifteen Paracoccidioides brasiliensis isolates were discriminated by the RAPD analysis into two groups with only 17% of genomic identity. The ability of P. brasiliensis isolates to invade tissues was studied in an experimental model using susceptible B10.A mice. The analysis was performed according to the severity of the lesions including the number and size of the granuloma, the number and dissemination of fungi to different organs. The isolates from two RAPD groups demonstrated a marked difference in their virulence patterns for B10.A mice. The isolates Pb S, 662, Bt and 166 (group I) elicited localized infection restricted to the liver showing compact epithelioid granuloma with few fungi in the early post-infection period (slightly virulent). On the other hand the isolates Pb 01 and 7455 (group II) elicited a disseminated infection with a mixed suppurative and looser granulomatous inflammation, showing extensive areas of necrosis and large numbers of viable fungal cells (highly virulent). These results are strong evidence for correlation between RAPD patterns and the virulence degree of P. brasiliensis.
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Chromoblastomycosis (CBM) is a disease caused by several dematiaceous fungi from different genera, and Fonsecaea is the most common which has been clinically isolated. Genetic transformation methods have recently been described; however,... more
Chromoblastomycosis (CBM) is a disease caused by several dematiaceous fungi from different genera, and Fonsecaea is the most common which has been clinically isolated. Genetic transformation methods have recently been described; however, molecular tools for the functional study of genes have been scarcely reported for those fungi. In this work, we demonstrated that gene deletion and generation of the null mutant by homologous recombination are achievable for Fonsecaea pedrosoi by the use of two approaches: use of double-joint PCR for cassette construction, followed by delivery of the split-marker by biolistic transformation. Through in silico analyses, we identified that F. pedrosoi presents the complete enzymatic apparatus required for tryptophan (trp) biosynthesis. The gene encoding a tryptophan synthase trpB —which converts chorismate to trp—was disrupted. The ΔtrpB auxotrophic mutant can grow with external trp supply, but germination, viability of conidia, and radial growth are ...
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Parkinson's disease (PD) is a progressive and chronic neurodegenerative disease of the central nervous system. Early treatment for PD is efficient; however, long-term systemic medication commonly leads to deleterious side-effects.... more
Parkinson's disease (PD) is a progressive and chronic neurodegenerative disease of the central nervous system. Early treatment for PD is efficient; however, long-term systemic medication commonly leads to deleterious side-effects. Strategies that enable more selective drug delivery to the brain using smaller dosages, while crossing the complex brain-blood barrier (BBB), are highly desirable to ensure treatment efficacy and decrease/avoid unwanted outcomes. Our goal was to design and test the neurotherapeutic potential of a forefront nanoparticle-based technology composed of albumin/PLGA nanosystems loaded with dopamine (ALNP-DA) in 6-OHDA PD mice model. ALNP-DA effectively crossed the BBB, replenishing dopamine at the nigrostriatal pathway, resulting in significant motor symptom improvement when compared to Lesioned and L-DOPA groups. Notably, ALNP-DA (20 mg/animal dose) additionally up-regulated and restored motor coordination, balance, and sensorimotor performance to non-lesio...
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Paracoccidioidesspp. are thermodimorphic pathogenic fungi endemic to Latin America. Predation is believed to drive the evolution of virulence for soil saprophytes. We evaluated the presence of environmental amoeboid predators in soil from... more
Paracoccidioidesspp. are thermodimorphic pathogenic fungi endemic to Latin America. Predation is believed to drive the evolution of virulence for soil saprophytes. We evaluated the presence of environmental amoeboid predators in soil from armadillo burrows whereParacoccidioideshad been previously detected and tested if interaction ofParacoccidioideswith amoebae increased fungal virulence. Nematodes, ciliates and amoebae – all potential predators of fungi – grew in cultures from soil samples. Microscopical observation and ITS sequencing identified the amoebae asAcanthamoebaspp,Allovahlkampfia spelaeaandVermamoeba vermiformis. These three amoebae efficiently ingested, killed and digestedParacoccidioidesspp. yeast cells, as did laboratory-adapted axenicAcanthamoeba castellanii. Sequential co-cultivation ofParacoccidioideswithA. castellaniiselected for phenotypical traits related to survival of the fungus within a natural predator as well as in murine macrophages and in vivo (Galleria m...
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Chromobacterium violaceumis one of millions of species of free-living microorganisms that populate the soil and water in the extant areas of tropical biodiversity around the world. Its complete genome sequence reveals (i) extensive... more
Chromobacterium violaceumis one of millions of species of free-living microorganisms that populate the soil and water in the extant areas of tropical biodiversity around the world. Its complete genome sequence reveals (i) extensive alternative pathways for energy generation, (ii) ≈500 ORFs for transport-related proteins, (iii) complex and extensive systems for stress adaptation and motility, and (iv) widespread utilization of quorum sensing for control of inducible systems, all of which underpin the versatility and adaptability of the organism. The genome also contains extensive but incomplete arrays of ORFs coding for proteins associated with mammalian pathogenicity, possibly involved in the occasional but often fatal cases of humanC. violaceuminfection. There is, in addition, a series of previously unknown but important enzymes and secondary metabolites including paraquat-inducible proteins, drug and heavy-metal-resistance proteins, multiple chitinases, and proteins for the detoxi...
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Research Interests: Microbiology, Immunology, Molecular Biology, Medical Microbiology, Biology, and 15 moreMedicine, Humans, Microbes, Escherichia coli, Gene, Molecular cloning, Genomic Library, Molecular Mass, Amino Acid Sequence, Base Sequence, Open Reading Frame, Membrane Protein, Paracoccidioides, Molecular Sequence Data, and Nucleotide sequence
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Acquired by inhalation of the thermal dimorphic fungi Paracoccidioides spp. conidia, paracoccidioidomycosis ranges from symptomatic to severe and potentially fatal disseminated disease. The main focus of this review is to highlight... more
Acquired by inhalation of the thermal dimorphic fungi Paracoccidioides spp. conidia, paracoccidioidomycosis ranges from symptomatic to severe and potentially fatal disseminated disease. The main focus of this review is to highlight clinical aspects of paracoccidioidomycosis and, its pathogens’ diversity ecology and particularities. In addition, we present strategies for therapy, including DNA vaccines and nanostructured drugs. Molecular and morphological data supported the split of the Paracoccidioides genus into two species, Paracoccidioides brasiliensis and Paracoccidioides lutzii. An acute form of the disease affects approximately 5% of cases and involves the phagocytic mononuclear system, resulting in progressive lymphadenopathy. The chronic form affects adult men and frequently involves lungs, skin and mucous membranes, lymph nodes, and adrenal glands. The clinical manifestations depend on the ability of the host to control the fungal multiplication and dissemination. The long ...
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Research Interests: Microbiology, Immunology, Data Analysis, Medical Microbiology, Biology, and 15 moreMedicine, Functional Genomics, Molecular Mechanics, Transcriptome, Latin America, Cell Differentiation, Humans, Expressed Sequence Tags, Cellular differentiation, Host Pathogen Interaction, Fungus, Paracoccidioides, paracoccidioidomycosis, Mycelium, and Cardiovascular medicine and haematology
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Research Interests: Microbiology, Computational Biology, Medical Microbiology, Biology, Medicine, and 13 moreMedical Mycology, Humans, Sequence alignment, Yeast, Two-Dimensional Gel Electrophoresis, Amino Acid Sequence, Two dimensional Gel Electrophoresis, Paracoccidioides, Medical, Differential expression, Molecular Sequence Data, paracoccidioidomycosis, and Mycelium
Research Interests: Biochemistry, Genetics, Microbiology, Plant Biology, Biology, and 15 moreOxidative Stress, Medicine, Antioxidants, Peroxidase, Humans, Antioxidant, Cytochrome C, Catalase, Hydrogen Peroxide, Antioxidant enzyme, Oxidative Damage, Enzymatic Activity, Paracoccidioides, paracoccidioidomycosis, and Human Immune System
... de Brasilia, 70910-900, Brasaia, DF, Brazil Accepted for publication July 27, 1994 SILVA, SP,FELIPE, MSS, PEREIRA, M., AZEVEDO, MO ... Paracoccidioides brasiliensis, the ethio-1986; Stambuck et al., 1988; Mendes-logical agent of... more
... de Brasilia, 70910-900, Brasaia, DF, Brazil Accepted for publication July 27, 1994 SILVA, SP,FELIPE, MSS, PEREIRA, M., AZEVEDO, MO ... Paracoccidioides brasiliensis, the ethio-1986; Stambuck et al., 1988; Mendes-logical agent of paracoccidioidomycosis, is Gianini et al ...