Polymorphism analysis of the CTLA-4 gene in paracoccidioidomycosis patients

The CTLA-4 protein is expressed in activated T cells and plays an essential role in the immune response through its regulatory effect on T cell activation. Polymorphisms of the CTLA-4 gene have been correlated with autoimmune, neoplastic and infectious illnesses. This work aimed to verify possible associations between single nucleotide polymorphisms (SNPs) in CTLA-4, -318C/T in the promoter and +49A/G in exon 1 and paracoccidioidomycosis (PCM) caused by Paracoccidioides brasiliensis. For this purpose, 66 chronic form PCM patients and 76 healthy controls had their allele, genotype and haplotype frequencies determined. The genetic admixture structure of the patients and controls was evaluated to eliminate ancestral bias. The comparison of frequencies indicated no significant differences between patients and controls that could link the SNPs to PCM. Groups were admixture matched with no difference observed in population ancestry inference, indicating that the absence of association between CTLA-4 polymorphisms and PCM could not be attributed to ancestral bias. This study showed that there was no association between the CTLA-4 SNPs -318 and +49 and the resistance or susceptibility to PCM.22022

Polymorphism analysis of the CTLA-4 gene in paracoccidioidomycosis patients

The CTLA-4 protein is expressed in activated T cells and plays an essential role in the immune response through its regulatory effect on T cell activation. Polymorphisms of the CTLA-4 gene have been correlated with autoimmune, neoplastic and infectious illnesses. This work aimed to verify possible associations between single nucleotide polymorphisms (SNPs) in CTLA-4, -318C/T in the promoter and +49A/G in exon 1 and paracoccidioidomycosis (PCM) caused by Paracoccidioides brasiliensis. For this purpose, 66 chronic form PCM patients and 76 healthy controls had their allele, genotype and haplotype frequencies determined. The genetic admixture structure of the patients and controls was evaluated to eliminate ancestral bias. The comparison of frequencies indicated no significant differences between patients and controls that could link the SNPs to PCM. Groups were admixture matched with no difference observed in population ancestry inference, indicating that the absence of association between CTLA-4 polymorphisms and PCM could not be attributed to ancestral bias. This study showed that there was no association between the CTLA-4 SNPs -318 and +49 and the resistance or susceptibility to PCM

DNAhsp65 vaccination induces protection in mice against Paracoccidioides brasiliensis infection

Heat-shock proteins are Molecules with extensive data showing their potential as immunomodulators of different types of diseases, The gene of HSP65 from Mycobacterium leprae has shown prophylactic and immunotherapeutic effects against a broad arrays of experimental models including tuberculosis, leishmaniasis, arthritis and diabetes. With this in mind, we tested the DNAhsp65 vaccine using an experimental model of Paraccocidiodomycosis, an important endemic mycosis in Latin America. The intramuscular immunization with DNAhsp65 induced, in BALB/c mice, an increase of Th1-levels cytokines and a reduction of fungal burdens resulted in a marked reduction of collagen and lung remodeling. DNAhsp65 may be an attractive candidate for prevention, therapy and as an adjuvant for mycosis treatment. (C) 2008 Elsevier Ltd. All rights reserved.Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq

In Vitro Antifungal Activity and Toxicity of Itraconazole in DMSA-PLGA Nanoparticles

Itraconazole (ITZ) is a drug used to treat various fungal infections and may cause side effects. The aim of this study was to develop and evaluate the in vitro activity of DMSA-PLGA nanoparticles loaded with ITZ against Paracoccidioides brasiliensis, as well as their cytotoxicity. Nanoparticles were prepared using the emulsification-evaporation technique and characterized by their encapsulation efficiency, morphology (TEM), size (Nanosight) and charge (zeta potential). Antifungal efficacy in P brasiliensis was determined by minimal inhibition concentration (MIC), and cytotoxicity using MU assay. ITZ was effectively incorporated in the PLGA-DMSA nanoparticles with a loading efficiency of 72.8 +/- 3.50%. The shape was round with a solid polymeric structure, and a size distribution of 174 +/- 86 nm (Average +/- SD). The particles were negatively charged. ITZ-NANO presented antifungal inhibition (MIC = 6.25 ug/mL) against P brasiliensis and showed lower in vitro cytotoxicity than free drug (ITZ).Brazilian agency MCT/CNPqBrazilian agency FINEPBrazilian agency FINATECBrazilian agency FAP-D

HSP65 DNA as therapeutic strategy to treat experimental paracoccidioidomycosis

The conventional treatment for paracoccidioidomycosis, the most prevalent mycosis in Latin America, involves long periods of therapy resulting in sequels and high frequency of relapses. The search for new alternatives of treatment is necessary. Previously, we have demonstrated that the hsp65 gene from Mycobacterium leprae shows prophylactic effects against murine paracoccidioidomycosis. Here, we tested the DNAhsp65 immunotherapy in BALB/c mice infected with Paracoccidioides brasiliensis, the agent of paracoccidioidomycosis. We observed an increase of Th1 cytokines accompanied by a reduction in fungal burden and pulmonary injury. These results provide new prospects for immunotherapy of paracoccidioidomycosis and other mycoses. (C) 2009 Elsevier Ltd. All rights reserved.Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)Conselho Nacional de Desenvolvimento Cientifico a Tecnologico (CNPq

Transcriptional response of murine macrophages upon infection with opsonized Paracoccidioides brasiliensis yeast cells

Paracoccidioides brasiliensis is the etiologic agent of the Paracoccidioidomycosis the most common systemic mycosis in Latin America. Little is known about the regulation of genes involved in the innate immune host response to P. brasiliensis. We therefore examined the kinetic profile of gene expression of peritoneal macrophage infected with P. brasiliensis. Total RNA from macrophages at 6, 24 and 48 h was extracted, hybridized onto nylon membranes and analyzed. An increase in the transcription of a number of pro-inflammatory molecules encoding membrane proteins, metalloproteases, involved in adhesion and phagocytosis, are described. We observed also the differential expression of genes whose products may cause apoptotic events induced at 24 h. In addition, considering the simultaneous analyses of differential gene expression for the pathogen reported before by our group, at six hours post infection, we propose a model at molecular level for the P. brasiliensis-macrophage early interaction. In this regard, P. brasiliensis regulates genes specially related to stress and macrophages, at the same time point, up-regulate genes related to inflammation and phagocytosis, probably as an effort to counteract infection by the fungus. (c) 2007 Elsevier Masson SAS. All fights reserved

Comparative Genomics of Sibling Species of Fonsecaea Associated with Human Chromoblastomycosis

Submitted by Manoel Barata ([email protected]) on 2018-02-09T13:19:12Z No. of bitstreams: 1 FaoroComparativ.pdf: 5399928 bytes, checksum: af51deed6d3e60618950577e576e6aad (MD5)Approved for entry into archive by Manoel Barata ([email protected]) on 2018-05-03T19:30:23Z (GMT) No. of bitstreams: 1 FaoroComparativ.pdf: 5399928 bytes, checksum: af51deed6d3e60618950577e576e6aad (MD5)Made available in DSpace on 2018-05-03T19:30:23Z (GMT). No. of bitstreams: 1 FaoroComparativ.pdf: 5399928 bytes, checksum: af51deed6d3e60618950577e576e6aad (MD5) Previous issue date: 2017Universidade Federal do Paraná. Departamento de Patologia Básica. Laboratório de Imunogenética e Histocompatibilidade. Curitiba, PR, Brasil / Universidade Federal do Paraná. Engenharia de Bioprocessos e Biotecnologia. Curitiba, PR, Brasil.Universidade Federal do Paraná. Setor de Educação Profissional e Tecnológica. Laboratório de Bioinformática. Curitiba, PR, Brasil / Universidade Federal do Paraná. Departamento de Bioquímica. Curitiba, PR, Brasil.Universidade Federal do Paraná. Departamento de Patologia Básica. Laboratório de Imunogenética e Histocompatibilidade. Curitiba, PR, Brasil.Universidade Federal do Paraná. Departamento de Patologia Básica. Laboratório de Imunogenética e Histocompatibilidade. Curitiba, PR, Brasil / CBS-KNAW Fungal Biodiversity Centre. Utrecht, Netherlands / University of Amsterdam. Institute for Biodiversity and Ecosystem Dynamics. Amsterdam, Netherlands.Universidade Federal do Paraná. Engenharia de Bioprocessos e Biotecnologia. Curitiba, PR, Brasil.Universidade Federal do Paraná. Setor de Educação Profissional e Tecnológica. Laboratório de Bioinformática. Curitiba, PR, Brasil.Universidade Federal do Paraná. Engenharia de Bioprocessos e Biotecnologia. Curitiba, PR, Brasil / Universidade Federal do Paraná. Setor de Educação Profissional e Tecnológica. Laboratório de Bioinformática. Curitiba, PR, Brasil / Universidade Federal do Paraná. Departamento de Bioquímica. Curitiba, PR, Brasil.Universidade Federal do Paraná. Departamento de Patologia Básica. Laboratório de Imunogenética e Histocompatibilidade. Curitiba, PR, Brasil.Universidade de Brasília. Departamento de Biologia Celular. Brasilia, Brasil.Universidade Federal do Paraná. Departamento de Patologia Básica. Laboratório de Imunogenética e Histocompatibilidade. Curitiba, PR, Brasil.Universidade de Brasília. Departamento de Biologia Celular. Brasilia, Brasil.Guangdong Provincial Center for Disease Control and Prevention. Guangdong Provincial Institute of Public Health, Guangzhou, China.Universidade Federal do Paraná. Departamento de Bioquímica. Curitiba, PR, Brasil / Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Universidade Federal do Paraná. Departamento de Bioquímica. Curitiba, PR, Brasil.Universidade Federal do Paraná. Departamento de Bioquímica. Curitiba, PR, Brasil.Universidade Federal do Paraná. Departamento de Bioquímica. Curitiba, PR, Brasil.Universidade de São Paulo. Faculdade de Ciências Farmacêuticas. Departamento de Análises Clínicas e Toxicológicas. São Paulo, SP, Brasil.Universidade de São Paulo. Faculdade de Ciências Farmacêuticas. Departamento de Análises Clínicas e Toxicológicas. São Paulo, SP, Brasil.Universidade de Brasília. Departamento de Biologia Celular. Brasilia, Brasil / Northern Arizona University. Pathogen and Microbiome Institute. Flagstaff, United States.Universidade Católica de Brasília. Departamento de Ciências Genômicas e Biotecnologia. Brasília, DF, Brasil.Universidade Federal do Paraná. Departamento de Patologia Básica. Laboratório de Imunogenética e Histocompatibilidade. Curitiba, PR, Brasil.Universidade Federal do Paraná. Departamento de Bioquímica. Curitiba, PR, Brasil.Universidade Federal do Paraná. Setor de Educação Profissional e Tecnológica. Laboratório de Bioinformática. Curitiba, PR, Brasil / Universidade Federal do Paraná. Departamento de Bioquímica. Curitiba, PR, Brasil.Universidade de Campinas. Divisão de Recursos Microbianos. Campinas, SP, Brasil.Mashhad University of Medical Sciences. School of Medicine. Department of Parasitology and Mycology. Mashhad, Iran.Universidade Federal do Paraná. Departamento de Patologia Básica. Laboratório de Imunogenética e Histocompatibilidade. Curitiba, PR, Brasil / Universidade Federal do Paraná. Hospital das Clínicas. Curitiba, PR, Brasil.Universidade Federal do Paraná. Setor de Educação Profissional e Tecnológica. Laboratório de Bioinformática. Curitiba, PR, Brasil / Universidade Federal do Paraná. Departamento de Bioquímica. Curitiba, PR, Brasil.Universidade Federal do Paraná. Departamento de Patologia Básica. Laboratório de Imunogenética e Histocompatibilidade. Curitiba, PR, Brasil / CBS-KNAW Fungal Biodiversity Centre. Utrecht, Netherlands / University of Amsterdam. Institute for Biodiversity and Ecosystem Dynamics. Amsterdam, Netherlands.Fonsecaea and Cladophialophora are genera of black yeast-like fungi harboring agents of a mutilating implantation disease in humans, along with strictly environmental species. The current hypothesis suggests that those species reside in somewhat adverse microhabitats, and pathogenic siblings share virulence factors enabling survival in mammal tissue after coincidental inoculation driven by pathogenic adaptation. A comparative genomic analysis of environmental and pathogenic siblings of Fonsecaea and Cladophialophora was undertaken, including de novo assembly of F. erecta from plant material. The genome size of Fonsecaea species varied between 33.39 and 35.23 Mb, and the core genomes of those species comprises almost 70% of the genes. Expansions of protein domains such as glyoxalases and peptidases suggested ability for pathogenicity in clinical agents, while the use of nitrogen and degradation of phenolic compounds was enriched in environmental species. The similarity of carbohydrate-active vs. protein-degrading enzymes associated with the occurrence of virulence factors suggested a general tolerance to extreme conditions, which might explain the opportunistic tendency of Fonsecaea sibling species. Virulence was tested in the Galleria mellonella model and immunological assays were performed in order to support this hypothesis. Larvae infected by environmental F. erecta had a lower survival. Fungal macrophage murine co-culture showed that F. erecta induced high levels of TNF-α contributing to macrophage activation that could increase the ability to control intracellular fungal growth although hyphal death were not observed, suggesting a higher level of extremotolerance of environmental species

Prognostic value of morphologic and clinical parameters in pT2 - pT3 prostate cancer

OBJECTIVES: Verify the efficacy of clinical and morphologic parameters currently applied, including an immunohistochemical panel, in the prognostic of prostate cancer, in specific stages of the disease MATERIALS AND METHODS: In the period from 2002 to 2005, 40 surgical specimens were selected from patients submitted to radical prostatectomy, with their respective diagnostic biopsies. Based on the pathological stage pT2 or pT3, the specimens were separated into two groups, each one with 20 specimens. The results were confronted with pre- and postoperative clinical data. Between the groups studied, the following was also analyzed: the profile of the expression of molecular markers such as PSA, E-caderin, chromogranin-A, synaptofisin, P53 and Ki-67, both in the material coming from the prostatic biopsy and from the surgical specimens of all patients RESULTS: Data showed that patients with prostate-confined disease (pT2) presented lower PSA and Gleason score rates, in relation to the group with extra-prostatic disease (pT3). Quantitative measures obtained for the percentage of positive fragments from the biopsy revealed that patients from the pT2 group presented a lower mean percentage when compared to the pT3 group. Positive margins of both groups influenced the need for complementary treatment before biochemical progression. The comparison of the molecular marker expression in both stages was not significantly different CONCLUSION: It is evident the need to improve new methods, predominantly morphologic and molecular, that are able to further exploit the study of the material from the prostatic biopsy. As to the profile of the molecular markers used in both studied groups, there was no significant difference in the sense of outlining an additional prognostic factor in the clinical practice

Eighty Years of Mycopathologia: A Retrospective Analysis of Progress Made in Understanding Human and Animal Fungal Pathogens

Mycopathologia was founded in 1938 to ‘diffuse the understanding of fungal diseases in man and animals among mycologists.’ This was an important mission considering that pathogenic fungi for humans and animals represent a tiny minority of the estimated 1.5–5 million fungal inhabitants on Earth. These pathogens have diverged from the usual saprotrophic lifestyles of most fungi to colonize and infect humans and animals. Medical and veterinary mycology is the subdiscipline of microbiology that dwells into the mysteries of parasitic, fungal lifestyles. Among the oldest continuing scientific publications on the subject, Mycopathologia had its share of ‘classic papers’ since the first issue was published in 1938. An analysis of the eight decades of notable contributions reveals many facets of host–pathogen interactions among 183 volumes comprising about 6885 articles. We have analyzed the impact and relevance of this body of work using a combination of citation tools (Google Scholar and Scopus) since no single citation metric gives an inclusive perspective. Among the highly cited Mycopathologia publications, those on experimental mycology accounted for the major part of the articles (36%), followed by diagnostic mycology (16%), ecology and epidemiology (15%), clinical mycology (14%), taxonomy and classification (10%), and veterinary mycology (9%). The first classic publication, collecting nearly 200 citations, appeared in 1957, while two articles published in 2010 received nearly 150 citations each, which is notable for a journal covering a highly specialized field of study. An empirical analysis of the publication trends suggests continuing interests in novel diagnostics, fungal pathogenesis, review of clinical diseases especially with relevance to the laboratory scientists, taxonomy and classification of fungal pathogens, fungal infections and carriage in pets and wildlife, and changing ecology and epidemiology of fungal diseases around the globe. We anticipate that emerging and re-emerging fungal pathogens will continue to cause significant health burden in the coming decades. It remains vital that scientists and physicians continue to collaborate by learning each other’s language for the study of fungal diseases, and Mycopathologia will strive to be their partner in this increasingly important endeavor to its 100th anniversary in 2038 and beyond