Development of a Multiplex qPCR Assay for Fast Detection and Differentiation of Paracoccidioidomycosis Agents.

Classic paracoccidioidomycosis (PCM) is a potentially deadly neglected tropical systemic mycosis caused by members of the complex (, , , and ) and . The laboratorial diagnosis of PCM relies on observing pathognomonic structures such as the "steering wheel" or "Mickey Mouse" shape in the direct mycological examination, fresh biopsied tissue in 10% KOH, histopathological analysis, and/or the isolation of the fungus in culture. However, these procedures are time-consuming and do not allow for the speciation of due to overlapping morphologies.

PbGP43 Genotyping Using Paraffin-Embedded Biopsies of Human Paracoccidioidomycosis Reveals a Genetically Distinct Lineage in the Paracoccidioides brasiliensis Complex.

Paracoccidioidomycosis (PCM) is a systemic mycosis caused by a group of cryptic species embedded in the Paracoccidioides brasiliensis complex and Paracoccidioides lutzii. Four species were recently inferred to belong to the P. brasiliensis complex, but the high genetic diversity found in both human and environmental samples have suggested that the number of lineages may be higher.

Therapeutic management of subcutaneous phaeohyphomycosis and chromoblastomycosis in kidney transplant recipients: a retrospective study of 82 cases in a single center.

Background: Melanized fungi are a distinct group of pathogens that cause infections like chromoblastomycosis and phaeohyphomycosis, especially in a state of immunosuppression including solid organ transplant recipients. Guidelines for the treatment of these infections are lacking, and there is no available standard recommendation.

Objective: To evaluate the therapeutic aspects of subcutaneous melanized fungal infections in kidney transplant recipients.

A New Duplex PCR-Assay for the Detection and Identification of Species.

Paracoccidioidomycosis (PCM) is a life-threatening systemic fungal infection caused by members of the complex and . Routine diagnoses of PCM down to the species level using classical mycological approaches are unspecific due to overlapping phenotypes. There is an urgent need for specific, sensitive, and cost-effective molecular tools to diagnose PCM.

Correlation between clinical outcome and tissue inflammatory response in kidney transplant recipients with cryptococcosis.

Cryptococcosis is the second most common invasive fungal infection reported in renal transplant recipients. Tissue granulomatous inflammation is necessary to contain Cryptococcus infection. This study aims to analyze the granuloma patterns and in situ expression of regulatory T (Treg) immune response in tissue samples from 12 renal transplant recipients with cryptococcosis.

Genetic diversity of Pneumocystis jirovecii from a cluster of cases of pneumonia in renal transplant patients: Cross-sectional study.

Pneumocystis jirovecii can cause severe potentially life-threatening pneumonia (PCP) in kidney transplant patients. Prophylaxis of patients against PCP in this setting is usually performed during 6 months after transplantation. The aim of this study is to describe the molecular epidemiology of a cluster of PCP in renal transplant recipients in Brazil.

Phenotypic and genotypic detection of Candida albicans and Candida dubliniensis strains isolated from oral mucosa of AIDS pediatric patients.

The aim of this study was to assess a collection of yeasts to verify the presence of Candida dubliniensis among strains isolated from the oral mucosa of AIDS pediatric patients which were initially characterized as Candida albicans by the traditional phenotypic method, as well as to evaluate the main phenotypic methods used in the discrimination between the two species and confirm the identification through genotypic techniques, i.e., DNA sequencing.

Spectral Manifestation of Melanized Fungal Infections in Kidney Transplant Recipients: Report of Six Cases.

Chromoblastomycosis and phaeohyphomycosis are melanized fungal infections, which affect skin and subcutaneous tissues in immunocompetent and immunosuppressed patients, as solid-organ transplant recipients, respectively. In this present study, we report six cases of melanized fungal infection in kidney transplant recipients. In five cases, culture of tissue specimens identified two cases of Exophiala spp.

Genetic diversity of medically important and emerging Candida species causing invasive infection.

Background: Genetic variation in the ribosomal DNA (rDNA) internal transcribed spacer (ITS) region has been studied among fungi. However, the numbers of ITS sequence polymorphisms in the various Candida species and their associations with sources of invasive fungal infections remain poorly investigated. Here, we characterized the intraspecific and interspecific ITS diversity of Candida spp.

International Society of Human and Animal Mycology (ISHAM)-ITS reference DNA barcoding database--the quality controlled standard tool for routine identification of human and animal pathogenic fungi.

Human and animal fungal pathogens are a growing threat worldwide leading to emerging infections and creating new risks for established ones. There is a growing need for a rapid and accurate identification of pathogens to enable early diagnosis and targeted antifungal therapy. Morphological and biochemical identification methods are time-consuming and require trained experts.

Fungemia caused by Candida species in a children's public hospital in the city of São Paulo, Brazil: study in the period 2007-2010.

Candidemia remains a major cause of morbidity and mortality in the health care environment. The epidemiology of Candida infection is changing, mainly in relation to the number of episodes caused by species C. non-albicans.

Trichosporon inkin: an uncommon agent of scalp white piedra. Report of four cases in Brazilian children.

We report four cases of scalp white piedra (SWP) in Brazilian female children. Morphological and physiological approaches gave inconsistent results for identifying Trichosporon to species level, while the sequencing of the intergenic spacer 1 region of ribosomal DNA accurately identified the agent of SWP as T. inkin.

Immunolocalization of IFN-gamma in the lesions of resistant and susceptible mice to Paracoccidioides brasiliensis infection.

The important role of interferon-gamma (IFN-γ) in protective immunity in mycosis is well established, except for its participation in fungal granulomas. Herein, we employ immunohistochemical reactions to describe the in situ localization of IFN-γ in granulomas of susceptible (B10.A) and resistant (A/J) mice to infection with Paracoccidioides brasiliensis (Pb).

Matrix metalloproteinases with gelatinolytic activity induced by Paracoccidioides brasiliensis infection.

Matrix metalloproteinases (MMPs) modulate extracellular matrix turnover, inflammation and immunity. We studied MMP-9 and MMP-2 in experimental paracoccidioidomycosis. At 15 and 120 days after infection (DAI) with virulent Paracoccidioides brasiliensis, MMP-9 was positive by immunohistochemistry in multinucleated giant cells, in mononuclear cells with macrophage and lymphocyte morphologies and also in fungal cells in the lesions of susceptible and resistant mice.

Nitric oxide participation in granulomatous response induced by Paracoccidioides brasiliensis infection in mice.

The role of nitric oxide (NO) in granulomas of Paracoccidioides brasiliensis-infected inducible NO synthase-deficient C57BL/6 mice (iNOS KO) and their wild-type counterparts and its association with osteopontin (OPN) and matrix metalloproteinases (MMPs) was studied. At 15 days after infection (DAI), iNOS KO mice showed compact and necrotic granulomas with OPN+ macrophages and multinucleated giant cells, whereas wild-type mice developed loose granulomas with many fungi and OPN+ cells distributed throughout the tissue. In addition, high OPN levels and fungal load were observed in iNOS KO mice.

Experimental infections with Paracoccidioides brasiliensis obtained from armadillos: comparison to clinical isolates.

Paracoccidioides brasiliensis causes paracoccidioidomycosis (PCM) that is one of the most prevalent systemic human mycoses in Latin America. Armadillos show a high incidence of PCM infection and could, therefore, be a natural reservoir for this fungus. In this study were compared the virulence profiles of isolates obtained from nine-banded armadillos (Dasypus novemcinctus) (PbT1 and PbT4) and isolates from PCM patients (Pb265 and Bt83).

Evaluation of fungal burden in experimental paracoccidioidomycosis by using the fluorescent dye Blankophor.

The fungal load in organs and blood of susceptible and resistant mice infected with Paracoccidioides brasiliensis was quantitated by using the optical brightener Blankophor and compared with CFU counts. Fluorescent staining of fungal cells proved to be a quick and easy procedure, suitable for evaluation of paracoccidioidomycotic infection.