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Id: biblio-957449 Autor: Nunes, Joslaine de Oliveira; Tsujisaki, Rosianne Assis de Sousa; Nunes, Maína de Oliveira; Lima, Gláucia Moreira Espíndola; Paniago, Anamaria Mello Miranda; Pontes, Elenir Rose Jardim Cury; Chang, Marilene Rodrigues. Título: Cryptococcal meningitis epidemiology: 17 years of experience in a State of the Brazilian Pantanal Fonte: Rev. Soc. Bras. Med. Trop;51(4):485-492, July-Aug. 2018. tab, graf. Idioma: en. Resumo: Abstract INTRODUCTION: This study aimed to describe cryptococcal meningitis (CM) cases and the associated demographic, clinical, and microbiological data obtained from cities in the State of Mato Grosso do Sul in the Midwestern region of Brazil. METHODS: The data from 129 patients with laboratory-confirmed CM admitted from 1997 to 2014 were retrospectively reviewed. The molecular types of Cryptococcus neoformans and Cryptococcus gattii isolated from cerebrospinal fluid were analyzed to determine their geographic distribution. RESULTS: The patients had a mean age of 37 years and consisted mostly of men (76.7%). Most of the Cryptococcus isolates were obtained from patients infected with human immunodeficiency virus (HIV) and included 105 (87.5%) and 5 (55.6%) isolates of C. neoformans and C. gattii complexes, respectively. A restriction fragment length polymorphism (RFLP) analysis of URA5 revealed that most of the isolates were C. neoformans molecular type VNI (89.1%), whereas the molecular types VGII (7%) and VNII (3.9%) were observed less frequently. Notably, 65% of the cases with a time from symptom onset to laboratory diagnosis of more than 60 days resulted in fatalities, and sequelae were observed among the patients who survived. CONCLUSIONS: The present study documents the occurrence of neurocryptococcosis, which is mainly caused by C. neoformans VNI, in Mato Grosso do Sul, Brazil, with probable autochthonous cases in the Brazilian Pantanal, the world's largest tropical wetland, and a biome where cryptococcosis has not yet been explored. Descritores: DNA Fúngico/análise Meningite Criptocócica/epidemiologia Cryptococcus neoformans/isolamento & purificação Cryptococcus gattii/isolamento & purificação -População Rural Fatores Socioeconômicos População Urbana Brasil/epidemiologia DNA Fúngico/líquido cefalorraquidiano Reação em Cadeia da Polimerase Estudos Retrospectivos Meningite Criptocócica/líquido cefalorraquidiano Meningite Criptocócica/microbiologia Cryptococcus neoformans/genética Cryptococcus gattii/genética Genótipo Pessoa de Meia-Idade Limites: Humanos Masculino Feminino Criança Adolescente Adulto Adulto Jovem Responsável: BR1.1 - BIREME

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Id: biblio-990445 Autor: Benedetti, Volmir Pitt; Savi, Daiani Cristina; Aluizio, Rodrigo; Adamoski, Douglas; Kava, Vanessa; Galli-Terasawa, Lygia Vitória; Glienke, Chirlei. Título: ERG11 gene polymorphisms and susceptibility to fluconazole in candida isolates from diabetic and kidney transplant patients Fonte: Rev. Soc. Bras. Med. Trop;52:e20180473, 2019. tab. Idioma: en. Resumo: Abstract INTRODUCTION: Candidiasis is the most frequent opportunistic mycosis in humans and can cause mortality, particularly in immunodeficient patients. One major concern is the increasing number of infections caused by drug-resistant Candidas trains, as these cannot be efficiently treated with standard therapeutics. The most common mechanism of fluconazole resistance in Candida is mutation of ERG11, a gene involved in the biosynthesis of ergosterol, a compound essential for cell integrity and membrane function. METHODS: Based on this knowledge, we investigated polymorphisms in the ERG11 gene of 3 Candida species isolated from immunocompromised and immunocompetent patients. In addition, we correlated the genetic data with the fluconazole susceptibility profile of the Candida isolates. RESULTS: A total of 80 Candida albicans, 8 Candida tropicalis and 6 Candida glabrata isolates were obtained from the saliva of diabetic, kidney transplant and immunocompetent patients. Isolates were considered susceptible to fluconazole if the minimum inhibitory concentration was lower than 8 μg/mL. The amino acid mutations F105L, D116E, K119N, S137L, and K128T were observed in C. albicans isolates, and T224C and G263A were found in C. tropicalis isolates. CONCLUSIONS: Despite the high number of polymorphisms observed, the mutations occurred in regions that are not predicted to interfere with ergosterol synthesis, and therefore are not related to fluconazole resistance. Descritores: Polimorfismo Genético/efeitos dos fármacos Candida/efeitos dos fármacos Candida/genética Fluconazol/farmacologia Transplante de Rim Diabetes Mellitus/microbiologia Antifúngicos/farmacologia -Valores de Referência Saliva/microbiologia Candida/isolamento & purificação DNA Fúngico/genética Testes de Sensibilidade Microbiana Reação em Cadeia da Polimerase Farmacorresistência Fúngica/genética Imunocompetência Pessoa de Meia-Idade Mutação/efeitos dos fármacos Limites: Humanos Masculino Feminino Adulto Idoso Responsável: BR1.1 - BIREME

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Id: biblio-978072 Autor: Delama, Ignacio; Legarraga, Paulette; González, Tamara; García, Patricia; Rabagliati, Ricardo. Título: Evaluación del Aspergillus lateral flow device para el diagnóstico de aspergilosis invasora, experiencia en un hospital universitario / Evaluation of the lateral flow Aspergillus assay for the diagnosis of invasive aspergillosis, experience in a university hospital Fonte: Rev. chil. infectol;35(5):574-579, 2018. tab, graf. Idioma: es. Resumo: Resumen Introducción: El diagnóstico de aspergilosis invasora (AI) se realiza mediante criterios clínicos y microbiológicos los que incluyen marcadores séricos. Recientemente, el test inmunocromatográfico Aspergillus lateral flow device (LFD), ha sido evaluado como método para diagnóstico de AI. Objetivo: Evaluar el desempeño de este test para el diagnóstico de AI. Material y Método: Estudio transversal en que se evaluaron muestras de suero y lavado bronco-alveolar (LBA) procesadas para galactomanano provenientes de pacientes adultos con sospecha de AI, atendidos en el Hospital Clínico de Red de Salud UCCHRISTUS. Resultados: Se procesó un total de 142 muestras de 98 pacientes, correspondientes a AI probada 5,6%, AI probable 41,5%, AI posible 12,7% y ausencia de AI 40,1%. Al confrontar los resultados con las categorías diagnósticas según criterios EORTC/MSG se obtuvo una sensibilidad y especificidad de LFD para diagnóstico de AI de 70,9 y 53,5% para muestras de suero y 83,3 y 38,5% para muestras de LBA. La concordancia entre galactomanano y LFD fue de 62,4% (54,1-69,9) con un índice Kappa de 0,202 (0,03682-0,3669). Conclusiones: Aspergillus LFD presentó una adecuada sensibilidad; sin embargo, la especificidad fue baja por lo que un resultado positivo requiere ser confirmado. Background: The incidence of invasive aspergillosis is increasing. Its diagnosis is based on clinical and microbiological criteria which include the determination of serological markers such as galactomannan. Recently, the Aspergillus lateral flow device, an inmunocromatograph assay has been described for its diagnosis. Aim: To evaluate the performance of the lateral flow device for the diagnosis of invasive aspergillosis (IA) in adult patients. Material and Method: In this cross-sectional study, frozen samples that had been previously evaluated for galactomannan from patients classified with proven/probable/possible or no AI according to the EORTC/MSG criteria were selected. Results: A total of 142 samples from 98 patients were processed, corresponding to proven AI 5.6%, probable IA 41.5%, possible IA 12.7% and no-IA 40.1%. The sensitivity and specificity of the Aspergillus lateral flow was 70.9% and 53.5% for serum samples and 83.3% and 38.5% for BAL samples. The concordance between the galactomannan and Aspergillus lateral flow was 62.4% (54.1 - 69.9) with a Kappa index of 0.202 (0.03682 - 0.3669). Conclusions: We observed a good sensitivity but low specificity, a positive result need a confirmatory test. Descritores: Aspergilose/diagnóstico Aspergillus/genética Aspergillus/imunologia DNA Fúngico/análise Líquido da Lavagem Broncoalveolar/microbiologia Mananas/análise -Estudos Transversais Cromatografia de Afinidade/métodos Sensibilidade e Especificidade Hospitais Universitários Limites: Humanos Masculino Feminino Adulto Pessoa de Meia-Idade Idoso Responsável: CL1.1 - Biblioteca Central

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Id: biblio-1057277 Autor: Damasceno, Lisandra Serra; Almeida Júnior, Antônio Mauro Barros; Aguiar, Bárbara de Oliveira; Muniz, Mauro de Medeiros; Almeida, Marcos de Abreu; Zancopé-Oliveira, Rosely Maria; Leitão, Terezinha do Menino Jesus Silva. Título: Postpartum histoplasmosis in an HIV-negative woman: a case report and phylogenetic characterization by internal transcribed spacer region analysis Fonte: Rev. Soc. Bras. Med. Trop;53:e20190364, 2020. tab, graf. Idioma: en. Projeto: Conselho Nacional de Desenvolvimento Científico e Tecnológico; . Fundação de Amparo à Pesquisa do Rio de Janeiro. Resumo: Abstract The present report describes the first case of postpartum disseminated histoplasmosis in a 24-year-old HIV-negative woman. On the tenth day after vaginal delivery, the patient presented with dyspnea, fever, hypotension, tachycardia, and painful hepatomegaly. Yeast-like Histoplasma capsulatum features were isolated in the buffy coat. The phylogenetic analysis demonstrated that the fungal isolate was similar to other H. capsulatum isolates identified in HIV patients from Ceará and Latin America. Thus, histoplasmosis development in individuals with transitory immunosuppression or during the period of immunological recovery should be carefully examined. Descritores: DNA Fúngico/análise DNA Espaçador Ribossômico/genética Período Pós-Parto Histoplasma/genética Histoplasmose/diagnóstico -Filogenia Reação em Cadeia da Polimerase Histoplasma/isolamento & purificação Histoplasmose/microbiologia Limites: Humanos Feminino Adulto Tipo de Publ: Relatos de Casos Revisão Responsável: BR1.1 - BIREME

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Id: biblio-1101256 Autor: EV, Laís Daniela; DAMÉ-TEIXEIRA, Nailê; DO, Thuy; MALTZ, Marisa; PAROLO, Clarissa Cavalcanti Fatturi. Título: The role of Candida albicans in root caries biofilms: an RNA-seq analysis Fonte: J. appl. oral sci;28:e20190578, 2020. tab, graf. Idioma: en. Projeto: Brazilian National Counsel of Technological and Scientific Development; . Coordination for the Improvement of Higher Level Education; . Rio Grande do Sul State Foundation for Research Support; . Leeds Teaching Hospitals Charitable Foundation; . Dunhill Medical Trust. Resumo: Abstract Objective This study sought to analyze the gene expression of Candida albicans in sound root surface and root caries lesions, exploring its role in root caries pathogenesis. Methodology The differential gene expression of C. albicans and the specific genes related to cariogenic traits were studied in association with samples of biofilm collected from exposed sound root surface (SRS, n=10) and from biofilm and carious dentin of active root carious lesions (RC, n=9). The total microbial RNA was extracted, and the cDNA libraries were prepared and sequenced on the Illumina Hi-Seq2500. Unique reads were mapped to 163 oral microbial reference genomes including two chromosomes of C. albicans SC5314 (14,217 genes). The putative presence of C. albicans was estimated (sum of reads/total number of genes≥1) in each sample. Count data were normalized (using the DESeq method package) to analyze differential gene expression (using the DESeq2R package) applying the Benjamini-Hochberg correction (FDR<0.05). Results Two genes (CaO19.610, FDR=0.009; CaO19.2506, FDR=0.018) were up-regulated on SRS, and their functions are related to biofilm formation. Seven genes ( UTP20 , FDR=0.018; ITR1 , FDR=0.036; DHN6 , FDR=0.046; CaO19.7197 , FDR=0.046; CaO19.7838 , FDR=0.046; STT4 , FDR=0.046; GUT1 , FDR=0.046) were up-regulated on RC and their functions are related to metabolic activity, sugar transport, stress tolerance, invasion and pH regulation. The use of alternative carbon sources, including lactate, and the ability to form hypha may be a unique trait of C. albicans influencing biofilm virulence. Conclusions C. albicans is metabolically active in SRS and RC biofilm, with different roles in health and disease. Descritores: Raiz Dentária/microbiologia Candida albicans/genética DNA Fúngico/genética Cárie Radicular/microbiologia Biofilmes/crescimento & desenvolvimento -Candida albicans/isolamento & purificação Candida albicans/crescimento & desenvolvimento Expressão Gênica Regulação Fúngica da Expressão Gênica Regulação para Cima Análise de Sequência de RNA Transcriptoma Morfogênese Limites: Humanos Responsável: BR28.1 - Serviço de Biblioteca e Documentação Professor Doutor Antônio Gabriel Atta

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Id: biblio-949893 Autor: Torres-Guerrero, Edoardo; Arenas, Roberto; Castro, Rigoberto Hernández. Título: Chromoblastomycosis due to Cladosporium langeronii. Molecular diagnosis of an agent previously diagnosed as Fonsecaea pedrosoi Fonte: An. bras. dermatol;93(3):475-476, May-June 2018. graf. Idioma: en. Descritores: Ascomicetos/isolamento & purificação Cromoblastomicose/microbiologia Cladosporium/isolamento & purificação -Ascomicetos/genética DNA Fúngico DNA Ribossômico Reação em Cadeia da Polimerase Cromoblastomicose/diagnóstico Cromoblastomicose/patologia Cladosporium/genética Técnicas de Diagnóstico Molecular Limites: Humanos Responsável: BR1.1 - BIREME

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Id: biblio-828187 Autor: Cheng, Fei; Hou, Lin; Woeste, Keith; Shang, Zhengchun; Peng, Xiaobang; Zhao, Peng; Zhang, Shuoxin. Título: Soil pretreatment and fast cell lysis for direct polymerase chain reaction from forest soils for terminal restriction fragment length polymorphism analysis of fungal communities Fonte: Braz. j. microbiol;47(4):817-827, Oct.-Dec. 2016. tab, graf. Idioma: en. Projeto: Technical management system for increasing the capacity of carbon sink and water regulation of mountain forests in the Qinling Mountains; . Detecting, simulating and applying techniques for coupling of carbon, nitrogen and water in forest ecosystems. Resumo: Abstract Humic substances in soil DNA samples can influence the assessment of microbial diversity and community composition. Using multiple steps during or after cell lysis adds expenses, is time-consuming, and causes DNA loss. A pretreatment of soil samples and a single step DNA extraction may improve experimental results. In order to optimize a protocol for obtaining high purity DNA from soil microbiota, five prewashing agents were compared in terms of their efficiency and effectiveness in removing soil contaminants. Residual contaminants were precipitated by adding 0.6 mL of 0.5 M CaCl2. Four cell lysis methods were applied to test their compatibility with the pretreatment (prewashing + Ca2+ flocculation) and to ultimately identify the optimal cell lysis method for analyzing fungal communities in forest soils. The results showed that pretreatment with TNP + Triton X-100 + skim milk (100 mM Tris, 100 mM Na4P2O7, 1% polyvinylpyrrolidone, 100 mM NaCl, 0.05% Triton X-100, 4% skim milk, pH 10.0) removed most soil humic contaminants. When the pretreatment was combined with Ca2+ flocculation, the purity of all soil DNA samples was further improved. DNA samples obtained by the fast glass bead-beating method (MethodFGB) had the highest purity. The resulting DNA was successfully used, without further purification steps, as a template for polymerase chain reaction targeting fungal internal transcribed spacer regions. The results obtained by terminal restriction fragment length polymorphism analysis indicated that the MethodFGB revealed greater fungal diversity and more distinctive community structure compared with the other methods tested. Our study provides a protocol for fungal cell lysis in soil, which is fast, convenient, and effective for analyzing fungal communities in forest soils. Descritores: Microbiologia do Solo Polimorfismo de Fragmento de Restrição Florestas Reação em Cadeia da Polimerase Microbiota Fungos/classificação Fungos/genética -Solo/química Cloreto de Cálcio DNA Bacteriano DNA Fúngico Fungos/isolamento & purificação Responsável: BR1.1 - BIREME

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Id: biblio-894939 Autor: Cuomo, Christina A; Rhodes, Johanna; Desjardins, Christopher A. Título: Advances in Cryptococcus genomics: insights into the evolution of pathogenesis Fonte: Mem. Inst. Oswaldo Cruz;113(7):e170473, 2018. tab. Idioma: en. Projeto: Broad Institute; . UK Medical Research Council. Resumo: Cryptococcus species are the causative agents of cryptococcal meningitis, a significant source of mortality in immunocompromised individuals. Initial work on the molecular epidemiology of this fungal pathogen utilized genotyping approaches to describe the genetic diversity and biogeography of two species, Cryptococcus neoformans and Cryptococcus gattii. Whole genome sequencing of representatives of both species resulted in reference assemblies enabling a wide array of downstream studies and genomic resources. With the increasing availability of whole genome sequencing, both species have now had hundreds of individual isolates sequenced, providing fine-scale insight into the evolution and diversification of Cryptococcus and allowing for the first genome-wide association studies to identify genetic variants associated with human virulence. Sequencing has also begun to examine the microevolution of isolates during prolonged infection and to identify variants specific to outbreak lineages, highlighting the potential role of hyper-mutation in evolving within short time scales. We can anticipate that further advances in sequencing technology and sequencing microbial genomes at scale, including metagenomics approaches, will continue to refine our view of how the evolution of Cryptococcus drives its success as a pathogen. Descritores: Filogenia Variação Genética Cryptococcus gattii/genética Tipagem de Sequências Multilocus Filogeografia -Filogenia DNA Fúngico Cryptococcus gattii/patogenicidade Filogeografia Genótipo Limites: Humanos Responsável: BR1.1 - BIREME

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Id: biblio-893617 Autor: GOMES, Cinthya Cristina; GUIMARÃES, Ludmila Silva; PINTO, Larissa Christina Costa; CAMARGO, Gabriela Alessandra da Cruz Galhardo; VALENTE, Maria Isabel Bastos; SARQUIS, Maria Inêz de Moura. Título: Investigations of the prevalence and virulence of Candida albicans in periodontal and endodontic lesions in diabetic and normoglycemic patients Fonte: J. appl. oral sci;25(3):274-281, May-June 2017. tab, graf. Idioma: en. Projeto: Rio de Janeiro Research Foundation. Resumo: Abstract Pulpal and periodontal tissues have similar microbiota that allows cross-contamination between the pulp and periodontal tissues. Objective The aim of this study was to investigate the prevalence of isolated Candida albicans from periodontal endodontic lesions in diabetic and normoglycemic patients, and the fungi's virulence in different atmospheric conditions. Material and Methods A case-control study was conducted on 15 patients with type 2 diabetes mellitus (G1) and 15 non-diabetics (G2) with periodontal endodontic lesions. Samples of root canals and periodontal pockets were plated on CHROMagar for later identification by polymerase chain reaction (PCR) and virulence test. Results C. albicans was identified in 79.2% and 20.8% of the 60 samples collected from diabetic and normoglycemic patients, respectively. Of the 30 samples collected from periodontal pockets, 13 showed a positive culture for C. albicans, with 77% belonging to G1 and 23% to G2. Of the 11 positive samples from root canals, 82% were from G1 and 18% from G2. Production of proteinase presented a precipitation zone Pz<0.63 of 100% in G1 and 72% in G2, in redox and negative (Pz=1), under anaerobic conditions in both groups. Hydrophobicity of the strains from G1 indicated 16.4% with low, 19.3% with moderate, and 64.3% with high hydrophobicity in redox. In G2, 42.2% had low, 39.8% had moderate, 18% had high hydrophobicity in redox. In anaerobic conditions, G1 showed 15.2% with low, 12.8% with moderate, and 72% with high hydrophobicity; in G2, 33.6% had low, 28.8% had moderate, and 37.6% had high hydrophobicity. There was statistical difference in the number of positive cultures between G1 and G2 (p<0.05) with predominance in G1. There was statistical difference for all virulence factors, except hemolysis (p=0.001). Conclusions Candida albicans was isolated more frequently and had higher virulence in diabetic patients. Descritores: Doenças Periodontais/microbiologia Candida albicans/isolamento & purificação Candida albicans/patogenicidade Doenças da Polpa Dentária/microbiologia Diabetes Mellitus Tipo 2/microbiologia -Oxirredução Peptídeo Hidrolases/análise Doenças Periodontais/fisiopatologia Doenças Periodontais/diagnóstico por imagem Bolsa Periodontal/microbiologia Fosfolipases/análise Virulência DNA Fúngico Radiografia Dentária Estudos de Casos e Controles Reação em Cadeia da Polimerase Estatísticas não Paramétricas Cavidade Pulpar/microbiologia Doenças da Polpa Dentária/fisiopatologia Doenças da Polpa Dentária/diagnóstico por imagem Diabetes Mellitus Tipo 2/fisiopatologia Eletroforese Interações Hidrofóbicas e Hidrofílicas Limites: Humanos Masculino Feminino Adulto Pessoa de Meia-Idade Idoso Responsável: BR1.1 - BIREME

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Id: biblio-841762 Autor: Arantes, Thales Domingos; Theodoro, Raquel Cordeiro; Teixeira, Marcus de Melo; Bagagli, Eduardo. Título: Use of fluorescent oligonucleotide probes for differentiation between Paracoccidioides brasiliensis and Paracoccidioides lutzii in yeast and mycelial phase Fonte: Mem. Inst. Oswaldo Cruz;112(2):140-145, Feb. 2017. graf. Idioma: en. Projeto: FAPESP; . FAPESP. Resumo: BACKGROUND Fluorescence in situ hybridisation (FISH) associated with Tyramide Signal Amplification (TSA) using oligonucleotides labeled with non-radioactive fluorophores is a promising technique for detection and differentiation of fungal species in environmental or clinical samples, being suitable for microorganisms which are difficult or even impossible to culture. OBJECTIVE In this study, we aimed to standardise an in situ hybridisation technique for the differentiation between the pathogenic species Paracoccidioides brasiliensis and Paracoccidioides lutzii, by using species-specific DNA probes targeting the internal transcribed spacer-1 (ITS-1) of the rRNA gene. METHODS Yeast and mycelial phase of each Paracoccidioides species, were tested by two different detection/differentiation techniques: TSA-FISH for P. brasiliensis with HRP (Horseradish Peroxidase) linked to the probe 5’ end; and FISH for P. lutzii with the fluorophore TEXAS RED-X® also linked to the probe 5’ end. After testing different protocols, the optimised procedure for both techniques was accomplished without cross-positivity with other pathogenic fungi. FINDINGS The in silico and in vitro tests show no reaction with controls, like Candida and Cryptococcus (in silico) and Histoplasma capsulatum and Aspergillus spp. (in vitro). For both phases (mycelial and yeast) the in situ hybridisation showed dots of hybridisation, with no cross-reaction between them, with a lower signal for Texas Red probe than HRP-TSA probe. The dots of hybridisation was confirmed with genetic material marked with 4’,6-diamidino-2-phenylindole (DAPI), visualised in a different filter (WU) on fluorescent microscopic. MAIN CONCLUSION Our results indicated that TSA-FISH and/or FISH are suitable for in situ detection and differentiation of Paracoccidioides species. This approach has the potential for future application in clinical samples for the improvement of paracoccidioidomycosis patients prognosis. Descritores: Paracoccidioides/classificação Paracoccidioides/genética DNA Fúngico DNA Espaçador Ribossômico -Especificidade da Espécie Sondas de Oligonucleotídeos Hibridização in Situ Fluorescente Fluorescência Corantes Fluorescentes Responsável: BR1.1 - BIREME

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