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Maltz, Marisa
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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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Id: biblio-841808
Autor: Yamashiro, Sandra; Fiuza, Vagner Ricardo da Silva; Teixeira, Ângela Therezinha Lauand Sampaio; Branco, Nilson; Levy, Carlos Emílio; Castro, Isabel Cristina Vidal Siqueira de; Franco, Regina Maura Bueno.
Título: Enterocytozoon bieneusi detected by molecular methods in raw sewage and treated effluent from a combined system in Brazil
Fonte: Mem. Inst. Oswaldo Cruz;112(6):403-410, June 2017. tab, graf.
Idioma: en.
Resumo: BACKGROUND Enterocytozoon bieneusi are the most common microsporidia associated with different clinical manifestations such as diarrhoea, respiratory tract inflammation and acalculous cholecystitis, especially in immunocompromised patients. Infection usually occurs by ingestion of food and water contaminated with spores, but can also result from direct contact with spores through broken skin, eye lesions, and sexual transmission, depending on the microsporidian species. Although there are reports of E. bieneusi found in humans and animals in Brazil, there are no published studies of environmental samples examined by molecular methods. OBJECTIVES The purpose of this study was to verify the presence of E. bieneusi in raw sewage and treated effluent from a combined system by molecular methods. METHODS Raw sewage and treated effluent samples collected from a combined system were analysed for the presence of E. bieneusi using the internal transcriber spacer (ITS) region of E. bieneusi by nested polymerase chain reaction. FINDINGS The analysis revealed E. bieneusi presence and a novel genotype (EbRB) in one raw sewage sample and one treated effluent. MAIN CONCLUSIONS The presence of E. bieneusi in final effluent indicates that the combined system may not remove microsporidian spores. This study is the first report of E. bieneusi in environmental samples in Brazil.
Descritores: DNA Fúngico/genética
Reação em Cadeia da Polimerase
Enterocytozoon/isolamento & purificação
Enterocytozoon/genética
-Filogenia
Brasil
Análise de Sequência
Genótipo
Responsável: BR1.1 - BIREME


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Id: biblio-842793
Autor: Aguiar, Paula Augusta Dias Fogaça de; Pedroso, Reginaldo dos Santos; Borges, Aércio Sebastião; Moreira, Tomaz de Aquino; Araújo, Lúcio Borges de; Röder, Denise Von Dolinger de Brito.
Título: The epidemiology of cryptococcosis and the characterization of Cryptococcus neoformans isolated in a Brazilian University Hospital
Fonte: Rev. Inst. Med. Trop. Säo Paulo;59:e13, 2017. tab, graf.
Idioma: en.
Resumo: ABSTRACT Cryptococcosis, a systemic disease caused by the fungus Cryptococcus neoformans/ Cryptococcus gattii is more severe in immunocompromised individuals. This study aimed to analyze the epidemiology of the disease, the molecular characteristics and the antifungal susceptibility of C. neoformans isolated from patients treated in a Brazilian university hospital. This retrospective study was conducted in the Clinical Hospital, Federal University of Uberlândia, and evaluated cases of cryptococcosis and strains of C. neoformans isolated from 2004 to 2013. We evaluated 41 patients, 85% of whom were diagnosed with AIDS. The fungus was isolated from the cerebrospinal fluid (CSF) of 21 patients (51%); 19.5% had fungemia and in 24% the agent was isolated from the CSF and blood, concurrently. Meningoencephalitis was the most frequent (75%) manifestation of infection. Despite adequate treatment, the mortality of the disease was 58.5%. Most isolates (97.5%) presented the VNI genotype (serotype A, var. grubii) and one isolate was genotyped as C. gattii (VGI); all the isolates were determined as mating type MATa and showed susceptibility to the tested antifungals (fluconazole, voriconazole, amphotericin B and 5-flucytosine). Although AIDS detection rates remain stable, opportunistic infections such as cryptococcosis remain as major causes of morbidity and mortality in these patients.
Descritores: Criptococose/mortalidade
Cryptococcus neoformans/isolamento & purificação
-Infecções Oportunistas Relacionadas com a AIDS/microbiologia
Infecções Oportunistas Relacionadas com a AIDS/mortalidade
Antifúngicos/farmacologia
Brasil/epidemiologia
Criptococose/microbiologia
Cryptococcus neoformans/efeitos dos fármacos
Cryptococcus neoformans/genética
DNA Fúngico/análise
Hospitais Universitários
Testes de Sensibilidade Microbiana
Estudos Retrospectivos
Limites: Humanos
Masculino
Feminino
Adulto
Pessoa de Meia-Idade
Idoso
Adulto Jovem
Responsável: BR1.1 - BIREME


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Id: biblio-842772
Autor: Livério, Harisson Oliveira; Ruiz, Luciana da Silva; Freitas, Roseli Santos de; Nishikaku, Angela; Souza, Ana Clara de; Paula, Claudete Rodrigues; Domaneschi, Carina.
Título: Phenotypic and genotypic detection of Candida albicans and Candida dubliniensis strains isolated from oral mucosa of AIDS pediatric patients
Fonte: Rev. Inst. Med. Trop. Säo Paulo;59:e14, 2017. tab.
Idioma: en.
Resumo: ABSTRACT 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. Twenty-nine samples of C. albicans isolated from this population and kept in a fungi collection were evaluated and re-characterized. In order to differentiate the two species, phenotypic tests (Thermotolerance tests, Chromogenic medium, Staib agar, Tobacco agar, Hypertonic medium) were performed and genotypic techniques using DNA sequencing were employed for confirmation of isolated species. Susceptibility and specificity were calculated for each test. No phenotypic test alone was sufficient to provide definitive identification of C. dubliniensis or C. albicans, as opposed to results of molecular tests. After amplification and sequencing of specific regions of the 29 studied strains, 93.1% of the isolates were identified as C. albicans and 6.9% as C. dubliniensis. The Staib agar assay showed a higher susceptibility (96.3%) in comparison with other phenotypic techniques. Therefore, genotypic methods are indispensable for the conclusive identification and differentiation between these species.
Descritores: Infecções Oportunistas Relacionadas com a AIDS/microbiologia
Candida/genética
Candidíase Bucal/microbiologia
DNA Fúngico/genética
-Candida albicans/genética
Candida albicans/isolamento & purificação
Candida/classificação
Candida/isolamento & purificação
Genótipo
Mucosa Bucal/microbiologia
Técnicas de Tipagem Micológica
Fenótipo
Reação em Cadeia da Polimerase
Limites: Humanos
Criança
Responsável: BR1.1 - BIREME


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Id: lil-796905
Autor: Vélez, Norida; Escandón, Patricia.
Título: Distribution and association between environmental and clinical isolates of Cryptococcus neoformans in Bogotá-Colombia, 2012-2015
Fonte: Mem. Inst. Oswaldo Cruz;111(10):642-648, Oct. 2016. tab, graf.
Idioma: en.
Projeto: Instituto Nacional de Salud.
Resumo: The propagules of the fungal species Cryptococcus neoformans and C. gattii, whose varieties are distributed world wide, are the primary cause of cryptococcosis, a life threatening disease. The study of environmental and clinical isolates of Cryptococcosis is an important contribution to the epidemiology and ecology of the fungus. The aim of this work was to determine the presence of C. neoformans and C. gattii in the environment in Bogotá, Colombia’s capital city and to establish the relation between clinical and environmental isolates in the period 2012-2015. From a total of 4.116 environmental samples collected between October 2012 - March 2014, 35 were positive for C. neoformans var. grubii. From 55 cryptococcosis cases reported in Bogotá during 2012-2015, 49 isolates were recovered. From those, 94% were identified as C. neoformans var. grubii molecular type VNI; 4% as VNII and 1,2% as C. neoformans var neoformans VNIV. The 84 detected clinical and environmental isolates studied had a similarity between 49-100% according with molecular typing. The correlation between environmental and clinical samples confirms the hypothesis that patients acquire the disease from environmental exposure to the fungal propagules.
Descritores: Criptococose/microbiologia
Cryptococcus gattii/isolamento & purificação
Cryptococcus neoformans/isolamento & purificação
Microbiologia Ambiental
-Cidades
Colômbia
Cryptococcus gattii/genética
Cryptococcus neoformans/genética
Impressões Digitais de DNA
DNA Fúngico
Genótipo
Tipagem Molecular
Técnicas de Tipagem Micológica
Limites: Humanos
Masculino
Feminino
Responsável: BR1.1 - BIREME



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