23 results on '"Melo, Analy Salles de Azevedo"'
Search Results
2. A consensus document for the clinical management of invasive fungal diseases in pediatric patients with hematologic cancer and/or undergoing hematopoietic stem cell transplantation in Brazilian medical centers
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Carlesse, Fabianne, Daudt, Liane Esteves, Seber, Adriana, Dutra, Álvaro Pimenta, Melo, Analy Salles de Azevedo, Simões, Belinda, Macedo, Carla Renata Donato, Bonfim, Carmem, Benites, Eliana, Gregianin, Lauro, Batista, Marjorie Vieira, Abramczyk, Marcelo, Tostes, Vivian, Lederman, Henrique Manoel, Lee, Maria Lúcia de Martino, Loggetto, Sandra, Galvão de Castro Junior, Cláudio, and Colombo, Arnaldo Lopes
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- 2019
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3. Emergence of cryptic species and clades of Meyerozyma guilliermondii species complex exhibiting limited in vitro susceptibility to antifungals in patients with candidemia
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Francisco, Elaine Cristina, primary, Ribeiro, Felipe de Camargo, additional, Almeida Junior, João Nobrega, additional, Pedoni, Diego Betto, additional, da Matta, Daniel Archimedes, additional, Dolande, Maribel, additional, Melo, Analy Salles de Azevedo, additional, Lima, Ricardo Ferreira, additional, Aquino, Valério Rodrigues, additional, Corzo-León, Dora E, additional, Zurita, Jeannete, additional, Cortes, Jorge Alberto, additional, Nucci, Marcio, additional, and Colombo, Arnaldo Lopes, additional
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- 2023
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4. Neonatal fungemia caused by Lodderomyces elongisporus: First case report in Latin America
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da Silva, Carolina Maria, primary, Jucá, Moacir Batista, additional, Melo, Analy Salles de Azevedo, additional, Lima, Soraia Lopes, additional, Galvão, Pauliana Valéria Machado, additional, Macêdo, Danielle Patrícia Cerqueira, additional, and Neves, Rejane Pereira, additional
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- 2023
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5. Increasing Prevalence of Multidrug-Resistant Candida haemulonii Species Complex among All Yeast Cultures Collected by a Reference Laboratory over the Past 11 Years
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Lima, Soraia Lopes, primary, Francisco, Elaine Cristina, additional, de Almeida Júnior, João Nóbrega, additional, Santos, Daniel Wagner de Castro Lima, additional, Carlesse, Fabiane, additional, Queiroz-Telles, Flávio, additional, Melo, Analy Salles de Azevedo, additional, and Colombo, Arnaldo Lopes, additional
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- 2020
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6. Evaluation of (1 → 3)‐β‐D‐glucan assay for diagnosing paracoccidioidomycosis
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Melo, Analy Salles de Azevedo, primary, Santos, Daniel Wagner de Castro Lima, additional, Lima, Soraia Lopes, additional, Rodrigues, Anderson Messias, additional, Camargo, Zoilo Pires, additional, Finkelman, Malcolm, additional, and Colombo, Arnaldo Lopes, additional
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- 2019
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7. Database establishment for the secondary fungal DNA barcode translational elongation factor 1 alpha (TEF1 alpha)
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Meyer, Wieland Irinyi, Laszlo Minh Thuy Vi Hoang Robert, Vincent Garcia-Hermoso, Dea Desnos-Ollivier, Marie Yurayart, Chompoonek Tsang, Chi-Ching Lee, Chun-Yi Woo, Patrick C. Y. and Pchelin, Ivan Mikhailovich Uhrlass, Silke Nenoff, Pietro and Chindamporn, Ariya Chen, Sharon Hebert, Paul D. N. Sorrell, Tania C. Halliday, Catriona Arthur, Ian Moretti, Maria Luiza and de Almeida Soares, Celia Maria Muniz, Mauro de Medeiros and Zancope-Oliveira, Rosely Maria Cruz, Fundacao Oswaldo Melo, Analy Salles de Azevedo Colombo, Arnaldo L. Nishikaku, Angela Satie Hendrickx, Marijke Stubbe, Dirk Normand, Anne-Cecile and Piarroux, Renaud Ranque, Stephane Dromer, Francoise and Arabatzis, Michael Velegraki, Aristea Cardinali, Gianluigi and Castanon, Laura Rosio Taylor, Maria Lucia Toriello, Conchita and de Hoog, Sybren Pais, Celia de Beer, Wilhelm Gryzenhout, Marieka Guarro, Josep Cano-Lira, Jose F. Robbertse, Barbara and Schoch, Conrad ISHAM Barcoding Pathogenic Fungi
- Abstract
With new or emerging fungal infections, human and animal fungal pathogens are a growing threat worldwide. Current diagnostic tools are slow, non-specific at the species and subspecies levels, and require specific morphological expertise to accurately identify pathogens from pure cultures. DNA barcodes are easily amplified, universal, short species-specific DNA sequences, which enable rapid identification by comparison with a well-curated reference sequence collection. The primary fungal DNA barcode, ITS region, was introduced in 2012 and is now routinely used in diagnostic laboratories. However, the ITS region only accurately identifies around 75% of all medically relevant fungal species, which has prompted the development of a secondary barcode to increase the resolution power and suitability of DNA barcoding for fungal disease diagnostics. The translational elongation factor 1 alpha (TEF1 alpha) was selected in 2015 as a secondary fungal DNA barcode, but it has not been implemented into practice, due to the absence of a reference database. Here, we have established a quality-controlled reference database for the secondary barcode that together with the ISHAM-ITS database, forms the ISHAM barcode database, available online at http://its.mycologylab.org/. We encourage the mycology community for active contributions.
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- 2019
8. Oral colonization by Candida spp. in liver transplant patients: Molecular identification and antifungal susceptibilityOral colonization by Candida spp. in liver transplant.
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Sabadin, Clarice Elvira Saggin, Lopes, Soraia Lima, Gompertz, Olga Fischmamm, Santana, Gilglécia Novaes Pereira, Melo, Analy Salles de Azevedo, Rigo, Lilian, Matta, Daniel Archimedes Da, and Barbosa, Dulce Aparecida
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Candida species are commensal to normal oral microbiota; however, they can cause infections if immune functions are reduced. The aim of this study was to investigate oral colonization, identify species, and test the susceptibility profile to antifungals. A descriptive study included 97 liver transplant patients who attended the transplant center of a referral hospital in southern Brazil. Two oral swab collections were performed, with a 6-month gap between collections. The samples were identified by sequencing the internal transcribed spacer ITS region of the ribosomal DNA. The sensitivity test was performed with fluconazole, amphotericin B, and micafungin using a broth microdilution method recommended by Clinical and Laboratory Standards Institute document M27-A4. Eighty-two patients were investigated and 15 were excluded for presenting clinical infection. The identification of yeasts showed colonization in 66% and 61.9% in collections A and B, respectively. Candida albicans was the most prevalent species in both collections (n = 29/50 and n = 27/49, respectively). In 31 (62%) patients, the yeast species remained the same for 6 months, and in 19 (38%) the colonizing species was substituted. Thirty-two isolates from collection A were sensitive (S) to Fluconazole, 13 sensitive dose-dependent (SDD), and five resistant (R). In collection B, 32 were S, 12 SDD, and 5 R. For amphotericin B and micafungin, all isolates were sensitive. With knowledge of the species and identification of strains resistant to fluconazole, useful information can be alerts about the emergence of antifungal resistance strains. [ABSTRACT FROM AUTHOR]
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- 2021
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9. Epidemiology and prognostic factors of nosocomial candidemia in Northeast Brazil: A six-year retrospective study
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Medeiros, Mariana Araújo Paulo de, primary, Melo, Ana Patrícia Vieira de, additional, Bento, Aurélio de Oliveira, additional, Souza, Luanda Bárbara Ferreira Canário de, additional, Neto, Francisco de Assis Bezerra, additional, Garcia, Jarmilla Bow-Ltaif, additional, Zuza-Alves, Diana Luzia, additional, Francisco, Elaine Cristina, additional, Melo, Analy Salles de Azevedo, additional, and Chaves, Guilherme Maranhão, additional
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- 2019
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10. Evaluation of (1 → 3)‐β‐D‐glucan assay for diagnosing paracoccidioidomycosis.
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Melo, Analy Salles de Azevedo, Santos, Daniel Wagner de Castro Lima, Lima, Soraia Lopes, Rodrigues, Anderson Messias, Camargo, Zoilo Pires, Finkelman, Malcolm, and Colombo, Arnaldo Lopes
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COCCIDIOIDOMYCOSIS , *MYCOSES , *AGGLUTINATION tests , *PARACOCCIDIOIDOMYCOSIS , *PULSE-code modulation - Abstract
Summary: Background: Paracoccidioidomycosis (PCM) is highly prevalent in Latin America, but no commercial system is available for diagnosing this endemic mycosis. Objectives: To check the performance of (1 → 3)‐β‐D‐glucan assay (BDG) for diagnosing PCM in 29 patients with proven fungal disease and compared with double immunodiffusion assay for detecting anti‐Paracoccidioides antibodies. Patients and Methods: We selected 52 serum samples sequentially obtained from 29 patients with active PCM (12 chronic and 17 acute form). Samples were collected at baseline, and for 16 patients, additional serum levels were obtained after 3 and 6 months of antifungal treatment. Detection of BDG in serum was performed by using the Fungitell® assay. For the double immunodiffusion assay, Paracoccidioides exoantigen was used in latex agglutination tests to detect serum anti‐Paracoccidioides antibodies. Results: Despite exhibiting good sensitivity in the diagnosis of patients with PCM, we failed to demonstrate any correlation between the postdiagnosis kinetic profile of BDG serum levels and clinical response to antifungal therapy. This finding may be related to the maintenance of quiescent foci of fungal infection in several organs and tissues, a phenomenon that has been previously reported by other authors and helps to understand why so many relapses are documented in patients treated for short periods of time. Finally, we did not find any correlation between BDG quantification and specific anti‐P brasiliensis antibodies serum titres in patients with PCM. Conclusions: In conclusion, BDG is detected in serum samples of most patients with PCM but is probably not useful for predicting clinical response to antifungal therapy. [ABSTRACT FROM AUTHOR]
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- 2020
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11. FUNGEMIA CAUSED BY Candida SPECIES IN A CHILDREN'S PUBLIC HOSPITAL IN THE CITY OF SÃO PAULO, BRAZIL: STUDY IN THE PERIOD 2007-2010
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Oliveira, Vanessa Kummer Perinazzo, primary, Ruiz, Luciana da Silva, additional, Oliveira, Nélio Alessandro Jesus, additional, Moreira, Débora, additional, Hahn, Rosane Christine, additional, Melo, Analy Salles de Azevedo, additional, Nishikaku, Angela Satie, additional, and Paula, Claudete Rodrigues, additional
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- 2014
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12. Identification of genes differentially expressed in hyphae of Candida albicans
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Melo,Analy Salles de Azevedo, Serafim,Rui Cosme, and Briones,Marcelo Ribeiro da Silva
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Differential screening ,fungi ,Candida albicans ,HXT7 ,YLL34 - Abstract
The ability to switch from yeast to hyphal forms is essential for Candida albicans virulence. This morphological switch involves the expression of hyphal-specific genes under the control of transcriptional factors. To contribute to the discovery of hyphal-specific genes, we used a differential screening method where clones of a genomic DNA library were hybridized with yeast and hyphal cDNA probes. Two clones with increased expression in hyphae were selected for study. Sequencing these clones, we found that they encoded two important metabolic genes, CaHXT7 (high-affinity hexose transporter) and CaYLL34 (member of the AAA ATPase family). CaHXT7 and CaYLL34 ORFs were completely determined. Analyses of the putative proteins show that: (1) CaHxt7p has one hexose transporter domain and (2) CaYll34p has two AAA ATPase domains. These results show, for the first time, increased expression of metabolic genes in C. albicans hyphae. Also, because the proteins encoded by CaHXT7 and CaYLL34 may be necessary for the switching to hyphae, they could be new targets for antifungal drugs.
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- 2003
13. Identification of genes differentially expressed in hyphae of Candida albicans
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Melo, Analy Salles de Azevedo, Serafim, Rui Cosme, and Briones, Marcelo Ribeiro da Silva
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Differential screening ,Candida albicans ,triagem diferencial ,HXT7 ,YLL34 - Abstract
The ability to switch from yeast to hyphal forms is essential for Candida albicans virulence. This morphological switch involves the expression of hyphal-specific genes under the control of transcriptional factors. To contribute to the discovery of hyphal-specific genes, we used a differential screening method where clones of a genomic DNA library were hybridized with yeast and hyphal cDNA probes. Two clones with increased expression in hyphae were selected for study. Sequencing these clones, we found that they encoded two important metabolic genes, CaHXT7 (high-affinity hexose transporter) and CaYLL34 (member of the AAA ATPase family). CaHXT7 and CaYLL34 ORFs were completely determined. Analyses of the putative proteins show that: (1) CaHxt7p has one hexose transporter domain and (2) CaYll34p has two AAA ATPase domains. These results show, for the first time, increased expression of metabolic genes in C. albicans hyphae. Also, because the proteins encoded by CaHXT7 and CaYLL34 may be necessary for the switching to hyphae, they could be new targets for antifungal drugs. A transição morfológica de levedura para hifa é essencial para a virulência de Candida albicans. Esta transição envolve a expressão de genes hifa-específicos que estão sob o controle de fatores transcricionais. Para descobrir genes hifa-específicos utilizamos um método de triagem diferencial, onde clones de biblioteca de DNA genômico foram hibridizados com sondas de cDNA de levedura e hifa. Dois clones com aumento de expressão em hifa foram selecionados. O sequenciamento dos insertos destes clones permitiu a identificação de dois genes metabólicos importantes: CaHXT7 (high-affinity hexose transporter) e CaYLL34 (da família AAA ATPase). As ORFs completas destes genes foram caracterizadas e a análise das proteínas hipotéticas revelou que: (1) CaHxt7p tem um domínio de transportador de hexose e (2) CaYll34 tem dois domínios AAA ATPase. Este é o primeiro estudo que demonstra aumento de expressão de genes metabólicos em hifas de C. albicans. Ainda, a associação dos produtos de CaHXT7 e CaYLL34 com a formação de hifas torna estas proteínas potenciais novos alvos para drogas antifúngicas.
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- 2003
14. Systematic review and new insights into the molecular characterization of the Candida rugosa species complex
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Padovan, Ana Carolina Barbosa, primary, Melo, Analy Salles de Azevedo, additional, and Colombo, Arnaldo Lopes, additional
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- 2013
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15. Echinocandin Resistance in Two Candida haemulonii Isolates from Pediatric Patients
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Dominguez Muro, Marisol, primary, Motta, Fábio de Araújo, additional, Burger, Marion, additional, Melo, Analy Salles de Azevedo, additional, and Dalla-Costa, Líbera Maria, additional
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- 2012
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16. Identification of genes differentially expressed in hyphae of Candida albicans
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Melo, Analy Salles de Azevedo, primary, Serafim, Rui Cosme, additional, and Briones, Marcelo Ribeiro da Silva, additional
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- 2003
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17. Echinocandin Resistance in Two Candida haemuloniiIsolates from Pediatric Patients
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Dominguez Muro, Marisol, Motta, Fábio de Araújo, Burger, Marion, Melo, Analy Salles de Azevedo, and Dalla-Costa, Líbera Maria
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ABSTRACTWe report 3 cases of patients with Candida haemuloniiisolates that were obtained from hemocultures. In 2 of the 3 cases, isolates exhibited resistance to echinocandins and fluconazole. This is the first report of an echinocandin-resistant species of this fungus in pediatric patients.
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- 2012
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18. Neonatal fungemia caused by Lodderomyces elongisporus: first case report in Latin America.
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Silva, Carolina Maria da, Jucá, Moacir Batista, Melo, Analy Salles de Azevedo, Lima, Soraia Lopes, Galvão, Pauliana Valéria Machado, Macêdo, Danielle Patrícia Cerqueira, and Neves, Rejane Pereira
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CANDIDEMIA , *FUNGEMIA , *NEONATAL intensive care units - Abstract
• Neonatal fungemia due to rare yeasts is associated with high mortality rates. • Fungemia by Lodderomyces elongisporus is usually associated with immunosuppressed adults. • We report a case of Lodderomyces elongisporus fungemia in a premature neonate. Premature hospitalized neonates have a greater risk for candidemia, however, fungemia due to rare opportunistic yeasts have been recently reported and is associated with high mortality rates. We herein report the first case in Latin America of Lodderomyces elongisporus fungemia in a premature neonate with a fatal outcome. [ABSTRACT FROM AUTHOR]
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- 2023
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19. Efeito da fração acetato de etila do extrato de Eugenia uniflora na expressão global de proteínas durante a morfogênese Candida albicans
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Silva, Walicyranison Plinio da, Melo, Analy Salles de Azevedo, Luchessi, André Ducati, Silva, Marcelo de Sousa da, Batista, Wagner Luiz, and Chaves, Guilherme Maranhão
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Modelo Murino ,Proteômica ,CIENCIAS DA SAUDE [CNPQ] ,Candida albicans ,Fagocitose ,Morfogênese ,Eugenia uniflora - Abstract
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Em certas circunstâncias, Candida albicans pode passar de colonizante para infectante e, como na candidíase oral. A morfogênese de C. albicans, bem como sua capacidade de combater o estresse oxidativo no interor de células fagocíticas, é um fator essencial para a invasão tecidual e estabelecimento da infecção. Devido ao baixo arsenal antifúngico disponível no mercado e o constante surgimento de cepas resistentes, faz-se necessária a pesquisa de novas fontes terapêuticas, principalmente oriundas de produtos naturais. No presente estudo foram selecionados 48 isolados clínicos de C. albicans oriundos da cavidade bucal de pacientes transplantados renais. A fração acetato de etila de Eugenia uniflora foi utilizada na concentração de 1000 μg/mL para avaliar a ação comparativa (tratada e não tratada com extrato) sobre a fagocitose e morfogênese de C. albicans. Foi realizado o ensaio de resistência ao ataque de neutrófilos polimorfonucleares. O isolado 111R, de alta capacidade filamentação foi utilizado para avaliação do perfil protéico por meio de análise proteômica, bem como da interação com proteínas diretamente associadas à morfogênese. A resposta à infecção foi observada em modelo murino de candidíase oral e a ação tóxica do extrato de E. uniflora foi observada em ensaio de MTT. O extrato de E. uniflora reduziu significativamente a capacidade de fagocitose de C. albicans (média total de 120.36 ± 36.71 vs. 44.68 ± 19.84). Trinta e nove proteínas foram identificadas na análise proteômica, relacionadas à geração de energia, metabolismo de proteínas e glicose, divisão celular, transporte citoplasmático, metabolismo de ácidos nucléicos, estrutura celular e resposta ao estresse. Importantes proteínas relacionadas com a formação do citoesqueleto foram reguladas negativamente nas células tratadas. Houve instalação de infecção na cavidade oral dos camundongos e a infecção foi atenuada quando C. albicans foi pré-incubada na presença do extrato de E. uniflora e quando o extrato foi aplicado na cavidade oral após a instalação da infecção. Este resultado foi condizente com a redução na contagem de UFC (2.36 vs. 1.85 Log10 CFU/ml) e a atenuação dos danos teciduais observados na análise histopatológica. O extrato de E. uniflora não foi tóxico para células humanas mesmo em concentrações 8x acima da utilizada nos experimentos. A fração acetato de etila de E. uniflora poderá causar danos à parede celular e proteínas essenciais ao metabolismo de C. albicans, afetando proteínas relacionadas à estrutura celular, reduzindo a capacidade plástica de filamentação, atenuando a ação invasiva em modelo animal, sem causar efeito tóxico em células humanas, podendo ser uma futura alternativa terapêutica para o tratamento de infecções por Candida. Under certain circumstances, Candida albicans may switch a colonizing to a infecting yeast, such as in oral candidiasis. Morphogenesis in C. albicans, besides its ability to combat the oxidative stress inside phagocytic cells is an essential step for tissue invasion and establishment of infection. Due to the reduced drug arsenal used for treatment of fungal infections and the constant emergence of resistant strains, it is mandatory to search for new therapeutic sources, mainly from natural products. In the present study, 48 clinical isolates of C. albicans from the oral cavity of renal transplant recipients were selected. In order to evaluate the comparative action (treated and untreated cellst) on phagocytosis and morphogenesis of C. albicans, the ethyl acetate fraction of Eugenia uniflora extract was used at a concentration of 1000 μg/mL. Resistance of C. albicans to polymorphonuclear neutrophils was carried out. The isolate 111R, a highly filamentous strain was used to evaluate protein profiling through proteomic analysis, as well as the interaction with proteins directly associated with morphogenesis. The in vivo response to infection was observed in murine model of oral candidiasis and the toxic action of E. uniflora extract was observed in the MTT assay. E. uniflora extract significantly reduced the phagocytosis of C. albicans (Mean 120.36 ± 36.71 vs. 44.68 ± 19.84). Thirty-nine proteins, related to energy generation, protein and glucose metabolism, cell division, cytoplasmic transport, nucleic acid metabolism, cell structure and stress response were identified with proteomics analysis. Important proteins directly related with cytoesqueleton formation were down regulated on treated cells. The infection in the oral cavity of the mice was established and it was attenuated when both C. albicans cells were either preincubated in the presence of E. uniflora extract or when the extract was applied to the surface of the oral cavity after infection. These results were consistent with the reduction in CFU countings (2.36 vs. 1.85 Log10 CFU/ml) and attenuation of tissue damages observed in the histopathologycal analysis. E. uniflora extract was non-toxic to human cells even at concentrations 8 fold hugher than the one used in the experiments. The ethyl acetate fraction of E. uniflora may act act damaging and metabolism essential proteinsmainly related to cellular structure, reducing the plastic capacity of filamentation and attenuating infection in a murine modell model, without causing any toxic effect on human cells, suggesting that it may be a future therapeutic alternative for the treatment of Candida infections.
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- 2017
20. Fatores de virulência, resistência ao estresse osmótico e susceptibilidade de isolados de Candida tropicalis oriundos de ambiente costeiro do nordeste brasileiro
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Alves, Diana Luzia Zuza, Pedrosa, Matheus de Freitas Fernandes, Melo, Analy Salles de Azevedo, and Chaves, Guilherme Maranhão
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Ambiente costeiro ,Fatores de virulência ,Susceptibilidade antifúngica ,Candida tropicalis ,CIENCIAS DA SAUDE::FARMACIA [CNPQ] - Abstract
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Vários estudos têm sido desenvolvidos com relação aos riscos à saúde humana associados ao uso recreativo de praias contaminadas com esgotos domésticos. Esses resíduos contêm vários micro-organismos, incluindo Candida tropicalis, agente etiológico tanto de infecções superficiais quanto sistêmicas, além de indicador de contaminação fecal do meio ambiente. Nesse contexto, o presente trabalho teve como objetivo caracterizar isolados de C. tropicalis oriundos da areia da Praia de Ponta Negra, Natal, Rio Grande do Norte, Brasil, quanto a expressão de fatores de virulência in vitro, a adaptação ao estresse osmótico e a susceptibilidade às fármacos antifúngicos. Foram analisados 62 isolados ambientais de C. tropicalis, observando-se grande variação entre os mesmos para os diversos fatores de virulência avaliados. Em geral, os isolados ambientais foram mais aderentes a células epiteliais bucais humanas (CEBH) do que a cepa de referência de C. tropicalis ATCC13803, além de serem altamente produtoras de biofilme. Em relação à morfogênese, a maioria dos isolados exibiu fenótipo rugoso em meio Spider (34 isolados, 54,8%). Na avaliação da atividade enzimática, a maioria dos isolados teve maior produção de proteinase do que a cepa de referência de C. tropicalis ATCC13803. Adicionalmente, 35 isolados (56,4%) tiveram alta atividade hemólitica (índice de hemólise > 55). Com relação à resistência de C. tropicalis ao estresse osmótico, 85,4% dos isolados foram resistentes em meio contendo 15% de cloreto de sódio, o que corrobora com a alta capacidade de sobrevivência descrita para essa levedura no ambiente marítimo. Finalmente, no que diz respeito à sensibilidade aos antifúngicos foi observada elevada resistência aos azólicos testados (fluconazol, voriconazol e itraconazol), com ocorrência do fenômeno “Low-high” e de efeito semelhante ao crescimento paradoxal que ocorre para as equinocandinas. As cepas resistentes aos três azólicos testados foram 15 (24,2%). Para a anfotericina B também ocorreu resistência (14 isolados, 22,6%), ao passo que para as equinocandinas todas as cepas foram sensíveis. Portanto, nossos resultados demonstram que isolados de C. tropicalis oriundos da areia de praia do nordeste brasileiro podem expressar plenamente atributos de virulência e apresentam alta capacidade de persistência no ambiente costeiro, além de serem significativamente resistentes aos antifúngicos mais empregados na prática clínica atual. Isso constitui potencial risco à saúde dos frequentadores desse ambiente, especialmente indivíduos imunocomprometidos e em extremos etários. Several studies have been developed regarding health risks associated with the recreational use of beaches contaminated with domestic sewage. These wastes contain various microorganisms, including Candida tropicalis, etiologic agent of both superficial infections such as systemic, as well as indicator of fecal contamination for the environment. In this context, the objective of this study was to characterize C. tropicalis isolates from the sandy beach of Ponta Negra, Natal, Rio Grande do Norte, Brazil, regarding the expression of in vitro virulence factors, adaptation to osmotic stress and susceptibility to antifungal drugs. We analyzed 62 environmental isolates of C. tropicalis and observed a great variation between them for the various virulence factors evaluated. In general, environmental isolates were more adherent to CEBH than C. tropicalis ATCC13803 reference strain, besides the fact they were also highly biofilm producers. In relation to morphogenesis, most isolates presented wrinkled phenotype in Spider medium (34 isolates, 54.8 %). When assessing enzyme activity, most isolates had higher proteinase production than C. tropicalis ATCC13803 reference strain. In addition, 35 isolates (56.4 %) had high hemolytic activity (hemolysis index > 55). With regard to C. tropicalis resistance to osmotic stress, 85.4% of the isolates were able to grow in a liquid medium containing 15% sodium chloride, corroborating to high survival capacity described for this yeast at marine environment. Finally, with regard to sensitivity to antifungal drugs, it was observed high resistance to the azoles tested, with the occurrence of the "Low-high" phenomenon and similar effect to the paradoxical growth which occurs to the echinocandins. For the three azoles tested we verified that 15 strains were resistant (24.2 %). Some strains were also resistant to amphotericin B (14 isolates, 22.6 %), while all of them were sensitive for the echinocandins tested. Therefore, our results demonstrate that C. tropicalis isolated from the sand of northeast of Brazil can fully express virulence attributes and showed a high persistence capacity on the coastal environment, in addition of being significantly resistant to most applied antifungals in current clinical practice. This constitutes a potential health risk to visitors of this environment, especially immunocompromised individuals and those with extreme age range.
- Published
- 2015
21. Characterization of Exophiala strains stored in the filamentous fungi culture collection of the Special Mycology Laboratory UNIFESP
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Silva, Wendy Colalto Rodrigues da [UNIFESP] and Melo, Analy Salles de Azevedo [UNIFESP]
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Exophiala ,Antifúngicos ,Exophiala/genética - Abstract
Coordenação e Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Introdução: Leveduras negras do gênero Exophiala respondem por número substancial de infecções humanas superficiais e invasivas causadas por fungos negros. A identificação acurada de espécies de Exophiala baseia-se em critérios morfológicos complementados por tipagem molecular. A infecção em humanos causada por Exophiala depende de três fatores principais: porta de entrada, estado imunológico do paciente e tolerância térmica do agente. A maioria destas infecções são cutâneas e superficiais, entretanto, infecções sistêmicas fatais já foram relatadas. Objetivos: Estudar as características fenotípicas, genotípicas e susceptibilidade a antifúngicos de 23 isolados clínicos, identificados fenotipicamente como Exophiala sp. armazenados no Banco de Fungos Filamentosos do LEMI. Métodos: Para a identificação molecular nos baseamos no sequenciamento da região ITS do rDNA, comparamos as sequências obtidas com sequências já depositadas em bancos genômicos (NCBI e CBS) e analizamos as sequências através da ferramenta BLASTn. Para a caracterização da macromorfologia e micromorfologia os isolados foram semeados em meio PDA e incubados a 25°C. Para análise de termotolerância, cada isolado foi semeado em 3 meios de cultura: PDA, SGA e MEA, e incubados em 4 temperaturas diferentes: 15°C, 25°C, 37°C e 42°C. Para cada um dos testes fenotípicos citados, a incubação foi de 14 dias nas devidas temperaturas para a realização das leituras. Os testes de susceptibilidade aos antifúngicos, anfotericina B (AMB), itraconazol (ITC), voriconazol (VRC), posaconazol (PSC), 5-fluorocitosina (5-FC), caspofungina (CFG) e terbinafina (TRB) foram realizados por microdiluição em caldo de acordo com o documento CLSI M38-A2. Resultados: A tipagem molecular teve sucesso na identificação de 100% das amostras que foram assim reconhecidas: E. dermatitidis (8), E. xenobiotica (6), E. bergeri (4), E. oligosperma (3), E. spinifera (1) e E. mesophila (1). A macromorfologia mostrou que as espécies de Exophiala são pleomórficas com relação ao aspecto e cor das colônias, mesmo considerando isolados da mesma espécie. Em relação à micromorfologia, observamos estruturas similares entre diferentes espécies do gênero, não possibilitando assim a identificação em nível de espécie, sendo necessário confirmação por método molecular. Na análise de termotolerância, todas as espécies cresceram a 25 e 37ºC com exceção de E. mesophila, que cresceu somente a 25oC. A 42ºC apenas as espécies E. dermatitidis, E. oligosperma e E. xenobiotica apresentaram crescimento, sendo as duas últimas com crescimento reduzido, não possibilitando caracterização. Em relação à susceptibilidade, as CIMs mais baixas foram verificadas para os azólicos, enquanto que as maiores foram observadas para 5-FC, CFG e TRB. Conclusões: (i) O sequenciamento da região ITS permitiu a correta identificação de espécie para os 23 isolados clínicos; (ii) As características observadas nos testes de macromorfologia, micromorfologia e termotolerância para os isolados de Exophiala spp. analisados, mostraram que não são suficientes para a identificação acurada de espécie; (iii) O teste de susceptibilidade para Exophiala spp, utilizando o método de microdiluição em caldo padronizado pelo CLSI (2008) documento M38-A2 para fungos negros, mostrou-se consistente e reprodutível; (iv) De forma geral, a grande maioria dos isolados testados “in vitro” apresentou CIM ≤0,5 μg/mL exceto para 5-FC e CFG; (v) Apesar das diretrizes não indicarem AMB para tratamento contra Exophiala sp., esta mostrou-se ativa com CIMs ≤1 μg/mL para todas as espécies deste estudo, com exceção de E. mesophila. Introduction: Black yeasts of Exophiala genus are responsible for a large number of superficial and invasive i infections. The identification of Exophiala at species level is based on morphological observation complemented by molecular typing. The human infection caused by Exophiala sp. depends on 3 main factors: fungus inoculation, patient’s immunologic status and thermotolerance of the agent. Most of these infections are superficial, but fatal systemic infections have been reported. Objective: The aim of this study was to identify Exophiala spp. by molecular method using internal transcript sequences (ITS) of rDNA gene of 23 clinical isolates of Exophiala deposited in our culture collection (LEMI). We evaluated macro and micromorphological characteristics, thermotolerance, and antifungal susceptibility to 7 different agents. Methods: Molecular identification was based on sequencing the ITS region and analyzed by BLASTn tool on GenBank (NCBI and CBS). For the macromorphology and micromorphology characterization, each isolate was streaked on PDA medium and incubated at 25ºC. For the thermotolerance analysis, each isolate was streaked in 3 different media: PDA, SGA and MEA, and incubated at 4 temperatures 15ºC, 25ºC, 37ºC and 42ºC, for 14 days. We used the CLSI M38-A2 broth microdilution method to test the 23 clinical isolates against 7 antifungal agents: amphotericin B (AMB), itraconazole (ITR), voriconazole (VOR), posaconazole (POS), caspofungin (CAS), terbinafine (TER) and 5-fluorocitosina (5-FC). Results: The BLASTn analysis of ITS sequences was able to identify 100% of the clinical isolates as follows: E. dermatitidis (8), E. xenobiotica (6), E. bergeri (4), E. oligosperma (3), E. spinifera (1), E. mesophila (1). The macromorphology showed that the Exophiala species have pleomorphic characteristics considering the colony aspect and color, even for isolates of the same species. On micromorphology analysis, we observed similar structures among different species of the genus, not allowing accurate species identification. The thermotolerance analysis showed that except for E. mesophila, all the isolates of different species grew at 25 and 37ºC. Only E. dermatitidis, E. oligosperma and E. xenobiotica grew as colonies at 42ºC, while the last two presented reduced growth, not allowing characterization. The antifungal susceptibility tests showed that triazols were the most active agents in vitro against Exophiala, while higher MICs were observed for 5-FC, CFG and TRB. Conclusions: (i) ITS sequencing allowed accurate identification of all the 23 clinical isolates tested; (ii) The characteristics observed on macromorphology, micromorphology and thermotolerance analysis for all the isolates showed that these tests are not sufficient for accurate species identification; (iii) The susceptibility tests using the microdilution method standardized by the CLSI broth (2008) document M38-A2 for black fungi, was consistent and reproducible for Exophiala spp; (iv) In general, the vast majority of isolates tested in vitro presented MIC ≤ 0.5 μg/mL for most of the antifungal agents, except for 5-FC and CFG; (v) Although the guidelines do not indicate AMB for treatment against Exophiala sp., this study showed activity with MICs ≤ 1 μg/mL for all the species tested, excepted for E. mesophila. BV UNIFESP: Teses e dissertações
- Published
- 2014
22. Fungal Peritonitis Due to Fusarium solani Species Complex Sequential Isolates Identified with DNA Sequencing in a Kidney Transplant Recipient in Brazil.
- Author
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da Silva-Rocha WP, Zuza-Alves DL, Melo AS, and Chaves GM
- Subjects
- Adult, Brazil, Enterobacter genetics, Female, Fusariosis microbiology, Fusariosis pathology, Fusarium genetics, Gram-Positive Bacterial Infections microbiology, Gram-Positive Bacterial Infections pathology, Humans, Kidney Transplantation, Molecular Sequence Data, Peritonitis microbiology, Peritonitis pathology, Sequence Analysis, DNA, Transplant Recipients, Enterobacter isolation & purification, Fusariosis complications, Fusariosis diagnosis, Fusarium isolation & purification, Gram-Positive Bacterial Infections complications, Gram-Positive Bacterial Infections diagnosis, Peritonitis diagnosis
- Abstract
Fungal peritonitis is a rare serious complication most commonly observed in immunocompromised patients under peritoneal dialysis. Nevertheless, this clinical condition is more difficult to treat than bacterial peritonitis. Bacterial peritonitis followed by the use of antibiotics is the main risk factor for developing fungal peritonitis. Candida spp. are more frequently isolated, and the isolation of filamentous fungi is only occasional. Here we describe a case of Fusarium solani species complex peritonitis associated with bacterial peritonitis in a female kidney transplant recipient with previous history of nephrotic syndrome. The patient has had Enterobacter sp. endocarditis and was hypertensive and diabetic. Two sequential isolates of F. solani were recovered from cultures and identified with different molecular techniques. She was successfully treated with 50 mg daily amphotericin B for 4 weeks.
- Published
- 2015
- Full Text
- View/download PDF
23. Echinocandin resistance in two Candida haemulonii isolates from pediatric patients.
- Author
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Muro MD, Motta Fde A, Burger M, Melo AS, and Dalla-Costa LM
- Subjects
- Adolescent, Child, Female, Fluconazole pharmacology, Humans, Infant, Antifungal Agents pharmacology, Candida drug effects, Candida isolation & purification, Candidemia microbiology, Drug Resistance, Fungal, Echinocandins pharmacology
- Abstract
We report 3 cases of patients with Candida haemulonii isolates that were obtained from hemocultures. In 2 of the 3 cases, isolates exhibited resistance to echinocandins and fluconazole. This is the first report of an echinocandin-resistant species of this fungus in pediatric patients.
- Published
- 2012
- Full Text
- View/download PDF
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