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1.
Virology ; 523: 41-51, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30077875

RESUMO

Several studies have related epigenetic mechanisms to HIV-1 latency. However, the epigenetic modifications of the host cell genome involved in the early stages of HIV-1 infection remain unclear. This study aimed to investigate epigenetic factors that are regulated at the beginning of HIV-1 infection in activated and resting CD4+ T cells. We analyzed the gene expression of 84 epigenetic targets, global DNA methylation, and HIV-1 replication kinetics for 36 h after infecting CD4+ T cells obtained from the blood of twelve healthy donors. The epigenetic targets aurora kinase B (AURKB), aurora kinase C (AURKC) and DNA methyltransferase 3B (DNMT3B), and the global DNA methylation profile are regulated during HIV-1 replication in CD4+ T cells, and this regulation can be influenced by the activation state of the cell at the time of infection. Approaches that affect the expression of these epigenetic targets could help current strategies to suppress HIV-1 replication.


Assuntos
Aurora Quinase B/genética , Aurora Quinase C/genética , Linfócitos T CD4-Positivos/metabolismo , DNA (Citosina-5-)-Metiltransferases/genética , Epigênese Genética , HIV-1/fisiologia , Interações Hospedeiro-Patógeno , Adulto , Aurora Quinase B/metabolismo , Aurora Quinase C/metabolismo , Linfócitos T CD4-Positivos/virologia , DNA (Citosina-5-)-Metiltransferases/metabolismo , Metilação de DNA , Perfilação da Expressão Gênica , Voluntários Saudáveis , Humanos , Ativação Linfocitária , Análise em Microsséries , Cultura Primária de Células , Transdução de Sinais , Internalização do Vírus , Latência Viral , Replicação Viral , DNA Metiltransferase 3B
2.
Antimicrob Agents Chemother ; 57(1): 382-9, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23114761

RESUMO

Rhodotorula species are emergent fungal pathogens capable of causing invasive infections, primarily fungemia. They are particularly problematic in immunosuppressed patients when using a central venous catheter. In this study, we evaluated the species distribution of 51 clinical and 8 environmental Rhodotorula species isolates using the ID32C system and internal transcribed spacer (ITS) sequencing. Antifungal susceptibility testing and biofilm formation capability using a crystal violet staining assay were performed. Using ITS sequencing as the gold standard, the clinical isolates were identified as follows: 44 R. mucilaginosa isolates, 2 R. glutinis isolates, 2 R. minuta isolates, 2 R. dairenensis isolates, and 1 Rhodosporidium fluviale isolate. The environmental isolates included 7 R. mucilaginosa isolates and 1 R. slooffiae isolate. Using the ID32C system, along with a nitrate assimilation test, only 90.3% of the isolates tested were correctly identified. In the biofilm formation assay, R. mucilaginosa and R. minuta exhibited greater biofilm formation ability compared to the other Rhodotorula species; the clinical isolates of R. mucilaginosa showed greater biofilm formation compared to the environmental isolates (P = 0.04). Amphotericin B showed good in vitro activity (MIC ≤ 1 µg/ml) against planktonic cells, whereas voriconazole and posaconazole showed poor activity (MIC(50)/MIC(90), 2/4 µg/ml). Caspofungin and fluconazole MICs were consistently high for all isolates tested (≥64 µg/ml and ≥ 4 µg/ml, respectively). In this study, we emphasized the importance of molecular methods to correctly identify Rhodotorula species isolates and non-R. mucilaginosa species in particular. The antifungal susceptibility profile reinforces amphotericin B as the antifungal drug of choice for the treatment of Rhodotorula infections. To our knowledge, this is the first study evaluating putative differences in the ability of biofilm formation among different Rhodotorula species.


Assuntos
Anfotericina B/farmacologia , Antifúngicos/farmacologia , DNA Espaçador Ribossômico/genética , Fungemia/microbiologia , Rhodotorula/efeitos dos fármacos , Rhodotorula/genética , Microbiologia do Solo , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , DNA Espaçador Ribossômico/classificação , Fungemia/tratamento farmacológico , Violeta Genciana , Humanos , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Microscopia Eletrônica de Varredura , Filogenia , Plâncton/efeitos dos fármacos , Plâncton/crescimento & desenvolvimento , Rhodotorula/classificação , Rhodotorula/isolamento & purificação , Análise de Sequência de DNA , Especificidade da Espécie
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