Species-Specific Differences in Rhodamine 6G Accumulation of Candida Isolates Detected by Flow Cytometric Analysis / 대한진단검사의학회지

BACKGROUND: Fluorescent dye Rhodamine 6G (R6G) is a substrate of multidrug resistance pumps and its accumulation is reduced in some azole-resistant Candida isolates with the upregulation of multidrug efflux transporter genes. Despite reports on species-specific differences in azole susceptibility in various Candida species, only a few studies have been reported on the R6G accumulation among clinical isolates of Candida species. In this study, we compared R6G accumulation between six different Candida species. METHODS: The intracellular accumulation of R6G and minimal inhibitory concentrations (MICs) of three triazole agents were investigated in 48 strains of six Candida species (14 C. albicans, 9 C. tropicalis, 8 C. glabrata, 8 C. krusei, 7 C. parapsilosis, and 2 C. haemulonii). R6G accumulation was measured by using flow cytometry and the geometric mean of the fluorescence intensity (GMF) was used to compare the accumulation between the Candida isolates. RESULTS: The GMF values for the C. tropicalis, C. albicans, C. krusei, C. parapsilosis, and C. glabrata isolates were 167.3+/-18.5, 126.9+/-6.6, 88.5+/-18.5, 50.8+/-7.0, and 38.1+/-3.9, respectively. C. glabrata had a significantly lower mean GMF than all the other Candida species (P<0.05). While some Candida strains with trailing growth phenomenon and increased fluconazole MIC did not have a reduced GMF, three Candida strains with increased MICs to all three triazole agents had a reduced GMF. CONCLUSIONS: This study found species-specific differences in R6G accumulation in Candida. In addition, the intracellular R6G accumulation can be used to investigate the drug efflux mechanism in azole-resistant Candida strains.

Dentin penetrability evaluation of three different dyes in root-end cavities filled with mineral trioxide aggregate (MTA)

A proposta deste trabalho foi avaliar a penetração de três corantes em retrocavidades obturadas com MTA. As retrocavidades foram confeccionadas com aparelho de ultra-som em 30 dentes unirradiculares e obturadas com MTA. Os espécimes foram divididos aleatoriamente em três grupos (n = 10) e imersos nas seguintes soluções corantes: azul de metileno a 2% (MET), nitrato de prata a 50% (NIT) e rodamina B a 0,2% (ROD). Duas fatias transversais (1 mm) da região retrobturada foram obtidas e avaliadas através do software Image Tool 3.0, objetivando quantificar a área (em mm2) de penetração do corante ao redor das retrobturações. Os dados foram submetidos à análise estatística, utilizando o teste t de Student. A menor área de penetração foi observada no grupo NIT, nas duas fatias (p < 0.05). A penetração de corante foi significativamente maior no grupo ROD quando comparado ao grupo NIT, nas duas fatias (p < 0.05), e ao grupo MET, somente na fatia 1 (p < 0.05). Dentro das limitações desta pesquisa, concluiu-se que a escolha da solução corante pode influenciar a avaliação da penetração em estudos sobre retrobturações e que o grupo NIT teve a menor capacidade de penetração na dentina apical.