An alternative method for the analysis of melanin production in Cryptococcus neoformans sensu lato and Cryptococcus gattii sensu lato

Melanin is an important virulence factor for several microorganisms, including Cryptococcus neoformans sensu lato and Cryptococcus gattii sensu lato, thus, the assessment of melanin production and its quantification may contribute to the understanding of microbial pathogenesis. The objective of this study was to standardise an alternative method for the production and indirect quantification of melanin in C. neoformans sensu lato and C. gattii sensu lato . Eight C. neoformans sensu lato and three C. gattii sensu lato, identified through URA5 methodology, Candida parapsilosis ATCC 22019 (negative control) and one Hortaea werneckii (positive control) were inoculated on minimal medium agar with or without L‐DOPA, in duplicate, and incubated at 35°C, for 7 days. Pictures were taken from the third to the seventh day, under standardised conditions in a photographic chamber. Then, photographs were analysed using grayscale images . All Cryptococcus spp. strains produced melanin after growth on minimal medium agar containing L‐DOPA. C. parapsilosis ATCC 22019 did not produce melanin on medium containing L‐DOPA, while H. werneckii presented the strongest pigmentation. This new method allows the indirect analysis of melanin production through pixel quantification in grayscale images, enabling the study of substances that can modulate melanin production.     

Marginal fit and photoelastic stress analysis of CAD-CAM and overcast 3-unit implant-supported frameworks

Statement of problem

Several studies have shown the superiority of computer-assisted design and computer-assisted manufacturing (CAD-CAM) technology compared with conventional casting. However, an advanced technology exists for casting procedures (the overcasting technique), which may serve as an acceptable and affordable alternative to CAD-CAM technology for fabricating 3-unit implant-supported fixed dental prostheses (FDPs).