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Determination of chitin content in fungal cell wall: an alternative flow cytome...

Costa-de-Oliveira, Sofia; Silva, Ana P.; Miranda, Isabel M.; Salvador, Alexandre; Azevedo, Maria M.; Munro, Carol A.; Rodrigues, Acácio G.

The conventional methods used to evaluate chitin content in fungi, such as biochemical assessment of glucosamine release after acid hydrolysis or epifluorescence microscopy, are low throughput, laborious, time-consuming, and cannot evaluate a large number of cells. We developed a flow cytometric assay, efficient, and fast, based on Calcofluor White staining to measure chitin content in yeast cells. A staining i...


A novel flow cytometric protocol for assessment of yeast cell adhesion

Silva-Dias, A.; Miranda, Isabel M.; Rocha, R.; Monteiro-Soares, M.; Salvador, Alexandre; Rodrigues, Acácio G.; Pina-Vaz, Cidália

Microbial adhesion is a field of recognized relevance and, as such, an impressive array of tools has been developed to understand its molecular mechanisms and ultimately for its quantification. Some of the major limitations found within these methodologies concern the incubation time, the small number of cells analyzed, and the operator's subjectivity. To overcome these aspects, we have developed a quantitative...


In vivo and in vitro effects of RAD001 on bladder cancer

Vasconcelos-Nóbrega, Carmen; Pinto-Leite, Rosário; Arantes-Rodrigues, Regina; Ferreira, Rita; Brochado, Paulo; Cardoso, Maria L.; Palmeira, Carlos

Objective: To evaluate the influence of Everolimus (RAD001) on chemically induced urothelial lesions in mice and its influence on in vitro human bladder cancer cell lines. Methods: ICR male mice were given N-butyl-N-(4-hydroxybutyl) nitrosamine in drinking water for a period of 12 weeks. Subsequently, RAD001 was administered via oral gavage, for 6 weeks. At the end of the experiment, all the animals were sacrif...


Novel method using a laser scanning cytometer for detection of Mycobacteria in ...

Pina-Vaz, Cidália; Costa-de-Oliveira, Sofia; Rodrigues, Acácio G.; Salvador, Alexandre

In order to evaluate the capacity of laser scanning cytometry (LSC) to detect acid-fast bacilli directly on clinical samples, a comparison between Kinyoun-stained smears analyzed under light microscopy and propidium iodide-auramine-stained smears analyzed by LSC was performed. The results were compared with those for culture on BACTEC MGIT 960. LSC is a new, reliable methodology to detect Mycobacteria.


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