Background: Penicillium marneffei is a pathogenic fungus that afflicts immunocompromised individuals having lived or traveled in Southeast Asia. This species is unique in that it is the only dimorphic member of the genus. Dimorphism results from a process, termed phase transition, which is regulated by temperature of incubation. At room temperature, the fungus grows filamentously (mould phase), but at body temperature (37°C), a uninucleate yeast form develops that reproduces by fission. Formation of the yeast phase appears to be a requisite for pathogenicity. To date, no genes have been identified in P. marneffei that strictly induce mould-to-yeast phase conversion. In an effort to help identify potential gene products associated with morphogenesis, protein profiles were generated from the yeast and mould phases of P. marneffei.
In this study, an in vitro synthetic gastrointestinal extraction protocol was used to estimate bioaccessibility of different arsenicals present in 17 rice samples of various grain types that were collected across the United States. The across matrix average for total arsenic was 209 ng/g±153 (\[xmacr]±2σ). The bioaccessibility estimate produced an across matrix average of 61%±19 (\[xmacr]±2σ). The across matrix average concentrations of inorganic arsenic (iAs) and dimethylarsinic acid (DMA) were 81 ng/g±67.7 and 41 ng/g±58.1 (\[xmacr]±2σ), respectively. This distribution of iAs concentrations in rice was combined with the distribution of consumption patterns (from WWEIA) in a Stochastic Human Exposure and Dose Simulator model to estimate population-based exposures. The mean consumption rate for the population as a whole was 15.7 g per day resulting in a 0.98 μg iAs per day exposure. The mean consumption rate for children 1-2 years old was 7 g per day resulting in a 0.48 μg iAs per day exposure. Presystemic biotransformation of DMA in rice was examined using an in vitro assay containing the anaerobic microbiota of mouse cecum. This assay indicated that DMA extracted from the rice was converted to dimethylthioarsinic acid, although a second oxygen-sulfur exchange to produce DMDTA was not observed.
Penicillium marneffei is an endemic, dimorphic fungus that exhibits very significant morbidity among immune compromised persons living or having traveled in Southeast Asia. The dimorphic nature of P. marneffei, which is believed to be a major contributing factor to infection by this fungus, is thermally regulated. At 25 °C, the fungus grows as a mold, but converts to a yeast phase when incubated at 37 °C. Hence, protein profiling of these developing forms will help ascertain the underpinning molecular mechanisms associated with this phase transition, and perhaps provide clues to virulence in this pathogenic fungus. This chapter outlines the basic procedures previously used to demonstrate distinct differences in protein expression between the mold and yeast phases of P. marneffei.
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