Dual candidemia detected by nested polymerase chain reaction in two critically ill children.

The use of improved microbiological procedures associated with molecular techniques has increased the identification of Candida bloodstream infections, even if the isolation of more than one species by culture methods remains uncommon. We report the cases of two children presenting with severe gastrointestinal disorders and other risk factors that contribute to Candida infections. In the first patient, C. albicans DNA was initially detected by a nested-amplification and C. tropicalis was found later during hospitalization, while blood cultures were persistently negative. In the second child, there was amplification of C. albicans and C. glabrata DNA in the same samples, but blood cultures yielded only C. albicans. Both patients received antifungal therapy but had unfavorable outcomes. These two cases illustrate that PCR was more successful than culture methods in detecting Candida in the bloodstream of high risk children, and was also able to detect the presence of more than one species in the same patient that might impact therapy when the fungi are resistant to azole compounds.

Effect of insulin-like growth factor-I on Leishmania amazonensis promastigote arginase activation and reciprocal inhibition of NOS2 pathway in macrophage in vitro.

We showed previously that insulin-like growth factor (IGF)-I induces an exacerbation of the lesion development in experimental cutaneous leishmaniasis favouring parasite growth within host macrophages. Here we studied the effect of IGF-I in vitro in BALB/c mouse peritoneal macrophages infected with stationary phase Leishmania amazonensis promastigotes. IGF-I was used to pre-incubate either macrophage or parasite before infection of the macrophages or adding it at the start of the Leishmania-macrophage culture and maintaining it throughout the experimental period. Independent of stimulation protocol, IGF-I induced significantly increased parasite growth within macrophages. Arginase activation considered as a key factor in Leishmania growth was studied, and its expression and activity were increased in Leishmania-infected macrophages but significantly more in infected cells upon IGF-I stimulus, an effect specifically inhibited by NOHA. Arginase known to be present on Leishmania was also studied, and its expression and activity were seen in the absence of any stimulus but significantly increased after 5 min of incubation with IGF-I. In addition, Leishmania was pre-incubated with NOHA for 5 min, washed, then macrophages infected observing a significantly reduced parasite burden in both IGF-I-stimulated and non-stimulated macrophages. Reciprocal decrease in the nitric oxide (NO) level and inhibition of nitric oxide synthase (NOS2) expression were also observed in IGF-I-stimulated infected macrophages. Our data strongly suggest that IGF-I induces preferential expression and activation of Leishmania promastigote arginase, contributes to the alternative activation of macrophages in the context of innate immunity and interferes with NOS pathway in infected macrophages probably as a reciprocal effect.