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28th November 2013 @ 02:06

Sandra Duffy from Vicky Avery's group at Griffith University has carried out a late stage gametocyte assay on two compounds, OSM-S-106 and OSM-S-111.

Uni Sydney14-06-13-Late stage GAM-SD OSM.xlsx


Comments on data from Sandra Duffy

"In the gametocyte late stage (IV-V) imaging assay, viable gametocytes have an elongated solid GFP fluorescent morphology and a MTR fluorescent staining pattern of a small intense spot within the GFP body of the gametocyte image. The MTR object is classified based on its relative fluorescent intensity, radius and contrast to background.  Gametocytes when treated with 5uM Puromycin loose the elongated morphology becoming a small disorganized object with no classified MTR objects. Essentially the gametocytes are destroyed in structure and have no centralized or recognizable MTR signal.

Gametocytes treated with OSM-S-111 demonstrate a different phenotype to puromycin treated gametocytes. The parasite morphology is still intact as demonstrated by the GFP fluorescence but the MTR signal has no concentrated spot of intensity. There is a small low fluorescent MTR staining pattern distributed throughout the parasite structure but there is no singular intense localized MTR signal to identify as an active mitochondria. Compounds with this phenotype may be determined as not active if geimsa staining and light microscopy were performed to determine compound action.

This phenotype is seen for small numbers of other compounds tested within this assay. At present we are unable to give a definitive reason for this different phenotype, but have hypothesized that it could be related to a slow acting mechanism where by the parasite slowly dies due to a direct effect on mitochondrial metabolism but not exclusively so. As the compound action is slow, the parasite structure is still intact within the assay time frame of a total of 96hours. We have not as yet tested compounds with this activity phenotype for longer treatment times as the total of 96 hours is already a significant activity time frame.'

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Uni Sydney14-06-13-Late stage GAM-SD OSM.xlsx