MbShea@PWP@English@UMCP

Leishmania alternate between an insect vector and a vertebrate host and these parasites differentiate from their avirulent form, promastigotes, into their virulent form, amastigotes, when they enter the phagolysosome of macrophages. Iron deprivation, which induces reactive oxygen species (ROS) accumulation in the mitochondria, can serve as a differentiation signal in Leishmania amazonensis. But little is known if this signal is widely conserved among Leishmania species. Here, we show that visceral species of Leishmania can be induced to differentiate into amastigotes after iron deprivation or ROS treatment. This treatment is much more efficient than traditional high temperature and low pH conditions. Treatment of Leishmania infantum with iron deprived media or ROS inducing drugs (H2O2 and menadione) caused them to lose their flagella and attenuate cell division, hallmarks of differentiation. qRT-PCR analysis confirms that amastigote specific genes were upregulated during ROS treatment, with the exception of electron transport chain (ETC) specific gene p27. This suggests that treatment of Leishmania with ROS bypasses the traditional upregulation of the ETC genes. L. donovani, another visceral Leishmania species, also showed a loss of flagella and attenuation of cell division after treatment with ROS. Scanning electron microscopy confirmed that both L. infantum and L. donovani underwent amastigote-like morphological changes after treatment with H2O2 or menadione. Therefore, this work confirms that ROS serves as a differentiation signal for multiple species of Leishmania and provides a plausible mechanism for how parasites treated with sub-lethal concentrations of menadione become highly infective.

Interesting work but I think you will need to do a very good job at briefing even non-parasitologist on background if you want a large readership. The way this is written, I feel like only people in your field will be able to grasp what you have a written. A good scientific article is accessible to both specialists and non-specialists in the field if it is written properly. Remember to use the wedge format with regard to details and always re-emphasize major themes.

Y! Like that you are thinking cognitive wedge for the I of IMRAD. Another reason we like IMRAD and related format conventions for common genres is that an experienced reader can skim or skip, leaving time for the complexity and analysis required of the methods, finding, and discussion portions.

You can also think of the methods place as of interest to other researchers who might appreciate thinking about ROS conditions as a method to be adapted or deployed to other study animals where gene conservation and differentiation help build knowledge.

Thank you for being in my class. Good luck to you. And, send a few pictures to me of YK in classic English garden settings. Will not be hard as is a national pastime.

:)