Changes in Lipid and Fatty Acid Composition During Intramacrophagic Transformation of Leishmania donovani Complex Promastigotes into Amastigotes.

Leishmania sp., are trypanosomatid parasites that are phagocytized by human and animal macrophages. Transformation from the vector promastigote stage to the intracellular amastigote host cell stage is mandatory, since development in the host depends on the internalization of the parasite. We identified and analyzed the lipids involved in the promastigote to amastigote transformation process in the Leishmania donovani complex. Four lipid classes, phospholipids, free fatty acids, triglycerides and sterols were studied. The derivatization method of Bligh and Dyer was used to establish the fatty acid composition in each stage of the parasite. To stay within the context of Leishmania infection, we used amastigotes extracted from macrophages after experimental in vitro infection. The purification process was checked by electronic microscopy, the absence of major contamination by host-cell debris and a correct purification yield validated our experimental model. Our results show that free fatty acids and cholesterol increased, whereas triglycerides and ergosterol decreased during the transition between promastigotes to amastigotes. With respect to phospholipid classes, we found increased proportion of sphingomyelin and phosphatidylserine and lowered proportion of phosphatidylinositol and lysophosphatidylethanolamine. Regarding fatty acid composition, a significant increase of n-7 fatty acids was observed in amastigotes. Overall, the total n-6 fatty acids were decreased in PL. Several of the changes were also observed in TG and free fatty acids. Particularly, n-7 fatty acids and 20:4n-6 were highly increased, whereas n-9 fatty acid and n-6 precursors decreased.

Examination of heterogeneous crossing sequences between toner and rollerball pen strokes by digital microscopy and 3-D laser profilometry.

The determination of line crossing sequences between rollerball pens and laser printers presents difficulties that may not be overcome using traditional techniques. This research aimed to study the potential of digital microscopy and 3-D laser profilometry to determine line crossing sequences between a toner and an aqueous ink line. Different paper types, rollerball pens, and writing pressure were tested. Correct opinions of the sequence were given for all case scenarios, using both techniques. When the toner was printed before the ink, a light reflection was observed in all crossing specimens, while this was never observed in the other sequence types. The 3-D laser profilometry, more time-consuming, presented the main advantage of providing quantitative results. The findings confirm the potential of the 3-D laser profilometry and demonstrate the efficiency of digital microscopy as a new technique for determining the sequence of line crossings involving rollerball pen ink and toner.