dAMD (BDgene Therapeutics)

The effect of the deacetylated (amine) metabolite of diamphenethide (DAMD, 10 μg mL^-1) on the vitelline cells of F. hepatica over an 18 h period in vitro has been determined by transmission electron microscopy.DAMD acts preferentially on the undifferentiated stem cells and the intermediate cells in the early stage of protein synthesis; it appears to prevent their continued development.In the stem cell the nucleolus is absent after 6 h.During the rest of the drug treatment period considerable clumping of heterochromatin takes place, the cells round up and become electron-dense.Signs of autophagy are also evident, and the mitochondria become swollen and disorganized.From 6 h onwards there are progressive changes in the It1 (intermediate type 1) cells, including clumping of the heterochromatin in the nucleus, a decrease in the number of egg-shell globules (and consequently a reduction in the number and size of the shell globule clusters), and a decrease in the number of ribosomes on the GER cisternae, although the GER system remains well-developed.By 18 h the nucleolus is absent, and the cells are very rounded and electron-dense; the mitochondria are swollen and disorganized.Similar changes are evident in the It2 (intermediate type 2) cells, so that by 18 h it is difficult to distinguish between the It1 and It2 cells.In the mature cells there is a progressive decrease in the number and size of the shell globule clusters from 9 h onwards.Glycogen synthesis and yolk formation may also be impaired and lipid droplets are present.Spaces begin to appear between the nurse cell cytoplasm and the vitelline cells after 9 h, and disruption of the nurse cell cytoplasm is evident after 12 h, becoming very severe by 18 h.By this time the follicle is very disorganized and empty-looking.In more severely affected follicles the mature cells are seen to be breaking down.Over the 18 h drug treatment period, a change in the cell population of the follicle takes place, with relatively more stem, early Ig1 and mature cells being present, while few if any characteristic It1 and It2 cells remain.The results are interpreted as being due to an inhibition of protein synthesis in the vitelline cells by DAMD.