Date of Award
Master of Science - Biotechnology
Rebecca D. Parr, PhD
Judith Ball, PhD
Josephine Taylor, PhD
Donald Pratt, PhD
Rotavirus (RV) causes severe, life-threatening diarrhea, in infants, young children and immunocompromised adults. There are several effective vaccines for young children, however they are strain specific and are not protective against many RV strains in developing countries. Therefore, it is important to investigate anti-RV therapeutic agents. Our laboratory has shown arachidin-1 (A1) and arachadin-3 (A3) significantly inhibit RV replication in two cell lines, however the molecular mechanism(s) of action are not known. A synthetic molecule of A3 (sA3) has been produced, but its’ antiviral effects have not been examined. Our hypothesis is that sA3 produces the same effects on RV-infected cells as natural A3. This study used plaque forming unit (PFU) assays to show a significant decrease in the amount of infectious RV particles released from arachidin treated cells, and tunable resistive pulse sensing technology (TRPS) revealed changes in the size distribution of released nanoparticles. Transmission electron microscopy (TEM) was utilized to observe alterations of nucleus to cytoplasm ratios which were confirmed with whole cell fluorescent staining techniques. This suggested that the arachidins modified the apoptosis and autophagy pathways. To support these observations, transcripts of initiator genes in both pathways were investigated using qRT-PCR, and the expression of two effector proteins in the apoptosis pathway were measured. Only small changes in the transcripts and proteins were detected which implied the regulation of other genes in the cell death signaling pathways that requires further examination. Both A3 and sA3 have similar antiviral activity that results in significant decreases in the production of infectious RV particles, thus revealing therapeutic potential for rotavirus infections.
Napier-Jameson, Rebekah, "THE COMPARISON OF EFFECTS OF SYNTHETIC AND NATURAL ARACHIDIN-3 ON ROTAVIRUS INFECTED CELLS" (2018). Electronic Theses and Dissertations. 195.
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