Date of Award

8-2018

Degree Type

Thesis

Degree Name

Master of Science - Biotechnology

Department

Biology

First Advisor

Rebecca D. Parr, PhD

Second Advisor

Judith Ball, PhD

Third Advisor

Josephine Taylor, PhD

Fourth Advisor

Donald Pratt, PhD

Abstract

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.

Creative Commons License

Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

Available for download on Friday, July 10, 2020

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