top of page

Research and Projects

Student-Faculty Research at Dickinson College – Autoinflammatory Disease Mechanisms with Dr. Tiffany Frey
Spring 2021, Fall 2021, and Spring 2022

Goal: Focusing on further characterizing the cellular location(s) of CD14 following isoprenoid depletion and investigating which isoprenoid compounds play a role in the trafficking of CD14.

​

Hypothesis: Isoprenoid depletion disrupts the ER to Golgi apparatus trafficking of CD14 and reduced Rab prenylation and/or sub-cellular cholesterol depletion causes altered trafficking of CD14 in MKD in RAW 264.7 macrophages

​

Specific aims:

  • ​Examine ER localization of CD14 following isoprenoid depletion using

    • ​Immunofluorescence co-localization of CD14 with the ER marker, calnexin.

      • ​If isoprenoid depletion causes the accumulation of CD14 in the ER, then we should see more co-localization of CD14 with calnexin in isoprenoid-depleted cells are compared to control cells.

    • Analysis of CD14 N-linked glycosylation patterns by western blot.

      • ​If isoprenoid depletion causes an accumulation of CD14 in the ER, then more CD14 will contain only high-mannose oligosaccharides and be sensitive to Endo H.

 

Experimental laboratory techniques: western blots, Bradford protein assays, polymerase chain reactions, cell splitting's and immunofluorescence staining's on an average of 3-6 hours per week with a mix of both in-person and remote work.

 

Results: Present findings in the Spring semester of 2022. 

American Heart Association Summer Research at Penn State Health and Penn State College of Medicine -  Impact of Cardiomyopathy Mutations on the Human Ventricular Light Chain with Dr. Christopher Yengo 
Summer 2021

Goal: Focused on the cardiomyopathy mutations in the human ventricular myosin essential light chain on the formation of the auto-inhibited super-relaxed (SRX) state of myosin, since many HCM mutations destabilize the SRX state and disrupt contractility.

​

Hypothesis: Point mutations A57G and E143K disrupt normal ELC structure that impairs stability and binding to the myosin heavy chain. We also hypothesized that the intrinsically disordered N-terminal region of the ELC binds directly to actin and plays a role in regulating the formation of the super-relaxed (SRX) state of myosin. Therefore, by removing the N-terminal region of the ELC, no ELC-actin-binding should occur. 

​

Specific aims:

  • Create mutations of interest, A57G, E143K and deletion of the N-terminal region​​

    • Site-directed mutagenesis was used to create the mutations of interest which were then expressed in bacteria​

  • Evaluate protein-expression levels of ELC using SDS-Page and Western Blots​

  • Analyze ELC-actin-binding through actin-cosedimentation assays

  • with the intrinsically disordered N-terminal region ​

​

Results: A57G and E143K had no impact on expression levels, purification, and degradation products compared to the WT control. Additionally, the ELC point mutations did not abolish actin binding. Additionally, there was ELC-actin binding even though the N-terminal region was deleted. Delivered and presented my research in the Hershey Summer Intern Symposium on Thursday, July 29th, 2021.  

 

Experimental laboratory techniques: Run mini preparations/madi preparations of plasmid DNA, bacterial transformations, QuikChange site-directed mutagenesis, gel electrophoresis, NanoDrop nucleic acid quantification, PCR purifications, and in-vitro motility analysis.

Baird Sustainability Fellows -  Cumberland County Vulnerability and Adaptation Planning Committee at Dickinson College Fall 2021

​​Goal: Assess climate change vulnerability and adaptation options in the county and produce a report that will be used by the Planning Department to inform further development of the county’s Climate Action Plan. The county’s current action plan, developed by the Planning Department with assistance from Dickinson College, Shippensburg University, ICLEI, and the Pennsylvania Department of Environmental Protection, addresses strategies for reducing emissions of greenhouse gases to mitigate climate change. As yet, the action plan gives little attention to strategies for reducing potential harm by adapting to a changing climate. Our task is to help the county fill this gap.

​

Specific aims:

  • My section on the Vulnerability and Adaptation assessment project focused on human health and safety in Cumberland County

  • Worked as a collaborative, interdisciplinary team

  • Collected and analyze information to assess who and what is vulnerable to changes in climate​Immunofluorescence co-localization

  • identify potential adaptation strategies, evaluate promising options and develop and support recommendations for action.

​

Results: Delivered and presented to the staff of the Planning Department on Thursday, December 9th, 2021. 

​

bottom of page