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Analysis of Inhibition of Influenza Replication via Synthetic Antibodies

Description
The influenza virus, also known as "the flu", is an infectious disease that has constantly affected the health of humanity. There is currently no known cure for Influenza. The Center for Innovations in Medicine at the Biodesign Institute located on campus at Arizona State University has been developing synbodies as…

The influenza virus, also known as "the flu", is an infectious disease that has constantly affected the health of humanity. There is currently no known cure for Influenza. The Center for Innovations in Medicine at the Biodesign Institute located on campus at Arizona State University has been developing synbodies as a possible Influenza therapeutic. Specifically, at CIM, we have attempted to design these initial synbodies to target the entire Influenza virus and preliminary data leads us to believe that these synbodies target Nucleoprotein (NP). Given that the synbody targets NP, the penetration of cells via synbody should also occur. Then by Western Blot analysis we evaluated for the diminution of NP level in treated cells versus untreated cells. The focus of my honors thesis is to explore how synthetic antibodies can potentially inhibit replication of the Influenza (H1N1) A/Puerto Rico/8/34 strain so that a therapeutic can be developed. A high affinity synbody for Influenza can be utilized to test for inhibition of Influenza as shown by preliminary data. The 5-5-3819 synthetic antibody's internalization in live cells was visualized with Madin-Darby Kidney Cells under a Confocal Microscope. Then by Western Blot analysis we evaluated for the diminution of NP level in treated cells versus untreated cells. Expression of NP over 8 hours time was analyzed via Western Blot Analysis, which showed NP accumulation was retarded in synbody treated cells. The data obtained from my honors thesis and preliminary data provided suggest that the synthetic antibody penetrates live cells and targets NP. The results of my thesis presents valuable information that can be utilized by other researchers so that future experiments can be performed, eventually leading to the creation of a more effective therapeutic for influenza.
ContributorsHayden, Joel James (Author) / Diehnelt, Chris (Thesis director) / Johnston, Stephen (Committee member) / Legutki, Bart (Committee member) / Barrett, The Honors College (Contributor) / Department of Psychology (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2014-05
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Structural-Based Study of Intact Human Particulate Guanylyl Cyclase Receptor A (pGC-A) and Biophysical Characterization

Description
Particulate Guanylyl Cyclase Receptor A (pGC-A) is an atrial natriuretic peptide receptor, which plays a vital role in controlling cardiovascular, renal, and endocrine functions. The extracellular domain of pGC-A interacts with natriuretic peptides and triggers the intracellular guanylyl cyclase domain to convert GTP to cGMP. To effectively develop a method…

Particulate Guanylyl Cyclase Receptor A (pGC-A) is an atrial natriuretic peptide receptor, which plays a vital role in controlling cardiovascular, renal, and endocrine functions. The extracellular domain of pGC-A interacts with natriuretic peptides and triggers the intracellular guanylyl cyclase domain to convert GTP to cGMP. To effectively develop a method that can regulate pGC-A, structural information regarding its intact form is necessary. Currently, only the extracellular domain structure of rat pGC-A has been determined. However, structural data regarding the transmembrane domain, as well as functional intracellular domain regions, need to be elucidated.This dissertation presents detailed information regarding pGC-A expression and optimization in the baculovirus expression vector system, along with the first purification method for purifying functional intact human pGC-A. The first in vitro evidence of a purified intact human pGC-A tetramer was detected in detergent micellar solution. Intact pGC-A is currently proposed to function as a homodimer. Upon analyzing my findings and acknowledging that dimer formation is required for pGC-A functionality, I proposed the first tetramer complex model composed of two functional subunits (homodimer). Forming tetramer complexes on the cell membrane increases pGC-A binding efficiency and ligand sensitivity. Currently, a two-step mechanism has been proposed for ATP-dependent pGC-A signal transduction. Based on cGMP functional assay results, it can be suggested that the binding ligand also moderately activates pGC-A, and that ATP is not crucial for the activation of guanylyl cyclase. Instead, three modulators can regulate different activation levels in intact pGC-A. Crystallization of purified intact pGC-A was performed to determine its structure. During the crystallization condition screening process, I successfully selected seven promising initial crystallization conditions for intact human pGC-A crystallization. One selected condition led to the formation of excellent needle-shaped crystals. During the serial crystallography diffraction experiment, five diffraction patterns were detected. The highest diffraction resolution spot reached 3 Å. This work will allow the determination of the intact human pGC-A structure while also providing structural information on the protein signal transduction mechanism. Further structural knowledge may potentially lead to improved drug design. More precise mutation experiments could help verify the current pGC-A signal transduction and activation mechanism.
ContributorsZhang, Shangji (Author) / Fromme, Petra (Thesis advisor) / Johnston, Stephen (Committee member) / Mazor, Yuval (Committee member) / Arizona State University (Publisher)
Created2021

Empathic Communication in Case Investigation: A Responding Guide for Difficult Situations

Description

Being prepared to respond to difficult situations that arise in public health practice is an essential skill for the public health workforce.This empathic responding guide was designed to train students, volunteers, and staff of the ASU COVID-19 Case Investigation Team. The guide provides an overview of empathic communication, walks through…

Being prepared to respond to difficult situations that arise in public health practice is an essential skill for the public health workforce.This empathic responding guide was designed to train students, volunteers, and staff of the ASU COVID-19 Case Investigation Team. The guide provides an overview of empathic communication, walks through a framework for responding with empathy, and outlines common difficult situations that arise in public health along with ways to respond with empathy to these situations. This guide can be adapted to a wide variety of settings and is meant to be used as a training tool for public health case investigators and other staff. This guide, available in a full and an abridged version, can be paired with hands-on workshops to provide engaging continuing education opportunities for public health teams.
ContributorsMeyer, Laura Grace (Author) / Arizona State University. AZ Infectious Disease Epi (AIDE) Lab (Contributor) / Arizona State University. Public Health Program (Contributor)
Created2021-07-12
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Empathic Communication in Public Health: A Responding Guide for Difficult Situations [Abridged]

Description

This communication guide outlines examples of specific situations that are difficult to respond to, and pairs them with examples of how to respond with empathy. This guide depicts these difficult case statements as rows with bold, italic text. Beneath each scenario is an example of an empathic response (underlined) that…

This communication guide outlines examples of specific situations that are difficult to respond to, and pairs them with examples of how to respond with empathy. This guide depicts these difficult case statements as rows with bold, italic text. Beneath each scenario is an example of an empathic response (underlined) that can lead to a factual response or survey prompt (Figure 1). The responses use empathic communication to show the case that you are witnessing the emotion, rather than moving to the survey without acknowledging emotion. There is no one right answer to any difficult case statement.
ContributorsMeyer, Laura Grace (Author) / Arizona State University. Public Health Program (Contributor) / Arizona State University. AZ Infectious Disease Epi (AIDE) Lab (Contributor)
Created2021-07-12
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Empathic Communication in Case Investigation: A Responding Guide for Difficult Situations [Full guide]

Description

Being prepared to respond to difficult situations that arise in public health practice is an essential skill for the public health workforce.This empathic responding guide was designed to train students, volunteers, and staff of the ASU COVID-19 Case Investigation Team. The guide provides an overview of empathic communication, walks through…

Being prepared to respond to difficult situations that arise in public health practice is an essential skill for the public health workforce.This empathic responding guide was designed to train students, volunteers, and staff of the ASU COVID-19 Case Investigation Team. The guide provides an overview of empathic communication, walks through a framework for responding with empathy, and outlines common difficult situations that arise in public health along with ways to respond with empathy to these situations. This guide can be adapted to a wide variety of settings and is meant to be used as a training tool for public health case investigators and other staff. This guide can be paired with hands-on workshops to provide engaging continuing education opportunities for public health teams.
ContributorsMeyer, Laura Grace (Author) / Arizona State University. Public Health Program (Contributor) / Arizona State University. AZ Infectious Disease Epi (AIDE) Lab (Contributor)
Created2021-07-12