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Characterizing the Mucin Layer in Eosinophilic Esophagitis

Description
The purpose of this project was to characterize the mucin layer in patients with eosinophilic esophagitis (EoE). EoE is a chronic disease that is characterized by eosinophilic inflammation in the esophagus. The current diagnosis and standard of care for patients with EoE is less than ideal. Diagnosis is highly invasive…

The purpose of this project was to characterize the mucin layer in patients with eosinophilic esophagitis (EoE). EoE is a chronic disease that is characterized by eosinophilic inflammation in the esophagus. The current diagnosis and standard of care for patients with EoE is less than ideal. Diagnosis is highly invasive as it requires histological confirmation of eosinophilic inflammation in the esophagus, the patient must undergo an upper endoscopy to obtain the tissue sample. The histology as determined by the pathologist is subjective not quantitative which causes significant error in diagnosis. The current treatment methods are dietary therapy or corticosteroids, which require significant cost and time. The pathology of EoE is largely unknown, though it is known to involve allergic inflammatory and type-2 cytokine-mediated responses. Past studies have determined the genetic expression of mucins to be varied in the esophagi of EoE patients using RNA sequencing techniques. The varied expression of mucins in the esophagi of EoE patients has not been validated at the protein level. This study sought to better define mucin protein expression, specifically that of MUC1, MUC4, and MUC7, in the esophagi of EoE patients (n=4) and control patients (n=3). This was accomplished using histological staining. The tissue samples were stained for eosinophil peroxidase (EPX) in order to visualize the eosinophils, which are a pathological marker of EoE. The results of this study showed a qualitative increase in the protein expression of MUC4 in patients with EoE, indicating that MUC4 may play a protective role in the body’s defense against EoE. MUC1 and MUC7 staining showed no pattern. This study defined the conditions necessary for precise staining of esophageal tissues with the MUC4 8G7 antibody. The orientation of the tissue samples on the slides and the small sample size created significant difficulty in analysis and inhibited quantitative analysis. Future studies with tissue orientation standardization and greater sample size are needed to confirm the findings of this study. If verified, the increase of MUC4 protein expression in patients with EoE has implications for EoE diagnostics and therapeutics.
ContributorsCall, Abigail (Author) / Plaisier, Christopher (Thesis director) / Jacobsen, Elizabeth (Committee member) / Chemical Engineering Program (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05

Optimizing Recombinant Protein Production for Domain Antibodies: Proof-of-Concept

Description
Recent studies in traumatic brain injury (TBI) have found a temporal window where therapeutics on the nanometer scale can cross the blood-brain barrier and enter the parenchyma. Developing protein-based therapeutics is attractive for a number of reasons, yet, the production pipeline for high yield and consistent bioactive recombinant proteins remains…

Recent studies in traumatic brain injury (TBI) have found a temporal window where therapeutics on the nanometer scale can cross the blood-brain barrier and enter the parenchyma. Developing protein-based therapeutics is attractive for a number of reasons, yet, the production pipeline for high yield and consistent bioactive recombinant proteins remains a major obstacle. Previous studies for recombinant protein production has utilized gram-negative hosts such as Escherichia coli (E. coli) due to its well-established genetics and fast growth for recombinant protein production. However, using gram-negative hosts require lysis that calls for additional optimization and also introduces endotoxins and proteases that contribute to protein degradation. This project directly addressed this issue and evaluated the potential to use a gram-positive host such as Brevibacillus choshinensis (Brevi) which does not require lysis as the proteins are expressed directly into the supernatant. This host was utilized to produce variants of Stock 11 (S11) protein as a proof-of-concept towards this methodology. Variants of S11 were synthesized using different restriction enzymes which will alter the location of protein tags that may affect production or purification. Factors such as incubation time, incubation temperature, and media were optimized for each variant of S11 using a robust design of experiments. All variants of S11 were grown using optimized parameters prior to purification via affinity chromatography. Results showed the efficiency of using Brevi as a potential host for domain antibody production in the Stabenfeldt lab. Future aims will focus on troubleshooting the purification process to optimize the protein production pipeline.
ContributorsEmbrador, Glenna Bea Rebano (Author) / Stabenfeldt, Sarah (Thesis director) / Plaisier, Christopher (Committee member) / Harrington Bioengineering Program (Contributor, Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
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Prototyping Flavors for Eosinophilic Esophagitis (EoE) Amino Acid Formulas (AAF) for Taste Enhancement & Evaluating Palatability

Description
Eosinophilic Esophagitis (EoE) is a chronic autoimmune disease that causes inflammation of the esophagus due to food allergy. In pediatric EoE, remission can be reduced by 95% through the use of hypoallergenic amino acid formulas (AAF), however its bitter taste gives it poor palatability, making nutrition difficult. This thesis highlights…

Eosinophilic Esophagitis (EoE) is a chronic autoimmune disease that causes inflammation of the esophagus due to food allergy. In pediatric EoE, remission can be reduced by 95% through the use of hypoallergenic amino acid formulas (AAF), however its bitter taste gives it poor palatability, making nutrition difficult. This thesis highlights the problem of poor palatability of AAF's and explores the idea of prototyping a new flavor enhanced recipe to minimize bitterness for EoE patients and how to evaluate it through sensory evaluation practices. Along the way, I also discovered that quail egg homogenate has novel therapeutic potential to reduce EoE symptoms.
ContributorsBorah, Priya Anjali (Co-author) / Holmes, Katherine (Co-author) / Serrano, Osvin (Co-author) / Spackman, Christy (Thesis director) / Schroeder, Shauna (Committee member) / Harrington Bioengineering Program (Contributor) / Barrett, The Honors College (Contributor)
Created2020-12
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Understanding Food Pathways and Their Association with Illness: An Examination of Eosinophilic Esophagitis (EoE) in College Students with a focus on Underserved Populations and Survey Development Techniques.

Description

Eosinophilic Esophagitis, EoE, is a newer autoimmune chronic esophageal inflammatory illness that results from food allergen triggers (Lucendo, 2017). EoE has created a wide range of diagnostic challenges for physicians because of the similarities that it has compared to gastroesophageal reflux disease, GERD. Current research indicates that EoE predominately impacts…

Eosinophilic Esophagitis, EoE, is a newer autoimmune chronic esophageal inflammatory illness that results from food allergen triggers (Lucendo, 2017). EoE has created a wide range of diagnostic challenges for physicians because of the similarities that it has compared to gastroesophageal reflux disease, GERD. Current research indicates that EoE predominately impacts males at higher rates compared to females, while White males are most affected by and at risk for developing EoE compared to people of other racial and ethnic backgrounds (Sperry, 2012). Despite these findings, there continues to be an absence of research on social factors and identities like gender, race, ethnicity, income, sexuality and more, that are neglected in understanding how these features interact with EoE. Thus, the purpose of this thesis was to provide more evidence on identity and social aspects in relation to EoE that remain ignored and explain why there might be difficulties in investigating these topics. Moreover, a survey was also developed in order to better understand the current EoE population between the ages of 18 to 26, in which the transition from pediatric to adult care occurs. Language has been shown to affect patient’s care and treatment regarding their health because of discriminations that patients have been subjected to in the past. Therefore, it is important to discuss the role played by language and political correctness, especially within surveys. This thesis will explore the depth of study that has been completed within neglected areas of research surrounding marginalized communities, while providing new insights and questions for future possible research regarding EoE. This will be accomplished by discussing the existing problems within each topic and draw conclusions as to why EoE research may have been hindered within these neglected subjects. This project will provide a literature review, survey development, and language strategies for conducting inclusive and diverse populations and research topics centered around EoE.
ContributorsSerrano, Osvin (Author) / Spackman, Christy (Thesis director) / Schroeder, Shauna (Committee member) / School of Art (Contributor) / School of Politics and Global Studies (Contributor) / School of Life Sciences (Contributor) / School of Human Evolution & Social Change (Contributor) / Barrett, The Honors College (Contributor)
Created2020-12
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The Association Between Identity and Illness: An Examination of Eosinophilic Esophagitis in Adults with an Emphasis on Assigned Sex and Social Factors through Survey Development

Description
With an ever-increasing diagnosis rate and no universal cure, eosinophilic esophagitis (EoE) lacks conclusive data regarding the onset of its autoimmune response and its preferred relation to assigned sex males. This thesis seeks to analyze the effects that assigned sex and determinants of health have on EoE diagnosis through previous…

With an ever-increasing diagnosis rate and no universal cure, eosinophilic esophagitis (EoE) lacks conclusive data regarding the onset of its autoimmune response and its preferred relation to assigned sex males. This thesis seeks to analyze the effects that assigned sex and determinants of health have on EoE diagnosis through previous research and survey development. Upon constructing a Qualtrics survey to collect data patterns and trends of subjects diagnosed with this autoimmune disorder, COVID-19 created a halt in its distribution and data collection. This unexpected event led to the collection and compilation of pre-existing research to be implemented into this thesis as a substitute. The key results of this thesis revealed that in twenty research studies regarding sex differences in EoE, at least 70% of patients diagnosed with EoE were male (Liacouras, 2005). With inconclusive results as to why males are diagnosed at a higher rate, results have not confirmed if this percentage ratio is due to social or genetic factors. To conclude, males are predominately diagnosed with EoE in comparison to the female and intersex population, however, with no data currently available on the effects that EoE has on intersex humans, it cannot be determined what factors create this recurring pattern.
ContributorsHolmes, Katherine Elizabeth (Co-author) / Serrano, Osvin (Co-author) / Borah, Priyah (Co-author) / Spackman, Christy (Thesis director) / Schroeder, Shauna (Committee member) / School of Human Evolution & Social Change (Contributor) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2020-12