EURJ Volume 18

EURJ

Emory Undergraduate Research Journal Volume XVIII | Spring 2022

Photo by Naomi Akram

about EURJ The Emory Undergraduate Research Journal (EURJ) is an annual print and online publication that accepts research manuscripts from all academic disciplines written by any undergraduate student. EURJ provides a venue for students to showcase their high-quality, original scholarship while fostering campus-wide interest in undergraduate research.

Students can submit research up to one year after they graduate. EURJ was founded with generous support by the Office of Undergraduate Education and is continuously supported by the Media Council. For more information on submisssions and to view past journals, please visit emoryurj.com.

Emory Undergraduate Research Journal Volume XVIII Spring 2022

1655 North Decatur Road, Atlanta, Georgia 30322

A Letter From Undergraduate Research Programs Undergraduate Research is one of the strategic priorities of the College of Arts and Sciences at Emory University. The College has a long history of supporting, promoting, and fostering a culture of undergraduate research across all disciplines. Our Emory undergraduate research scholars are collaborators with our world-class faculty mentors. As the Senior Associate Director of Undergraduate Research Programs, I am thrilled to be positioned to support Emory student’s exploration in scholarly inquiry and experiential research and my office is committed to expanding research opportunities and lowering barriers to increase student engagement in this high impact practice.

As the faculty advisor for the Emory Undergraduate research Journal (EURJ), I am excited to support the mission of EURJ though highlighting the multidisciplinary and interdisciplinary research by undergraduates by Emory and other institutions. This volume brings together diverse voices and narratives that are a great representation of student undergraduate research journeys from questions to publication. The issue of EURJ will further the conversation of what defines research and highlight a broader community of scholars.

The journal editors have captured the true essence of research scholarship; energized by topics that inspired student creativity, exploration, and project development. These articles demonstrate how Emory undergraduates are going beyond the classrooms to generate new knowledge in laboratories, libraries, studios, and on stages across our institution.

We at Undergraduate Research Programs are proud of the efforts of EURJ’s editorial team. We hope this issue provides insight to broaden your understanding of research at Emory.

Timothy A. Raines, PhD Senior Associate Director | Undergraduate Research Programs (URP) Undergraduate Research Programs Division Chair | Council on Undergraduate Research (CUR)

Editorial Board 2021-2022 Editor-in-Chief Caroline Lee

Natural Sciences Editors Lizzy Wagman * Mario Sanchez del Campo Managing Editors Priscilla Cho Ben Thomas Jojo Liu Vishal Shankar Ishan Nadkarni Jay Patel Treasurer Shreya Ramanathan Megan Yang Kriti Rastogi Landi White Programming Director Jisu Yang Garrett George Social Sciences Editors Web Director Esther Seo * Eddy Pineda Bryn Evohr Sarah Kim Publicity Committee Isha Konety Srijita Nandy * Jordan Leslie Harry Li * Natalie McGrath Rishab Bhatt Greer Spralding Graphics and Layout Caroline Lee

Photo by Sivaram Gunisetty

Humanities Editors Ellie Coe * Justin Bright Harrison Helms Rosa Kim Chloe Kuo Ava Lewandowski Special Features Sayli Sonsurkar * Editors Emily Choi Elise Etrheim Jackson Gardner Saketh Kollipara Sabrina Li Ana Rivadeneira Web Writers Sarah Abraham Oanh Nguyen Disha Srivastava Ananya Talanki Mirielle Ma * Head Editors

A Letter from the Editor To our readers, We are excited to present Volume 18 of the Emory Undergraduate Research Journal (EURJ). This year’s edition continues with our tradition of providing an interdisciplinary platform for undergraduates to showcase their research in the natural sciences, social sciences, arts, and humanities. It is apropos to acknowledge the resilience and initiative of undergraduate researchers featured in this year's issue of EURJ, as worldwide hardships due to the pandemic have presented various obstacles to accessing and pursuing research opportunities. However, the pandemic also fostered initiative among numerous undergraduate research journals nationwide, forming the National Undergraduate Consortium for Science Journalism. With increased intercollegiate communication, we were fortunate to recruit submissions from brilliant undergraduate researchers from beyond the Emory community as well. It is our hope that EURJ will continue to highlight the intellectual vitality of the undergraduate students within and beyond Emory University, and recognize their scholarly work in a manner that fosters discussion. The articles in this issue were selected to provide a small glimpse into the variety and breadth of the research opportunities and topics undertaken by students.

We would like to express our gratitude to all of the the people whose work and support have made this journal possible. In particular, we would like to recognize our faculty advisor and senior associate director of the Undergraduate Research Programs, Dr. Tim Raines, who has provided us with the resources and insight to ensure the success of the journal. We would also like to extend a huge thank you to all the authors and their mentors, whose research propels this journal and inspires us all. Lastly, we would like to thank our incredibly talented editorial staff. Together, your creativity and dedication brought hours of research to life, and made this journal a possibility despite the circumstances.

On behalf of the entire 2021-2022 EURJ staff, we would like to thank you for reading the journal and we hope you enjoy the exceptional work featured in our annual issue.

Stay safe, and happy reading!

Hyung Seo (Caroline) Lee Editor-in-Chief

Photo by Leo Huang

Table of Contents

8

Examining the incredible claims of EMDR

Information Sharing by the EEOC during Conciliation to Settle Employment Discrimination Charges: a Proposed Approach post-Mach Mining

26

Nutrition Education to the Public

Interaction between Tau and U1-70k Proteins in Alzheimer's Disease – Liquid-Liquid Phase Separation

44

46

Applying Neuropsychological Aspects of Psychopathy to a Dictator

Addressing Food Insecurity in Appalachia: An Anthropological Public Health Assessment and Recommendations

Photo by Jae Sun Kim

34

Special Features: Brains in a Dish

Emotional Pictures: The Psychological Effects of the Color Yellow

54

12

59

Examining the Incredible Claims of EMDR CLINICAL PSYCHOLOGY

Katerina Rinaldi

Imagine hearing about a new, nontraditional therapy that can greatly alleviate symptoms of trauma and mental distress in just a few sessions. Imagine reading interviews with celebrities who promote this new technique in widely read sources like The Guardian (Saner, 2018).

This therapy is currently endorsed by the American Psychological Association, The Department of Defense, and the World Health Organization, and is described as a quick and effective intervention for PTSD, anxiety, depression, and even chronic pain, with success rates at 90100%. All you would have to do to start feeling better is follow someone’s finger.

Such an incredible therapy exists in reality, according to claims made by its proponents. EMDR, or Eye Movement Desensitization and Reprocessing, is a newly popular and relatively brief therapeutic intervention centered on eye movement as a way to change psychological function. The EMDR Institute claims that, to date, over 100,000 clinicians have been trained in the technique.

Do its claims sound a bit too good to be true? Perhaps unsurprisingly, many argue that they are.

EMDR was developed in the 1980s by a psychologist named Francine Shapiro (EMDR Institute). Dr. Shapiro published a small study about the effects of an eye movement procedure on patients diagnosed with PTSD and found that their distress was reduced without requiring exposure to their anxiety-provoking stimuli (Shapiro, 1989). Shapiro wrote that her own distressing thoughts went away when she moved her eyes in a “multi saccadic pattern” and further claimed that only one session was “sufficient to desensitize completely the subject’s traumatic memories and dramatically alter their cognitive assessments” (Shapiro, 1989). Such results are surprising given the evidence that PTSD can have wide variation in presentation, often presenting with co-morbid disorders like depression or anxiety that make it difficult to treat with any single therapeutic modality (Rauch, Eftekhari, & Ruzek, 2012). Additionally, when Rauch et al. examined Cognitive Behavioral Therapy, an existing “gold standard” treatment for PTSD, they found that CBT had a significantly smaller effect on veterans with PTSD than for non-veterans whose PTSD resulted from a sexual asssault. And although veterans have better responses to Exposure Therapy, another standard treatment, this the8 | Emory Undergraduate Research Journal

-rapy actually has a specific form for victims of sexual assault that is different than the Exposure Therapy veterans receive (Rauch, 2012). In other words, the precipitating event or events for PTSD affect therapy’s efficacy.

So how, then, could EMDR perform so invariably well for such a variable illness? We might remember Carl Sagan’s saying that extraordinary claims require extraordinary evidence to prove them. Does EMDR measure up?

EMDR, or Eye Movement Desensitization and Reprocessing, is a newly popular and relatively brief therapeutic intervention centered on eye movement as a way to change psychological function.

Practitioners of EMDR say making horizontal eye movements while recalling traumatic memories results in successful therapy by enhancing one’s information processing ability and allowing the brain to process the traumatic event (Shapiro, 2001). These eye movements can be guided by a finger or by intermittent lights or beeps (Van den Hout & Engelhard, 2012). Although the exact mechanism by which eye movements alter the brain or induce a more receptive emotional state is unclear, Shapiro argues that psychopathology results from dysfunctional storage of memories, kept “frozen” in a neuro network, and that therapeutic progress can be made when those memories are moved into a more adaptive processing network (Shapiro, 2001).

Eye movements supposedly allow the necessary transmutation of memory and also in lowered physiological arousal, which means the client can focus on processing the difficult memories without having an overwhelming physical reaction to fear and distress.

Several studies looking at EMDR found it to be an equally effective treatment for PTSD compared with Cognitive Behavioral Therapy (Siedler & Wagner, 2006).

In these studies, EMDR began by asking the patients to imagine the traumatic event as vividly as possible before

used in therapy? In addition, the sample consisted entirely of undergraduate psychology students who were not actually diagnosed with PTSD. These students were receiving credit for their major for participating. It is possible that they might have shown demand characteristics, an effect that occurs when participants catch on (or think they catch on) to what the experimenter is trying to measure. These students might have been studying EMDR themselves and wanted to try to “assist” the researchers in proving their hypothesis. We cannot know for sure, but it is worth entertaining the possibility.

Relaxation therapy may be the best control to choose for EMDR, since it involves doing small activities to reduce symptoms of hyperarousal and is also often employed to treat PTSD (Taylor et al., 2003). EMDR, while effective in these trials, did not show greater effects than relaxation therapy, and the greatest symptom reduction was in Exposure Therapy which is still considered the standard for treatment (Taylor et al., 2003). The authors of this study concluded that EMDR’s efficacy was due to the imaginal exposure, not the eye movements.

Media coverage of EMDR often does not paint the full and nuanced picture of what actually makes it effective. It is easy enough to understand why popular news sources might include some missteps in evaluating its claims, since descriptions of how the therapy works are seriously complicated. Psychology Today, a source the public often turns to in order to better understand therapy and mental health, actually gives a valiant effort at presenting both sides of the EMDR debate. A 2014 article titled “Why are Deepak Chopra and Sanjay Gupta important for science and life?” pointed out how the claims that EMDR shows efficacy for a range of conditions such as anxiety, cancer, eating disorders, and the obscure language and shifting standards of proof of the claims make it a prime example of pseudoscience (Kashdan, 2014). While Kashdan’s criticism was valid, one could argue that his tone might have been a little strong to handle a delicate topic like trauma therapy. It is unlikely that anyone who supports EMDR or is currently receiving it as a treatment would be convinced that a therapy which can alleviate devastating symptoms is totally pseudoscience. Most other articles on Psychology Today focusing on EMDR support its claims seemingly unconditionally, so it also seems doubtful that people would weigh the information they find on that site equally.

Volume XVIII, Spring 2022 | 9

CLINICAL PSYCHOLOGY

starting bilateral stimulation, usually through the therapist’s finger movement. CBT was practiced similarly, with the patients reliving the event in session; however, it involved listening to video recordings of their own narrative after each session to introduce continual exposure. Meta-analysis of the studies suggests that while posttreatment symptom differences were comparable for the EMDR and CBT groups, EMDR may just be another form of exposure therapy and that it is impossible to tell if eye movements made any contribution. Additionally, the sample sizes in the reviewed studies were quite small, with each treatment group having under twenty participants. The authors of the meta-analysis justify the choice to include studies with so few particiapnts because of the “small number of studies available that directly compare EMDR and CBT” (Siedler &Wagner, 2006). They continue with a vague statement: “It therefore did not seem advisable to include only those studies that satisfy high methodological standards” (Siedler, 2006). If there were other methodological concerns besides small sample sizes in these studies, the authors unfortunately do not elaborate.

In order to come to any conclusion about EMDR’s efficacy, it is necessary to isolate the eye movement as the variable of interest as that is what makes EMDR different from other forms of exposure therapy.

Researchers at Murdoch University in Australia conducted an experiment by assigning students with distressing memories into three groups: EMDR with eye movements guided by a fixed protocol, EMDR with random, varied eye movements, and lastly, EMDR with no eye movements at all. The findings showed that the group that received EMDR with any guided eye movement had lower self-reported ratings of distress and lower physiological signs of other distress such as skin conductance compared with the group who simply talked through their experience (Schubert, Lee & Drummond, 2011).

The evidence is compelling, but there was no significant difference between the group with eye movements adhering to the typical EMDR protocol and the group with random eye movements. So, it is possible that eye movements adhering to the typical EMDR protocol and the group with random eye movements. So, is it possible that eye movements serve merely as a source of distraction and relief from the distress, similar to how mindful breathing or muscle relaxation might be

CLINICAL PSYCHOLOGY

We might expect “pop” sources like Psychology Today to have some bias, as they want to publish articles that will gain many views. Most people want to find out how a therapy will make them feel better, not about the uncertainty of evidence supporting it. Though we would expect scholarly sources like peer reviewed journals on Emory’s databases to not have a bias, searching “EMDR” on PsychInfo yields mostly articles from the EMDR Institute’s own journal. Many of the remaining articles were written by the same groups of authors. It is concerning that limited access to unbiased information is available to students of psychology, clinicians, and other researchers who use these scholarly sources. Why would the Journal of EMDR Research publish a study that refutes its efficacy? Why would the same group of researchers who have been making the same claims for years suddenly change their minds? Confirmation bias, or our tendency to take in information that further proves our existing ideas and ignore opposing evidence, is easy to fall into even at this scholarly level.

For all its claims as a “paradigm shift” for therapy, EMDR is a disappointment. It is effective at reducing symptoms of PTSD, but meta analyses consistently show that its effects are not different from exposure therapy, and that eye movements make no impact on symptom reduction (Davidson & Parker, 2001). It does not actually introduce anything new at all to PTSD treatment, since we know imaginal exposure is already part of exposure therapy. It does meet criteria for pseudoscience, like proponents who fall back on ad hoc hypotheses, claiming faulty methods when EMDR studies don’t prove their points (Kashdan, 2014). There are also no studies that prove its efficacy against a control group for the incredible range of mental and physical illnesses it claims to treat. Shapiro even claims EMDR can be used for children as young as two years (Shapiro, 2001).

There do not seem to be any boundary conditions.

All this considered, it is necessary to remember who the real audience of this research is. It is not enough to write off EMDR as pure pseudoscience and denounce its practitioners as “quacks”. Doing so would likely just ruffle some feathers and further the gap that already exists between research and therapists who are trying to alleviate suffering. PTSD is a debilitating illness, and EMDR can alleviate its symptoms. Trauma is such a sensitive issue, and it does not seem helpful to tell people they are healing in the “wrong” way. Rather, we might do a better job at publicizing results of valid, controlled 10 | Emory Undergraduate Research Journal

studies of EMDR and disseminating that information to the people who are seeking help. The causes and boundaries of EMDR’s efficacy are just simply not what it claims. Knowledge is power, and hopefully people could use that knowledge for healing, too.

References

Davidson, P. & Parker, K. (2001). EMDR: A meta-analysis.

Journal of Counseling and Clinical Psychology.

69(2).

Kashdan, T. (2014). Why are Deepak Chopra and EMDR important for life and science? Psychology Today.

Rauch, S., Eftekhari, A., & Ruzek, J. (2012). Review of exposure therapy: a gold standard for PTSD treat ment. Journal of rehabilitation research and devel opment, 49(5). 679-87.

Saner, E. (September, 2018). Mel B is watching flashing lights to help with trauma. But does EMDR really work? The Guardian.

Schubert, S., Lee, C., & Drummond, P. (2011). The efficacy and psychophysiological correlation of dual atten tion tasks in Eye Movement Desensitization and Reprocessing. Journal of Anxiety Disorders, 25(1).

1-11.

Shapiro, F. (1989). Eye movement desensitization: A new treatment for post-traumatic stress disorder.

Journal of Behavior Therapy and Experimental Psychiatry, 20(3), 211–217.

Shapiro, F. (2001). Eye movement desensitization and re processing: Basic principles, protocols, and proce dures. Guilford Press.

Siedler, G., & Wagner, F. (2006). Comparing the efficacy of EMDR and trauma focused cognitive behavioral therapy in the treatment of PTSD: a meta-analytic study. Psychological Medicine, 36, 1515-1522.

Taylor, S., Thordarson, D., Federoff, I., Maxfield, L., Lovell, K., & Ogrodnizcuk, J. (2003). Comparative effi cacy, speed, and adverse effects of three PTSD treatments: exposure therapy, EMDR, and relax ation training. Journal of Consulting and Clincial Psychology, 71(2), 330-338.

Van den Hout, M., & Engelhard, I. (2012). How does EMDR work? Journal of Experimental Psychopa thology, 3(5), 724-738.

Katerina Rinaldi is a 2021 graduate of Emory University. During her time at Emory, she has worked as a research assistant and as a peer mentor at the Emory Autism Center. This fall, she will begin a doctorate program in clinical psychology, with a special focus on eating disorders. She works as a mental health technician in an inpatient adolescent unit and is excited to become a clinician in the future.

Volume XVIII, Spring 2022 | 11

CLINICAL PSYCHOLOGY

Photo by Jessie Ni

Information Sharing by the EEOC during Conciliation to Settle Employment Discrimination Charges: a Proposed Approach post-Mach Mining David Miller

POLITICAL SCIENCE

Abstract

This paper summarizes the evolution of the standard for what type of information the EEOC is required to provide to respondent employers during conciliation to settle charges. Title VII of the Civil Rights Act requires the EEOC to “endeavor to eliminate [the] alleged unlawful employment practice by informal methods of conciliation” before suing an employer for employment discrimination. However, consistent with the theory of Macro Politics, the legal standard for what qualifies as “endeavor” has changed over time with new electorates, influenced by a Supreme Court case, formal rulemaking, and Congressional review and oversight.

At present, the Supreme Court has established a general standard for what type of information the EEOC must provide to respondents in conciliation, but the EEOC does not have any official interpretation of how that requirement will translate to their information-sharing, nor has it obligated itself in any official way to follow the standards set forth by the Supreme Court.

From a policy standpoint, it is argued that requiring the EEOC to provide more, detailed information about a charge during conciliation will bring more respondent employers to the table. Currently 33% of respondents who are offered conciliation decline it, many of whom cite the lack of information about the charge and the resulting inability to weigh risk as the reason. EEOC staff argues that, from a policy perspective, more conciliations are ultimately successfully settled when they are given flexibility to decide how much information to provide in any given case and tailor the negotiations accordingly.

The policy question of whether requiring the EEOC to provide more information to respondents will result in more or less successful settlements is empirical. The goal of the EEOC’s policy should be to uphold Title VII’s intent by encouraging more conciliation and less time-consuming, costly litigation. Therefore, this paper argues that, while it is advisable for the EEOC to ultimately take some action to formally obligate itself to more specific information-sharing requirements, a more clear determination of what types of information-sharing will maximize successful conciliations is needed before the EEOC does so. This paper proposes some methods for gaining that clear determination. Furthermore, viewing through a Macro-Politics lens, there is little downside to the EEOC waiting for more empirical data before taking action.

I. Introduction: The EEOC I. A. History of the EEOC The Equal Employment Opportunity Commission (“EEOC” or “Commission”) was created by the Title VII of the Civil Rights Act of 1964--the section that makes it illegal to discriminate in employment on the basis of race, religion, color, national origin, or sex; it also makes it illegal for employers to retaliate if an employee complains about discrimination or participates in an employment discrimination proceeding (Laws Enforced by EEOC). The Commission was created to enforce Title VII, but now also acts as the enforcement body for many more federal employment laws, including the Pregnancy Discrimination Act, the Equal Pay Act of 1963, Title I of the Americans with Disabilites Act, Sections 501 and 505 of the Rehabilitation Act of 1973, the Age Discrimination in Employment Act of 1967 (ADEA), and Title 12 | Emory Undergraduate Research Journal

II of The Genetic Information Nondiscrimination Act of 2008 (GINA) (Laws Enforced by EEOC).

I. B. The EEOC Complaint Process When an employee or job applicant believes they have been the victim of discrimination protected by any of the above-referenced laws, they can file a charge with the EEOC. They then enter into the EEOC’s administrative process. They file a charge of discrimination--a summary or story of the unfair or disparate treatment that occurred from their point of view--and thereafter are referred to as the Charging Party (What You Can Expect After a Charge is Filed). As the EEOC outlines in “What You Can Expect After a Charge is Filed,” the EEOC makes an initial determination; if the charge lacks merit completely the EEOC will provide a Dismissal and Notice of Rights Letter, which allows the Charging Party (What You Can Expect After a

I. C. The Conciliation Requirement When discussing the remedy for discriminatory practices in employment, the statutory language of Title VII makes clear that the EEOC “must first ‘endeavor to eliminate [the] alleged unlawful employment practice by informal methods of conference, conciliation, and persuasion’” before suing an employer for employment discrimination (Title VII of the Civil Rights Act of 1964). The Supreme Court made clear that one of the policy goals of encouraging voluntary conciliation is to end harmful discrimination rapidly when it said that Title VII was designed to encourage “`voluntary compliance' and ending discrimination far more quickly than could litigation proceeding at its often ponderous pace” (Ford Motor Co. v. EEOC 458 US 219 (1982) at 228 qtd. in “Update of Commission's Conciliation Procedures: A Rule by the Equal Employment Opportunity Commission” 2975). According to the Court, “[d] elays in litigation unfortunately are now commonplace, forcing the victims of discrimination to suffer years of underemployment or unemployment before they can obtain a court order awarding them the jobs unlawfully denied them” (Ford Motor Co. v. EEOC 458 US 219 (1982) at 228). The Commission itself recognizes that an effective conciliation program makes relief for victims

of discrimination possible without the added time, expense, and uncertainty of litigation (Update of Commission's Conciliation Procedures: A Rule by the Equal Employment Opportunity Commission 2975). It is clear that conciliation was designed to be the centerpiece of the EEOC's mission to eliminate discrimination in employment without the time and expense of litigation.

II. The EEOC's Conciliation Process II.A. The Statistics As discussed above, voluntary resolution of cases through conciliation is Congress's preferred method for resolving claims of employment discrimination.

However, despite Congress’s intent that the EEOC attempt to resolve the charges voluntarily with the respondent employer before filing a lawsuit on behalf of the Charging Party, conciliation actually results in successful settlement in only a small number of cases. According to written testimony given by Andrew Maunz, former Legal Counsel of the EEOC, at an August 18, 2020 Commission meeting, the EEOC has only succeeded in resolving about 40% of the cases for which it finds reasonable cause to believe that discrimination occurred in the past eight years. Furthermore, since the Commission only finds reasonable cause in 2.8% of all the cases filed, the 40% number represents a miniscule fraction of all the cases alleging discrimination that are filed (Statistics, Title VII of the Civil Rights Act of 1964 Charges). This means that, even for cases deemed to have the most merit, the EEOC reaches a voluntary resolution less than half of the time. According to Maunz, one of the reasons the EEOC’s conciliation rate is so low is the fact that one-third of respondent employers who have received a reasonable cause finding decide not to participate in conciliation, indicating a lack of trust in and significant rejection of the conciliation system. Because the EEOC only has resources to pursue litigation itself in about 200 cases per year, thousands of charging parties with valid claims of discrimation are left on their own to pursue litigation (Written Testimony of Andrew Maunz, EEOC Legal Counsel).

II. B. Evolving Standards for Conciliation One factor contributing to the failure of the EEOC’s conciliation program is the fact that the Commission does not often communicate with respondent employers about the factual basis for the discrimination claims made by the charging party. Over the past ten years, Volume XVIII, Spring 2022 | 13

POLITICAL SCIENCE

Charge is Filed). As the EEOC outlines in “What You Can Expect After a Charge is Filed,” the EEOC makes an initial determination; if the charge lacks merit completely the EEOC will provide a Dismissal and Notice of Rights Letter, which allows the Charging Party to pursue their case on their own via a civil suit in court.

If the EEOC views the charge as having any merit, it will investigate further. The next step is, if deemed an appropriate charge, for the EEOC to offer use of their mediation program to resolve the charge voluntarily.

Many charges are resolved by this method, however if a charge is not identified for mediation, or either party declines mediation, the EEOC begins an investigation of the charge. If the EEOC determines that there is no reasonable cause for discrimination, they provide a Dismissal and Notice of Rights Letter similarly to how they would if the charge were dismissed initially. If the EEOC does find reasonable cause, they provide both parties with a Letter of Determination--a document that states the EEOC’s position and that the Commission will attempt to resolve the dispute through an informal and voluntary process called conciliation.

1963 Congress passes the Equal Pay Act of 1963, which protects men and women who perform substantially equal work in the same establishment from sex-based wage discrimination. The Department of Labor is given authority to enforce the new law.

1964 President Lyndon B. Johnson signs into law the Civil Rights Act of 1964. One section of the Act, referred to as Title VII, prohibits employment discrimination based on race, sex, color, religion and national origin.

The Act applies to private employers, labor unions and employment agencies. The Act also creates the Equal Employment Opportunity Commission (EEOC) to enforce Title VII and eliminate unlawful employment discrimination.

1973

Congress passes the Rehabilitation Act of 1973. Section 501 prohibits the Federal Government from discriminating against qualified individuals with disabilities.

1978 Congress amends Title VII by passing the Pregnancy Discrimination Act of 1978 to make clear that discrimination based on pregnancy is unlawful sex discrimination. In the same year, President Jimmy Carter transfers responsibility for enforcing the Equal Pay Act and the Age Discrimination in Employment Act from the Department of Labor to EEOC.

2008 President George W. Bush signs into law the Genetic Information Nondiscrimination Act of 2008. EEOC is given authority to enforce Title II of the Act, which prohibits employment discrimination based on genetic information.

2009

President Barack H. Obama signs into law the Lilly Ledbetter Fair Pay Act of 2009. The Act addresses when pay discrimination charges can be filed with EEOC. This Act was the first bill signed into law by President Obama.

Source: Timeline of Important EEOC Events, U.S. Equal Employment Opportunity Commission

many stakeholders representing respondent employers have complained that the EEOC’s procedures prevent the communication that is necessary to conciliate charges (Update of Commission's Conciliation Procedures: A Rule by the Equal Employment Opportunity Commission 2975). Specifically, respondents have argued that lack of information from the EEOC about the details of the charges prevents them from evaluating and weighing the merit of the Commission’s charges or taking action to address the alleged discriminatory practices (Freedman and Lorber). Respondents say that the EEOC’s practice of not providing basic factual information about charges is so commonplace that it is “deeply engrained in the agency’s culture” (Update of Commission's Conciliation Procedures: A Rule by the Equal Employment Opportunity Commission 2975).

According to many respondents, lack of information about the factual and legal basis for a claim also reduces their ability to provide information to the EEOC that could help facilitate a settlement (Meeting of March 20, 2013-Development of a Quality Control Plan for Private Sector Investigations and Conciliations).

EEOC staff argues that respondents use the conciliation requirement to delay the investigation and make the controversy about the EEOC’s conciliatory compliance rather than the respondents’ discriminatory behavior. Staff believes that if the Commission required itself to disclose certain information about each case, respondents would file ‘collateral litigation’ suits alleging that the Commission failed to comply with its self-imposed information requirements (Update of Commission's Conciliation Procedures: A Rule by the Equal Employment Opportunity Commission 2981-2982). In addition, Commission staff assert that they are able to reach more settlements specifically because of the high level of flexibility permitted to them; they believe that their ability to tailor the exact type and amount of information they share with respondents in each case is crucial to reaching a settlement (Update of Commission's Conciliation Procedures: A Rule by the Equal Employment Opportunity Commission 2981-2982).

II. B. 1. Theoretical Lens: Macro Politics As discussed below, the standard for how much and what type of information the EEOC is required to provide to respondents has bounced back and forth over recent years. Before discussing this evolution of the standards on conciliation information requirements below,

II. B. 2. Mach Mining The complaints by respondent employers about lack of information related to charges during EEOC conciliation came to a head in a court case involving a mining company in Illinois called Mach Mining -- litigation that ultimately made its way to the Supreme Court. In the years leading up to the Supreme Court case, a number of respondent companies in various cases had argued for dismissal of charges against them due to the fact that the EEOC failed to meet its statutory requirement to conciliate, arguing either that the EEOC made no effort at all to conciliate or that it did not provide sufficient information to the companies during conciliation (Mach Mining LLC v. EEOC 4). In some instances, companies were successful in getting cases dismissed on the basis that the EEOC failed to conciliate. However, the 2015 Supreme Court case Mach Mining LLC v. EEOC set the standards for what was expected of the EEOC during conciliation going forward.

In the Mach Mining case, a woman filed a charge with the EEOC, asserting that Mach Mining had not hired her as a coal miner due to her gender (in fact Mach Mining had never hired a woman miner). After determining there was reasonable cause to believe discrimination had occurred, the EEOC sent a letter to Mach Mining inviting them to participate in informal conciliation proceedings, and letting them know that a different EEOC representative would follow up to begin the conciliation process. The next correspondence Mach Mining received from the EEOC was one year later: a letter informing them that conciliation had been attempted and failed, and the EEOC planned to move forward with the litigation, representing the charging

party in court.

Mach Mining argued that the EEOC failed to meet the Title VII requirement that the Commission must first “endeavor to eliminate [the] alleged unlawful employment practice by informal methods of conference, conciliation, and persuasion” (Title VII of the Civil Rights Act of 1964, qtd. in Mach Mining 1). Specifically, Mach Mining stated that the EEOC had not attempted to conciliate in good faith. The EEOC, on the other hand, argued that the two letters it had sent to Mach Mining, on their face, provided enough proof that the Commission had fulfilled its statutory obligation and that federal courts do not have the authority to review their conciliation efforts. According to the Commission, Title VII grants them broad authority relating to the conciliation requirement, and federal courts cannot examine the reasonableness or good faith of such efforts.

The Supreme Court ruled in favor of Mach Mining on the question of whether courts can review EEOC’s conciliation process, but refused to establish a good faith standard. The Court found that since the statute did not explicitly prohibit judicial review, EEOC actions under the statute were reviewable. However, its ruling was friendly to the Commission because it granted broad discretion for the Commission to dictate its own conciliation process. The Court established a new standard for evaluating whether the Commission has met its obligation to conciliate, but that standard was extremely deferential to the EEOC. According to the Court, the EEOC’s obligation is met so long as it: 1) Informs the respondent employer about the claim, providing a description of what the employer has done and which employees (or class of employees) has been harmed, and 2) Provides the respondent employer the chance to achieve voluntary compliance by discussing the matter with them.

The Court emphasized the deference it applies to the EEOC when evaluating whether it has met the standard for attempting conciliation, saying every aspect of Title VII “smacks of flexibility” for EEOC staff in settlement discussions (Mach Mining 11). In addition to granting the Commission fairly broad discretion, other aspects of the Mach Mining ruling favored the EEOC as well. First, the Court established that the only appropriate remedy for a failure to conciliate is that the EEOC go back and engage in conciliation that they originally failed to do.

Volume XVIII, Spring 2022 | 15

POLITICAL SCIENCE

it is important to make clear that the Macro Politics theoretical framework will underlie the analysis. According to this theory, the best explanation for policy changes is an electoral shift, often driven by public opinion: “[w]hen new party majorities or political leaders with different ideologies are elected, new policy can pass” (Grossman 20). Although there are other theories that could explain the recent rapid shifts in policy relating to conciliation information requirements, the Macro Politics theory seems to provide the best explanation; as discussed below, these ‘boomerang’ policy changes were only possible because of changes in majority parties in Congress and the Presidential transition from Donald Trump to Joe Biden.

POLITICAL SCIENCE

This means the respondent employer will no longer be able to use failure to conciliate as an affirmative defense to a charge, and thus will no longer be able have a charge completely dismissed on that basis. The policy significance of this is to deter respondents from using failure to conciliate as a strategy to have the case dismissed and prolong litigation, and therefore creates more of an incentive for respondents to engage in conciliation. Additionally, the Court ruled that subsequent reviewing courts could not consider evidence of what was said or done during conciliation due to the confidentiality provisions found in Title VII of the Civil Rights Act. If this proves to be enforceable, it could create an incentive for the EEOC to disclose more information during conciliation, thereby facilitating more trust from respondents in conciliation and thus resulting in a higher rate of respondents accepting conciliation and reaching a successful settlement.

The broad standard of the Mach Mining decision preserved some flexibility for EEOC staff to decide the amount of detail to include in the information it provides to respondents, thus enabling staff to tailor the information exchange to maximize the chances of a favorable settlement (Update of Commission's Conciliation Procedures: A Rule by the Equal Employment Opportunity Commission 2980). Consistent with the Macro Politics theory mentioned above, a year after the Mach Mining decision, upon the change in Presidential administration from Barack Obama to Donald Trump in 2016, the standard was poised to become less friendly to EEOC staff and more favorable to respondent businesses. When Trump assumed office, one of his first actions was issuing Executive Order 13771, the so-called TwoFor-One Rule, which required each federal agency to repeal two pre-existing regulations for every new regulation it made final. This Order captured the pro-business zeitgeist of the new administration; any government action or interference that impacted the private sector was automatically suspicious.

II. B. 3. The 2021 Rulemaking Reflecting the Trump administration’s pro-business posture, political appointees within the EEOC initiated formal rulemaking in which the Commission obligated itself to provide more detailed information to respondent employers than the Mach Mining Court standards required for attempting conciliation (Update of Commission's Conciliation Procedures: A Rule by the Equal 16 | Emory Undergraduate Research Journal

Employment Opportunity Commission). These new EEOC conciliation regulations became final on January 14, 2021 and effective on February 16, 2021 after a Notice of Proposed Rulemaking on October 9, 2020.

Although the new rulemaking purported to bring the EEOC’s conciliation process up to what was deemed necessary by Mach Mining--and was pitched as simply obligating EEOC staff to comply with the dictates of Mach Mining--in reality it required the EEOC to provide more information to respondents than Mach Mining did. In writing the new rules, the Commission interpreted Mach Mining’s requirement that the EEOC “must inform the employer about the specific allegation” (13) as obligating the Commission to provide the following specific information to respondent employers in any conciliation: 1) a written summary of the facts on which the Com mission based its reasonable cause finding 2) a summary of the legal basis used by the Commis sion to determine reasonable cause--this includes ex plaining how the law was applied to the facts of the case.

The Mach Mining requirement that the EEOC must give respondent a description of what the employer has done and who has been harmed provides more flexibility to staff than the 2021 rules’ requirements that the EEOC provide the specific legal and factual basis for the discrimination claim. Specifically, while it is reasonable to interpret Mach Mining requirements to mean the EEOC must give their factual basis for the reasonable cause finding, the rules go further by forcing the EEOC to provide their legal argument, potentially putting the Commission at a disadvantage if there is subsequent litigation.

The policy argument in favor of the new rule is that specific information about the factual and legal basis of the reasonable cause finding enables the respondents to identify the specific discriminatory practices being alleged in the charge, thus providing them more information on which to focus discussions and make discussions productive, gauge potential liability, and base their conciliation posture or attempt to remedy the alleged discrimination (Update of Commission's Conciliation Procedures: A Rule by the Equal Employment Opportunity Commission). The theory is that respondents are more likely to agree to participate in conciliation discussions if they have information on which to weigh the

II. B. 4. Congressional Repeal of the 2021 Rulemaking While the conciliation regulations became effective in February 2021, their existence was short-lived as they were repealed just four months later (Gibbs and Bates).

Consistent with Macro Politics Theory (discussed above), an electoral shift brought about change in the EEOC’s conciliation policy shortly after President Biden took office and both houses of Congress became majority Democrat. Congress repealed the conciliation rules on June 24th, 2021, using the Congressional Review Act--only the seventeenth use of the Act in its history (Congressional Review Act Overview and Tracking).

The votes were directly along party lines.

According to EEOC Chair Charlotte Burrows, a Democrat who was elevated to the chairmanship under President Biden, the repeal of the rulemaking was necessary to ensure that more conciliation attempts are successful: “the action by Congress restores the Commission’s flexibility to tailor the conciliation process to the facts and circumstances of each case, thus increasing the likelihood of a successful resolution” (Congress Acts to Restore Flexibility to EEOC’s Conciliation Process).

II. B. 5. Current Standards for Information Sharing During Conciliation: Rules and Guidance With the 2021 rulemaking repealed, the EEOC’s public pronouncements on the standards it will follow in conciliation are few and barebones. The Commission has some rulemaking related to conciliation, but it was issued long before the Mach Mining decision. The rules that originally established the EEOC’s standards for conciliation were published in 1977 (Equal Employment Opportunity Commission Procedural Regulations).

There have been only minor amendments to these rules since they were published (Update of Commission's

The arguments in favor and against the Trump-era rulemaking provisions assume knowledge about whether providing more detailed information to respondent employers is likely to result in more or fewer cases being conciliated and/or successfully settled. This is an empirical question that has not been definitively answered by the Commission, legislators, or stakeholders on either side of the debate.

Volume XVIII, Spring 2022 | 17

POLITICAL SCIENCE

potential liability of the claims, thus the 33% of respondents who decline conciliation could be reduced.

There are two primary policy arguments against this Trump-era rule, which are revealed in the comments to the rule when it was still in its proposal stage (Update of Commission's Conciliation Procedures: A Rule by the Equal Employment Opportunity Commission). First, by placing more specific requirements on EEOC staff, the rule opens up more possibilities for finding that staff has not met its conciliation obligation and therefore invites collateral litigation (lawsuits about whether the EEOC followed its regulation-mandated procedures as opposed to addressing the underlying claim of discrimination). By setting more specific standards for EEOC staff to follow in conciliation, the EEOC opens itself to counter-litigation about its own processes--this distraction slows down the ability of the EEOC to successfully pursue its primary goal of eliminating discrimination in the workplace. Second, critics argue that more rigid requirements for information sharing leads to less flexibility for EEOC investigators to use their expertise to make strategic decisions regarding the type and amount of information that will make conciliation attempts more likely to result in successful settlement (Update of Commission's Conciliation Procedures: A Rule by the Equal Employment Opportunity Commission). The theory is that with less flexibility, fewer of the conciliation attempts will be successful.

The arguments in favor and against the Trump-era rulemaking provisions assume knowledge about whether providing more detailed information to respondent employers is likely to result in more or fewer cases being conciliated and/or successfully settled. This is an empirical question that has not yet been definitively answered by the Commission, legislators, or stakeholders on either side of the debate. This knowledge gap and potential ways to address it will be addressed in Section IV below.

POLITICAL SCIENCE

conciliation Procedures: A Rule by the Equal Employ- -he absence of any official guidance that incorporates ment Opportunity Commission). The 1977 procedural Mach Mining requirements, the only public statement regulations on conciliation provide general procedure by the Commission obligating itself to follow the stanand authority for the Commission’s conciliation pro- dards for information sharing that came out of the Mach cess, and mirror the Title VII statutory language by re- Mining decision is in a News Release issued on June 24, quiring that the EEOC “shall endeavor to eliminate such 2021. In the news release announcing the repeal of the practice by informal methods of conference, conicila- 2021 rulemaking, EEOC Chairwoman Charlotte A. tion, and persuasion” (Equal Employment Opportunity Burrows is quoted as saying that the EEOC will now reCommission Procedural Regulations 55388). The reg- turn to the standard provided by Mach Mining. ulations do not provide any examples or specific inforBecause the statement by Chairwoman Burrows in mation that must be shared with respondents to meet the news release is so general--and does not interpret the requirement of endeavoring exactly how much and what to eliminate discriminatory practypes of information Mach Mintices through informal methods, ing requires the Commission to ... issuing a public pronuncestating that “...the Commission give respondents--the question shall attempt to achieve a just ment of the Commission's inremains whether the EEOC resolution of all violations found should act in some way to clarand to obtain agreement that the terpretation of Mach Mining's ify its obligations post-Mach respondent will eliminate the un- information-sharing requireMining. Binding itself explicitly lawful employment practice and to Mach Mining’s requirement ments, and its intent to comprovide appropriate affirmative that it provide respondents with relief ” (Equal Employment Opa description of what the emply with those requirements, portunity Commission Proceployer has done and which emdural Regulations 55388). ployees (or class of employees) would be a good strategy for The current sub-regulatory has been harmed would have guidance on conciliation was also the Commission to avoid inthe benefit of providing clearer written before the Mach Mining terference from Congress or guidance for charging parties decision in 2015. The first piece and respondent employers as the Judiciary and provide of guidance is a publication titled they evaluate settlement and “Quality Practices for Effective would help insulate the EEOC clearer guidance to respondent Investigations and Conciliations,” from future Congressional or based on a Quality Control Plan employers.

Judicial interference. However, that the EEOC conducted with as discussed below, there is litrepresentatives of respondents, tle cost to waiting at least until EEOC staff representatives, and after midterm elections to take other stakeholders beginning in 2012. This guidance action in the form of public, binding pronouncements, only requires that “the conciliation request is based on and a potential benefit: the time and opportunity to the findings of the investigation and informs the parties study the exact causes of unsuccessful conciliations and of the relief sought.” Unlike the 2021 rulemaking, this nonparticipation in conciliation by 33% of respondent guidance does not obligate the EEOC to provide any employers. specific information to respondent employers about the III. Proposed actions the EEOC could take factual or legal basis of the case. It also does not require as much information to the respondent as Mach Mining; As mentioned in the previous section, issuing a public rather than requiring that the EEOC tell respondents pronouncement of the Commission’s interpretation of the practices that harmed a specific person or class, it Mach Mining’s information-sharing requirements, and only goes so far as to obligate the EEOC to inform the its intent to comply with those requirements, and its parties that the request is based upon the findings of the intent to comply with those requirements, would be a investigation. The other guidance publicly available on good strategy for the Commission to avoid interference 18 | Emory Undergraduate Research Journal

III. A. Option 1: Do not clarify interpretation of Mach Mining requirements through guidance or rulemaking As discussed in section II.5 above, the EEOC has two guidance documents on their website currently that inform the public of the amount and type of information the EEOC will provide respondents during conciliation; however, neither of these guidances specifically require the Commission to provide respondents with information that reaches the level of the standard laid out by Mach Mining. Shortly after the 2021 rules were repealed by the Congressional Review Act, the EEOC issued a

press release that states the “EEOC’s conciliation process will again be governed by the standards set forth by the Supreme Court in Mach Mining” (Congress Acts to Restore Flexibility to EEOC’s Conciliation Process).

However, the rules that were repealed were originally purported to address the standards set forth by Mach Mining and actually bring the Commission up to those standards. This seeming inconsistency can be explained using a Macro Politics lens, which emphasizes the significance of electoral changes in catalyzing policy results.

While the same organization published these seemingly contradictory documents, it is logical given that the Trump-ruled EEOC had different goals and interpretations than the Biden-ruled EEOC currently does.

The main positive of a wait-and-see plan is that it maintains more flexibility for EEOC staff to tailor the type of information they provide to each case, using their judgment to maximize the chances that a settlement will be successful. While Chairwoman Burrows has publicly stated that staff will provide the information required by Mach Mining, the public has no knowledge of how the Commission interprets the Mach Mining requirement that the EEOC “must inform the employer about the specific allegation” (Mach Mining 13). By simply saying they will abide by this standard in conciliations, without actually endeavoring to define the standard itself, the EEOC allows its investigators a very high level of discretion in how to handle each particular case.

While the main drawback of a wait-and-see plan is that Congress or the Judiciary might decide to check the power of the Commission and reduce its autonomy, judicial or legislative interference is unlikely under current circumstances. First, it is unlikely the Supreme Court would grant certiorari to hear a case involving EEOC conciliation procedures so soon after resolving the questions addressed in Mach Mining. Second, even if it agreed to hear a case involving EEOC conciliation procedures, it would be unlikely to impose specific information-sharing requirements on the Commission; to do so, would require it to ignore precedent and partially overturn the part of Mach Mining that emphasized the high level of flexibility staff should have when negotiating settlements. Thus, if the Commission refuses to provide further conciliation guidance, its main concern would not be Judicial Review--it would be much more concerned about Congressional action.

There are several checks that Congress can exercise over agencies, and they have already exercised their moVolume XVIII, Spring 2022 | 19

POLITICAL SCIENCE

from Congress or the Judiciary and provide clearer guidance to respondent employers. However, as discussed below, this is one of three options that the EEOC has moving forward.

First, the EEOC could do nothing (for now). If they see the likelihood that Congress would spend its time on EEOC oversight and the likelihood that another Supreme Court case obligates them further than Mach Mining as relatively low, they could decide to not act. This would keep the vagueness about their information-sharing requirements that currently exists, in which they purport to follow the standard of Mach Mining without laying out how they interpret it.

The second option would be to issue new rulemaking regarding the EEOC’s conciliation process. This option is arguably the most complex since the Commission would be issuing new rules on top of rules repealed by the Congressional Review Act. This means the new rules could not be “of substantially the same form” as the Trump-era rules (Small Business Regulatory Enforcement Fairness Act). While this may be the case naturally, since the rules would be implemented by a Biden administration, there is a severe lack of precedent regarding how “substantially of the same form” is defined.

The last option, as previously discussed, would be to issue further sub-regulatory guidance (published guidelines, speeches, press releases, etc.) to inform the public how they interpret Mach Mining. Due to the far-reaching requirements of the Trump-era rules, it is likely that this guidance would interpret Mach Mining in a significantly different manner than the 2021 rules did--in fact it is very unlikely the EEOC would choose to obligate itself to share both the factual and legal basis for their case with respondents. Each of the three options laid out will be discussed further in their own subsection below.

POLITICAL SCIENCE

-st direct and extreme check: using the Congressional Review Act to repeal a rule. In repealing the rigid rules, Congress indicated that they preferred the EEOC to have a high level of flexibility in conciliations, which makes current Congressional interference unlikely.

Apart from using the Congressional Review Act, Congress has a few main ways of exerting control over agencies: appointment politics, the power of the purse, and oversight. Appointment politics often play an important role in how agencies function, however it is unlikely to affect conciliation procedures because an EEOC Commissioner nomination will not be necessary for several years.

The power of the purse--which refers to Congress’ ability to clip agencies’ wings through the appropriations process--is also a fairly minor concern in this situation.

The Commission’s budget does not change drastically from year to year, and appropriating less money to the EEOC overall would not force the Commission to act more strictly in regards to conciliation specifically (Fiscal Year 2021 Congressional Budget Justification). Additionally, Congress is poorly positioned to exert influence over the Commission with the power of the purse because a common way for them to do this is through limitation riders: requirements built into appropriation legislation banning agencies from spending money on certain things (MacDonald 766). MacDonald explains that limitation riders are a mechanism that helps Congress exert influence over policy decisions in periods of divided government (773).

While appointment politics and the power of the purse are not likely to be sources of Congressional interference if the Commission decides to not further clarify its interpretation of its obligations under Mach Mining, Congressional oversight could be. There are many ways that Congress can conduct oversight over agencies: committee hearings and investigations, informal contacts between staff members and agency officials, information from affected parties, through reporting requirements and procedures enhancing transparency, special investigatory panels commissioned by Congress, investigations and studies by congressional agencies, and through the Inspector General (Bolton). Consistent with Macro Politics theory, one explanation of how and when Congress decides to conduct oversight is based on elections (Lowande and Peck). Lowande and Peck use oversight data from before and after members of the Senate were directly elected to argue that members of 20 | Emory Undergraduate Research Journal

Congress decide to conduct oversight solely as a political tool to discredit the opposition, motivated more by re-election than anything (1). This typically happens in periods of divided government, since members of Congress of the opposite party of the President use oversight as an opportunity to show the deficiencies of the President’s administration before elections (4). Thus, in the current state of unified government, Congressional oversight of the EEOC is less likely than normal.

Throughout this section, a pattern has been present: unified governments, such as the one we have now, are far less likely to exert their influence over agencies.

This means that while the EEOC would likely be safe in choosing to do nothing for now, it should consider a plan for how to protect itself from other-branch interference in the future when faced with a divided government; the two options for doing so are discussed below.

III. B. Option 2: Issue new rulemaking Another option by which the Commission could bind itself to a public interpretation of what Mach Mining requires is to issue new rulemaking to replace the repealed Trump-era rules. The benefits to this are the same benefits for rulemaking in all cases across all agencies: in theory, published rules that have gone through notice-and-comment should hold more weight and permanence than any guidance would. Conversely, the main drawback is that it is more difficult and more expensive for the agency to engage in notice-and-comment rulemaking than it is to issue guidance. The pros and cons of guidance and rulemaking will be further discussed in the following subsection on guidance.

Despite the benefits of rulemaking, it is likely not the best choice in this case. The main drawback to issuing new rulemaking that is unique to this circumstance is that the Commission would be making new rules on top of Congressionally-repealed rules; of the seventeen uses of the Congressional Review Act, this has never been done before (Chen). There is language in the statute that suggests that issuing a new rule would be complicated and not necessarily permissible. The statute states that a rule “may not be reissued in substantially the same form, and a new rule that is substantially the same as such a rule may not be issued” (Small Business Regulatory Enforcement Fairness Act; Congressional Research Service). Even if the EEOC publishes new rules that set a different standard for how much information is necessary to share with respondents (i.e. must provide the

III. C. Option 3: Sub-regulatory guidance Sub-regulatory guidance is an informal, supposedly non-binding form of policymaking in which agencies create standards, interpret their own rules, and impose obligations on regulated parties. Because guidance is not subject to the same notice-and-comment procedural requirements as formal rulemaking, it is quicker and requires fewer resources. Despite this, guidance can bind regulated entities in practice just as formal rulemaking can because agencies follow it as closely as notice-and-comment rulemaking, and thus regulated parties must follow it as well in fear of agency enforcement (Parrillo; Potter). One drawback to guidance is that many see the in-practice binding effect that guidance has as a Constitutional problem, since agencies are promulgating ‘rules’ that they were never delegated the power to make without due process in the form of notice and comment (Parrillo). However, Parrillo suggests that fixing these Constitutional concerns with guidance are not a responsibility of agencies themselves, since the issues arise from institutional/structural problems with the government as opposed to bad-faith acting on the part of agencies.

One positive to guidance is that agencies issue it mostly to help regulated parties understand how to comply with rules--thus regulated parties often appreciate the clarity and detail that guidance provides (Potter).

Understanding that a) the EEOC is not responsible for the Constitutional concerns with guidance, b) guidance allows agencies to act more swiftly and efficiently than engaging in notice-and-comment rulemaking, and c) as discussed above, rulemaking would be a particularly risky endeavor in this circumstance, the EEOC should opt to engage in guidance over rulemaking in regards to issuing interpretations of their information-sharing requirements post-Mach Mining, when the time comes to clarify their position.

Another positive to guidance is that issuing a public pronouncement of its interpretation of the information-sharing requirements of Mach Mining would align with Congress’s underlying premise when it delegates to agencies: that agencies have “unique expertise” about their subject matter area (Kisor 10). The EEOC is in the best position to make policy judgments about the conciliation process because of its comparative advantage in substantive expertise, so the EEOC should issue guidance on information sharing at some point.

Kisor v. Wilkie, a recent Supreme Court case, establishes the types of subregulatory guidance that do and do not warrant deference by the Judiciary; those that warrant deference should reflect the “agency’s authoritative, expertise-based, fair [and] considered judgment” (12).

First, in order to be authoritative, the guidance must come from Chairwoman Charlotte Burrows because it must be the agency’s official position, “rather than [an] ad hoc statement not reflecting the [full] agency’s views” (Kisor 15-16). Under this analysis, the news release alone would not meet the Kisor standard, because it is not signed by Chairwoman Burrows, was not published in the Federal Register, and was not presented as an official agency pronouncement. To be ‘expertise-based,’ the guidance must “in some way implicate its substantive expertise” (Kisor 16). When discussing what qualifies as ‘fair and considered judgment,’ the Court established that they refuse “to defer to a merely convenient litigating position,” meaning the EEOC must have substantive reasons for providing less information to respondents as opposed to doing so to help its case if it reaches litigation (Kisor 17).

IV. My recommendation My recommendation is for the EEOC to wait and do nothing for now, while conducting research/analysis to answer the empirical question of whether providing more information to respondents will ultimately result Volume XVIII, Spring 2022 | 21

POLITICAL SCIENCE

factual basis for the claim, but legal basis/strategy is not necessary), it is possible that this would be viewed as substantially the same as the Trump-era rules and thus would be struck down because both interpret Mach Mining requirements.

Regardless of whether new rules would ultimately be found permissible, the ambiguity of the ‘substantially the same form’ standard alone makes rulemaking a time-and-resource-consuming and potentially wasteful option. The statute itself does little to define this standard, and the legislative history does little to provide additional guidance (Chen). To make matters worse, lower courts are split on the issue of whether the ‘substantially the same’ standard is reviewable in courts (Chen). The ambiguity of the Congressional Review Act’s ‘substantially the same’ standard has deterred agencies from issuing new rulemaking on the same subject in all seventeen previous repeals under the Act. The EEOC should follow in line with these agencies and not consider rulemaking as a main option.

in more settlements.

As established in section III.B. above, issuing new rulemaking is not advisable because it would take significant Commission time and resources and risk being struck down due to the nature of the Congressional Review Act. The particular circumstances of this case mean that the best solution may be a combination of Option 1 and Option 3. The option to do nothing right away maintains the Commission's flexibility for tailoring the amount of information shared with respondents to each case. Therefore, as long as Congressional interference continues to remain unlikely, this course of action is advisable. As of now, the possibility of Congressional interference is the lowest it could possibly be due to the current state of a united Democratic government. Although this indicates that the do-nothing plan is safe for now, that will not necessarily remain the case. As soon as the 2022 midterms, the circumstances could change regarding how interested Congress is in clipping the wings of the EEOC. This means that the EEOC should be prepared by then to issue guidance that defines their view of the information-sharing requirements from Mach Mining.

The best way for the Commission to be prepared to issue guidance when the time comes is to have a better empirical understanding of the policy issue itself. The most logical way to do this is by conducting a study that investigates the extent to which sharing more information with respondents would help or hurt the Commission successfully settle more cases. The Trump-era rules claimed that by increasing the amount of information required to share with respondent companies, they would improve the EEOC’s conciliation process by decreasing the 33% of respondents who choose not to participate in conciliation. Part of the Commission’s study should endeavor to find out if, and to what extent, this is true. On the other side, EEOC staff assert that by not requiring such a large and clearly defined amount of information to respondents, they have flexibility that allows them to successfully settle a higher percentage of cases in conciliation. The Commission’s study should endeavor to find out to what extent this is true. The two sides are not necessarily contradictory; it could be true .1 2

POLITICAL SCIENCE

1

that by requiring that more information be shared with respondents, more than 67% of them would agree to conciliation, but fewer of the cases that came to conciliation would actually be successfully settled. However, if the EEOC can obtain a better understanding of the extent to which each claim is true, it can decide what level of information should be required to provide to respondents based on what would result in the most total cases being successfully resolved. After learning more, the Commission will be in a position to issue informed guidance that actually represents its own “unique expertise” and “considered judgment” (Kisor 10, 17) when it becomes appropriate to do so.

The goal of the study should be to find the point that would maximize successfully conciliated cases. Assuming arguments on both sides are correct at least to an extent, providing a higher level of information to respondents will increase the number of attempted conciliations, but will decrease the rate of successful conciliations of those attempted. Thus, there is an ideal point of information shared with respondent companies that would result in the highest total number of conciliated cases. In attempting to find this point, the Commission can lay out a three-part study, as discussed below.2 The first part would attempt to find out the extent to which it is true that providing a higher level of information to respondents will result in more attempted conciliations.

The second part would endeavor to find out the extent to which it is true that providing more information to respondents about the case will result in fewer successful settlements of the conciliated cases. The third part would compare the information found in the first two parts and use mathematical calculations to find the ideal point.

IV. A. Studying why respondents decline conciliation This subsection discusses potential ideas and suggestions for how the EEOC may go about learning the extent to which the basis for the Trump-era rules are valid. Is it true that providing more information to respondents will cut into the 33% that are uninterested in conciliation? To answer this question, the Commission

1 My recommendation assumes that the goal is to maximize the number of successful settlements that come out of conciliation, because Title VII -- the legislation upon which any EEOC guidance would be based -- clearly favors conciliation (Title VII of the Civil Rights Act of 1964).

2 Writing the basis for a full study is beyond the scope of this paper; the following can be considered an outline of how the EEOC might consider approaching this study.

22 | Emory Undergraduate Research Journal

You were provided with [insert information given to respondent for each case]. Please indicate (by circling Y/N) if the following information were provided, it would have changed your company's decision to engage in conciliation: •

•

•

Name of Charging Party/class of Charging Parties [only include if this was not provided to respondents] [Y/N] Further factual basis about the claim [Y/N] (if yes, please indicate what additional information would have led you to accept conciliation) The EEOC's legal basis for the reasonable cause finding [Y/N]

By collecting and compiling data on these questions, the Commission could learn what percentage of respondents that decline conciliation would agree to it at each additional level of information shared. Some qualitative work will be necessary, but these data should give the Commission an idea of the level to which it is possible to cut down on the 33% who decline conciliation. It is possible that the Trump-era rules were purported to decrease this 33% figure, but this was actually a post-hoc reasoning as the intended goal was simply to help businesses. If this were the case, many respondents would answer ‘no’ to each of the questions. Alternatively, it is possible that respondents actually do perceive that a dearth of information harms their chances in conciliation, and a much higher percentage of companies would conciliate with no information asymmetry. Regardless, this information would be useful to the Commission, particularly when measured against the information that would come from the study outlined in the following subsection.

IV. B. Studying the link between the flexibility of information shared with respondents and successful conciliations

The next part of the study, which would be conducted concurrently with the first part, would attempt to learn the extent to which EEOC investigators rely on the lack of requirement that they share information with respondents to tailor each case towards settlement. Is it true that requiring investigators to share the factual and legal basis for the reasonable cause finding would decrease their percentage of successful settlements? If so, by how much? In an attempt to answer these questions, the Commission could collect data in two main ways: interviews with investigators and case studies of successful and unsuccessful conciliations.

Interviews with investigators would serve the purpose of understanding how they tailor the information provided to respondents to successfully conciliate, and determine how important this actually is to investigators. Questions could begin generally and get more specific over the interview: •

• •

Do you actually need the current level of flexibility to reach a settlement, or does it simply allow you to reach a more favorable settlement for charging parties if one is reached? (if the answer to this question is the latter, then the interview can end there).

How do you decide which information to give to respondents when attempting conciliation?

Do you ever see it as necessary or helpful to provide respondents with the EEOC’s legal basis for the reasonable cause finding?

These questions are just examples; in an informal, qualitative interview, the interviewer could tailor the questions as the interview progresses to best understand investigators’ processes and how many fewer cases might be successfully settled if investigators were forced to abide by higher standards of sharing information with respondents.

Another strategy would be to conduct studies of successful and unsuccessful conciliation attempts.

Those conducting the study could sort conciliations into groups of cases that are similar to one another in terms of how much information the investigator chose to share with the respondent, and run regression analyses to determine if this is as significant a factor in reaching settlement as EEOC staff claims. Combined with a solid understanding of how and why investigators choose to Volume XVIII, Spring 2022 | 23

POLITICAL SCIENCE

could attach a mandatory survey with the offer to conciliate that they send to each respondent, to be filled out by respondents who decline conciliation. The survey would essentially find out whether or not providing more information than they did about the case would have changed the respondent’s mind about conciliation.

The survey could be a single question, as follows:

POLITICAL SCIENCE

provide levels of information in each case, the Commission could use this data to determine the extent to which staff requires this high level of flexibility to reach as many successful settlements as they do. Combining the results of the regression analysis with the qualitative context would allow the Commission to make an educated guess as to how many fewer cases would reach settlement if there was a more specific standard for what information must be provided to respondents.

IV. C. Combining data to reach a conclusion The third part of the study would take place after the first two have been completed. It would aim to use the data collected in the first two sections to answer questions such as whether an increased requirement for information sharing would result in a higher enough percentage of respondents agreeing to conciliation to make the drop in percentage of successful settlements worth it? Another question that could be answered by combining data is: what type and level of sharing information with respondents is the ideal point to reach the most total successful settlements? By knowing the answer to these questions, the EEOC would be well-positioned to issue legitimate guidance that reflects “fair and considered judgment” based on its “unique expertise” (Kisor 10, 17) as soon as macro-political factors prompt it.

V. Conclusion Right now, there is a void in clearly defined conciliation guidance. The EEOC was provided standards for how much information is necessary to share with respondents in 2015; under President Trump it issued strict rulemaking that purported to bring the standards up to those laid out by Mach Mining, but shortly thereafter those rules were repealed only for the EEOC to state that it would return to the standard that was laid out by Mach Mining. This void in clear definitions and practices needs to be filled. However, the current circumstances of unified and Democratic government mean that the EEOC need not fill the void immediately; instead, the Commission can use this time to gain empirical knowledge on its best conciliation practices.

This is important not only because the EEOC’s authorizing statute demonstrates a preference for informal methods of settlement such as conciliation, but also because conciliation helps those that Title VII was designed to protect: employees who have suffered from workplace discrimination. Conciliation is the last step 24 | Emory Undergraduate Research Journal

before litigation, so when the EEOC does not meaningfully engage in conciliation, it harms aggrieved individuals because more often than not, when conciliation fails the Commission will abandon the case and issue the individual a Right to Sue letter. This leaves individuals feeling confused and abandoned as they were told that the Commission had determined there was a reasonable basis to believe they were discriminated against, but then left to pursue the case by themselves--often without the resources to do so. Conciliation is not just a statutory requirement for the EEOC; it is also an opportunity to fulfill its mission of stopping and remedying unlawful employment discrimination in the workplace by enforcing Federal laws that prohibit employment discrimination. By gaining empirical knowledge of which practices would lead to successfully settling the most possible cases in conciliation, the EEOC would better fulfill its own obligations, and would help more Americans who face discrimination.

Works Cited

ACUS Guidance documents. Guidance Documents | Ad ministrative Conference of the United States.

Bolton, A. (2020, March). Congressional Oversight. POLS 362: Executive Branch Governance.

Chen, K. (2020, May 15). The Congressional Review Act's Legal Uncertainties. Harvard Law School Environ mental and Energy Law Program.

Congress Acts to Restore Flexibility to EEOC's Conciliation Process. (2021, June 24). EEOC News.

Congressional Review Act: Overview and Tracking. Nation al Conference of State Legislatures. (2021, April 15).

Congressional Research Service, Carey, M. P., &; Davis, C. M., The Congressional Review Act: Frequently Asked Questions (2020).

EEOC Guidance, “Quality Practices for Effective Investiga tions and Conciliations” (September 30, 2015).

Equal Employment Opportunity Commission Procedural Regulations. 29 CFR § 1601.24 (1977). Exec. Order No. 13771, 82 FR 9339 (2017).

Fiscal Year 2021 Congressional Budget Justification. U.S. Equal Employment Opportunity Commission. (n.d.).

Freedman, M., & Lorber, L. (2020, November 9). Comments to EEOC re: Conciliation Procedures. U.S. Chamber of Commerce.

Gibbs, S. R., & Bates, R. M. (2021). Trump Era EEOC Con ciliation Rule Repealed. National Law Review, XI (222).

Statistics, Title VII of the Civil Rights Act of 1964 Charges (charges filed with EEOC) Fy 1997 - FY 2020. U.S. Equal Employment Opportunity Commission.

(2020).

Update of Commission's Conciliation Procedures: A Rule by the Equal Employment Opportunity Commis sion on 01/14/2021, 86 FR 2974 (2021) What You Can Expect After a Charge is Filed. U.S. Equal Employment Opportunity Commission.(n.d.).

What You Should Know: The EEOC, Conciliation, and Lit igation. U.S. Equal Employment Opportunity Com mission. (n.d.).

Written Testimony of Andrew F. Maunz, Legal Counsel, for August 18, 2020 Commission Meeting. U.S. Equal Employment Opportunity Commission.

Additional Sources Consulted

Morris, F. C., & Glasser, N. M. (2015). Supreme Court's Decision in Mach Mining Impacts Employers' Approach to Conciliation with the EEOC. The Na tional Law Review, 5(125).

Office of Management and Budget, & Vought, R. T., Mem orandum for the Heads of Executive Departments and Agencies: Guidance on Compliance with the Congressional Review Act (2019).

Shapiro, S. (2021). The Impossibility of Legislative Regulato ry Reform and the Futility of Executive Regulatory Reform. George Mason Law Review, 28(3).

Vaas, F. J. (1966). Title VII: Legislative History. Boston Col lege Law Review, 7(3).

Written testimony Of Lorene F. Schaefer, Esq. Workplace In vestigations Group One Mediation, Inc. U.S. Equal Employment Opportunity Commission. (2013, March 20).

David was born in Washington, D.C. and grew up in Falls Church, Virginia. David is pre-law at Emory, majoring in Spanish and Political Science. At Emory, David is the President of the Club Baseball team. Outside of Emory, David is also an avid sports fan, cheering for his home teams of the Nationals, Wizards, Capitals, and Commanders.

Volume XVIII, Spring 2022 | 25

POLITICAL SCIENCE

Grossmann, M. (2014). Artists of the possible: Governing networks and American policy change since 1945.

Oxford Univ. Press.

Kisor v. Wilkie, 588 U.S. __ (2019) Laws enforced by EEOC. U.S. Equal Employment Opportu nity Commission. (n.d.).

Lowande, K., & Peck, J. (2016). Congressional investigations and the Electoral Connection. Journal of Law, Eco nomics, and Organization, 33(1).

MacDonald, J. A. (2010). Limitation riders and congres sional influence over bureaucratic policy decisions.

American Political Science Review, 104(4), 766– 782.

Mach Mining, LLC v. EEOC, 575 US 480 (2015) Meeting of March 20, 2013 - Development of a Quality Con trol Plan for Private Sector Investigations and Con ciliations. U.S. Equal Employment Opportunity Commission.

Parrillo, N. R. (2019). Federal Agency Guidance and the Power to Bind: An Empirical Study of Agencies and Industries. Yale Journal on Regulation, 36(1), 165271.

Potter, R. A. (2021, July 8). Improving White House Review of Agency Guidance. Brookings.

Potter, R. A. (2020). Providing political guidance? Agency politicization and "as if " policymaking. Journal of Political Institutions and Political Economy, 1(1), 1–26.

Small Business Regulatory Enforcement Fairness Act, Pub. L. No. 104-121, 110 Stat. 857 (1996) (5 U.S.C. §§ 601, 631, 648(c)(3), 657) Title VII of the Civil R ights Act of 1964, 78 Stat. 241, 42 U.S.C. § 2000e-5(b).

NUTRITION

Nutrition Education to the Public Introduction

Role of the Private Sector in Nutrition Nichole Stettner

Throughout the world, and specifically in the United States, malnutrition –including under and overnutrition– is its own pandemic. Many of the foods in the mainstream market are filled with more than double the recommended daily sugar intake, have more preservatives than actual ingredients, and contain genetically modified foods. There is also not enough education within food labeling, which may also be correlated with the high prevalence of malnutrition in the United States.

Additionally, food labels with inaccurate ingredient lists and potentially misleading nutritional information have a negative impact on the health of consumers. Through a collaborative effort between the government, the people, and –most importantly– the private sector, there can be a fundamental change in the nutritional status in the United States.

Background The history behind nutritional information spans many decades and has many aspects with designated purposes. The “inclusion of [nutrition information on packaged food labels] was to be voluntary, except when nutrition claims were made on the label, in labeling, or in advertising, or when nutrients were added to the food.”1(p1) These guidelines, therefore, could allow for much flexibility between products. Around the same period, these manufacturers began making vague claims to increase sales.1 This correlates with the idea that profit is the primary factor when making the products. To address these concerns and the formatting of the nutrition label, public hearings were held throughout the United States.1 Once the FDA had created the Daily Reference Values, this was one step closer to having the label of the present day.

Since 1990, there have been some adjustments to the nutrition label, such as the addition of the Added Sugars label and Daily Reference Values. After about thirty years, it seems appropriate to have a major adjustment to the nutrition food label in America.

Another factor to consider is how Americans were living their daily lives at the time. For example, “Until the 1960’s, most Americans prepared the majority of their meals at home, but with a shift in consumer demand for 26 | Emory Undergraduate Research Journal

prepared products came a public demand for detailed production information.”2(p1) The food and nutrition industry has been changing with the times: making “onthe-go” features more in demand when that shift may have occurred.

Nutrition Facts: The Specifics Because of the larger issues surrounding malnutrition, the nutrition labels were altered. Many guidelines have been developed by the FDA for claims that are allowed on packaged sources. The FDA indicates that “A “good source” claim may be made when a food contains 1019% of the RDI or DRV (both declared on the label as the % Daily Value (%DV)). A “high” claim may be made when a food contains at least 20% of the DV.”3(p74) Thus, there is a specific percentage that qualifies a product as a “good source” of a nutrient, which is not always indicated to the public. When shopping in a grocery store, it is rare to find detailed information on how to use Daily Reference Values and, more importantly, how they impact one’s everyday eating.

The wide breadth of information to give about health claims on food claims is too much to give in an overview. One type of claim is described as “qualified health claims.” 4(p4) These are health claims that can be (and must be) verified by the FDA if there is an association between the disease and an ingredient in the product or the product itself, the company must go through a petition process to allow for that claim to be made.4 These guidelines for nutrient content claims are, although slightly confusing, fair in the information required to have them. For example, the FDA mentioned that putting “only 200 mg of sodium” on the package would imply that the product is “low” in sodium according to the health standards, so the companies must ensure that their claims are correct.

4(p5) The issue with all these guidelines, however, is that they are not being publicly addressed to consumers.

The nutrition facts label has been updated to have a larger font size for the information concerning the serving size and the calories.5 Added sugars have now also been added to the label beneath the total sugars.6 This new labeling allows for the consumers to be aware of what sugars have been added to the product during pro-

The purpose behind nutritional information is that the public should be provided truthful information, not tricked into thinking that a product is healthy.

New terms, same problems As previously noted, many packaged foods use vague promotions, such as a good source of Vitamin D. Since these terms are indistinct, at first it seems that there is no way for them to have any numerical meaning. Despite this, studies have indicated that the “FDA and USDA should define descriptors (e.g., “high,” “good source of ”) for the content of nutrients such as fat, cholesterol, sodium, and micronutrients.”1(p5) Ambiguous labeling made for the sole purpose of advertising is defeating the consumer out of their health. People put their trust in these corporations to provide them with accurate claims regarding the nutrition of their foods. Ambiguous claims combined with the complex Daily Reference Values equates to a confusing way of interpreting presented nutrition information. Why are the companies not indicating the ingredients clearly and concisely? The purpose behind nutritional information is that the public should be provided truthful information, not tricked into thinking that a product is healthy.

How True are Organic Claims?

An example of the challenge to accurately label nutritional claims to consumers is demonstrated by organic food claims. Products are marketed as organic, which leaves consumers with the idea that their food is safer, even when it is not. By definition, organic food is grown in soil without the use of artificial pesticides.8 All organic foods may not necessarily be organic by definition; this is due to previous farming with pesticides on the same land that organic foods are now being planted on.9 Thus, there is still a chance that the organic food that we have in the stores may be affected by harsh chemicals.

this, organic foods do have lower levels of chemicals than other non-organic products.9 Thus, in deciding between purchasing organic food versus non-organic food, completed studies demonstrate that it may serve to be more beneficial to consume organic foods. Another way to ensure the validity and quality of the produce is to consider buying locally grown and in-season foods. If organic food is simply a marketing ploy, then the people have a right to know; however, there is more to marketing when it comes to organic food. Whether or not organic food is better, it still costs more for the corporations to not put pesticides and preservatives in the making of the food. This in turn leads consumers to believe that the companies are trying to find new ways to make profits instead of helping people.

Can We Trust the Companies?

Because of advanced advertising strategies, many companies are able to market products that are in no way nutritious. These companies are doing a great job at marketing otherwise unmarketable products. The corporations can market to all age ranges through the use of cartoon figures, exploitation of emotions, and the vague and misuse of terminology. Advertisements for products high in sugar can be seen as comparable to those for cigarettes in the 1950’s.10 Even though healthy food can be properly and effectively marketed, it is still the “bad” or unhealthy food that is in the mainstream market.

As previously mentioned, organic is a term that can be used as a marketing tool to gain more customers.11 These companies are using human health as a marketing tool, which, again, begs the question of whether they care more about nutrition or profits. There is also often a higher price on organic foods, so consumers must determine whether the purchase is worth the extra expense.

11 In many cases, the added prices may be present in addition to the health benefits of the food. It is important, as a consumer, to research products before purchasing to determine whether they align with one’s beliefs. Many true and ethical companies want consumers to have organic and healthy food. There is also a demonstrated difference between organic shoppers and non-organic shoppers.11 Products that promote a certain vitamin Volume XVIII, Spring 2022 | 27

NUTRITION

-cessing.6 Additionally, the Daily Values have been updated to new standards.7 Daily Values can help consumers become more aware of how much nutrients they are consuming in their average diet. So, the work that has been done as of now (according to the information provided by the FDA) has managed to provide these general rules to predominantly all packaged and processed foods, which is a much needed first step; despite this, there are still many lingering problems.

NUTRITION

without considering a missed nutrient or bad side effects can create more damage than intended.

It may not come as a surprise that most adults in the United States are not consuming enough fruits and vegetables as they should be.12 In a study conducted in 2011, it was noted how “orange juice was the largest contributor to overall fruit intake in the USA.”12(p8) Are people simply obtaining all of their necessary vitamin C through juice, or are they also consuming a variety of whole fruits? Fruit juices do not have the amounts of fiber that we need in them that we get from whole fruits. It may come as a second thought to how people are receiving their necessary nutrients. The question that needs to be answered is should companies be required to indicate the missing –yet implied– nutrients in their products?

One of the main reasons that unhealthy foods can have better marketing is because of the added value of the product.13 The unhealthy foods can “appeal to consumer preferences for salt, sugar, fat, and convenience.”13(p2) Poor nutrition marketing takes an even worse role in the form of advertisements geared to children. Cartoon characters and mythical settings create the illusion that the feelings advertised with those products are what will occur when consuming them. It is more difficult to create an appeal for ordinary fruits and vegetables. Henceforth, when you go to the store, there will be sliced apples in decorated plastic bags and spring mixes in their “washed and ready to eat” containers. The convenience factor is being sold, as it is simply a ploy to allow more people to purchase those items.

to have before working out, but people should be informed about the types of supplements they are taking and whether it even has sufficient evidence to provide those uses. In addition to this, all people should take caution with the number of supplements that they are consuming to ensure that they do not suffer malnutrition.

When discussing supplementation and sport, it may serve important to differentiate between “elite” and “not-elite” athletes.15(p1) Those who are considered “elite” athletes may be more likely to take supplements.15 Additionally, following the implementation of the Dietary Supplement Health and Education Act (1994), supplement sales had skyrocketed.15 The types of dietary supplements that “elite” athletes are more likely to take include vitamins, minerals, multivitamins, amino acid/ protein supplementation, creatine, sports drinks, sports bars, caffeine, and energy drinks.15 Without a nutritionist or trainer to guide, it may be difficult to determine how to take supplements. Thus, it is important that athletes do not take supplements “on an “ad hoc basis”, unaware of what the active ingredient(s) are or of the amount of those ingredients.”16(p133) When various types of supplements are consumed at a time, there may be an increased “risk of exposure to harmful levels of specific substances or interactions causing adverse health outcomes (e.g., calcium and interference of iron uptake).”16(p133) It is very unlikely that consuming more supplements is equivalent to being healthier or stronger.16 There are proposed pathways that may determine whether or not an athlete should even be taking a supplement, whether the supplement has accurate scientific evidence, and more.16 To How True are Organic Claims? reiterate, it must be understood how a product’s ingrediMany types of dietary supplements have been developed ents affect the consumer and their health. and are common for athletes to take.14 Despite this, are Who should be responsible? all supplements necessary and/or healthy to take? Not necessarily. Creatine, for example, is popular amongst Consumers individuals who participate in weightlifting.14 Many Many organizations and corporations have indicated college athletes, in particular, consume creatine to help that it is the consumers’ responsibility to maintain a them build muscle. An adverse effect it has is “Weight healthy and balanced diet. Despite this, “Consumers are gain due to water retention,” which can lead to false as- more likely to purchase good quality products that help sumptions as body mass is not defined by what you see them to meet dietary recommendations as they become on the scale.14(p5) Many people also consume caffeine pri- more informed,” in addition to other attributes.17(p2) or to a workout to try to energize themselves.14 Adverse Since there is a lack of food education among consumeffects of consuming caffeine are restlessness, arrhyth- ers, nutrition education about food products must be mias, and insomnia.14 Caffeine and the workout would better communicated with the public. thus promote a very accelerated heart rate. This is not to Guidelines have been proposed to help consumers in say that all creatine or every caffeinated substance is bad remembering to eat healthily and committing to it thr28 | Emory Undergraduate Research Journal

-ough their actions.18 However, the methods seemed contradictory as individuals had difficulty remembering what all of the various nutritional information and serving sizes meant to themselves.18 Nutrition facts labels are typically based on a 2,000-calorie diet. They do not account for weight, height, chronic condition, family history, and so much more. Without a nutritionist or physician to guide every food decision a person makes, how must they be expected to stay healthy?

Government The government is often one of the first places that people go to for health information. A 2016 paper indicated that “Viewing food labels as a market trust device leads to two challenges for proponents: how to establish a credible system of institutional accounting to win consumers’ trust in a certification scheme… and how to address the perennial moral hazard of cheating or of ‘parasitic’ reappropriations of labels (i.e. cheap copies) that undermine credibility.”19(p148) The U.S. government does not necessarily manage its food services in this way. Since there is so much confusion surrounding the labeling and a lack of education in it, that does not lead to trust in the programs that are required to regulate it. How can we put our full trust into an organization that will often

take and validate the cheaper option despite the health consequences? Consumers need to understand and be educated on what goes into their food products and how that is affecting their health.

One of the government's main ways of establishing health among the citizens is through passing legislation.

In general, three federal laws that control food and nutrition labeling: FD&C Act, The Nutrition and Labeling and Education Act, and The Fair Packaging and Labeling Act of 1966.19 The last time the nutrition label was minimally adjusted was in 2016; the updates to the nutrition label allowed for information to be read more readily, the addition of added sugars, change in how serving size is measured, and “A new footnote [explaining] what percentage of the Daily Value (DV) means.”20(p1) Yes, these changes are a step in the right direction; however, as previously noted, these lead to significant miscommunication. A lawsuit case involved Kellogg’s paying $20,250,000 to settle a lawsuit because of their claims that their sugar-cereals are healthy.21 In the report, Kellogg’s also had to remove the “heart healthy” promotion that was on some of their products.21(pp3-4) Another case involved Frito-Lay and the fake promotions they constructed concerning their salt and vinegar chips: “containing only natural ingredients.”20(p1) Despite their clai-

Source: U.S. Food and Drug Administration, Creative Commons

NUTRITION

-ms, the chips also contained malic acid, a food preservative.22 The case was dismissed as there needed to be crystal clear evidence that the company was wrong.22 A third court case concerned a “lawsuit against Barlean’s Organic Oils alleged the company engaged in misleading marketing by [calling] its coconut oil “healthy” when it contained high levels of saturated fats.”20(p1) A point in the case was that the consumers would not have used the products if they were aware of the harmful effects.23 As previously stated, many varied health phrases and claims can appear on a food product, and how it is defined and whether it is regulated may differ from what is implied.

to be excluded in academic public health discourse.”26(p1) Many companies have mainstream advertisements for unhealthy foods, which means that there is potential for nutritional foods. Large corporations have a large influence on public health. It is a “shared responsibility” between the public and private sector that can encourage proper health.26(p1) There are many reasons why there is poor nutrition amongst individuals (i.e. the social determinants of health), which is why “collective action” is needed.26(p2) A way in which the private sector is regulated by the government helps to combat terrorism, which is a point that is rarely focused on.27 To fight against terrorism in the food market, food that gets imported into Private Sector the United States must be approved and recorded under Food marketing helps assist in building relationships the FDA.27 A strict record must also be kept of the cuswith customers and builds up their brand.13 This, in tomers of the product.27 turn, can help to pay the grocery stores more for access to better advertising locations throughout the store.13 When considering food labeling and nutrition, a Johns Hopkins article posed, “To what degree are individuals responsible for their own food choices, versus the com- There are many reasons why there is poor panies that leverage their knowledge of human psychology to promote sales of their products? And what, if any, nutrition amongst individuals (i.e. the sois the role of [the] government in regulating food marcial determinants of health), which is why keting, particularly online?”13(p2) An interrelationship between the private sector, government, and consumer "collective action" is needed. can be developed.

People across the United States and the world are experiencing chronic malnutrition.24 The private sector has the “potential” to “address food security and malnutrition worldwide through nutrition-sensitive inter- How Can the Private Sector Assist with Public ventions.”24(p2) This is especially important in the United Nutrition?

States because of the limited funding that public health is allocated.25 Investing in local, specialized practices is Although the private sector is a part of the nutritioncritical to counteract corporate domination.24 This may al crisis, they can still take action.28 The private sector lead to a similar system to that of the present, in that currently assists by “investing in agriculture,” “improvthere would be little room for change and only one solu- ing the social, economic and environmental practices tion. For private sector engagement, local solutions can in farming and the supply chain,” and “mobilizing, inbe adopted and can provide “nutrition education in [the] novating, and finally delivering agricultural products workplace.”24(p3) These solutions, amongst many more, and food.”29(p1) However, one company cannot provide can help proper nutrition become normalized. The most the end-all solutions to malnutrition across the Unitimportant aspect that the private sector has when trying ed States and the world.29 One of the most significant to communicate nutritional information to consumers aspects of improving malnutrition through the private is to connect with them.24 If individuals do not feel like sector is to “[increase] trade, particularly in the agriculit matters and do not understand the complex terminol- ture and food industry, [which can raise] the standard of ogy of the labeling, it will be more difficult to put nu- living in developing countries and [improve] the perfortritional education into action. The private sector has a mance of national economies.”29(p2) When these underde“corporate responsibility for public health [that] seems veloped economies and communities are invested into, 30 | Emory Undergraduate Research Journal

Discussion How to engage the customers in nutritional information When trying to eat healthily, it is difficult to keep track of all your vitamins, added sugars, daily values, and more.

A way to implement all these ideas is to have messages that are easy to remember and relatable to the individual.18 A proposed method to promote nutrition and heart health to the people was through individuals’ hairdressers.18 Although uncommon, this method would allow the person to understand what is being said and feel a drive to listen to the information.18 Most people develop their eating patterns during childhood, so focusing on adolescents can help shape a greater future. A study that focused on high school athletes and their nutrition noted that many students regularly skipped meals.30 These students also had poor meal choices in addition to their irregular scheduling.30 The high school students were then given small-group nutritional education.30 The small-group nutritional education is a way to connect with the students one-on-one to allow the bad habits to dissipate before adulthood.30 A school environment, therefore, has a significant effect on adolescent nutrition. Although adolescents and children can learn to eat healthier through school programs, the ultimate decision-makers for the food that enters a household are often the parents.

There has been a lack of nationwide nutritional programs in the United States, so tackling the issues at the source and engaging individuals directly can lead to the most benefits. Since the private sector cannot bring na-

-tionwide nutritional success by itself, it may help if the government provides some aid as well. The food label that was previously discussed can undergo major adjustments to help consumers understand the products they are purchasing. For instance, in France, the food label has been updated to be color-coded, so customers can better manage their nutrition.31 This idea is especially helpful for quick and on-the-go purchases, for the consumer would be able to simply look at the colors of the label and better understand if that food would exceed that individual’s daily limits. Since there is a lack of nutritional education and intervention within the United States, interventions promoting these would be most successful.

Corporations need to become more responsible for their products by informing the public of how each of the ingredients in the product affects people’s health and the purpose of each ingredient in the products.

Conclusion Throughout its history, the nutrition label has had (and still has) many controversial points. It is essential that the private sector and government collaborate to ensure that the public is getting the most accurate information possible about the food they are purchasing and consuming.

Successful nutrition education can be achieved through joint campaigns and educational forums. As the private sector is able to connect more with its consumers, it has an opportunity to give consumers accurate information about products’ ingredients, nutrition information, and conditions they are grown in. It is necessary that the marketing of foods begins to acknowledge the harmfulness of some products, which can allow for nutrition to finally be a part in considerations of profits.

References

1. Institute of Medicine (US) Committee on Examination of Front-of-Package Nutrition Rating Systems and Symbols, Wartella EA, Lichtenstein AH, Boon CS. History of Nutri -tion Labeling. National Academies Press; 2010.

2. Factual food labels: A closer look at the history. Utexas. edu. Accessed April 18, 2021.

3. A Food Labeling Guide. Fda.gov. Published January 2013.

4. Center for Food Safety, Applied Nutrition. Label claims for conventional foods and dietary supplements. Fda.gov.

Published May 2, 2020.

5. Center for Food Safety, Applied Nutrition. The new Nu trition Facts label. Fda.gov. Published 2020.

6. Center for Food Safety, Applied Nutrition. Added sugars

Volume XVIII, Spring 2022 | 31

NUTRITION

that in turn will allow for more resources and education in those regions.29 Committing to the investments poses a financial risk.28 Some ways to increase the likelihood of success in engaging the private sector in these matters include “facilitating the supply of nutritious foods by reducing production costs,” “increasing the demand for nutritious food and marketing,” and “accessing [the] funds and investments.”28(p5) In addition to investing in agriculture, companies in the private sector can also work to create products that are affordable and healthy for all socioeconomic classes.28 Despite setbacks that may occur when forming partnerships, the private sector can allow for significant nutritional growth due to its sturdy foundation in business.28

NUTRITION

on the new Nutrition Facts label. Fda.gov. Published August 1, 2021.

7. Center for Food Safety, Applied Nutrition. The lows and highs of percent Daily Value on the label. Fda.gov. Pub lished August 1, 2021.

8. Organic 101: What the USDA organic label means. Usda. gov.

9. Magkos F, Arvaniti F, Zampelas A. Organic food: buying more safety or just peace of mind? A critical review of the literature. Crit Rev Food Sci Nutr. 2006;46(1):23-56.

10. Little B. When cigarette companies used doctors to push smoking. HISTORY. Published September 13, 2018.

11. Sinclair AM, Smith RR. The Hochschild Cohomology Problem for Von Neumann Algebras. Proc Natl Acad Sci U S A. 1998;95(7):3376-3379.

12. Demydas T. Consumer segmentation based on the level and structure of fruit and vegetable intake: an empirical evidence for US adults from the National Health and Nu trition Examination Survey (NHANES) 2005-2006. Pub lic Health Nutr. 2011;14(6):1088-1095.

13. Driver K, JH Bloomberg School of Public Health. Food Marketing and Labeling. Foodsystemprimer.org. Pub lished August 5, 2016.

14. Dietary supplements for exercise and athletic performance. Nih.gov.

15. Knapik JJ, Steelman RA, Hoedebecke SS, Austin KG, Farina EK, Lieberman HR. Prevalence of dietary supple ment use by athletes: Systematic review and meta-analy sis. Sports Med. 2016;46(1):103-123.

16. Garthe I, Maughan RJ. Athletes and supplements: Prevalence and perspectives. Int J Sport Nutr Exerc Me tab. 2018;28(2):126-138.

17. Institute of Medicine (US) Committee on Dietary Guide lines Implementation, Thomas PR. Private Sector. Strat egies and Actions for Implementation. National Acade mies Press; 1991.

18. Promoting Food Literacy: Communication Tools and Strategies. The National Academies Press.

19. Frohlich X. The informational turn in food politics: The US FDA’s nutrition label as information infrastructure.

Soc Stud Sci. 2017;47(2):145-171.

20. Legal guide to health claims on food. Udayton.edu. Pub lished March 12, 2021.

21. $20 million class action settlement reached with Kellogg’s over sugary cereal health claims. Classaction.org.

22. Allred v. Frito-lay N. am., Inc., Case No.: 17-CV-1345 JLS (BGS). Casetext.com. Published March 5, 2019.

23. Class action claims Barlean’s misrepresents coconut oil p-

32 | Emory Undergraduate Research Journal

-roducts as healthy. Classaction.org.

24. Global lessons learned. The role of the private sector in food and nutrition security. Alliance2015.org.

25. Himmelstein DU, Woolhandler S. Public health’s fall ing share of US health spending. Am J Public Health.

2016;106(1):56-57.

26. Tempels T, Verweij M, Blok V. Big food’s ambivalence: Seeking profit and responsibility for health. Am J Public Health. 2017;107(3):402-406.

27. Meadows M. A Century of Ensuring Safe Foods and Cosmetics. Fda.gov.

28. Navigating the labyrinth of public-private engagements for Global Nutrition. Csis.org.

29. Staff Contact: Brian Lowry. Global nutrition: What is the private sector doing? Uscib.org. Published March 22, 2016.

30. Abbey E. EXPANDING THE ARENA OF SPORT NU TRITION. Lww.com.

31. France becomes one of the first countries in Region to recommend colour-coded front-of-pack nutrition label ling system. Published online 2017.

Photo by Naomi Akram

NUTRITION

Nichole Stettner is a third year undergrad at Emory College studying chemistry and human health. After graduating, she hopes to become a physician and focus on preventive care.

Outside of academics, Nichole is involved in research, connecting with hospice patients, and mentoring for various organizations. In her free time, she enjoys going on runs through Lullwater and singing in Emory’s University Chorus.

Volume XVIII, Spring 2022 | 33

Interaction between Tau and U1-70k Proteins in Alzheimer's Disease – Liquid-Liquid Phase Separation

BIOCHEMISTRY

Abstract

Yingrong Chen, Brianna Hammond, Joan Adebowale, Bo Liang Department of Biochemistry, Emory University School of Medicine

Tau is a microtubule-associated protein playing important roles in neuron microtubule stabilization and axonal transport. Its abnormal aggregation – tau neurofibrillary tangles – is one of the pathological hallmarks of Alzheimer's disease. Studies have shown that tau coacervates with several RNA-binding proteins and disrupts the RNA regulation process. In particular, U1 small nuclear ribonucleoprotein 70 kDa (U1-70k) is an RNA-binding protein found in the U1 snRNP component of the spliceosome. In Alzheimer's disease, U1-70k aggregates and colocalizes with tau neurofibrillary tangles, a unique association absent in any other tauopathies and spliceosome subunits. The interaction between tau and U1-70k may explain the impairing pre-mRNA regulation and further pathological symptoms in Alzheimer's disease; however, its molecular basis remains unclear. This paper reviews the physical chemistry, physiologic functions, and aberrant activities of tau and U1-70k and proposes an interacting mechanism. Tau and U1-70k may form complex coacervates via liquid-liquid phase separation, a biophysical process when one homogenous liquid phase separates into dense aggregates and dilute surroundings. Learning the mechanism underlying tau and U1-70k association sheds light on Alzheimer's disease pathogenesis.

Keywords: Alzheimer's Disease, Tau, U1-70k, Liquid-liquid phase separation, Hyperphosphorylation

Introduction Alzheimer's disease (AD) is a progressive neurodegenerative disease responsible for most dementia and cognitive decline medical cases today. The hallmarks of AD pathology are amyloid-β plaques (Aβ) and tau neurofibrillary tangles (NFTs), along with neuronal cell death and atrophy. Tau malfunction is associated with both the onset and progression of AD; therefore, learning the pathological mechanism of tau can provide significant insights for AD therapy. Tau-targeted immunology has received increasing attention from the scientific community recently due to its effectiveness in attenuating cognitive decline and pathology propagation in AD.1 Tau is a microtubule-associated protein found in the neuronal axons, cytosols, and plasma membranes. It has six isoforms due to the alternative splicing of the microtubule-associated protein tau gene (MAPT). The physiological activity of tau is predominantly associated with microtubule formation and dynamics, but it is also involved in neuronal development, axonal transport, and synaptic functions. Aberrant tau activities are indicators for a set of neurodegenerative diseases called tauopathies, including Alzheimer's disease (AD), Pick's disease (PiD), frontotemporal dementia (FTD), argyrophilic grain disease, and progressive supranuclear palsy.2-4 34 | Emory Undergraduate Research Journal

Tau has been found to colocalize with RNA-binding proteins (RBPs) and facilitate important membrane-less organelle formations in RNA processing, such as cytoplasmic stress granules and nuclear speckles.5 In neurodegenerative diseases, the pathological coaggregation between tau and RBPs is responsible for deleterious cell splicing and translation. In particular, U1 small nuclear ribonucleoprotein 70kDa (U1-70k) is an RBP in the U1 snRNP component of spliceosome. During mRNA splicing, it facilitates the recognition of the 5' splice site of introns on the pre-mRNA. Recent studies show that U1-70k aggregates and colocalizes with NFTs in AD pathogenesis.6-8 However, the underlying mechanism between U1-70k and tau association remains unclear.

One potential explanation for the interaction between tau and U1-70k is liquid-liquid phase separation (LLPS). LLPS is a biophysical process when one homogenous fluid phase de-mixes into separate compartments with different concentrations – the solutes condense into coacervates (the dense aqueous droplets rich in macromolecules) and the surrounding environments become a dilute phase. This creates a chemically distinct compartment so that molecules separated from their surroundings have higher local concentrations, interact more efficiently, and undergo specific reactions. The

molecular base of LLPS is the intrinsically disordered regions, in other words the low-complexity domains (LCDs), in proteins. Since the LCDs prefer to interact with each other over surrounding solvents, proteins with such characteristics can easily co-aggregate. Both tau and U1-70k have LCDs and can undergo LLPS. They may coacervate via lateral coaggregation. 9-11 This review summarizes the physical chemistry, physiological functions, and aberrant activities of tau and U170k in relation to AD. Potential interaction mechanisms between tau and U1-70k are reviewed, and the role of LLPS is emphasized.

1. Tau 1.1 Physical Chemistry of Tau From N-terminus to C-terminus, tau is mainly composed of a projection domain (1–151 aa), a proline-rich domain (PRD, 151–244 aa), a microtubule-binding region (MTBR, 245–368 aa), and a C-terminal domain (369–441 aa) (Figure 1). Tau forms secondary structures in specific regions – the β-strands in the MTBR and polyproline II helices in the PRD. It is suggested that the two terminals of tau can fold back to the central PRD and MTBR.12 The PRD contains the most phosphorylation sites of tau, and it is also responsible for regulating the electrostatic interaction between two oppositely charged termini in tau.3 Tau is innately disordered since it does not possess well-defined tertiary conformations. Its intrinsically disordered domain is able to form weak, transient intermolecular interactions, such as electrostatics, pi-pi stacking, and hydrophobic interactions.3 Therefore, tau is able to adopt various structures and engage in numerous biological functions such as microtubule stabilization, cell signaling, axonal transport, and so forth.

Different tau species have different microtubule-binding affinities and aggregating rates, so they function differently in the human brain. In human beings, the aternative splicing of exons 2, 3, 10 on the MAPT gene on chromosome 17 produces six tau isoforms named

Different tau species have different microtubule-binding affinities and aggregating rates, so they function differently in the human brain. In human beings, the aternative splicing of exons 2, 3, 10 on the MAPT gene on chromosome 17 produces six tau isoforms named 0N3R, 0N4R, 1N3R, 1N4R, 2N3R, and 2N4R (352, 381, 383, 410, 412 and 441) (Figure 1). These isoforms are differentiated by the number of inserts (N0, N1, N2) in the projection domain and the number of imperfect repeat sequences (3R, 4R) in MTBR. Generally, the 4R tau is the fast-aggregating species compared to 3R tau. In early developmental stages, only 0N3R tau is expressed along with an elevated level of tau phosphorylation, which enables high brain plasticity with less aggregating tau and more dynamic microtubules. The ratios of 4R to 3R tau isoforms are responsible for the phenotypic variability of different tauopathies. The NFTs deposits in AD are made of both 3R and 4R tau. The progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), and argyrophilic grain disease (AGD) all have upregulated 4R isoforms; and the Pick's disease is characterized by 3R isoforms.10 It is found that tau aggregation can be regulated by the recruitment of different forms of tau (mutants or isoforms) that “dilute” the aggregation-prone tau species and lower the fibrillation rate.13 There are four assembly states of tau with distinctive physicochemical and pathological characteristics found in the neurons: soluble monomers, liquid or gel-like condensates, soluble oligomers, and insoluble fibrillary aggregates (Figure 2).4 The soluble monomeric tau is inherently disordered and adopts many different molecular structures. The liquid or gel-like state is a dense liquid-like condensate formed by the reversible LLPS of tau monomers. There is higher tau concentration and amyloid-promoting factors inside the liquid condensate, which optimizes the intermolecular interaction and continuing aggregation of tau.10 Thus, the liquid condensate serves as an intermediate from monomeric tau to oligomeric tau.14 Volume XVIII, Spring 2022 | 35

BIOCHEMISTRY

Figure 1. Amino acid sequence and important domains of the longest Tau isoform (2N4R; 441). There are two N-terminal inserts (N1 and N2) in the projection domain and four pseudo-repeats (R1-4) in MTBR.4

BIOCHEMISTRY

Formation of NFTs in AD starts when hyperphosphorylated tau accumulates into oligomeric tau. Although NFTs are the main pathological manifests of neurological decline, studies show that the tau oligomers prior to NFT formation are the toxic species responsible for aberrant axonal transport, synaptic loss, and mitochondrial abnormalities. Tau oligomers can induce abberant endogenous tau folding and propagate in unaffected brains via a prion-like pattern.15 The NFTs are made of ordered, filamentous hyperphosphorylated tau. The formation of NFTs is correlated with the degree of neuron loss and cognitive impairment in AD cases.

However, instead of causing the disease as most people assume, NFTs may serve as a protective role in response to pathological conditions.16 Typical tau post-translational modifications include phosphorylation, acetylation, truncation, and ubiquitination, which influence the tau activity by regulating its net charge, local structure, and intermolecular interaction. In the normal brain, tau is mostly phosphorylated; there are 2–3 moles of phosphates on each tau molecule.17 Physiologic tau phosphorylation is a beneficial response to various environmental stressors, such as insulin dysfunction and glucose deprivation.18 By adding negative charges and lowering the isoelectric point (pI) of the protein, phosphorylation favors tau LLPS. Phosphorylation regulates the microtubule-binding behavior of tau and therefore the imperative cytoskeletal integrity and axonal transport efficiency.

In acetylation, an acetyl group is added to the amide group of lysine residues on tau. This neutralizes the positive charges on the side chains of tau and increases hydrophobicity. Thus, by disrupting the electrostatic interactions mediated by lysine residues and the hydrogen bonds between lysine side chains and RNA, acetylation can attenuate tau LLPS.3 Although both phosphoryla-

tion and acetylation decrease the positive charge of tau, they affect tau LLPS and pathological aggregation differently.

1.2 Physiological activities of Tau Due to its extensive distribution in the body, tau participates in a wide range of physiological activities. Tau has been known to bind with the microtubule since 1975 and plays an important role in microtubule formation and dynamics.19 Its proline-rich region and pseudo-repeat domain in the C-terminal half are associated with tubulin and microtubule filaments. Tau serves as a nucleation center and decreases the equilibrium concentration needed in tubulin polymerization. It also binds to the tubulin heterodimer interface in microtubules, decreases the tubulin dissociation rate, and stabilizes microtubule filaments. Tau condensates (tau islands) formed on the microtubule surface can protect microtubules from damaging enzymes such as katanin and spastin. Moreover, since the microtubule is an important structure for organelle transports, axonal growth, neuronal development, and cytoskeleton arrangement, tau can contribute to these activities via its interaction with the microtubule. For instance, by interacting with the motor proteins on microtubules, such as kinesin-1 motor protein and dynactin complex, tau can regulate axonal transport.

Besides axons, tau can be found in dendritic and synaptic compartments of healthy neurons. It interacts with a number of molecules in the signal transduction cascades, such as the synaptic receptors muscarinic acetylcholine receptors (mAChRs), N-methyl-d-aspartate receptors (NMDARs), post-synaptic enzymes glycogen synthase kinase–3β (GSK-3β), and p38 mitogen-activated protein kinase (p38MAPK). The post-synaptic tau phosphorylation is shown to act as a "synaptic brake".20

Figure 2. Four assembly states of Tau. Soluble tau monomers can condensate into liquid droplets via reversible liquid-liquid phase separation. Soluble tau oligomers can either be built from liquid droplets or from monomers and can further aggregate into neurofibrillary tangles. These four states of tau have distinctive physicochemical and pathological characteristics.4 36 | Emory Undergraduate Research Journal

This suggests that tau can regulate the synaptic function as well as neuronal plasticity.

Tau is also found in the nucleus and centrosome, so it may also play roles in nucleus-cytoplasm communication and cell cycle regulation. Under oxidative stress conditions, tau can even bind to DNA with its proline-rich domain and R2 repeat in the MTBR. This interaction may contribute to the nucleolar organization and protect genomic DNA.21 Although tau is mainly found in the central neuron system, researchers have identified some of its peripheral functions as well. For example, tau is found in cardiac tissue. The gene knockout of tau can cause deteriorated cardiovascular performance such as increased systolic blood pressure and cardiac hypertrophy.22

1.3 Pathological Tau in AD As a neurodegenerative trademark of tau aggregates, hyperphosphorylation of tau involves 39 phosphorylation sites. There are about 6–8 moles of phosphates per tau molecule, and the total phosphate content is three times more than that in the normal brain.18 However, instead of increasing the number of phosphorylated sites, hyperphosphorylation increases the level of phosphorylation at given residues. Thus, compared to healthy phosphorylation, hyperphosphorylation is an over-active reaction rather than arbitrarily adding phosphate at extra residues.

Prominent pathogenic tau mutations include glycine residue 272 to valine (G272V), asparagine residue 279 to lysine (N279K), proline residue 301 to leucine (P301L), valine residue 337 to methionine (V337M), and arginine residue 406 to tryptophan (R406W). These mutations typically promote the hyperphosphorylation

and self-coacervation of tau into amyloid fibrils.23 In particular, these mutations do not affect the propensity of tau to undergo LLPS; instead, they accelerate the liquid-to-solid phase transition, leading to rapid formation of fibrillar condensates from liquid droplets.24 Kinases is enzymes responsible for attaching phosphate groups to a protein, and phosphatase is enzymes responsible for removeing phosphate groups. Together, kinases and phosphatases can regulate tau’s phosphorylation status, while abnormal kinases and phosphatases lead to pathological hyperphosphorylation. The inhibition of specific kinases, such as such as glycogen synthase kinase-3 (GSK-3) and microtubule affinity regulating kinase 2 (MARK2), reduces the level of tau aggregation and pathological degeneration.25 On the other hand, the downregulation of phosphatases, like protein phosphatase 2 (PP2A), damages the neurons' ability to remove the phosphate group and leads to phosphate accumulation.26 Abnormal hyperphosphorylation of tau is also induced by other overlying processes, including Aß accumulation, impaired brain glucose metabolism, and inflammation. Hyperphosphorylation at Thr231, Ser262, and Ser356 sites can inhibit tau's normal microtubule binding, while that at PRD will promote tau self-aggregation into tangles.

Typical morphologies of pathological tau fibrils are the paired-helical filaments (PHFs), the straight filaments (SFs), and the twisted ribbons (TRs). A hypothetical pathway of tau fibrillization is through spherical nucleation units (SNU), which then aggregate into nascent fibrils. The protofibrils merge and stretch to form the striated ribbons, which further evolve into different types of tau filaments.27 In AD, tau aggregates first appear at the entorhinal cortex (EC) and eventually Volume XVIII, Spring 2022 | 37

BIOCHEMISTRY

Figure 3. Comparison between the healthy brain and Alzheimer’s brain.

Hyperphosphorylated Tau aggregates into neurofibrillary tangles and disintegrates microtubules. As a result, the axonal transport of neurons is impaired.

BIOCHEMISTRY

propagate to the hippocampus, limbic cortex, and other neocortical areas. The trans-synaptic spread hypothesis argues that pathologic tau festers through anatomically connected neuronal networks.28 Based on the physiological activities of tau, abnormal tau aggregation in AD impairs microtubule, synaptic and mitochondrial functions (Figure 3). The altered microtubule dynamics further cause cytoskeletal dysfunction, defective axonal transport, and faulty signaling. Impairment of nucleocytoplasmic transport accumulates RNA, RNA-binding proteins, and nucleoporins such as Nup98 in the soma and promotes them to further interact with mislocalized hyperphosphorylated tau, perhaps by LLPS. Tau condensates also act as templates for pathological stress granules and toxic fibrils formation. By aggregating membrane-less organelles that have critical roles in RNA processing, such as cytoplasmic RNA stress granules and nuclear speckle, pathological tau condensates cause impaired RNA processing.

The presence of pathological tau is correlated with synaptic weakening and long-term depression. Typical AD symptoms such as progressive memory loss, cognitive failure, and learning problems are strongly associated with the synaptic dysfunction and irreparable synaptic loss. Additionally, tau oligomers may lead to mitochondrial damage. That is, when the exogenous tau is internalized by brain cells, it binds to the dynamin-related protein 1 (DRP1) and stimulates mitochondrial fusion and fission imbalance.15

arginine/serine-rich domain (RS). The N-terminal region is highly conserved and interacts with other U1 snRNPs. The RBD domain binds to the stem-loop of the RNA in U1 snRNP assembly (Figure 4). The RS domain is essential for the interaction between U1-70k and regulatory RNA-binding factors, post-translational modification, and abnormal aggregation. In particular, there are two disordered low-complexity domains LC1 (231– 308 aa) and LC2 (317–407 aa) in the RS domain that are necessary for U1-70k aggregation in AD. The LC1 domain has highly repetitive basic (Arg/Lys) and acidic (Asp/Glu) motifs, so it is referred to as a basic-acidic dipeptide (BAD) domain.

2.2 Physiological activities of U1-70k

Spliceosome is a catalytic ribonucleoprotein complex responsible for removing introns on precursor messenger RNA (pre-mRNA) and rejoining exons during RNA splicing. Instead of a preassembled complex, the spliceosome is composed of five small nuclear RNAs (snRNA) – U1, U2, U4, U5, and U6 – and around 100 associated proteins. Among them, U1 small nuclear ribonucleoprotein (U1 snRNP) is an RNA-protein complex that can recognize from −2 to +6 sequence along with the 5'-splice site of the intron and initiate the spliceosome assembly by forming a pre-spliceosome. It is composed of U1 snRNA, seven Sm proteins U1A, U170k, and other U1 specific proteins.31 U1-70k catalyzes the interaction between the U1 snRNP and regulatory RNA-binding factors. A typical family of RNA-binding factors is the serine/arginine 2. U1-70k (SR) proteins, including ASF/SF2 and SC35, which 2.1 Physical Chemistry of U1-70k binds to the exonic splicing enhancer (ESE) near the U1-70k protein is composed of an N-terminus (1-60 pre-mRNA splice site. The arginine/serine-rich LC1 aa), an RNA binding domain (RBD, 92-202 aa), and an in U1-70k can bind to the arginine/serine domain of

Figure 4. Interaction between U170k protein and U1 snRNA. The RBD domain and N-terminus of U1-70k bind to the stem-loop of the RNA in U1 snRNP assembly.12,30

38 | Emory Undergraduate Research Journal

nuclear aggregation of U1 snRNP is detected in 67% of sporadic AD cases and 100% of familial AD cases.6,8 Not all sporadic AD cases demonstrate snRNA aggregates, suggesting the heterogeneity in AD subgroup cases. The redistribution and aggregation of U1-70k is unique to AD pathology. The tangle-like structures of U1-70k are found only in the cytoplasm of AD brain but are absent in other neurodegenerative disorders like corticobasal degeneration (CBD), frontotemporal disorders (FTD), and amyotrophic lateral sclerosis (ALS). Other U1 proteins also accumulate in AD but do not demonstrate the same tangle-like structure as U1-70k does. U1 snRNP alteration occurs early in AD development. Abnormal U1-70k can be detected in the prodromal stage of AD with mild cognitive impairment when cells with U1-70k cytoplasmic aggregates remain intact and functional.6 Based on the role of U1-70k in spliceosome, the major dysfunction caused by abnormal U1-70k is the disruption of RNA regulation – the impaired RNA splicing as well as the premature cleavage and polyadenylation (PCPA). In vitro experiments show that U1 snRNP knockdown causes un-spliced pre-mRNA accumulation and PCPA in pre-mRNAs. The RNA-sequencing analysis of neurons from AD brains shows disease-related transcripts in un-spliced pre-mRNA accumulation. Experiments also show that U1-70k knockdown increases the expression of amyloid precursor protein (APP) by altering protein traffic and facilitating its intracellular cumulation. U1-70k also reduces the amount of Nicastrin, a component of the γ- secretase balancing Aβ production. Therefore, abnormal U1-70k is correlated with the Aβ protein accumulation in AD.37 Similarly, the expression of GSK-3β – an important kinase responsible for tau phosphorylation – is increased in cells with aberrant U1-70k activities, while the overall level of phosphorylation of tau decreases. The impairment of U1 snRNP is associated with tau phosphorylation but not pathological tau hyperphosphorylation, consistent with the fact that U1-70k redistribution and tau hyperphosphorylation do not always coexist.37

3. Possible Interaction Mechanisms 3.1 Tau LLPS

The lysine-rich microtubule-binding repeat domain (MTBR) of tau drives its LLPS; thus, 3R and 4R tau iso2.3 Pathological U1-70k in AD forms with a different number of inserts at MTBR will The level of U1-70k raises significantly in the AD pro- have different de-mixing abilities and form tau compartteome, indicating its pathological deposition. The extra- ments with different structures.10 On the other hand, the Volume XVIII, Spring 2022 | 39

BIOCHEMISTRY

ASF/SF2. This provides additional specificity for the 5' splice-site assignment and enhances the U1 snRNP's ability to recognize the right intron. Such interaction also facilitates the subsequent formation of the E complex, the first intermediate in the assembly process of the mature spliceosome (C complex).32 The phosphorylation of serine residues improves the binding between ASF/SF2 and U1 70k and is necessary for accurate splicing.16 Important enzymes in this process include the Cdc2-like kinase 1 (CLK1) that phosphorylates the Ser226 in the C terminal half. This interrupts the intercellular interaction between the C terminus and the RBD and releases the RBD motif.

Then, the RBD motif can bind to SRSF1, an RNA-binding factor. The SR protein kinase 1 (SRPK1) dissociates CLK1 from U1-70k and recycles the kinase for future use.33 Inhibition of U1-70k leads to embryonic lethality in Drosophila, which suggests that U1-70k is essential to RNA splicing and cell viability. However, in vivo, the interactions between ASF/SF2 and U1-70k are not as important as they are in vitro. Most of the time, the absence of the RS domain does not significantly influence U1-70k viability and function. The role of U1- 70K in facilitating the U1 snRNP and 5' splicing site interaction is not unique nor indispensable; instead, there is a synergistic network of interactions to support such recognition.34 The survival of motor neurons (SMN) proteins can be found in all metazoan cells and play an essential role in spliceosome assembly. U1-70k interacts with SMN complex by its unstructured N-terminal tail in an RNA-independent manner. The SMN complex promotes the Sm protein formation and facilitates snRNP biogenesis in the cytoplasm. In the nucleus, the SMN complex can colocalize to a non-membranous body called nuclear gem, the loss of which can be found in several neurodegenerative diseases. Since it is shown that U1-70k colocalizes with nuclear gems, it may act as an SMN-complex-associating protein, facilitating gems formation and integrity.35 Furthermore, U1 snRNP and U1-70k protect pre-mRNAs from premature cleavage and polyadenylation (PCPA) by binding to their putative poly-A sites.36

BIOCHEMISTRY

proline-rich domain (PRD) regulates LLPS by its phosphorylation state. The aggregation‐prone hexapeptides VQIINK and VQIVYK at the beginning of R2 and R3 regions as well as regulatory KXGS motifs in tau serve as local nucleation sites converting the MTBR from its scattered monomeric state to liquid-like droplets.38 There are two models of tau LLPS based on dominant intermolecular interactions: hydrophobically driven LLPS induced by high salt concentration (LLPS-HS) and electrostatically driven LLPS along with tau-RNA/ heparin complex coacervation (LLPS-ED). The hydrophobic interaction between tau proteins occurs when the hydrophobic polypeptide backbones favor chain‐ chain interactions with one another over solvation with solvents. It is favored by aromatic amino acids, crowding agent polyethyleneglycol (PEG), and Ficoll due to the volume exclusion effect, when the crowding agents occupy the solvent space that are otherwise accessible volume to the protein. A solution of very high ionic strength can also reduced solubility of proteins due to the salting-out Hofmeister effect and facilitate the hydrophobic interaction.3 On the other hand, tau is amphipathic. Its charged residues spread across the polypeptide chain. In the LLPS-ED model, the negatively charged N‐terminal projection domain (1–150 aa) and the slightly positive mid‐region (244–372 aa) are attracted to each other, anchored by the PRD.3 The LLPS-ED and LLPS-HS serve separate but complementary roles in tau LLPS. LLPS-ED is hydrated and reversible, so it does not promote tau aggregation, while the LLPS-HS promotes tau dehydration and fibrillation. This explains the difference between aggregation-prone and aggregation-independent LLPS.39 Previous studies show that tau LLPS is mainly driven by electrostatics, whereas hydrophobic interactions play a surprisingly small role. 40 Tau LLPS can be regulated by various factors, such as pH, ionic strength, protein concentrations, mutations, and post-translational modifications. Strongly phosphorylated full-length tau demonstrates high LLPS propensity in many studies.

tion with nucleic acids. RNAs, especially those involved in RNA modification like small nuclear RNAs (snRNAs) or small nucleolar RNAs (snoRNAs), are found in both cytosolic and nuclear tau aggregates.41 The negatively charged RNA can bind to the positively charged lysine residues of tau, and the lysine and arginine residues in tau can establish hydrogen bonds with RNA. RNA can induce and regulate tau LLPS by functioning as a scaffold platform for the protein-protein interactions. The negatively charged phosphate of RNA can neutralize the positively charged repeat domain of tau, allow tighter packing of tau molecules, and facilitate fibril formation.

Like RNA, other polyanionic molecules, such as DNA and heparin, also induce tau LLPS. Tau can also associate with ligands via LLPS. There are multiple zinc-binding sites on tau. The zinc (II) concentration is highly elevated in AD brains. Adding zinc(II) in the reaction can reduce the critical concentration of tau or crowding agent, facilitating tau LLPS.42 Tau and other proteins can synergistically aggregate by recruiting one protein into the condensates of another protein. Proteins in neurodegenerative diseases such as α-synuclein, Aβ, and TDP-43 can all co-aggregate with tau. Two critical proteins in AD, Aβ and tau, are found cross-seeding with each other. The amyloid core of Aβ plaques can form intermolecular β-sheet interactions with aggregation-prone hexapeptide sequences VQIINK or VQIVYK of tau. This interaction promotes tau aggregation and tangle formation in AD.43 Moreover, tau can colocalize with various RBPs, including hnRNPA0, DDX6, eIF2α, and PABP. In turn, RBPs can regulate the splicing of MAPT, which affects tau splicing and aggregation.5 Since U1-70k is a type of RBPs in the spliceosome, it is expected to show similar interaction with tau as other RBPs do.

The protein-amyloid interactions are classified into four types: titration, sequestration, axial coaggregation, and lateral coaggregation. In lateral coaggregation, two proteins aggregate into amyloids with similar structures; thus, they tend to colocalize in the same portion 3.2 Complex coacervation of Tau: RNA and Proteins of the cell and co-aggregate by sticking to each other by their sides. The interactions between tau and α-Syn Tau can undergo LLPS either via homotypic interac- aggregates, or tau and Aβ aggregates, are classified into tion with itself or via heterotypic association with other lateral coaggregation.44 molecules in the cells. Complex coacervation is an asso3.3 U1-70k LLPS ciative LLPS between oppositely charged species, such as proteins, polymers, and colloids.

A number of RBPs have been found to undergo LLPS Tau can undergo charge‐driven complex coacerva- with their conserved domains: hydrophobic glycine-rich 40 | Emory Undergraduate Research Journal

3.4 U1-70k and Tau association U1-70k forms unique cytoplasmic tangle-like aggregates in AD. In the cytoplasm, the U1-70k aggregates are mis-localized to the filamentous structures of NFTs and spread across the brain with the same propagating pattern of NFTs; in other words, the U1 snRNP aggregates and tau tangles show coincidental localization.

Moreover, the Sarkosyl-insoluble proteins including U1-70k and tau aggregates in AD homogenates can sequester the soluble U1-70k into insoluble condensates.45 The BAD regions in U1-70k and tau aberrantly interact in the cytoplasm to form aggregates under pathological conditions. Such cooperative protein-protein interaction is unique with the pathological tau species from AD but not tau from other tauopathies.5 However, although both of them mostly colocalize in AD brains, the levels of tau and U1-70k seem to be independent of each other. Furthermore, U1-70k and tau show the discordant distribution in the hippocampus.

The U1-70k cytoplasmic aggregates are detected during the early disease development stage when the NFTs remain absent. In the mouse model of tauopathy, U1-70k colocalizes with oligomeric tau, but is repelled from more mature, fibrillar aggregates. The U1-70k and tau aggregates show inconsistency in certain brain areas, disease stages, and animal experiments, so further research is needed to accurately describe their association.

Discussion Overall, tau and U1-70k aggregate and colocalize with each other in AD, which can be explained by their liquid-liquid phase separation and complex coacervation. Although it is shown that the U1-70k can interact with tau through its BAD domain, the type of intermolecular forces and tau respective regions of interest are still speculative.5 Since electrostatically-driven LLPS (LLPS-ED) is the major driving force of tau complex coacervations, it is hypothesized that the charged re-

Further investigations are needed to identify the hot-spot areas of Tau that interact with U1-70k. Based on the typical interaction mechanism between Tau and other proteins, Tau and U1-70k are expected to associate through lateral coaggregation. regions and proline‐rich domain of tau will interact with the BAD domain on U1-70k. On the other hand, since these aggregates are disease-related and the hydrophobically driven LLPS (LLPS-HS) of tau is responsible for protein dehydration and fibrillation, tau may also interact with U1-70k via its hydrophobic regions. Further investigations are needed to identify the hot-spot areas of tau that interact with U1-70k.

Based on the typical interaction mechanism between tau and other proteins, tau and U1-70k are expected to associate through lateral coaggregation.44 Since both can form aggregates independently, this coaggregation occurs between existing protein amyloids. Besides, tau has been shown to go through lateral coaggregation with other protein aggregates.

Volume XVIII, Spring 2022 | 41

BIOCHEMISTRY

domains and RBDs. Secondary structures like the helix-helix interactions of TDP-43 also drive LLPS. Arginine is rich in many RBPs, such as FUS, and drives LLPS through electrostatic interactions. LLPS allows specific RBPs to self-associate and form increasing higher-order structures, such as oligomers, granules, and ultimately aggregates. RNA granules are complexes made of RBPs and mRNA through protein-protein interactions mediated by glycine-rich domains and protein-mRNA coacervation mediated by RBDs. Typical RNA granules include the P body, neuronal transport granule, stress granule, and nuclear speckle. The reversible, regulated RBPs aggregation facilitates mRNA processing. However, many aberrant RBPs aggregations are detected in several neurodegenerative diseases.

As an RBP found in spliceosome, U1-70k undergoes LLPS mainly with its two low-complexity domains (LC1 and LC2) in the C terminus.11 The insoluble U1-70k aggregates directly interact with soluble U1-70k's LC1 domain in a process called homotypic trapping. Different from the prion-like Gln/Asn-rich LC domains in TDP43 and FUS, the LC1 domain of U1-70k is composed of highly repetitive basic-acidic dipeptide (BAD) domain.

The BAD domain can self-assemble through polar zippers formation and form oligomers (mostly dimers and tetramers than trimers). Thus, LC1/BAD influences U170k’s subcellular nuclear localization and colocalizes with other U1-70k in nuclear granules.

BIOCHEMISTRY

Other possible explanations for the tau and U170k association includes the stress granule (SG) theory and the cell cycle reentry theory. Stress granule (SG) is a dense RNA and protein complex formed in response to stressful cellular conditions. Its formation is vulnerable to abnormal protein aggregations in neurodegenerative disease that can serve as a cross-seeding nidus.46 In turn, the overactive SG can stimulate the phosphorylated tau aggregation and redistribution to the soma and dendrite.47 In particular, T-cell intracellular antigen 1 (TIA1) is the core protein in stress granule and it can colocalize with tau. The knockdown of TIA1 reduces tau misfolding and toxicity, while its overexpressing can induce tau misfolding.48 On the other hand, the cell cycle reentry hypothesis regards U1 snRNP redistribution and tau tangle formation as two independent events. U1 snRNPs distributes diffusively and form filamentous structure8, which is not a typical result of apoptosis. Instead, such pathological redistribution resembles the cytoplasmic movement of U1 snRNPs in mitosis. Thus, this theory argues that although most neurons are postmitotic, neurons in the neurodegenerative disease reenter the cell cycle. The upregulated expression of several cell-cycle-related molecules detected by previous studies strongly supports this hypothesis. The neuronal death in AD can therefore be explained by mitotic catastrophe (MC), a type of neuronal death caused by failed completion of mitosis. This hypothesis correlates AD and tumorigenesis as two degeneration pathways all caused by the loss of control in cell cycle, driven by inflammation and strongly associated with age 49.

Pivotal biophysical tools that can be used in studying protein coaggregation include co-immunoprecipitation, colocalization, affinity chromatography, gel filtration, cross-seeding, and co-incubation. Traditional techniques like fluorescence recovery after photo-bleaching (FRAP), electron paramagnetic resonance (EPR), and nuclear magnetic resonance (NMR) spectroscopy can also provide integral information on protein structure and dynamics. Given the complexity of the physiological system, bioinformatic software like CatGranule (calculating LLPS propensity), PONDR-FIT (characterizing intrinsically disordered proteins), and prion-like amino acid composition (PLAAC) are shown to be very effective in analyzing the protein composition and phase separation propensity. However, the in silico, in vitro, and in vivo experiments are sometimes inconsistent with one 42 | Emory Undergraduate Research Journal

another due to the dynamic structure change. Thus, future reviews of tau and U1-70k interaction should include a critical evaluation of the techniques used and limitations involved in the experiments.

Conclusion This paper discusses the physiological and pathological roles of tau and U1-70k in Alzhemer’s Disease. The coaggregation of tau and U170k opens up a new approach to explore the pathological symptons in AD that are not yet understood. While the correlation between liquid-liquid phase separation and protein biophysical changes has just been widely noticed, in the case of tau and U170K, the roles of LLPS and resulting dense coacervates remain rather unexplored. This calls for future research to experimentally or computationally simulate the tau and U1-70k LLPS and to evaluate their roles in the diseased brains.

Acknowledgements I would like to express my gratitude to the Liang Lab at the Department of Biochemistry, Emory University School of Medicine. The completion of this paper could not have been possible without the summer research opportunity they provided and the guidance of my mentor Brianna Hammond and my PI Dr. Bo Liang. Figures, unless otherwise specified, are created with Biorender (BioRender.com) or VMD (http://www.ks.uiuc.edu/Research/vmd/).

References

1.

Bittar A, Bhatt N, Kayed R. Advances and considerations in AD tau-targeted immunotherapy. Neurobiol Dis. Feb 2020;134:104707. doi:10.1016/j.nbd.2019.104707 2.

Chong FP, Ng KY, Koh RY, Chye SM. Tau Proteins and Tauopathies in Alzheimer’s Disease. Cellular and Molecular Neurobiology. 2018-07-01 2018;38(5):965-980. doi:10.1007/s10571-017-0574-1 3.

Rai SK, Savastano A, Singh P, Mukhopadhyay S, Zweckstetter M. Liquid–liquid phase separation of tau: From molecular biophysics to physiology and disease. Protein Science. 2021-05-14 2021;doi:10.1002/pro.4093 4.

Wegmann S. Liquid-Liquid Phase Separation of Tau Protein in Neurobiology and Pathology. Advances in Experimental Medicine and Biology. Springer Singapore; 2019:341357.

5.

Maziuk BF, Apicco DJ, Cruz AL, et al. RNA binding proteins co-localize with small tau inclusions in tauopathy. Acta Neuropathologica Communications. 2018-12-01 2018;6(1)doi:10.1186/s40478-018-0574-5 6.

Bai B, Hales CM, Chen PC, et al. U1 small nuclear

MW. A protein factor essential for microtubule assembly. Proc Natl Acad Sci U S A. May 1975;72(5):1858-62. doi:10.1073/ pnas.72.5.1858 20.

Regan P, Whitcomb DJ, Cho K. Physiological and Pathophysiological Implications of Synaptic Tau. The Neuroscientist. 2017-04-01 2017;23(2):137-151. doi:10.1177/1073858416633439 21.

Qi H, Cantrelle F-X, Benhelli-Mokrani H, et al.

Nuclear Magnetic Resonance Spectroscopy Characterization of Interaction of Tau with DNA and Its Regulation by Phosphorylation. Biochemistry. 2015-02-24 2015;54(7):15251533. doi:10.1021/bi5014613 22.

Betrie AH, Ayton S, Bush AI, Angus JA, Lei P, Wright CE. Evidence of a Cardiovascular Function for Microtubule-Associated Protein Tau. J Alzheimers Dis. 2017;56(2):849-860. doi:10.3233/JAD-161093 23.

Wolfe MS. Tau Mutations in Neurodegenerative Diseases. Journal of Biological Chemistry. 2009-03-01 2009;284(10):6021-6025. doi:10.1074/jbc.r800013200 24.

Iqbal K, Liu F, Gong CX, Grundke-Iqbal I. Tau in Alzheimer disease and related tauopathies. Curr Alzheimer Res. Dec 2010;7(8):656-64. doi:10.2174/156720510793611592 25.

Noble W, Planel E, Zehr C, et al. Inhibition of glycogen synthase kinase-3 by lithium correlates with reduced tauopathy and degeneration in vivo. Proceedings of the National Academy of Sciences. 2005-05-10 2005;102(19):69906995. doi:10.1073/pnas.0500466102 26.

Sontag JM, Sontag E. Protein phosphatase 2A dysfunction in Alzheimer's disease. Front Mol Neurosci.

2014;7:16. doi:10.3389/fnmol.2014.00016 27.

Xu S, Brunden KR, Trojanowski JQ, Lee VM-Y. Characterization of tau fibrillization in vitro. Alzheimer's & Dementia. 2010-03-01 2010;6(2):110-117. doi:10.1016/j. jalz.2009.06.002 28.

Liu L, Drouet V, Wu JW, et al. Trans-Synaptic Spread of Tau Pathology In Vivo. PLoS ONE. 2012-02-01 2012;7(2):e31302. doi:10.1371/journal.pone.0031302 29.

Adapted from "Pathology of Alzheimer's Disease", by BioRender.com (2022). Retrieved from https://app.biorender.com/biorender-templates 30.

Humphrey W, Dalke A, Schulten K. VMD - Visual Molecular Dynamics (Graphics). J Molec. 1996;14:33-38.

31.

Wilkinson ME, Charenton C, Nagai K. RNA Splicing by the Spliceosome. Annual Review of Biochemistry.

2020-06-20 2020;89(1):359-388. doi:10.1146/annurev-biochem-091719-064225 32.

Cho S, Hoang A, Sinha R, et al. Interaction between the RNA binding domains of Ser-Arg splicing factor 1 and U1-70K snRNP protein determines early spliceosome assembly. Proceedings of the National Academy of Sciences. 2011-0517 2011;108(20):8233-8238. doi:10.1073/pnas.1017700108 33.

Aubol BE, Wozniak JM, Fattet L, Gonzalez DJ, Adams JA. CLK1 reorganizes the splicing factor U1-70K for early spliceosomal protein assembly. Proceedings of the National Academy of Sciences. 2021-04-06 2021;118(14):e2018251118.

Volume XVIII, Spring 2022 | 43

BIOCHEMISTRY

alterations in Alzheimer's disease. Proc Natl Acad Sci U S A. Oct 8 2013;110(41):16562-7. doi:10.1073/pnas.1310249110 7.

Hales CM, Dammer EB, Diner I, et al. Aggregates of Small Nuclear Ribonucleic Acids (snRNAs) in Alzheimer's Disease. Brain Pathology. 2014-07-01 2014;24(4):344-351. doi:10.1111/bpa.12133 8.

Hales CM, Seyfried NT, Dammer EB, et al. U1 small nuclear ribonucleoproteins (snRNPs) aggregate in Alzheimer’s disease due to autosomal dominant genetic mutations and trisomy 21. Molecular Neurodegeneration. 2014-01-01 2014;9(1):15. doi:10.1186/1750-1326-9-15 9.

Bishof I, Dammer EB, Duong DM, et al. RNA-binding proteins with basic-acidic dipeptide (BAD) domains self-assemble and aggregate in Alzheimer's disease. J Biol Chem. Jul 13 2018;293(28):11047-11066. doi:10.1074/jbc.

RA118.001747 10.

Ambadipudi S, Biernat J, Riedel D, Mandelkow E, Zweckstetter M. Liquid-liquid phase separation of the microtubule-binding repeats of the Alzheimer-related protein Tau. Nat Commun. Aug 17 2017;8(1):275. doi:10.1038/s41467017-00480-0 11.

Xue S, Gong R, He F, et al. Low-complexity domain of U1-70K modulates phase separation and aggregation through distinctive basic-acidic motifs. Sci Adv. Nov 2019;5(11):eaax5349. doi:10.1126/sciadv.aax5349 12.

Kondo Y, Oubridge C, Van Roon A-MM, Nagai K. Crystal structure of human U1 snRNP, a small nuclear ribonucleoprotein particle, reveals the mechanism of 5' splice site recognition. eLife. 2015-01-02 2015;4doi:10.7554/elife.04986 13.

Boyko S, Surewicz K, Surewicz WK. Regulatory mechanisms of tau protein fibrillation under the conditions of liquid–liquid phase separation. Proceedings of the National Academy of Sciences. 2020-12-15 2020;117(50):31882-31890. doi:10.1073/pnas.2012460117 14.

Wegmann S, Eftekharzadeh B, Tepper K, et al. Tau protein liquid–liquid phase separation can initiate tau aggregation. The EMBO Journal. 2018-04-03 2018;37(7):e98049. doi:10.15252/embj.201798049 15.

Shafiei SS, Guerrero-Munoz MJ, Castillo-Carranza DL. Tau Oligomers: Cytotoxicity, Propagation, and Mitochondrial Damage. Front Aging Neurosci. 2017;9:83. doi:10.3389/fnagi.2017.00083 16.

Cao W, Garcia-Blanco MA. A Serine/Arginine-rich Domain in the Human U1 70k Protein Is Necessary and Sufficient for ASF/SF2 Binding. Journal of Biological Chemistry. 1998-08-01 1998;273(32):20629-20635. doi:10.1074/ jbc.273.32.20629 17.

Ksiezak-Reding H, Liu WK, Yen SH. Phosphate analysis and dephosphorylation of modified tau associated with paired helical filaments. Brain Res. Dec 4 1992;597(2):209-19. doi:10.1016/0006-8993(92)91476-u 18.

Castellani RJ, Perry G. Tau Biology, Tauopathy, Traumatic Brain Injury, and Diagnostic Challenges. J Alzheimers Dis. 2019;67(2):447-467. doi:10.3233/JAD-180721 19.

Weingarten MD, Lockwood AH, Hwo SY, Kirschner

BIOCHEMISTRY

doi:10.1073/pnas.2018251118 34.

Salz HK, Mancebo RSY, Nagengast AA, Speck O, Psotka M, Mount SM. The Drosophila U1-70K Protein Is Required for Viability, but Its Arginine-Rich Domain Is Dispensable. Genetics. 2004-12-01 2004;168(4):2059-2065. doi:10.1534/genetics.104.032532 35.

Stejskalová E, Staněk D. Splicing factor U1-70K interacts with the SMN complex and is required for nuclear Gem integrity. Journal of Cell Science. 2014-01-01 2014;127(18):3909-3915. doi:10.1242/jcs.155838 36.

Kaida D, Berg MG, Younis I, et al. U1 snRNP protects pre-mRNAs from premature cleavage and polyadenylation.

Nature. 2010-12-01 2010;468(7324):664-668. doi:10.1038/ nature09479 37.

Zhu W, Wei X, Wang Y, et al. Effects of U1 Small Nuclear Ribonucleoprotein Inhibition on the Expression of Genes Involved in Alzheimer’s Disease. ACS Omega. 2020-1006 2020;5(39):25306-25311. doi:10.1021/acsomega.0c03568 38.

Ambadipudi S, Reddy JG, Biernat J, Mandelkow E, Zweckstetter M. Residue-specific identification of phase separation hot spots of Alzheimer's-related protein tau. Chemical Science. 2019-01-01 2019;10(26):6503-6507. doi:10.1039/ c9sc00531e 39.

Lin Y, Fichou Y, Longhini AP, et al. Liquid-Liquid Phase Separation of Tau Driven by Hydrophobic Interaction Facilitates Fibrillization of Tau. J Mol Biol. Jan 22 2021;433(2):166731. doi:10.1016/j.jmb.2020.166731 40.

Boyko S, Qi X, Chen T-H, Surewicz K, Surewicz WK. Liquid–liquid phase separation of tau protein: The crucial role of electrostatic interactions. Journal of Biological Chemistry. 2019-07-01 2019;294(29):11054-11059. doi:10.1074/jbc. ac119.009198 41.

Lester E, Ooi FK, Bakkar N, et al. Tau aggregates are RNA-protein assemblies that mislocalize multiple nuclear speckle components. Neuron. 2021-05-01 2021;109(10):16751691.e9. doi:10.1016/j.neuron.2021.03.026 42.

Singh V, Xu L, Boyko S, Surewicz K, Surewicz WK. Zinc promotes liquid–liquid phase separation of tau protein. Journal of Biological Chemistry. 2020-05-01 2020;295(18):5850-5856. doi:10.1074/jbc.ac120.013166 43.

Manczak M, Reddy PH. Abnormal Interaction of Oligomeric Amyloid-β with Phosphorylated Tau: Implications to Synaptic Dysfunction and Neuronal Damage. Journal of Alzheimer's Disease. 2013-06-18 2013;36(2):285-295. doi:10.3233/jad-130275 44.

Bondarev S, Antonets K, Kajava A, Nizhnikov A, Zhouravleva G. Protein Co-Aggregation Related to Amyloids: Methods of Investigation, Diversity, and Classification.

International Journal of Molecular Sciences. 2018-08-04 2018;19(8):2292. doi:10.3390/ijms19082292 45.

Diner I, Hales CM, Bishof I, et al. Aggregation properties of the small nuclear ribonucleoprotein U1-70K in Alzheimer disease. J Biol Chem. Dec 19 2014;289(51):35296313. doi:10.1074/jbc.M114.562959 46.

Wolozin B. Regulated protein aggregation: stress

44 | Emory Undergraduate Research Journal

granules and neurodegeneration. Molecular Neurodegeneration. 2012-01-01 2012;7(1):56. doi:10.1186/1750-1326-7-56 47.

Vanderweyde T, Yu H, Varnum M, et al. Contrasting Pathology of the Stress Granule Proteins TIA-1 and G3BP in Tauopathies. Journal of Neuroscience. 2012-06-13 2012;32(24):8270-8283. doi:10.1523/jneurosci.1592-12.2012 48.

Wolozin B, Ivanov P. Stress granules and neurodegeneration. Nature Reviews Neuroscience. 2019-11-01 2019;20(11):649-666. doi:10.1038/s41583-019-0222-5 49.

Bai B. U1 snRNP Alteration and Neuronal Cell Cycle Reentry in Alzheimer Disease. Front Aging Neurosci.

2018;10:75. doi:10.3389/fnagi.2018.00075

BIOCHEMISTRY Photo by Naomi Akram

Yingrong is a sophomore at Emory Oxford College majoring in Chemistry and Applied Math. This article comes from her summer research on the interaction of U1-70k protein and tau in Alzheimer’s Disease at Dr. Liang’s Laboratory of Structural Cell Biology, Emory School of Medicine. Currently, she is an Oxford research scholar of computational chemistry and studies the effect of oxidative environment on psoriasin protein structure. Other than biochemistry, Yingrong is also passionate about engineering and astronomy.

Volume XVII, Spring 2021 | 45

Special Features Brains in a Dish

by Sayli Sonsurkar

Dr. Steven Sloan (M.D, Ph.D.)., assistant professor in the Department of Genetics at Emory School of Medicine, has long been fascinated by the unknown. To him, science truly revolves around “thinking of things you don’t know and deliberately trying to answer them.” Dedicated to this sentiment, his research revolves around trying to understand how the human brain develops with all of its intensely unique complexity. Brain development has been a fascinating area of study considering “what were just a few thousand cells in a small embryo become so intricate with all the correct instructions and cues.” By focusing on brain development, the Sloan Lab hopes to better understand neurodevelopmental disorders such as autism, schizophrenia, ADHD, learning disabilities, and more.

Dr. Steven Sloan, Assistant Professor, Department of Human Genetics, Emory University School of Medicine

What is most exciting about Dr. Sloan’s research is that the field of research did not really exist as of a few years ago. He explains that “five years ago, it was not possible to answer specific questions about how the human brain develops” due to a lack of precision and accuracy in neural imaging technology. Researchers did not have the necessary techniques to observe early development in a crucial time frame, during which a couple thousand cells lead to trillions of cells. They could get insight from looking at worms, flies, mice, and even monkeys, but the question of brain development could not be answered in a specific human platform. The research of Dr. Sloan and others in this field have transformed both the foundation of knowledge and the mechanisms of getting said knowledge.

Dr. Sloan’s lab has found “ways to use patient derived stem cells to turn back time, go back to the stage of a couple thousand cells, and watch the process happen in a dish.” The Sloan lab takes reprogrammed stem cells, places them in a dish, and uses them to make 3-D tissue structures, called embryoids, in a dish. These structures recapitulate many aspects of the human brain. This field is still at its beginning, and thus the embryoids have limitations as embryonic brain development involves so many complex signaling pathways that allow for cells to become the right kind of cells in the right time, place, and part of the brain. Dr. Sloan explains that you do not have this kind of control in the lab when you have the cells floating in a dish.

This experimentation and limitation bring up the question of how a stem cell knows how to become each specific kind of brain cell and how can that be controlled. To target this issue, the Sloan Lab has started collaborating with faculty at both Emory University and Georgia Institute of Technology to bring in engineering as a method of answering this question. Together, they have developed 3-D bioprinting techniques using hydrogels, which are gelatin like material in which cells can grow and survive. 3-D bioprinting gives you the flexibility to create any desired architecture. They are now printing scaffolds, cube-like structures, and placing the organoids inside.

This technique allows them to control exactly what is happening in the cube. They can now precisely control the spatial environment in which these stem cells grow to better mimic the environment of an embryo.

The process by which the Sloan lab is able to take stem cells and create a brain-like structure is fascinating. The process takes three to four weeks to make “baby neurons,” and by three to four months, you begin to have a hefty population of neurons. By four to six months, you begin to have other types of brain cells such as glial cells. These structures can then be maintained for years. The process is quite analogous to the nine-month gestation of normal human development.

46 | Emory Undergraduate Research Journal

Overview of 3D Bioprinting for Neural Tissue Engineering.

Computer aided design models use patient derived images to mimic the specific geometry of tissues of interest. The printing bioinks may contain a combination of biomaterials, bioactive molecules, or cells to create functionalized and personalized scaffolds. Scaffolds are then printed using the computer aided design and desired bioink(s). An extrusion bioprinter (BioAssemblyBot) is shown in this schematic. Applications for bioprinted scaffolds include high throughput drug screening, in vitro modeling, or in vivo repair and regeneration.1

The advancement of these “brain-in-a-dish” techniques have created a new window to understanding development. They serve as a platform to better understand many aspects of the brain. For example, the Sloan Lab was curious about the process of the maturation of astrocytes, which area type of glial cell in the brain. Dr. Sloan explains that astrocytes are like the “puppet master of neural circuit function.” With this model, they are able to directly watch these cells develop in the dish and then watch as they get older and compare the two. They found that the two actually are not the same at all and undergo huge transformation in terms of their gene expression and in how they function.

There are still many questions being asked about brain development and research still has a long way to go.

Current brain-in-a-dish models still have some limitations, and there are still questions on how to include blood and other kinds of brain cells into the models. Despite these future challenges, the recent developments and innovative techniques from the Sloan Lab are truly fascinating and have greatly transformed the way we approach and understand human conditions. The group also hopes to gain insight on how abnormal development contributes to neurodevelopmental disorders through studying normal brain cellular development and function. By combining state-of-the-art genome engineering, stem cell biology, imaging, and neurobiological approaches they can hopefully continue to discover new mechanisms and therapeutic targets to improve human health.

Melissa Cadena is a Ph.D. student in the joint Georgia Institute of Technology and Emory University Biomedical Engineering Program. She received her B.S.E. in biomedical engineering from the University of Michigan (2018). Her research focuses on using 3D bioprinting and brain organoids to model neurodevelopment.

Vahid Serpooshan is an assistant professor of biomedical engineering and pediatrics at Emory University School of Medicine and Georgia Institute of Technology.

He received his B.Sc. and M.Sc. degrees in materials science from Sharif University and his Ph.D. in tissue engineering from McGill University (2011). Following Ph.D., he worked as a postdoctoral fellow and then as an instructor at Stanford University. His multidisciplinary research team at Emory-Georgia Tech works on a variety of 3D bioprinting-based approaches for in vivo regenerative therapies and in vitro disease modeling and drug screening applications.

word1 1

Cadena, M., Ning, L.., King, A., Hwang, B., Jin, L,, Serpooshan, V*., Sloan, S.A.* 3D Bioprinting of Neural Tissues (2020), Advanced Healthcare Materials. *Co-corresponding authors

Volume XVIII, Spring 2022 | 47

AFFECTIVE SCIENCE

Emotional Pictures: The Psychological Effects of the Color Yellow Introduction

Olivia He, Rhea Mishra, Madi Herndon

Chromatic vision and color perception are some of the most telling aspects of the human experience. People tend to attach emotional meanings to particular hues and show consistent preferences for certain colors (Palmer and Schloss, 2010), suggesting that color has a non-negligible impact on affect, cognition, and behavior (Elliot & Maier, 2007). This has implications for our everyday interactions with our environment , whether that involves the spaces we pass through, the product advertisements we see, our friends’ social media posts, or the change in screen display when our iPhones switch to “night shift” mode. Importantly, many scholars in both psychology and the arts agree that color is contextual, with regard to factors such as physical surface properties and illumination (Olkkonen & Ekroll, 2016) as well as larger-scale aesthetic considerations like product design (Labrecque & Milne, 2013). In the literal sense, colors are illusionary because human perceptual processes do not necessarily provide us with a veridical representation of our surroundings (Locatelli & Wilson, 2017). Color experts Eiseman and Recker have correspondingly stated: What starts as a signal passing along the optic nerve quickly develops into an emotional, social, and spatial phenomenon that carries many layers of vivid meaning. Light with a wavelength of 650 nanometers or so is seen as red. But it is experienced as warmth or danger, romance or revolution, heroism or evil, depending on the cultural and personal matrix in which it appears (Eiseman & Recker, 2011).

Until recently, substantive investigation of color was fairly restricted to philosophy and visual culture studies, or to the technical side of physics and physiology. The

color-in-context theory (Elliot & Maier, 2012) is one of the first analyses of color through psychology, and explicates the umbrella of what is being studied here. Given the intuitive inclination for humans to assign meaning using color associations (Elliot & Maier, 2007), it is implied that a color stimulus deemed as hospitable incites a positive and appetitive effect (excitement, flexibility, etc.), while one that is deemed as hostile incites an aversive effect (anxiety, rigidity, etc.). These emotional effects of color are important to consider because all of the content that is shared on various media platforms has become increasingly obtrusive and influential on society; pictures in the media are largely unavoidable and reach large populations in small amounts of time (Akram & Kumar, 2017). By honing in on a potential cause of emotional impact in current technology, a negative valence can potentially be identified and perhaps partially alleviated in the media.

In this study, yellow tint was added to stimulus photos to identify any variation in emotional response compared to untinted photos. The color yellow was chosen because application of a yellow hue to a photo has been found to induce a negative valence (Valdez and Mehrabian, 1994), but the color itself is still widely associated with both negative and positive feelings. Its implications are ambiguous — it can be hospitable or hostile, as it can be associated with warmth and cheerfulness, or danger and caution (Elliot & Maier, 2012). Brands that use yellow packaging such as McDonalds, Nesquik, and Cheerios may be seen as highly identifiable and memorable.

Yellow is often used in safety products as well, such as caution tape and other advisory road signs. Considering yellow’s psychological association with both appetitive and aversive feelings, we hypothesize that yellow tint would augment emotional properties independent of photo valence.

People tend to attach emotional meanings to particular hues and show consistent preferences for certain colors, suggesting that color has a non-negligible impact on affect, cognition, and behavior.

48 | Emory Undergraduate Research Journal

AFFECTIVE SCIENCE

Method

Photo by Jae Sun Kim

Participants. Forty-six undergraduate students (28 Stimuli. women and 18 men) from universities in the U.S. served Emotional Pictures. Ten photos were chosen at the as voluntary subjects. The age range was between 18 and discretion of the researchers conducting the survey; four 22 years old. of the photos had subject matters typically deemed as positive and four photos with subject matters typically Setting. All subjects were instructed on what environ- deemed as negative were chosen. Two additional photos ment the survey should be taken in. The following in- with neither positive nor negative subject matter served structions were given prior to beginning the survey: as controls. The photos with ‘negative’ subject matters included: a burn victim, a car accident victim, a hospitalized patient, and an elderly man who almost lost his “This survey is part of a research project conductspouse. The photos with ‘positive’ subject matters ined by undergraduate students at Emory University. cluded: a laughing infant, a victorious sports team, a dog The purpose of this survey is to measure the emoand her puppies, and a skydiver with a joyful expression. tional impact of certain photos on various individThe two controls were photos of nature scenery: a treeuals. lined path and a lake surrounded by mountains. Each stimulus photo originated from a different source found Please take this survey alone, in a private room through Google search. with minimal distractions and natural light. Record only your immediate reactions to the photos; try Yellow Tint. All 10 photos were then edited using not to overthink your response or consequently the photo and design software Adobe Photoshop. The scroll back to change answers. “yellow boost” setting was applied to adjust the yellow If a photo does not make you feel any particular emotion, keep in mind that "neutral" or N/A is an option.”

hue by +10. By increasing “yellow boost,” the hue of the non-yellow pixels become more yellow (there is a shift towards yellow on the color wheel), while the existing yellow pixels remain unchanged. Additionally, saturation was increased by +20. Saturation adjustment entails Volume XVIII, Spring 2022 | 49

Figure 1. Stimulus photos used in experimental survey without (top) and with (bottom) yellow tint.

First column: negative; Second column: neutral; Third column: positive.

Figure 2. Statistically significant findings: Car Accident (top) and World Cup (bottom).

First column: without tint; Second column: with tint.

changing the chroma and vibrancy of the colors within the photo, and making them less gray. This change was made to increase the intensity of the existing yellow hues in the photos that were unchanged by the “yellow boost,” which further intensifies the new yellow tints. The yellow hue was increased by +10 and the saturation by +20 so that the adjustments remained subtle enough that participants would not become consciously aware of the disparities in the hues of the photos.

Procedure. The stimuli were compiled into a survey administered on Google Forms. Identity-based questions regarding age and gender preceded the initial instructions. With a total of 20 original photos and edited photos, a randomized mix of 10 of these photos was randomly placed into one survey, while the other 10 were placed into the second survey. Each subject took one of the two surveys, the assignment of which was randomized on the basis of the subject’s selection of multiple-choice options. The edited and unedited, and positive, neutral, and negative photos were arranged randomly and without order as well, to prevent the subject from identifying themes in the nature of the subject matter.

50 | Emory Undergraduate Research Journal

The following instructions were then given prior to initiating the survey: Record only your immediate reactions to the photos; try not to overthink your response or consequently scroll back to change answers.

These instructions were given to preface that the subjects should have recorded their immediate reactions upon seeing each photo, rather than dwelling on possible implications or examining the discrepancies amongst the photos. It was important that they did not force an inorganic or inauthentic reaction. The subject was then given the ten stimulus photos individually with the following prompts for each: In one word, describe what emotion you feel when you look at this photo. [free response] How would you classify this emotion? [multiple choice: positive, neutral, negative] Then, rate how intensely you feel this emotion (1— very slightly, 10 — very intensely) [scale of 1-10]

Data Collection & Analysis

Five-number summaries. The survey responses were initially organized by valence, utilizing the question regarding classification of the emotional response as positive or negative. The valence was then applied to the quantitative intensity ratings by adjusting the corresponding numbers to be positive or negative (a negative response that was assigned an intensity of 5 becomes -5). These response categories were then assessed simultaneously using JMP statistical software to produce five-number summaries in the form of box plots, consisting of the maximum, minimum, upper quartile, lower quartile, and implied median values.

Statistics. A Wilcoxon Rank Sums test on all the ratings was also conducted using the same software; the resulting mean of ranks, p-values, and z-scores were used to evaluate whether the differences between the edited and unedited versions of a given photo were statistically significant or not. Due to the possibility of a Type 1 error, a Bonferroni correction (and a p-value threshold of 0.005) was used for the 10 images.

Results

The results of the two statistical analyses are as follows:

Figure 3. Emotional response intensities of unedited vs. yellow-tinted photos. * p < 0.05 Volume XVIII, Spring 2022 | 51

AFFECTIVE SCIENCE

If a photo does not make you feel any particular emotion, keep in mind that "neutral" or N/A is an option.

AFFECTIVE SCIENCE

Wilcoxon Rank Sums Test p-value

Z-score

Burn Victim

0.6068

0.5146

23.8182

Foliage

0.9401

-0.0751

22.5682

25.2600

Baby

0.5040

-0.6682

Car Accident

33.7400

12.9318

Car Accident

<0.001

-5.2218

Skydiver

26.0909

22.1600

Skydiver

0.3299

0.9743

Lake

25.8182

22.4000

Lake

0.3943

0.8518

World Cup

18.2727

29.0400

World Cup

0.0068

-2.7049

Puppies

27.6000

19.9091

Puppies

0.0525

-1.9387

Hospital

21.9600

26.3182

Hospital

0.2784

1.0840

Thankful Man

27.8200

19.6591

Thankful Man

0.0403

-2.5112

Burn Victim

Mean of Ranks Unedited 23.0200

Mean of Ranks Edited 25.1136

Foliage

24.1600

Baby

Figure 4. Mean of Ranks for unedited vs. yellow-tinted photos table found using the Wilcoxon Rank Sums Test, where a larger difference in magnitude across rows indicates difference in subject intensity

Figure 5. p-value and z-score derived from unedited vs. yellow-tinted photos table found using the Wilcoxon Rank Sums Test.

Photo by Jae Sun Kim

52 | Emory Undergraduate Research Journal

Discussion The hypothesis that the addition of a yellow hue to photographs would elicit a more intensified emotional response than would occur without the hue was suppo-

-rted, although a larger set of stimulus photos as well as more participants would lend itself to more conclusive data.

Yellow was chosen because there is not a rigid emotional idea associated with emotion—yellow can be eerie or joyful. Therefore, the color was chosen because it was thought that the existing emotional implication of the photo, whatever that may be, would be amplified by adding the hue.

There were many sources of variability in this survey that could explain the inconclusiveness of the data.

Firstly, the photos could have been evaluated for their preexisting dominant hues, as adding yellow has a different effect on different colors (For example, adding yellow hue to red intensifies it to become more orange, while adding a yellow hue to blue lessens its intensity and makes it greener, which could prompt opposing emotional reactions to the subconscious associations participants might have with these new colors.) Photos with similar color compositions could have produced more consistent results. Additionally, although the yellow hue was meant to be subtle and aimed to trigger a subconscious reaction to the photos, a stronger yellow hue could have been applied to prompt more significant differences in emotional response that would make distinctions in reaction more apparent.

There were also inevitable errors with the mechanics of the survey. Due to the COVID-19 pandemic, the survey had to be administered online on an electronic device so participants could take it individually in their own spaces. Therefore, there was variability in the environment where the survey was taken. It could not be ensured that each participant completed it thoughtfully, attentively, and without distraction. There could also have been variability in the brightness and hue of the lighting of the room, color and atmosphere of the room, brightness of the subjects’ laptop, amongst other factors. Another limitation of unmonitored, voluntary participation was that the survey had to be intentionally brief due to fear of losing engagement from the subject or repelling potential subjects who would not have had incentive to engage in a lengthy or tedious survey. The data would likely be more conclusive if a larger number of photos were evaluated. A flaw of the survey mechanics that did not affect the data but was convoluted was how the assignment of the emotion being positive, negative, and neutral was separate from the intensity rank ng, when the rankings should have just included “0” for Volume XVIII, Spring 2022 | 53

AFFECTIVE SCIENCE

The present study investigated whether or not students reported an overall more intense positive or negative emotional response to either the edited (“Yellow Boost” applied) or unedited version of each photo. Each of the 46 responses to the randomly assigned photos, rated based on valence and intensity, allowed us to evaluate the differences between edited/unedited photo sets.

In correspondence with the Wilcoxon Rank Sums test conducted, the car accident, thankful man at the hospital, and celebratory World Cup photos had the largest deviations from their means (Figure 4), giving them the largest z-scores (Figure 5). Accordingly, these three photos have significant p-values less than 0.05, indicating that there is indeed a meaningful difference in the intensity of emotional response to these three photos based on their yellow hues.

On average, the unedited “Thankful Man” photo had a positive valence with an emotional intensity rating of 9, while the edited photo had a positive valence with an emotional intensity rating of 8. The unedited “World Cup” photo had an average positive valence with an emotional intensity rating of 7, while the edited photo had a positive valence with an emotional intensity rating of 8.5. The emotional intensity of the unedited “Car Accident” photo had a negative valence and was rated at -7, while that of the edited version of the photo was -6. All three of these photos showed relatively large differences between the edited and unedited photos in terms of emotional intensity rating, relative to the results of the other stimulus photos. Correspondingly, the “Thankful Man” photo had a p-value of 0.403, the “World Cup” photos had a p-value of 0.0068, and the “Car Accident” photo had a p-value of <0.001. Regarding the Bonferroni Correction, only the “Car Accident” photo had a significant p-value less than 0.005.

There were no significant differences observed amongst the remaining seven photos. One exception is that the magnitude of emotional intensity in response to the edited photo of the skydiver was most frequently 9 (out of 10), while those to the unedited photo were most frequently 6/10. In both cases, the valence of the majority of responses to both photos were positive.

AFFECTIVE SCIENCE

neutral and a negative side.

Additionally, the participants also could have irregular vision, such as a color deficiency, which they were not asked to disclose. Finally, the survey was only given to college students, so all the participants were in the same age range and therefore had similar socialization regarding color association. This survey does not address disparities between generations caused shifts in color psychology that may have occurred (i.e., changes in media and perception of general aesthetics).

Future experiments, taking these factors into consideration, could be useful for advertising and the overall marketing field, as the deliberate usage of color could be powerful in subconsciously manipulating the response an audience may have to a stimulus. Psychiatrist Carl Jung called color the “mother tongue of the subconscious;” intentional utilization can influence the decision of a consumer. In marketing today, yellow is often used to provoke feelings of anxiety and therefore impulsiveness, which is why it is regularly used for clearance signs in retail stores. On the other hand, it is used to incite creativity and happiness. A simple example of this might include the color-scheme of the Snapchat app, a social media platform used to casually engage with friends. Taken together, the inconclusiveness of the data may suggest that yellow is best suited as a “helping” color, one that can be used to amplify the other hues and tones present that have a more particular emotional association.

Works Cited

Akram, W., & Kumar, R. (2017). A study on positive and negative effects of social media on society. International Journal of Computer Sciences and Engineering, 5(10), 351-354.

Eiseman, L., & Recker, K. (2011). Pantone: The 20th Cen tury in Color. San Francisco, California: Chronicle Books LLC. Elliot, A. J. & Maier, M. A. (2012). Color-in-context the ory. Advances in Experimental Social Psychology, 61– 125. https://doi.org/10.1016/b978-0-12-394286-9.0000 2-0 Elliot, A. J., Maier, M. A., Moller, A. C., Friedman, R., & Meinhardt, J. (2007). Color and psychological function ing: The effect of red on performance attainment. Jour nal of Experimental Psychology: General, 136(1), 154–168. https://doi.org/10.1037/0096-3445.136.1.154 Labrecque, L.I., Patrick, V.M. and Milne, G.R. (2013), The Marketers’ Prismatic Palette: A Review of Color Resear-

54 | Emory Undergraduate Research Journal

ch and Future Directions. Psycholology and Marketing, 30, 187-202. https://doi.org/10.1002/mar.20597 Locatelli, R., Wilson, K.A. (2017). Introduction: Perception Without Representation. Topoi 36, 197–212. https://doi. org/10.1007/s11245-017-9460-1 Olkkonen, M., & Ekroll, V. (2016). Color constancy and contextual effects on color appearance. In Human color vision (pp. 159-188). Springer, Cham.

Palmer, S. E. & Schloss, K. B. (2010). An ecological valence theory of human color preference. Proceedings of the National Academy of Sciences, 107(19), 8877–8882. https://doi.org/10.1073/pnas.0906172107 Valdez, P. & Mehrabian, A. (1994). Effects of color on emo tions. Journal of Experimental Psychology: Gen eral, 123(4), 394–409. https://doi.org/10.1037/00963445.123.4.394

Rhea Mishra is a sophomore tentatively majoring in Marketing & Art History. Her research was conducted for her Psychobiology & Cognition class with Professor Andrew Neff. She was eager to explore the cognitive side of the aesthetic experience and hopes to apply her newfound understanding of color theory to her majors. In her free time, she enjoys spending time with friends and listening to music.

Madi Herndon is a sophomore from Raleigh, North Carolina. She is currently majoring in Human Health with a minor in Neuroethics on the pre-medicine track at Oxford College of Emory University.

While her main focus is academics, she also enjoys cheerleading and dancing in her free time.

Volume XVIII, Spring 2022 | 55

AFFECTIVE SCIENCE

Olivia is a third-year from NYC studying Neuroscience & Behavioral Biology with a minor in Dance & Movement Studies. She loves learning about all of the intricate ways through which we interact with our world, and using that knowledge as a lens to make daily life seem just a little bit more interesting. Her drive to understand mechanisms underlying the human experience give rise to her interest in drawing connections between neuroscience and various other disciplines of study. When she's not studying at Emory, she loves choreographing and taking tons of dance classes with her friends in NYC.

Applying Neuropsychological Aspects of Psychopathy to a Dictator Ernest Murphy

POLITICAL PSYCHOLOGY

Abstract

The crimes against humanity that Saddam Hussein was responsible for cemented him as one of the most infamous leaders in political history. This article aims to explain the possible correlates of his cruel behavior through the interdisciplinary application of neuroscience and psychology. Discussion of biological and environmental factors that contribute to violence are applied to Hussein's known biographical information. The paper shows that Hussein would likely be considered a psychopath based on the Psychopathy Checklist Revised (PCL-R) criteria (Hare et al., 1990). Future research on ‘psychological profiles’ of world leaders may be important for the decision making of political advisors and diplomats, as well as for education of the public.

Introduction

Neuroscience, the study of the nervous system, has revolutionized our understanding of the causes of certain human behaviors. The thought patterns of murderous psychopaths, for example, can be explained by abnormal brain anatomy, which can be shaped by both genetics and upbringing (Raine, 2013). Neuroscience, specifically the discussion of brain functions, genes, and environmental factors, may be applied to a group of people who share many common characteristics with psychopaths: dictators.

During the July Revolution of 1968 the Ba’ath Socialist Party overthrew the Iraqi government, which led to the beginning of Saddam Hussein’s long political reign (Karsh & Rautsi, 2002). Thanks to advancements in neuroscience, the behavior of Hussein, who is widely regarded as a ruthless, power-hungry leader, may be explained through analysis of biological factors, such as atypical development of brain regions, and gene-environment interplay.

Biography To understand the mind of Hussein, it is essential to first look at his biography, starting with his birth. He was born in a village in Northern Iraq in 1937 to a poor became an important member of the party, aiding in coups and eventually helping the Ba’ath Party rise to power in Iraq.

Hussein found endless ways to abuse his power after becoming the president of Iraq, including starting wars and using chemical weapons on innocent people (Al-Ani, 2010; Bureau of Public Affairs, 2003). Soon after he rose to power, he ordered the execution of 21 senior officials of the party because he questioned their loyalty. He was known to arrest or execute dissidents and people who simply criticized him (Post, 1991). Furthermore, Hussein ordered the invasion of neighboring Iran in September of 1980, the second year of his presidency (Al-Ani, 2010). The Iran-Iraq War was a conflict that lasted eight years, resulting in countless casualties on both sides. Immediately following the war with Iran, Hussein ordered the invasion of Kuwait in 1990 for reasons that are not fully understood (Al-Ani, 2010). It may have been a display to assert dominance in the Middle East or to capture oil reserves.

Hussein was also well known for committing merciless acts of genocide. He was a proponent of the ethnic cleansing of people who were not of native Iraqi origin. He ordered an infamous chemical weapons attack on Halabja, a city in Northern Iraq, where he targeted non-native Kurdish people with mustard gas and other chemical agents. It is estimated that roughly 5,000 people died from the attack, with another 10,000 being injured or permanently disfigured (Bureau of Public Affairs, 2003). Hussein used chemical warfare on other

1990 invasion of Kuwait, Creative Commons

56 | Emory Undergraduate Research Journal

POLITICAL PSYCHOLOGY

Figure 1. Brain regions included in the empathy circuit (Decety et al., 2020).

Kurdish and Iranian populations during his reign. Analysis of Hussein’s reign raises questions, such as: how could a country’s leader purposefully commit so much evil? Could genetics or environmental influences play a role in Hussein’s evil behaviors?

Biological Factors of Violence Someone with the inclination to perform such horrible acts as Hussein likely has dysfunction in particular areas of the brain. The functioning of brain regions belonging to the empathy circuit can explain differing empathy levels among individuals. The empathy circuit comprises over ten regions: the medial prefrontal cortex, orbito-frontal cortex, frontal operculum, inferior frontal gyrus, caudal anterior cingulate cortex, anterior insula, temporoparietal junction, superior temporal sulcus, somatosensory cortex, inferior parietal lobule, inferior parietal sulcus, and the amygdala (Baron-Cohen, 2011).

The medial prefrontal cortex, specifically the anterior rostral region, is used for social cognition, allowing one to compare their perspectives and knowledge with that of somebody else (Amodio & Frith, 2006). Hence, someone with subnormal development of this area of the brain will have reduced empathetic capability, as they may have trouble understanding others. The inferior frontal gyrus allows for the emotional recognition and regulation of fear and stress (Baron-Cohen, 2011).

It is likely that Hussein has decreased functionality in his brain’s empathy circuit. This could be in the form of low level of activation in the brain region or decreased volume density. In Hussein, low empathy is easily observable. If he had normal empathy levels, certainly he would not have considered the unprovoked invasions of

countries, acts of genocide, and merciless brutalizing of dissidents.

Differences in brain structure and function are not the only biological factors that are associated with more violence. The monoamine oxidase A (MAOA) gene, also known as the warrior gene, breaks down several neurotransmitters, including dopamine, serotonin, and norepinephrine. An abnormal variant of the gene causes these neurotransmitters to accumulate, which has been correlated to aggressive behavior (Garcia-Arocena, 2015). Other factors such as birth complications, maternal smoking during pregnancy, and high levels of testosterone may also increase the risk of committing violence due to their effects on human physiology (Batrinos, 2012; Rantakallio et al., 1992). Additionally, the biological marker of a low resting heart rate has been linked with proactive (i.e., cold and calculative) aggression in psychopaths. Low cardiac arousal may be associated with stimulation-seeking behaviors, including risk-taking behaviors such as violence (Raine, 2013).

Environmental Effects on Development In addition to genetics, environmental factors are thought to play an important role in the development of violent tendencies and behaviors. As discussed previously, Hussein grew up in a small village and lived in poverty. With poverty often comes malnutrition, and thus subnormal physical development, which could inhibit the ability of the brain to grow properly during childhood, including regions of the empathy circuit.

Malnourished children may also be more likely to engage in aggressive behaviors (Liu et al., 2004). Specifically, Omega 3 deficiency is correlated with violence Volume XVIII, Spring 2022 | 57

POLITICAL PSYCHOLOGY

Table 1. Applying the Psychopathy Checklist, Revised (PCL-R) to Saddam Hussein. Based on the revised 1990 version (Hare et al., 1990).

Trait

Score

Rationale

Glibness and Superficial Charm

2

Was very popular among the Iraqi people in the early years of his reign.

Grandiose Sense of Self Worth

2

Claimed to have descended from the Prophet Muhammad. Lived a lavish lifestyle.

Pathological Lying

2

Plead not guilty after being charged with crimes against humanity.

Manipulative Behavior

2

Manipulated the media and Iraqi people with charisma and denying of crime.

Lack of Remorse or Guilt

2

Mercilessly committed acts of genocide and war.

Shallow Affect

1

Had low emotional empathy, but appeared charming and responsive.

Lack of Empathy

2

Lack of regard for others, especially for those he saw as inferior.

Failure to Accept Responsibility

2

Plead not guilty in trial.

Need for Stimulation

2

Owned multiple palaces and slept with many women.

Parasitic Lifestyle

2

Treated other people as tools for personal gain.

Poor Behavioral Controls

2

Punished those who criticized him, often with torture or death.

Lack of Goals

0

His high ambition led him to him becoming Iraq’s president.

Impulsivity

2

Exhibited unpredictable behavior.

Irresponsibility

2

Abused his power as the ruler of Iraq.

Juvenile Delinquency

0

No evidence of arrests as a juvenile.

Revocation of Conditional Release

N/A

Captured and arrested once, led to his execution.

Sexual Promiscuity

2

Reportedly slept with multiple women every night during his reign.

Many Short-Term Relationships

2

Had multiple marital relationships and mistresses.

Criminal Versatility

2

Charged with crimes against humanity. Responsible for acts of war and genocide.

Early Behavior Problems

2

Reportedly tortured small animals as a child.

58 | Emory Undergraduate Research Journal

Was Hussein a Psychopath?

Although not all psychopaths are killers, there are a series of traits that are homologous among them. These traits make up the PCL-R, or the Psychopathy Checklist, Revised. It consists of 20 items which are scored based on the presence of the characteristic, allowing for accurate identification of psychopaths (Hare et al., 1990).

Subjects who score at least 30 are normally considered to be psychopaths.

Many would consider Hussein a psychopath solely because of his crimes against humanity, but only if he shares a large number of PCL-R traits is it reasonable to make this claim. This paper demonstrates a possible result of administration of the PCL-R test to Saddam Hussein, with a score of 33 (see Table 1). This is above the cutoff score required to be clinically referred to as a psychopath.

The possibility that Hussein may have been a psychopath is significant; it would suggest that he likely had abnormal empathy circuit functionality, which could largely be explained by genetic and environmental factors.

Conclusion A man such as Hussein makes for an interesting case study. He had impressive cold cognition abilities of decision making and planning, allowing him to move up in rank within the Ba’ath Party, and eventually became Iraq’s president. At the same time, his impulsivity and hotheadedness caused him to make rash decisions, often at the expense of the lives of others. Whether or not Hussein was a psychopath, it is worth considering the effects of the environment on brain development and emotional responses, as well as the possibility of genetic predisposition to violence in certain people. Education on how social relationships, diet, and parenting practices can affect the brain is crucial in ensuring healthy development of populations. From an international relations perspective, having knowledge of other world leaders’ psychological profiles is imperative in building relationships, or at the very least, preventing conflicts between countries.

References

Ainsworth, M. D. S., Bell, S. M., Stayton, D. J. (1971). Indi vidual differences in strange-situation behavior of one-year-olds. In H.R. Schaffer (Ed.), The origins of human social relations (pp.17-57). London: Academic Press.

Al-Ani, A. H. (2010). The trial of Saddam Hussein. SCB Distributors.

Amodio, D. M., Frith, C. D. (2006). Meeting of minds: The medial frontal cortex and social cognition. Nature Reviews Neuroscience, 7, 268-277. https://doi. org/10.1038/nrn1884 The Attachment Project. (2021). Disorganized attachment: Causes & symptoms. https://www.attachmentproject. com/blog/disorganized-attachment/ Baron-Cohen, S. (2011). The science of evil: On empathy and the origins of cruelty. Basic Books.

Batrinos, M. L. (2012). Testosterone and aggressive be havior in man. International Journal of Endocrinology and Metabolism, 10(3), 563-568. https://doi. org/10.5812/ijem.3661 Bureau of Public Affairs. (2003). Saddam’s chemical weap ons campaign: Halabja, March 16, 1988. https://2001-2009.state.gov/r/pa/ei/rls/18714.htm Decety J. (2020). Empathy in Medicine: What It Is, and How Much We Really Need It. The American journal of medicine, 133(5), 561–566. https://doi. org/10.1016/j.amjmed.2019.12.012

Volume XVIII, Spring 2022 | 59

POLITICAL PSYCHOLOGY

prevalence as evidenced by higher homicide rates in countries with less seafood consumption (Hibbeln, 2001). Hussein also lacked a normal family structure: his mother rejected him and his father and brother died before Hussein’s birth. Maternal rejection, along with poverty and poor education, are considered social risk factors for future violent behavior (Raine, 2013). It has also been shown that unsatisfactory interpersonal experiences with a rejecting mother affect the development and behaviors of children (Ainsworth et al., 1971). Hussein may have developed what is called a disorganized attachment style as a result of the lack of attention he received as a child. Children with this attachment style often grow up to be distrusting of others and live in fear of rejection and abandonment (The Attachment Project, 2021).

It is likely that being in Iraq during a time of political instability and rampant violence is detrimental to development. Regular exposure to violence can cause desensitization, or a diminished emotional response, to events that normally cause stimulation. Research demonstrates that exposure to violence at a young age is correlated with a greater incidence of violence in late adolescence (Mrug et al., 2016). Hence, violence exposure paired with Hussein’s family structure possibly affected his psychological development, resulting in diminished empathy and a reduced fear response.

POLITICAL PSYCHOLOGY

Garcia-Arocena, D. (2015). The genetics of violent behavior.

21(6), 1106-1113. https://doi.org/10.1093/ The Jackson Laboratory. https://www.jax.org/newsije/21.6.1106 and-insights/jax-blog/2015/december/the-genet Shamay-Tsoory, S. G., Aharon-Peretz, J., Perry, D. (2009). ics-of-violent-behavior# Two systems for empathy: A double dissociation be Hare, R. D., Harpur, T. J., Hakstian, A. R., Forth, A. E., tween emotional and cognitive empathy in inferior Hart, S. D., Newman, J. P. (1990). The revised Psy frontal gyrus versus ventromedial prefrontal lesions. chopathy Checklist: Reliability and factor struc Brain, 132(3), 617-627. https://doi.org/10.1093/ ture. Psychological Assessment: A Journal of Con brain/awn279 sulting and Clinical Psychology, 2(3), 338–341. https://doi.org/10.1037/1040-3590.2.3.338 Hibbeln, J. R. (2001). Seafood consumption and homi cide mortality. A cross-national ecological analysis.

World Review of Nutrition and Dietetics, 88, 41-46. https://doi.org/10.1159/000059747 Karsh, E., Rautsi, I. (2002). Saddam Hussein: A political biography. Grove Press.

Liu, J., Raine, A., Venables, P. H., Mednick, S. A. (2004).

Malnutrition at age 3 years and externalizing be havior problems at ages 8, 11 and 17 years. The American Journal of Psychiatry, 161(11), 2005-2013. https://doi.org/10.1176/appi.ajp.161.11.2005 Mrug, S., Madan, A., Windle, M. (2016). Emotional desen sitization to violence contributes to adolescents’ violent behavior. Journal of Abnormal Child Psy chology, 44(1), 75-86. https://doi.org/10.1007/ s10802-015-9986-x Post, J. M. (1991). Saddam Hussein of Iraq: A political psychology profile. Political Psychology, 12(2), 279289. https://doi.org/10.2307/3791465 Raine, A. (2013). The anatomy of violence: The biological roots of crime. Vintage Books.

Rantakallio, P., Läärä, E., Isohanni, M., Moilanen, I. (1992). Maternal smoking during pregnancy and delinquency of the offspring: An association without causation? International Journal of Epidemiology,

Ernest Murphy is a junior at the University of Lynchburg majoring in Biology. He is an EMT and volunteer Crisis Counselor, and plans to pursue PA school for his master’s after graduating from Lynchburg. His article in this issue is based on research he conducted while in Dr. Ei Hlaing’s honors psychology course.

His non-academic interests include playing music, making art, and going to the gym.

60 | Emory Undergraduate Research Journal

Addressing Food Insecurity in Appalachia: An Anthropological Public Health Assessment and Recommendations Katherine Morgan

With a dramatic increase in noncommunicable, food-related diseases throughout the United States, rural areas have demonstrated significant rates of obesity. Confronted with one of the highest obesity rates among all regions in the United States, there exists an urgent need to investigate the variables and themes contributing to diet-related disease and food scarcity in Appalachia. Dialogue about Appalachian lifestyle and culture has inundated popular culture with the publication of "Hillbilly Elegy" and subsequent movie release. However, highly personalized qualitative accounts leave much to be inferred for the rest of the world, often painting only a partial picture of what life is like in modern Appalachia.

This paper considers the current institutions, cultural factors, and historical themes that contribute to adverse diet-related health outcomes through the cross-comparison of quantitative public health studies, qualitative anthropological research, and by analyzing primary accounts of Appalachian life. The paper concludes with a short research proposal designed to fill existing gaps in data on food insecurity in the region.

Overview The U.S. Department of Agriculture reports that 10.5% of all U.S. households experience food insecurity, defined through the following definition: "At times during the year, these households were uncertain of having, or unable to acquire, enough food to meet the needs of all their mem bers because they had insufficient money or

other resources for food." (USDA Measurement) This research explores how income and region impact access to nutritious food. As an individual with Appalachian roots, I am interested in determining how food insecurity affects poor, rural populations throughout the expansive territory.

The geographic area in discussion, according to the Appalachian Regional Commission, "is made up of 420 counties across 13 states and spans 205,000 square miles, from southern New York to northern Mississippi."

(About the Appalachian Region, 2021) Food insecurity is a multifaceted variable that is measured according to the USDA by assessing the likelihood that a household lacks consistent funds to keep healthy and nutritious food accessible the majority of the time. Appalachia is a region long known for endemic poverty, food deserts, and high rates of food-related disease. Research on food insecurity is essential to understand and address the scope of the issue for residents in the region. By analyzing Appalachian culture, history, and lifeways, themes may be identified to aid public health professionals in understanding the scope of food insecurity and how to improve health status through community-based interventions.

The Problem: Poverty, Food Choice, and Accessibility There is much discussion concerning food accessibility, health literacy, and eating practices in Appalachia.

Public health data suggests that there is limited access

Southern Appalachia, Creative Commons

Volume XVIII, Spring 2022 | 61

ANTHROPOLOGY

Introduction

ANTHROPOLOGY

Although it is easy to view obesity statistics as indicators of food insecurity, farmer's markets, individual gardens, and other means allow for the incorporation of produce and other goods to be available in local food systems in Applachia. to food retailers in general and that food insecurity still manifests in areas where food retailers are abundant.

This need calls researchers to address the differences between food choice and food scarcity. Qualitative foodways research in Appalachia suggests that people living in Appalachia are generally knowledgeable about food and health literacy. Individuals may still favor calorie-dense foods over more nutritious alternatives in specific contexts, even with healthy alternatives available. It is crucial to address the role that health literacy plays in contributing to obesity. One qualitative research study found that community stigma may decrease an individual's willingness to consume healthy foods. (Sharaievska, 2018) However, there is still a need for educational interventions to improve overall access, willingness, and ability to purchase and consume healthy foods.

As previously discussed, poor economic conditions throughout Appalachia contribute to food insecurity.

Not only does food insecurity concern a deficit of nutritionally adequate foods, but it also extends to limited or uncertain ability to acquire nutritious foods, meaning that even if healthy options are available, they may also be inaccessible to consumers with decreased economic means. (Andersen, 2020) With the loss of traditional mining and manufacturing jobs, there is little money to spend; thus, businesses leave communities, meaning that grocery stores, in particular, may be more spread out, less accessible, and more likely to sell pre-packaged foods with long shelf lives. Alternatively, non-local fresh produce is more expensive concerning high transportation costs incurred from shipping to remote areas. Appalachian children and adults experience food insecurity at a higher rate than the national average, as "rural and economically challenged areas seem to be at higher risk." (Chubinski, 2012) Endemic poverty and decreased purchasing power indicate that healthy foods are less desirable due to higher fresh produce prices in rural areas. As Elizabeth Catt discusses in "What You Are Getting Wrong About Appalachia," the poor economic conditions throughout the region can be attribut62 | Emory Undergraduate Research Journal

-ed to limited jobs, which act as a spiral effect in increasing rates of food insecurity and disease.

A Solution: Historical Context of Local Food Production Despite poor modern economic conditions, it would be a great oversight to not acknowledge a rich history of self-sufficiency and gardening throughout the regionand the role of long-established local food production in feeding Appalachians. Although it is easy to view obesity statistics as indicators of food insecurity, farmers' markets, individual gardens, and other means allow for the incorporation of produce and other goods to be available in local food systems in Appalachia. Historically, and to a lesser extent in modern times, many Appalachians subsisted mainly from family farms and gardens. Local foodstuffs, such as pawpaws, squirrel, pike, morel mushrooms, black walnuts, corn, beans, and wild greens, were common and used in many ways. Another source found that the four largest categories in the historical Traditional Southern Appalachian Diet pyramid were "homegrown produce (34%), added fats and sugars (18%), sources of protein (17%), and sources of carbohydrates (16%)." (Visocky, 2016) The tradition of growing produce hasn't disappeared in the region. For example, the USDA's StrikeForce for Rural Growth and Opportunity Initiative bolsters local efforts to decrease food insecurity through gardening.

The Grow Appalachia program supports 91 community gardens throughout the region. Endemic poverty often prevents fresh produce from being accessible to families.

Local gardening projects have the unique opportunity to grow sustainable produce and foster healthy eating practices in rural areas. (USDA's strikeforce Initiative).

Conceptualizing a Solution: Foodways and Culture To understand the full scope of food insecurity in Appalachia, we must acknowledge the ethnic diversity and ranging socioeconomic conditions in the region. Anthropologists rely on ethnographic data to understand

Intervention Recommendations: Making healthy food accessible, affordable, and attractive As previously discussed, healthy food is hard to access, costly, and not necessarily a primary choice. A solution may be found from the purview of the social determi nants of health, especially concerning environmental

and socioeconomic status. Affordability is comparatively one of the greatest barriers to achieving food security as food cost disparities adversely affect rural counties with high poverty rates. Research confined to four counties in Kentucky found low poverty urban counties had lower food costs than high poverty rural counties. Importantly, a significant cost disparity was found among foods with a higher nutritional value between impoverished rural counties and urban counties as the per-serving cost of highly nutritious foods was significantly higher in the aforementioned rural groceries.

(Hardin-Fanning 2015) Accessibility is also a concern. Beyond the fact that healthy food is expensive, it oftentimes may not be available at all. People living below the poverty line often rely on charitable organizations. Research has found that community-level interventions were less impactful, namely that food banks, although common in high-income countries, fall short in efficacy due to reliance on donations and volunteer support. Community shops or kitchens may also be limited in effectiveness. Thus community and household-based interventions fall short in addressing the dire need to prevent food insecurity in high-income countries. (Loopstra, 2018) Making healthy food accessible and affordable in Appalachian populations will likely require government-mandated interventions beyond pre-existing programs such as the food stamp system in the United States, which enforces people's dependency on cheap, low nutrient-dense foods. Unhealthy foods disrupted local food systems where for generations, Appalachians independently grew their produce. Implementing programs to encourage local gardening and food production to supplement additional government interventions could significantly improve access to healthy food in rural areas. One example of an effective program that could be reproduced throughout the region is the Grow Your Health Program, implemented in an Appalachian Ohio food desert region. The Putting Healthy Food on the Table program enabled residents to grow fruits and vegetables through community and container gardens.

(Rural Information Hub, 2020) Aggregating findings from multiple published research sources, Loopstraet al. indicated that Governmental efforts and policy interventions are impactful in reducing food insecurity in high-income countries.

Government interventions that support community and household gardens may be the key to addressing the Volume XVIII, Spring 2022 | 63

ANTHROPOLOGY

culture through scientific documentation of customs and habits. The problem of food insecurity will not be solved without delving into the belief systems and varying cultural contexts; thus, Appalachians must be thoroughly understood to serve their needs effectively. Although this analysis does not provide a solution to addressing this gap, it does acknowledge the shortcomings of previous assertions and identifies important considerations for future research agendas.

Understanding foodways and food-related health outcomes in Appalachia requires careful consideration of quantitative research conducted on health metrics and qualitative research to understand how culture and lifestyle impact people's food consumption choices and the richly diverse cultural legacy of the Appalachian region at large.

"Hillbilly Elegy" asserts the poor, white descendants of Scotch-Irish immigrants make up the Appalachian ethnic group. However, there is wide socioeconomic and ethnic diversity throughout the region. Such a vast geographic region demands that researchers look at the microcosm of community culture and individual choice within the macrocosm of the geopolitical and cultural influences on diet and food accessibility. The rich food cultures of the region incorporate cuisine of Native American, African, and Indo-European origin. From twelve thousand years ago, Native Americans were cultivating food in the region, with traditional foodstuffs limited to locally grown and sourced foods. (Dragovich, 2020) Qualitative accounts such as "What You Are Getting Wrong About Appalachia" are in direct contrast to the assertion of a single, homogenous Appalachian culture as indicated in "Hillbilly Elegy."

By accumulating a socioeconomic, gender, and racially diverse focus group of Eastern Kentucky residents, a research study investigating obesity and perspectives on healthy eating noted the importance of utilizing a community purview to understand how Appalachians relate to and consume food. (Shoenberg, 2013) Multiple voices should be listened to in addressing the obesity problem rampant in the region.

ANTHROPOLOGY

Photo by Leo Huang

affordability and accessibility of fresh produce. In order to do this, evidence-based and community-engaged research must be implemented so that government-mandated public health interventions may be targeted to improve the food supply chain to rural areas and improve the health literacy and behaviors of individuals in these communities.

Reducing dependency on cheap, low nutrient foods can also be attained through healthy eating educational initiatives. Increasing the attractiveness and willingness to consume healthful food options will likely depend on public health interventions to change existing eating behaviors, such as the USDA Supplemental Nutrition Assistance Program has a SNAP-Ed initiative that teaches low-income families eligible for SNAP benefits to shop for and prepare healthy meals. By supporting evidence-based nutrition programming, SNAP-Ed partners with community organizations to improve people's health and wellbeing. Although programming exists to address Appalachians' needs, these programs need to improve their efficacy as research indicates that meal preparation and facilitating making healthy eat64 | Emory Undergraduate Research Journal

-ing choices are key areas interventions should address.

(Supplemental Nutrition Assistance Program (SNAP)) Ethnographic research provides an invaluable perspective on the foodways of Appalachians. As Sharaievska et al. found, many participants in ethnographic interviews about diet indicated an understanding of what it means to eat healthily. A different study found through a series of interviews that "At the individual level, participants described the need to increase knowledge to develop healthier eating habits, emphasizing nutrition and cooking classes." Schoenberg et al. Both research articles suggest that an important factor in improving the quality of food consumption is teaching Appalachians healthy cooking and meal preparation strategies through community-based interventions.

Encouraging local food production and the integration of locally grown foods into grocery stores has the potential to greatly reduce food security throughout the region. Grocery stores are key community assets, particularly in rural, spread-out areas where food access is disrupted by geography, endemic poverty, and high food costs, among other factors. Supporting regionally

Qualitative Data: Twentieth Century Diet and Food Cultivation More data must be collected through focus groups, pilot programs, and historical analysis if a solution to food insecurity were to be found by encouraging household models of small-scale food production. As previously discussed, subsistence farming long dominated Appalachian foodways. Aptly stated in The New Encyclopedia of Southern Culture, "Farming activities there not only supported the traditional economy but also formed the backdrop for much cultural distinctiveness." However, following World War II, there was a great decline in subsistence agriculture, jointly due to The Great Migration and industrialization of farming. (Edge, 2007) With the rising popularity of processed foods in the 1950s due to affordability and long shelf lives, multiple factors may be attributed to the decline of small-scale food production in Appalachia and the shift from dependency on smallscale food production to grocery stores.

One qualitative interview with a 90 year female in Southern Appalachia brings to light many of the changes in subsistence patterns throughout the region. The research participant was raised on a farm and spent adulthood living in urban/peri-urban areas where she and her late husband cultivated gardens throughout their lives. When prompted about her gardening experience,

she shared, "Well, I've been gardening all my life. I've probably been working in the garden since I was five years old, so that's 85 years." When questioned about the foods she had cultivated and eaten throughout her life, she noted: • "We had peaches, apples, plums, and figs when we grew up."

• "Well, you know, corn and beans and peas.

It's mostly that's what southerners grow. And they grow okra and tomatoes. When we grew up, everyone had lard, even townsfolk who had the money."

• "In the springtime, it would be cabbage, onions, squash, and uh when I was small, my parents grew cucumbers, green beans, butterbeans, field peas, radishes, turnips, and corn. That is typical of a southern garden."

Interestingly, the interviewee felt that older traditional cooking methods had improved for the better during her lifetime. "When I was growing up, it [traditional food] wasn't healthy because you used lard in it. You seasoned it in lard, and you fried it in lard. I would think it's healthier now because we don't do that. Southerners don't use lard anymore, and they don't use as much salt as they did when I was growing." After World War II, she noted that there was a shift in rural food consumption patterns, noting that people began to utilize grocery stores more and rely less on home food production. In particular, the interviewee shared a story that highlights the shift from consuming fresh, homegrown foods to reliance on grocery store processed goods.

"Back then, the U.S. government would send out applications to ask how much you spent at the grocery store. They wanted to know how much I spent per week on groceries. I think I spent about $15.

They sent a letter back and said I could apply for food stamps and assistance. I don't remember if food stamps was available in the '60s. I guess they were. But anyway, they suggested I apply for assistance because that wasn't enough money to feed four people. We were eating high on the hog because we were eating all these nice vegetables. I was feeding the family good because I was growing it. I had to buy milk, meat, and meal [cornmeal]. They didn't ask if I had a garden, so I just filled it out. We laughed about it. We thought it was funny. They Volume XVIII, Spring 2022 | 65

ANTHROPOLOGY

oriented food systems has the potential to cut costs of produce in rural areas, reinvigorate long-standing traditions of gardening and food production in the region, and improve local economic conditions.

As previously mentioned, the historical isolation of the Appalachian region necessitated home gardening and farming as a key subsistence method for people throughout the region. The study "Perspectives on Healthy Eating Among Appalachian Residents" found through qualitative interviews that participants identified gardening as the "backbone" of the Appalachian diet. Multiple interviewees highlighted their inability to afford or access high-quality foods due to low socioeconomic status, insufficient transportation, and geographic isolation. Many informants suggested community gardening as a critical intervention method to increase healthy eating in Appalachia. (Schoenberg, 2013) Another study focused on youth perceptions of healthful eating and food choice found that interview participants responded positively to prompts about gardening.

(Swanson, 2013)

ANTHROPOLOGY

didn't ask what we were growing. Sometimes we spent even less than 15. $15 went a long, long way back then. We could get a bag of meal for about 39-40, average about 50 cents. And flour about the same. We grew our garden, and we were eating better than most people who didn't have gardens."

This single interview in no way serves to generalize the experience of all individuals in the vast Appalachian region. However, it provides cultural context to traditionally consumed southern Appalachian produce and insight into the transition to a difference in food consumption patterns following the rise of reliance on grocery stores and processed foods. Many people often consider traditional cuisine to be fatty, greasy, and unhealthy. However, as this personalized historical account demonstrated, traditional Appalachian staples are highly nutritious, easy to grow, and native to the region for thousands of years. Traditional gardening can serve as an effective public health intervention by increasing access to fresh produce in a culturally relevant manner for Appalachians.

Comparative Qualitative Literature The findings of the ethnographic interview conducted for this research paper share key similarities with other ethnographic research on farming in Appalachia.

However, it is important to note that these qualitative interviews are not representative of the diverse population and only included interviews with white females between the ages of 90 - 100+ years old.

Growing up on an Appalachian Farm (Holston, 2017) shares key similarities with the findings of the ethnographic interview conducted for this research paper.

This qualitative descriptive study used a convenience sample to identify Appalachian women that grew up in a farm setting. All participants "ate only what was healthy," and their diets consisted of "the things we grew on the farm." Key crops include tomatoes, okra, green beans, pinto beans, and corn.

Another similarity lies in the self-sufficiency lifestyle modality, particularly regarding household staples.

It was commonplace to rely on farm foods for the bulk of a family's diet, and only certain staples or production methods would be outsourced from the farm. One interviewee shared: "My daddy would take the cornmeal to town and have it ground and sold." A parallel story was shared by the interviewee of this qualitative study.

66 | Emory Undergraduate Research Journal

An important aspect of farm life noted in Holston, 2017 is community solidarity despite relative isolation between homesteads. Participants highlighted that farm life allowed for a strong sense of community due to the isolative nature of farmstead lifestyles. This rural upbringing likely provided the physical activity and nutrition literacy to engage in a healthy lifestyle that supported sustained longevity.

Qualitative findings from these comparative research studies highlight the role that land cultivation played in the Appalachian lifestyle over the last century. Activities that promoted individual and community wellbeing in the past could effectively target food insecurity and health concerns throughout the Appalachian region.

Supporting Local Food Production to Reduce Food Insecurity In a qualitative analysis of traditional Appalachian Foodways, Visocky found that across various historical documents, cookbooks, and memoirs, foods most prevalent in these texts include: corn, wheat, green beans, cabbage, greens, peas, tree fruit, squash, tomatoes, and okra. These foods mirror the insight gained from the qualitative interview found in the previous section.

Local food production provides a means to build upon historical and cultural legacies in land cultivation to build a promising future. A long-term solution to tackling food insecurity entails sustainable solutions where communities have the necessary skills to continue the intervention efforts for generations to come. In rural populations, there have been efforts to revitalize local food systems through farming. Community orchards and local farming initiatives could save the region from endemic poverty and, importantly, food security. Aptly stated in Community Orchards and Food Security in Appalachia, the Appalachian "region's robust agricultural past and promise seems incongruous with its present lack of food security." Local food production is a means through which the Appalachian region can be nutritionally and economically revitalized.

According to Agriculture and Food System Trends in the Appalachian Region: 2007-2012, "Farm size has particular relevance for local food production and system building." In Appalachia, the majority of farms selling to local markets are small farms (Low et al., 2015). However, in 2012, merely 5.2% of farmers in Appalachia were under the age of 35. Successful community crop cultivation will depend on providing young Appalachians with

farming know-how, technical literacy, and economic means to agriculturally revitalize the region. (Jackson, Perrett, & Descieux, 2015) Due to specific geographic concerns, it is important to establish local food systems that prioritize access to affordable, healthy, and fresh foods. Local agriculture fosters economic growth in rural areas with limited industry, particularly in areas where biodiversity has been greatly damaged by coal production. The reduction of 858,858 farmland acres in Appalachia between 2007 and 2012 highlights the dire need for more community farming and land cultivation initiatives, starting with fruit and vegetable cultivation as a fundamental basis for local food system development. Currently, monoculture crop production has saturated the region, and produce is a small component of overall agricultural production in the region. But community agriculture has strong prospects in being a successful means of reducing food insecurity and improving community health. Existing infrastructure for community-supported agriculture exists, as a total of 1,461 farms (0.57%) in Appalachia sell CSA. (Jackson, Perrett, & Descieux, 2015) In areas such as the Blue Ridge mountains, the terrain is limiting to agricultural success, and mountainous terrain is not conducive to plant cultivation. Although it can be challenging to reclaim mining areas for land cultivation due to soil quality (Helmer, 2019), successful results at cultivating land have developed from aquaponics facilities. The organization Refresh Appalachia provides training initiatives for locals to learn the trade of farming and partners with local producers to supply restaurants, stores, and schools with fresh produce. Successful interventions should incorporate economic opportunity through sustainable, local food production initiatives.

Considerations for replication of gardening interventions across Appalachia One qualitative research study, Perspectives on Healthy Eating Among Appalachian Residents, noted residents' perspectives on barriers to healthy eating included: low socioeconomic status, loss of gardening as a traditional Appalachian subsistence method, and geographic isolation among other factors. (Schoenberg, 2013) The Grow Your Health intervention cited the need for considering space to grow gardens, soil quality, water access, and general flexibility in reacting to the influence of weather on growing produce. Additionally, the REACH curriculum empowered participants to alter shopping, meal preparation, and food consumption patterns, utilizing the programming knowledge to make healthier eating

Case Study Example 1 The Grow Your Health intervention was established through Ohio State University to support the reduction of chronic disease in Appalachia. By establishing community and container gardens and classes to teach residents about healthy cooking and canning techniques, practitioners hoped to increase the consumption of fruits and vegetables in food-insecure Vinton County, Ohio. Boasting a population of 13,000, the county has one full-service grocery store to support residents. With a focus on disease prevention and increasing the accessibility of fresh foods, Grow Your Health provides community members with instruction on gardening, healthy eating, food budgeting, meal planning, and canning and preserving fresh foods. Thus far, the program has seen many successes, including increasing the number of local gardeners and empowering residents to grow their own healthy foods. Challenges associated with this program include the local soil quality was not conducive to gardening and rural residents' ability to access and find transportation to the garden and courses offered by the program.

Case Study Example 2 In conjunction with local county offices and the University of Kentucky College of Nursing and the U.K. College of Agriculture, Food and Environment Department of Family and Consumer Sciences collaborated with 180 cooks in six Eastern Kentucky counties in the Rural Eating and Healthy Cooking program. This program was created in response to vast food deserts and limited socioeconomic means throughout the region. The goal of the program is to teach participants how to cook healthfully with limited means. In order to better understand strategies that support healthy eating behaviors, researchers from the University of Kentucky are collecting qualitative and quantitative data on this program. This 12-month study includes cooking classes, health coaching and evaluates participants' progress in improving healthy eating.

Consisting of cooking classes, participants are provided with cooking tools to take home and continue meal preparation as taught in the REACH curriculum. Participants take meals and recipes home after classes, and researchers measure the consumption patterns of families through their monthly grocery store receipts. Participants noted that participation in the classes was an enjoyable experience and that they gained a strong understanding of how to shop and prepare healthy meals.

Volume XVIII, Spring 2022 | 67

ANTHROPOLOGY

choices. Hardin-Fanning, the lead researcher of the program states, "many participants discovered a new favorite vegetable or ended the bad habit of walking down the snack aisle of their grocery store."

Previous research indicates that many people living in Appalachia understand healthy eating patterns but face systemic barriers and accessing and preparing healthy meals. Initiatives that are place-based and relevant to the needs of Appalachians both address the cultivation of fresh foods and educational interventions that teach people to utilize limited resources in preparing healthy meals.

Mixed-methods Research Model to Gauge Food Insecurity Levels As mentioned above under "Local Food Production," the organization Grow Appalachia takes a holistic, multifaceted approach to target food insecurity. Beyond a mere intervention, this organization works to create sustained change throughout the region, with successful initiative models that could be built upon for greater impact.

The following projected research model aims to understand the health status of low-income households that experience food insecurity in the region of Appalachia. Relying on quantitative and qualitative data collection, the purpose of this mixed-method research is to allow future public health practitioners to better understand the relationship between income, food insecurity, and health status so that they may design effective health interventions.

Food insecurity is a prevalent issue in Appalachia.

Food security and income are independent variables in analyzing health status. According to the USDA, food insecurity necessitates that households lack the means to acquire or experience uncertainty in food availability to meet nutritional needs. This phenomenon is measured as reduced quality and quantity in the nutritional content of the standard American diet recommended by the USDA, including minimized variety and desirability of available food. (USDA, 2021) Food insecurity is a variable to be assessed by the lack of available resources to access food within a household.

Participants in government nutrition and food programs, such as SNAP or WIC, are ideal candidates to make up the research sample. They meet USDA definitions of food insecurity and income level to receive government benefits. Income is an ordinal variable. The USDA defines annual household income as "the total 68 | Emory Undergraduate Research Journal

projected household income." (USDA Income Types, 2021) According to the U.S. Census Bureau, a household is defined as "the related family members and all the unrelated people, if any, such as lodgers, foster children, wards, or employees who share the housing unit." Health status is "the level of health of the individual, group, or population as subjectively assessed by the individual or by more objective measures." (U.S. Census Bureau, 2021) As an outcome, health status can be ascribed through the purview of income and food security by understanding how access to healthy food or the lack thereof influences Appalachian wellbeing.

Face-to-face qualitative, ethnographic interviews will be conducted with participants to further gauge the relationship between income and food insecurity in affecting health status. Data will be collected from a systematic random sample of rural households throughout the Appalachian region. This method incorporates Cluster Sampling, where the research includes different sampling stages, concluding with State data on WIC and SNAP-eligible households. The first stage begins by isolating the states within the greater Appalachian region.

Simple random samples will be conducted within the states to select specific counties and their health departments. These health departments will be contacted and asked to provide an anonymous list with phone numbers of the local households that receive SNAP and WIC benefits. From these lists, investigators will conduct a systematic random sample to select the households to be interviewed.

Roughly 4,075,000 people across 14 Appalachian states live in poverty. It is fair to assume that a large portion of Appalachia meets this study's sample requirements as SNAP and WIC eligibility are determined by poverty levels. Of those living in poverty, 10% of the population would yield a 407,500 person research sample. A systematic sampling of every 100th household would create a smaller sample pool for this study. With a skip number of 100, the qualitative research would have a sample size of 82. Due to the nature of ethnographic interviewing, a small pool is acceptable. These interviews provide a more holistic and detailed scope of health status related to income and food insecurity. This research will be conducted cross-sectionally. This research is meant to inform public health practitioners of the current state of food access as it relates to health status. Thus, a longitudinal study is unnecessary as this research does not aim to follow participants over time.

Conclusions Not only is there food scarcity in Appalachia due to rural supply chain shortages, but oftentimes healthy foods and produce are considerably more expensive due to high transportation costs. In an area plagued with endemic poverty, Appalachians are more likely to rely on low nutrient-dense foods, such as processed goods with long shelf lives. Through careful analysis of historical data, public health practitioners may seek to encourage community gardening initiatives that hearken back to

old ways of homestead food cultivation. The region has supported a millennium of food cultivation, and before the rise of post-war industrial farming during the 1950s, many families successfully relied on home gardens.

Supporting community-level food cultivation could be a critical move toward greatly reducing food insecurity in the region. However, more data must be collected to truly gauge the persistence and gravity of food insecurity for Appalachians among various demographic backgrounds.

Bibliography

About the Appalachian region. (2021, February 03). Re trieved February 08, 2021, from https://www.arc.gov/about-the-appalachian-region/ About the Appalachian region. (2021, February 03).

Retrieved February 15, 2021, from https://www.arc. gov/about-the-appalachian-region/ An in-depth look At USDA Income Types. (n.d.). Retrieved March 28, 2021, from https://www.usdaloans.com/ articles/usda-income-types/?go=10 Appalachian Residents Adopt Heart-Healthy Eating Hab its through Cooking Intervention | Appalachia. https://www.uky.edu/appalachia/articles/appa lachian-residents-adopt-heart-healthy-eating-hab its-through-cooking-intervention.

Bureau, U. (2020, August 27). Subject definitions. Re trieved March 28, 2021, from https://www.census. gov/programs-surveys/cps/technical-documenta tion/subject-definitions.html#:text=Household,-A%20 household%20consists&amp;text=A%20house hold%20includes%20the%20related,who%20

Volume XVIII, Spring 2022 | 69

ANTHROPOLOGY

Randomly sampled SNAP and WIC recipients will be contacted to schedule in-person interviews to understand participant experiences in accessing healthy food, gauging how long a household has received government benefits, what type of food they purchase with these benefits, and their perceived relationship between diet and health status will be gauged through detailed qualitative interviews. The in-depth interviews will follow a discussion guide with prepared questions and trained interviewers. The coding methodology will be inductive to allow researchers greater flexibility in interpreting and forming conclusions from the data.

This research project's healthy budget allows for incorporating in-person interviews, a generally expensive research methodology. Qualitative data provides rich insight and supplements the shortcomings of the quantitative phone interview by providing a more holistic picture of how income and food insecurity determine health status. However, it is not a statistically representative form of data collection and only provides research data from• participant perspectives and life experiences. However, with a cross-sectional research design, the lack of rigidity in cross-sectional data collection is acceptable. Since the qualitative data is only reliable at the time it is gathered, it will provide a snapshot understanding of the re-• lationship between income and food insecurity, allowing future public health practitioners to draw conclusions from the data and design impactful health interventions.

This is also a very time-consuming methodology that requires experienced interviewers with the ability to travel and interact with interviewees. One shortfall of the qualitative arms concerns the logistical difficulty and relative expense of conducting extended, personal interviews. Due to the time-consuming nature of qualitative interviews, with a sample size of 82 households, there may be difficulty in conducting, analyzing, and coding a vast pool of qualitative data.

ANTHROPOLOGY

share%20the%20housing%20unit Chubinski, J., & Carrozza, M. (2012). Obesity and Food Insecurity. In Couto R. (Author) & Ludke R. & Obermiller P. (Eds.), Appalachian Health and Well-Being (pp. 149-166).

Lexington, Kentucky: University Press of Kentucky. doi:10.2307/j.ctt2jcssn.12 Dragovich, S. (2020). Telling the story of Appalachia one food at a time. Retrieved February 15, 2021, from https://www.virginia.org/appalachiafoodstory/ Definitions of food security. (n.d.). Retrieved March 28, 2021, from https://www.ers.usda.gov/topics/ food-nutrition-assistance/food-security-in-the-us/ definitions-of-food-security.aspx#:text=Food%20inse curity%E2%80%94the%20condition%20assessed, may%20result%20from%20food%20insecurity.

Hardin-Fanning, F., & Rayens, M. (2015). Food Cost Dis parities in Rural Communities. Health Promotion Prac tice, 16(3), 383-391. Retrieved February 15, 2021, from http://www.jstor.org/stable/26741229 Helmer, J. (2019, May 16). Can farming save Appalachia?

Retrieved March 25, 2021, from https://modernfarmer. com/2019/05/can-farming-save-appalachia/ Holston, E. C., & Callen, B. (2017). Growing Up on an Appalachian Farm: A Centenarian Perspective. Inter national journal of aging & human development, 85(3), 289–304. https://doi.org/10.1177/0091415016688306 Jackson, C., Perrett, A., & Descieux, K. (2015). Agricul ture and Food System Trends in the Appalachian Re gion: 2007-2012 (pp. 4-21) (United States, Appalachian Regional Commission, Appalachian Sustainable Agri culture Project). Washington, DC: Agriculture and Food System Trends in the Appalachian Region: 20072012.

John T. Edge. (2007). The New Encyclopedia of Southern Culture : Volume 7: Foodways. The University of North Carolina Press.

Life Sciences Research Office, S.A. Andersen, ed., "Core Indicators of Nutritional State for Difficult to Sample Populations," The Journal of Nutrition 120:1557S-1600S, 1990.

Rural Health Information Hub, 2020. Grow Your Health [online]. Rural Health Information Hub. Available at: https://www.ruralhealthinfo.org/project-examples/912 Loopstra R. (2018). Interventions to address household food insecurity in high-income countries.

The Proceedings of the Nutrition Society, 77(3), 270–281.

70 | Emory Undergraduate Research Journal

https://doi.org/10.1017/S002966511800006X Pheley, A. M., Holben, D. H., Graham, A. S., & Simp son, C. (2002). Food Security and Perceptions of Health Status: A Preliminary Study in Rural Appala chia. The Journal of Rural Health, 18(3), 447–453. https://doi.org/10.1111/j.1748-0361.2002.tb00909.x Ramsey, Community Orchards and Food Security in Appalachia, 120 W. Va. L. Rev. (2018). Available at: https://researchrepository.wvu.edu/wvlr/vol120/ iss3/16 Rural Health Information Hub. Rural Project Summary: Grow Your Health. https://www.ruralhealthinfo.org/ project-examples/912 Schoenberg, N.E., Howell, B.M., Swanson, M., Grosh, C. and Bardach, S. (2013), Perspectives on Healthy Eating Among Appalachian Residents. The Journal of Rural Health, 29:s25-s34. https://doi.org/10.1111/jrh.12009 Sharaievska, Iryna Dr.; West, Stephanie Dr.; and Wed dell, Melissa Dr. (2018) "The Privilege of Healthy Eat ing: A Qualitative Study Exploring the Local Food Choices of Low-Income Families from Appalachia," Journal of Health Disparities Research and Practice: Vol. 11: Iss. 3 , Article 10. Avail able at: https://digitalscholarship.unlv.edu/jhdrp/ vol11/iss3/10 Shaefer, H., Edin, K., & Talbert, E. (2015). Understanding the dynamics Of $2-a-Day poverty in the United States.

RSF: The Russell Sage Foundation Journal of the Social Sciences, 1(1), 120. https://doi.org/10.7758/ rsf.2015.1.1.07 Supplemental Nutrition Assistance Program (SNAP). (n.d.). Retrieved February 15, 2021, from https://www. fns.usda.gov/snap/supplemental-nutrition-assis tance-program USDA's strikeforce Initiative INVESTS $23.5 billion in rural Communities, expands to four new states. (n.d.).

Retrieved February 15, 2021, from https://www. usda.gov/media/press-releases/2016/01/15/us das-strikeforce-initiative-invests-235-billion-ru ral-communities USDA Measurement. (n.d.).

Vance, J. D. (2020). Hillbilly Elegy. Harper Collins UK. Visocky, S. (2016). APPALACHIAN FOODWAYS FROM THEN TO NOW: USING TRADITIONAL FOODS TO ENHANCE DIETETIC PRACTICE. Appalachian State University. doi:http://libres.uncg.edu/ir/asu/f/ Visocky,%20Sarah2016Thesis.pdf

ANTHROPOLOGY Photo by Sivaram Gunisetty

Kat Morgan is a 2021 graduate of Emory University and incoming Master of Public Health candidate at Columbia University. Finishing her undergraduate studies in three years, Kat was honored with the Emory Department of Anthropology Outstanding Senior Award and admitted into the Phi Beta Kappa, Omicron Delta Kappa, and Lambda Alpha honor societies. With a holistic view of health and diverse academic background, her professional interests lie in health promotion, cultural competency, and food systems.

Volume XVIII, Spring 2022 | 71

Photographers Jessie (Xuetong) Ni is a member of the Emory class of 2025. She was born and raised in Shanghai, China. She discovered her interest in photography during the pandemic in 2019. She also loves to be behind the camera and to preserves precious moments. Through the viewfinder, she constructs a more personal version of reality. She intends to major in Philosophy and BBA in film and media management concentration.

Naomi is an NBB major at Emory. Her photos focus on portraits and nature photography, and her passion for photography began with a camera she borrowed from her middle school where she recreated famous portraits with friends and captured school trips. Photography allows her to appreciate the beauty in this world and highlight the little things that often go unnoticed, and she finds joy in sharing them with others. Naomi took the first photo featured at a lagoon in California and captured the second photo in a high-speed train going through the countryside in France.

Leopold (Leo) is from Jackson, New Jersey, and is planning to major in business and East Asian studies. Leo is also a member of the Emory class of 2025. Aside from studying, photography has been one of my recent hobbies. The types of photography he enjoys taking are portraits, street photography, landscape photography, etc. As a beginning photographer, taking pictures is his way of recording the bits and pieces happening in his daily life. Feel free to follow his photography page on Instagram (@lhuang_photos) for more pictures like the ones featured in this issue!

72 | Emory Undergraduate Research Journal

Sivaram Gunisetty is an Assistant Scientist at Emory Vaccine Center, Department of Microbiology and Immunology, Emory University.

His research work contributes to vaccine research and research relevant in answering questions that are of public health importance. He is an amateur photographer; photography is his passion and he loves capturing the beauty of nature. He also loves traveling and exploring new places.

Jae Sun is from South Korea, but grew up in Mozambique and Swaziland. Although she is a Chemistry major, she has loved photography since high school and loves taking pictures of different school events and landscapes. She always thought that the only thing we can remember from an event is by leaving evidence such as photographs or videos, so we are given a chance to relive the moments.

Volume XVIII, Spring 2022 | 73

74 | Emory Undergraduate Research Journal

Photo by Leo Huang