This document summarizes a proposed study examining the effect of obesity on nonalcoholic fatty liver disease (NAFLD) and whether nutritional interventions can help. Specifically, it will investigate if obesity increases fat cell (adipocyte) infiltration and inflammation in the liver, and if polyphenolic compounds can inhibit these obesity-induced effects. The study will use a cell culture model mimicking the obese environment to test the impacts of serum from obese individuals on liver cells, with and without polyphenols. The goal is to better understand how obesity promotes NAFLD and potentially identify new treatment options through nutrition.

1. THE EFFECT OF OBESITY ON THE PROGRESSION OF NONALCOHOLIC FATTY
LIVER DISEASE AND NUTRITIONAL INTERVENTION
Hallie Nix, A04282861, hmn9@txstate.edu, Senior, College of Family and Consumer Sciences,
Department of Nutrition and Foods, PI.
Ana M. Hernandez, RDN, LDN, A04478943, amh219@txstate.edu, Graduate Research Assistant,
College of Family and Consumer Sciences, Department of Nutrition and Foods
Ramona Salcedo Price, PhD., Associate Professor, Department of Nutrition and Foods,
rsp44@txstate.edu, Faculty Advisor
IRB/IACUC reviews are not needed for this project.
Table of Contents
Title, PI, Faculty Advisor, Table of Contents 1
Abstract, Statement of Significance 2
Project Description and Narrative 3
Resume 5
References 6
Endorsement Statement 7
Budget Narrative 8
Project Budget 9

2. 2
Abstract
Understanding the impact of obesity on health is a significant public health issue. The prevalence of
obesity is major health concern because it is associated with co-morbidities such as diabetes, metabolic
disorders, liver disease, and cancer. Specifically, obesity is associated with nonalcoholic liver disease
(NAFLD). NAFLD is characterized by an increase in adipose tissue inflammation, and fat accumulation
in liver cells. The mechanisms linking obesity, inflammation, adipocytes and its role in transforming
liver cells remain unclear. The local inflammatory microenvironment has been linked to the progression
of NAFLD to liver cancer. Obesity is characterized by increased adipocyte (fat cell) accumulation and can
result in chronic inflammation and metabolic dysfunction. Interestingly, dietary polyphenols (chemicals
found in plants) can decrease inflammation and improve the health of the liver. Therefore, we hypothesize
that in the context of obesity, dietary polyphenols may reduce adipocyte infiltration and inflammation in
the liver. To test this hypothesis, we will use a novel cell culture approach that mimics the obese
environment. Briefly, studies will aim to address the following: determine if obesity increases adipocyte
infiltration and inflammation, and whether polyphenolic compounds can inhibit obesity-induced
adipocyte infiltration and inflammation.
Statement of Significance
Due in part to the rising obesity epidemic and increase in NAFLD, there is a critical need to understand
the mechanisms by which obesity promotes liver disease. The outcomes of this study would contribute to
designing studies that could be funded by external grants. In addition, this study could have an impact in
the local community. Hays County has twice the incidence of liver cancer compared to the average liver
cancer incidence in Texas.1
The information gained in this study would be used in designing clinical
studies with a local hospital to reverse the impact of obesity on liver disease.

3. 3
Project Description and Narrative:
Introduction: Obesity is a health concern in the U.S., showing no sign of declining. With obesity come
several comorbidities such as diabetes, hypertension and other metabolic disorders.2
Obesity increases an
individual’s chance of developing nonalcoholic fatty liver disease (NAFLD).2,3
NAFLD and obesity are
relevant due to the high prevalence in the U.S., 30% and 36%, respectively.4
NAFLD is not a stagnant
disease, if left untreated; it can progress to steatohepatitis, cirrhosis or liver cancer.2,4
Advanced NAFLD
can be characterized by fat accumulation in liver cells and adipocyte (fat cell) inflammation. It has been
shown that obesity increases fatty liver disease and progression to liver cancer but the mechanisms by
which adipocytes accumulate in the liver are unclear. Therefore, it is necessary to understand the
biological impact obesity has on the progression of NAFLD.
Problem Statement: Studies have investigated the correlation of adipocytes and the progression of
NAFLD. The problem lies in the fact that there have been few studies that have tested nutrition
intervention to reverse the obesity-related advancement of NAFLD. To date, there is no known
pharmacologic treatment for NAFLD to reverse its progression,2
which leads us to search for nutritional
intervention to decrease the inflammatory havoc brought by adipocyte infiltration in the liver.
Project Description: First, we will characterize human serum, isolated from blood, for obesogenic
hormones and proteins by immunoblot adipokine protein array. To investigate the obesity-induced
adipocyte recruitment by liver cells, liver cells will be exposed to obese serum. After 24 hours,
conditioned media from liver cells will be collected and used as a chemoattractant to determine
differences in adipocyte infiltration. Adipocyte infiltration will be measured by counting cells that invade
a basement membrane. Next, we will determine whether polyphenols inhibit obesity-induced
inflammation and adipocyte infiltration. A cytokine protein panel will be used measure the pro-
inflammatory environment that is induced by obesity.
Goals and Objectives: The goal of this study is to find a break in the obesity-associated progression of
NAFLD. This will be accomplished through testing the effects of dietary polyphenolic compounds on the

4. 4
hepatocytes exposed to serum from obese, or overweight individuals. Ultimately, the goal is to find new
treatment options for the growing population of NAFLD.
Dissemination Plan: The data collected from this study will be available to the public through peer-
reviewed, scientific journals.
Evaluation Plan: For validity purposes, experiments will be repeated three times. I will participate in
weekly lab meetings to assess progress in my experimental findings.
Continuation Plan: The data will be used to further the study of the impact of polyphenols as an
intervention for NAFLD in obese individuals.
Management Plan: I will conduct experiments in consultation with Dr. Salcedo and Ana Hernandez.
Timeline: The three components of this study will come to completion after 3 months.
Personnel: Hallie Nix, PI, is a senior undergraduate in the Department of Nutrition and Foods. Upon
completion of her degree, she plans on pursuing a career educating the value of nutrition intervention of
disease in developing countries. Hallie will commit 11 hours per week during the funding period.
Ramona Salcedo Price, PhD., Faculty Advisor.
Dr. Salcedo is an Assistant Professor in the Nutrition and Foods Program. She has published research
related to obesity and cancer. Her laboratory is equipped to perform the experiments in this proposal.
Ana M. Hernandez, RDN, LDN, Graduate Research Assistant.
Ms. Hernández-Rosa is a Graduate Research Assistant in the Nutrition and Foods Program. After
completion of her M.S. she plans to obtain a Ph.D. in Clinical Nutrition. She aspires to become a board
certified specialist in Oncology Nutrition. She will optimize the experiments described in this proposal.
3.11 Enhancing Knowledge Through Innovation
Dr. Salcedo developed and published a novel cell culture technique that mimics the effect of obesity.5
This technique will provide insight into how obesity promotes NALFD and if this association can be
inhibited by dietary intervention. Dr. Salcedo is working with a local hospital for clinical research and the
results from this study will aid in designing clinical studies in collaboration with hospital.
3.12 Funding Resources This study is not funded by another source.

5. 5
Hallie Nix
2211 Timberway Court, Magnolia, Texas, 77355
(281) 475-6311
EDUCATION
Bachelors of Science in Nutrition and Foods Dec. 2016
Texas State University: San Marcos, Texas
• Concentration: Dietetics
• Institution GPA: 3.97
HONORS AND AWARDS
Deans List: 4 semesters, Texas State University.
Family Consumer Science Endowment: $950, Texas State University. Apr. 2015
Honors College: 4 semesters, Texas State University.
EXPERIENCE
Undergraduate Research Assistant, Nutrition and Foods Nov. 2015 – Present
Texas State University: San Marcos, Texas.
• Gained skills in cell culture, including growing advanced stage prostate cancer cells.
• Became competent in using PCR and western blot techniques to detect cell viability.
Resident Assistant Feb. 2015 - Present
Texas State University: Housing and Residential Life. San Marcos, Texas.
• Taught day seminars to 20-50 students on topics such as diversity and nutrition.
• Counseled 50 girls on peer conflict and career paths.
Student Grader, Nutrition and Foods Jan. 2015- May 2015
Texas State University: Nutrition and Foods. San Marcos, Texas.
• Analyzed and graded Dietary Analysis Projects.
• Provided quality feedback on the academic work of 200 undergraduate students.
Hostess, Waitress, Expediter, Job Trainer Apr. 2010-Jan. 2014
The Whistle Stop Tea Room: Tomball, Texas.
• Gained knowledge in many areas of foodservice including excellent customer service and
measures to ensure food safety.
• Trained 15 new employees in all areas of food service.
INVOLVEMENT/CERTIFICATIONS
CITI Program Certified Jan. 2016-Present
Member of the Student Nutrition Organization 2014-Present
• Secretary 2014-2015
Member of the Academy of Nutrition and Dietetics 2014-Present

6. 6
References
1. State Cancer Profiles. National Cancer Institute. Available at:
http://statecancerprofiles.cancer.gov/incidencerates/index.php?statefips=48&cancer=035&race=00
&sex=0&age=001&type=incd&sortvariablename=name&sortorder=asc#result. Accessed
February 25, 2016
2. Leamy AK, Egnatchik RA, Young JD. Molecular mechanisms and the role of saturated fatty acids
in the progression of non-alcoholic fatty liver disease. Prog Lipid Res. 2013;52(1):165-174.
doi:10.1016/j.plipres.2012.10.004.
3. Xu L, Kitade H, Ni Y, Ota T. Roles of Chemokines and Chemokine Receptors in Obesity-
Associated Insulin Resistance and Nonalcoholic Fatty Liver Disease. Biomolecules.
2015;5(3):1563-1579. doi:10.3390/biom5031563.
4. Musso G, Gambino R, Cassader M. Cholesterol metabolism and the pathogenesis of non-alcoholic
steatohepatitis. Prog Lipid Res. 2013;52(1):175-191. doi:10.1016/j.plipres.2012.11.002.
5. Price RS, Cavazos DA, deAngel RA, Hursting S, deGraffenried. Obesity-related system factors
promote an invasive phenotype in prostate cancer cells. Prostate Cancer and Prostatic Diseases.
2012:15(2):135-43.

7. 7

8. 8
Budget Narrative
Materials and supplies listed in the budget are core items for the study. Hallie will need liver cells (line
item 12) to perform her dietary intervention experiments. The inflammatory protein panel (line item 13)
will help identify the protein characteristics of factors secreted by the liver cells. The invasion chambers
(line item 14) are necessary to measure the infiltration of adipocytes (fat cells). Dr. Salcedo’s laboratory
will provide the equipment, serum and common chemicals that are necessary to conduct the experiments
in Hallie’s proposal.

9. 9
Budget
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