Running Head: RESEARCH PROPOSAL TESTING THE IMPACT OF DIET ON

DISEASE

Research Proposal Testing the Impact of Diet on Disease

Karthik Sridasyam

Wayne State University

RESEARCH PROPOSAL TESTING THE IMPACT OF DIET ON DISEASE

Abstract

Though numerous research studies exist testing the effects of diet on the health of individuals,

there seems to be very little improvement in the overall health of the American people despite

the enormous efforts of countless organizations. With the arising interest in the involvement of

gut bacteria in daily functions, and even in their link to diseases, there appears to be a vacuum in

the study of the foods we eat to the bacterial involvement of disease. The objective of this paper

is to provide a sample research proposal on a hypothetical research study which would attempt to

understand the link between diet and disease, through the involvement of the gut bacteria. This

research can aid in both the benefit of federal organizations by helping them target more

proactive methods of intervention, as well as aid in the general population's knowledge of how

the food they eat is affecting them.

Keywords: Disease, Bacteria, Microbiome, Diet, Proposal

RESEARCH PROPOSAL TESTING THE IMPACT OF DIET ON DISEASE

Introduction

In spite of heavy intervention from numerous health organizations such as the World

Health Organization, and the Center for Disease Control, there was once again a sharp increase

in the number of obese Americans in 2016. Currently, over 40% of all the American adults are

obese and in severe risk of developing diabetes, cancers and heart disease (Richtel, Jacobs).

Despite large-scale federal agencies increasing the awareness of the public to the risk of being

obese, the trend did not negate from nearly a decade earlier. In addition to adults, children ages 2

through 5 had obesity rates increase to nearly 14 percent in 2016, in comparison to the prior 10

percent in 2008 (Richtel, Jacobs). It is clear that our efforts to abate this issue are futile, and it

became crucial that new methods have to be implemented in order to understand and solve this

issue.

Though there has been ample research done by Universities (i.e. Harvard, Princeton, etc.)

on the link between the foods we eat, and their caloric and nutritional benefits and downfalls,

there has not been much research on the influence of the foods we eat on the individual

organisms living inside of us. Every human being carries inside of them, a mirage of bacterial

cells, which work in a symbiotic relationship with us in order to allow us to digest our meals and

carry on our daily functions. It has only recently been identified as an organ (Baquero),

demonstrating the importance of it within our body.

Prior research studies such as The Effect of Diet on the Human Gut Microbiome by Peter

J. Turnbaugh, have identified that the there is a significant link between the foods we eat and the

composition of the bacteria within our gut. There were several bacteria’s which were more

prevalent in individuals with diets that were high in triglycerides, commonly associated with

obesity and certain bacterium which were specifically prevalent in individuals who engaged in a
RESEARCH PROPOSAL TESTING THE IMPACT OF DIET ON DISEASE

diet composed of mostly fresh vegetables. Needless to say, it became clear that the food we eat

impacts the organisms in our gut. Another research study attempted to understand the reciprocal

relationship between the gut bacteria and our desire to eat, by using a stool transplant procedure.

Their results showed that there was indeed a significant influence of the bacteria on the foods we

crave, which further promotes the hypothesis that maybe the lack of improvement in average

American health was not due to pure ignorance. Further studies have also correlated the bacterial

composition to the prevalence of diseases, such as liver cirrhosis, which is commonly associated

with bacterial translocation from the colon.

Though there have been several small steps taken within the overall realm of food to

physical health, there has not been a research study which specifically targeted its focus on the

effects of individual foods, and their correlation to health disorders through the bacterial

environment. Although many nutritionists would suggest that it is simply a case of “calories in vs

calories out” (Howell), the research that I have identified seems to show a much more significant

cause. One where the foods they eat, physically influence what is viewed to be completely free-

willed decisions to further their binge. It is crucial that we focus our research on the influence of

certain foods, such as fast food for example, which may give a key into understanding, and even

perhaps solve, this global crisis.

In my research, I will focus my efforts on understanding the common dietary choices of

the average American and use that information in addition to samples of their gut bacterial

composition and medical records in order to identify correlations within foods and their

influence in the disease prevalence through bacterium. As a result, I will be able to conclude the

true reason why despite the copious amounts of resources being put into the awareness of the

public about proper diet, and the importance of personal health, obesity and cardiovascular
RESEARCH PROPOSAL TESTING THE IMPACT OF DIET ON DISEASE

disease is continuously rising. Programs such as this can help federal agencies make better use of

their efforts and provide people with a better way to understand how their food affects them, and

maybe even rid the world, of this arising epidemic.

Literature Review

Though food is primarily a means of sustenance for all people, in recent years it has

supplanted into its own culture. Individuals from across the globe are able to interact through the

food that they eat, and it is now clear that food impacts our everyday lives more now than ever

before (Stajcic). Today, the food industry has boomed into a multibillion-dollar industry, from

magazines to books to movies, it has taken over the pop culture industry1, so it is by no means

surprising that certain behaviors for food have cemented themselves in our lives.

For most of the population, individual food habits are fairly consistent. The desire to eat

certain foods often is correlated to their personal upbringing, and individual culture. Any

attempts to alter their diet is often met with near immediate revulsion and an inevitable relapse to

my prior behavior. It is clear that food habits are much more secure in our lives than we may

hope to believe. Modern research tells us that our divulgence in certain foods may not even stem

from our own innate desire for them, but merely a result of the decisions made by

microorganisms located within us (Alcock). Microbes under selective pressure can manipulate

host eating habits in order to increase their own fitness, sometimes at the risk of their host’s

fitness as well. Though our reluctance to engage in certain food habits may be coined “self-

control” by most individuals, its there seems to exist a reciprocal interaction between the foods

we have eaten and the foods we want to eat.

RESEARCH PROPOSAL TESTING THE IMPACT OF DIET ON DISEASE

Gut Bacteria and Diet Choices

The human body is the living ground of trillions of microorganisms within the gut, and

“long-term diet has been noted as one of the significant factors linked to gut bacterial

composition” (Knights et al.) While most modern diets focus on enhancing the diversity of gut

flora, in a study done to compare Western- and Mediterranean diet in their impact on bacterial

levels within the gut, it was demonstrated that while there were marginal changes in the

diversity, there were dramatic changes in the composition of each type of bacterium (Nagpal).

For example, due to the high animal fat and protein diets of American and European populations,

there were “significantly lower abundances of Bacteroidetes and Prevotella bacterium, when

compared to the diets of Egyptian and rural African population” (Jain). Though both still existed

within the individual, the particular quantity of each was influenced by their dietary choices.

These particular types of bacteria have shown to be significant biomarkers when making

interpretations on host diet choices as well, particularly “Prevotella was associated with non-

Western, rural communities with a plant-based rich in polysaccharides and fiber" while

Bacteroidetes were more associated with samples from US subjects eating a Western diet, richer

in protein and fat” (Gorvitovskaia). These results are consistent when compared to individuals

who have engaged in the more Indian dietary choices, which consists of largely whole grains,

vegetables, and simple carbohydrates like potatoes. These individuals have shown much larger

populations of the Prevotella bacteria (Jain). While it is supported that our choices in the food

significantly influence the bacteria that live in our gut, the inverse relationship is also believed to

be true by new experiments testing behavior in response to gut flora. The experiments done to

study the effects of gut bacteria are commonly done through a technique called Fecal Microbiota

Transplantation or FMT, which is the administration of a solution of fecal matter from one donor
RESEARCH PROPOSAL TESTING THE IMPACT OF DIET ON DISEASE

into the gastrointestinal tract of a recipient done to directly change the recipients gut bacterial

composition and to study the health benefits (Bakken et al).

In a study to understand the effects of gut bacteria on mood, mice were fed different feces

from other mice donors that showed a certain innate behavior categorized by a behavior chart.

“When the germ-free mice were fed a fecal pellet from a control mouse, the inoculated mice

exhibited behavior that similar to the behaviors of the fecal donors (Bercik et al). In addition to

mood, microbes have also shown that they can influence their hosts through neural pathways,

causing changes in behavior which the hosts can mistakenly believe to be native. The primary

interaction between bacteria and the brain occurs through the Vagus nerve, commonly known as

the “central communication axis between the 100 million neurons of the enteric nervous system

in the gut to the base of the brain at the medulla” (Mayer). Research studies have shown that the

Vagus nerve activity significantly impacts eating behaviors in mice, specifically when the

production of adrenergic neurochemicals by the microbes in the gut may contribute to over-

eating behavior of mice (Sawchenko). In light of these experiments, we can infer that there is a

significant correlation between the food we eat, and the bacteria we have living inside of us.

Intestinal Bacteria and Disease

We have obtained copious evidence which points to the coevolution of human beings

with their internal microorganisms, the transplant of these organisms to different hosts elicits a

different type of symbiotic relationship which thereby alters their own body in ways that are not

completely clear to us today (Rawls, Mahowald, Ley, Gordon). One of the most significant roles

of the gut microbiota is their role in the intestinal mucosal and systemic immune systems within

the body, which is hardly surprising when you consider that the mucosa represents the largest

surface area which is in direct interaction with the antigens of the external environment
RESEARCH PROPOSAL TESTING THE IMPACT OF DIET ON DISEASE

(Ishiguro). In an experiment done on intestinal bacterial deficient animals, there were significant

immune deficiencies exhibited by the animals which were reversed by their treatment with a

particularly prominent bacteria Bacteroides fragilis (Mazmanian). B. Fragilis induces the

proliferation of CD4+ T cells (main) which are one of the primary cells for fighting infections.

Removal of this bacterium from the initial animal caused a drastic increase of inflammation

within the gut, possibly leading to Inflammatory Bowel Syndrome (Rawls, Mahowald, Ley,

Gordon).

Though there are severe immune influences from the deficiency of a certain bacterium,

there are also health deficits caused by an increase in certain bacteria that may be harmful to the

body if made contact. Bacterial translocations through the hepatic portal system, which links the

gastrointestinal tract to the liver, could cause an accumulation of neurotoxins within the liver

leading to cirrhosis, and liver disease (Lorenzo-Zuniga et al). Studies done on rats have shown

that treatments of a growth factor responsible for the gut barrier maintenance were able to reduce

the development of liver disease in the animals by "enhancing the intestinal barrier and reducing

levels of bacterial translocation" (Ley et al). In addition to clinical diseases, conditions common

to the Western world like obesity have also been shown to be strongly correlated to certain

bacteria within the gut. Specifically, studies which studied the bacterial makeup of humans who

were lean and those who were obese showed significant similarities in bacterial makeup. When

the obese individuals were placed onto a diet similar to that of the lean individual, the change in

their gut bacteria makeup was significant (Ley et al). Additionally, bacterially sterile mice which

were fed a common western diet consisting heavily of animal fat and protein resulted in high

obesity, while those that had similar bacterial makeups as those who regularly engaged in the
RESEARCH PROPOSAL TESTING THE IMPACT OF DIET ON DISEASE

western diet showed a reduction in their body composition when the bacteria were removed.

(Turnbaugh).

Conclusion

The goal of this research paper was the investigate the relationship between our diet and

diseases which influence our lives. Analysis of countless research studies has demonstrated

individual relationships between host dietary choices and their gut bacteria, and the influence of

that bacteria in disease manifestation. As food becomes a larger and larger part of our modern

lives, influencing everything from pop culture, to interactions with other members of society, we

can conclude that the study on its influence on our disease is very important. Though we can

influence disease prognosis in a myriad of ways from surgical intervention to pharmaceutical

products, the influence of diet can allow us to realize that there are changes in eating habits may

provide alternative routes of therapy. With the easily changeable and highly influencing

characteristics of diet, accompanies with the rising numbers of obesity and human illness, it has

never been more crucial to understand the trillions of organisms which live inside of us and

influence our everyday activities.

RESEARCH PROPOSAL TESTING THE IMPACT OF DIET ON DISEASE

Methodology

The research conducted will need a large sample size, near 100 individuals in order to

reduce as much error as possible. Individuals will be divided according to their dietary choices

based on a food frequency and sleep questionnaires, the individual’s cardiovascular strength and

endurance will need to be tested in order to set a baseline. In addition, a psychological analysis is

required to understand the individual’s mental states prior to the study and ensure that they are

capable of undertaking the study. A medical consultation will also be taken by the physician

involved in the study, as well as fecal samples in order to check for bacterial composition.

Consent forms will be given out explaining the entire procedure and verifying their

conformation.

After the individuals have been divided into their prospective groups and rated based on their

test results, they will be separated into three groups. One group will be instructed to engage in a

standard Mediterranean diet consisting heavily of legumes, nuts, and vegetables, with small

protein, while another group will be instructed to take upon a more western diet of high fat, and

protein, with less emphasis on vegetables. The third group will be instructed to carry forth their

present diet and will act as the control. Each of their group analysis will be cross-checked with

certified dieticians to ensure that their food choice places them at no risk of illness. All

individuals will be instructed to engage in similar levels of physical activity, cross-checked with

a pedometer given to them, in order to ensure that physical activity may not affect the results.

Every week, the individuals will receive a full physical examination, with a fecal sample

taken as well. The individuals will repeat this process for 4 weeks, and at the end of the four

weeks, they will be retested on all the initial data. Cardiovascular strength and endurance

(through VO2 max), medical check, a fecal sample, a full psychological profile, and food
RESEARCH PROPOSAL TESTING THE IMPACT OF DIET ON DISEASE

frequency and sleep questionnaires once again. Upon completion of this study, extensive data

analysis will be required in order to understand the complete extent of the results. Test subjects

will be asked to come in one month after their completion for a reevaluation as well.

Discussion

The data collected from this study will be invaluable in understanding the effects of diet

on the body. The fecal samples taken will allow us to see the progression of the make-up of the

various bacteria within the gut, and how they were influenced by the diets of the individuals. The

psychological analysis allows us to check for their emotional states and how they may have

improved/ regressed through the study. In addition to simply testing its effects on disease risk,

we can also use it to understand how the foods we eat may even influence our behavior and our

emotions, leading to a new line of treatment methods. Food is something that is required by

every organism to survive and therefore is a valuable evolutionary tool. There are countless

biological systems in place which allow our bodies to utilize the stored energy we have, which

even influences our neurological function at a point to prevent us from feeling hunger, in order to

function (Nagpal). This alone shows that there is much about our diet that we take for granted.

This research project is by no means the final frontier in the realm of understanding the

extent of gut microbiota in our bodies, but its results could spark the next age in medical

research. There are several limitations to this study as well, such as the lack of a longitudinal

study, which considers years and years of data and analyzes it. In addition, there are several

control variables that are difficult to account for, such as age and body composition. Attempting

to create a completely controlled data collection would require an enormous sample size, which

is not practical in a preliminary study. In regards to sources of error, one would include
RESEARCH PROPOSAL TESTING THE IMPACT OF DIET ON DISEASE

individuals not abiding by the guidelines of the study and consuming foods outside of their

dietary guidelines, as well as those who simply leave the program.

The results of this study, if they were to show that diet indeed does have an impact on

disease, will revolutionize the way medicine is given to individuals. Doctors will likely be more

open to directing their focus onto osteopathic medicine and relinquishing their dependence using

pharmaceutical drugs. It will allow individuals to realize the true impact that their diet has and

could be used to promote the importance of a healthy diet in the world, and thereby possibly

reducing the incidence of heart disease, diabetes, and childhood obesity. The benefits of this

study are limitless, and more research needs to be done in order to not only confirm any results

but further it by testing the individual nuances within the system of gut microbiota.
RESEARCH PROPOSAL TESTING THE IMPACT OF DIET ON DISEASE

Bibliography

Alcock, Joe, et al. “Is Eating Behavior Manipulated by the Gastrointestinal Microbiota?

Evolutionary Pressures and Potential Mechanisms.” BioEssays, vol. 36, no. 10, 2014, pp.

940–949., doi:10.1002/bies.201400071.

Bakken, J., Borody, T., Brandt, L., Brill, J., Demarco, D., Franzos, M. et al. (2011) Treating

Clostridium difficile infection with fecal microbiota transplantation. Clin Gastroenterol

Hepatol 9: 1044–1049.

Baquero, F, and C Nombela. “The Microbiome as a Human Organ.” Advances in Pediatrics.,

U.S. National Library of Medicine, July 2012, www.ncbi.nlm.nih.gov/pubmed/22647038.

Bercik P, Denou E, Collins J, Jackson W, et al. 2011. The intestinal microbiota affects central

levels of brain-derived neurotropic factor and behavior in mice. Gastroenterology 141:

599–609 e1-3.

Gorvitovskaia, Anastassia, et al. “Interpreting Prevotella and Bacteroides as Biomarkers of Diet

and Lifestyle.” Microbiome, vol. 4, no. 1, 2016, doi:10.1186/s40168-016-0160-7.

Howell, S, and R Kones. “‘Calories in, Calories out’ and Macronutrient Intake: the Hope, Hype,

and Science of Calories.” Advances in Pediatrics., U.S. National Library of Medicine, 1

Nov. 2017, www.ncbi.nlm.nih.gov/pubmed/28765272.

Ishiguro, Edward, et al. “Gut Microbiota in Health and Disease.” Gut Microbiota, 2018, pp. 57–

90., doi:10.1016/b978-0-12-810541-2.00004-x.

Jain, Abhishek, et al. “Similarities and Differences in Gut Microbiome Composition Correlate

with Dietary Patterns of Indian and Chinese Adults.” AMB Express, vol. 8, no. 1, 2018
RESEARCH PROPOSAL TESTING THE IMPACT OF DIET ON DISEASE

Knights D, Ward TL, McKinlay CE, Miller H, Gonzalez A, McDonald D, et al. Rethinking

“Enterotypes”. Cell Host & Microbe. 2014;16(4):433–7. http://dx.

doi.org/10.1016/j.chom.2014.09.013.

Ley RE, Backhed F, Turnbaugh P, Lozupone CA, Knight RD, Gordon JI. Obesity alters gut

microbial ecology. Proc Natl Acad Sci USA 102: 11070–11075, 2005.

Ley RE, Turnbaugh PJ, Klein S, Gordon JI. Microbial ecology: human gut microbes associated

with obesity. Nature 444: 1022– 1023, 2006.

Lorenzo-Zuniga V, Rodriguez-Ortigosa CM, Bartoli R, Mar- tinez-Chantar ML, Martinez-

Peralta L, Pardo A, Ojanguren I, Quiroga J, Planas R, Prieto J. Insulin-like growth factor

I im- proves intestinal barrier function in cirrhotic rats. Gut 55: 1306– 1312, 2006.

Mayer EA. 2011. Gut feelings: the emerging biology of gut-brain communication. Nat Rev

Neurosci 12: 453–66.

Mazmanian SK, Liu CH, Tzianabos AO, Kasper DL. An immu-nomodulatory molecule of

symbiotic bacteria directs maturation of the host immune system. Cell 122: 107–118,

2005.

Nagpal, Ravinder, et al. “Gut Microbiome Composition in Non-Human Primates Consuming a

Western or Mediterranean Diet.” Frontiers in Nutrition, vol. 5, 2018,

doi:10.3389/fnut.2018.00028.

Rawls JF, Mahowald MA, Ley RE, Gordon JI. Reciprocal gut microbiota transplants from

zebrafish and mice to germ-free recip- ients reveal host habitat selection. Cell 127: 423–

433, 2006.
RESEARCH PROPOSAL TESTING THE IMPACT OF DIET ON DISEASE

Rawls JF, Mahowald MA, Ley RE, Gordon JI. Reciprocal gut microbiota transplants from

zebrafish and mice to germ-free recip- ients reveal host habitat selection. Cell 127: 423–

433, 2006.

Richtel, Matt, and Andrew Jacobs. “American Adults Just Keep Getting Fatter.” The New York

Times, The New York Times, 23 Mar. 2018,

www.nytimes.com/2018/03/23/health/obesity-us-adults.html.

Sawchenko PE, Gold RM, Leibowitz SF. 1981. Evidence for vagal involvement in the eating

elicited by adrenergic stimulation of the paraventricular nucleus. Brain Res 225: 249–69.

Stajcic, Nevana. Understanding Culture: Food as a Means of Communication. 2013,

yadda.icm.edu.pl/yadda/element/bwmeta1.element.desklight-3b71a1ed-2c03-499a-8884-

ab5ec51e1f56/c/05-Stajcic_v01.pdf.

Turnbaugh PJ, Backhed F, Fulton L, Gordon JI. Diet-induced obesity is linked to marked but

reversible alterations in the mouse distal gut microbiome. Cell Host Microbe 3: 213–223,

2008.