Johanna Muller

University of Maine
168 College Ave
Orono, ME 04469
johanna.muller@maine.edu

March 20th, 2022

Griffin P. Rodgers, M.D., M.A.C.P.

NIDDK Director
9000 Rockville Pike
Bethesda, MD 20892

Re: Diabetes Research Centers Grant RFD-DK-21-029

Dear Griffin P. Rodgers,

I am writing you this letter to declare my intention to apply for the Diabetes Research Centers
Grant RFD-DK-21-029. I am a graduate student at the University of Maine, an R1 Carnegie
research university, in the Food Science and Human Nutrition program.

The objective of applying for this grant is to aid in the research on the effect of resistant starches
on postprandial blood glucose, specifically the ones in cooked and cooled pasta.

With this grant, this very important research could provide an affordable and simple diet change
that could aid in preventing and controlling diabetes. I will be submitting a full proposal through
the NIH ASSIST program, and I look forward to hearing from you and your colleagues.

Sincerely,

Johanna Muller
University of Maine
National Institute of Health
National Institute of Diabetes and Digestive and Kidney Diseases

Name: Johanna Muller

Contact Information: 123 Fake Address Way
Pretend, NY, 12345
(888)123-4567
johanna.muller@maine.edu

Grant Title: Diabetes Research Centers Grant

RFA-DK-21-029

Amount Requested: $50,000

Proposed Start Date: September 2022

Duration: 2022/2023 Academic Year

Johanna Muller
Graduate Student
University of Maine
 The Effect of Resistant Starch in Cooked and Cooled Pasta on
Postprandial Blood Glucose Levels

Abstract
Dietary fiber has a profound impact on the digestion of sugar and simple carbohydrates, which
are known to increase blood glucose. Slowing the digestion and subsequent intake of
carbohydrates with dietary fiber can significantly reduce postprandial blood glucose response.
Frequent hyperglycemia, or high blood sugar, can lead to reduced insulin sensitivity, causing
Type 2 diabetes. Type 2 diabetes is the most common form of diabetes and is one of the leading
causes of death in the US. Researchers have performed many studies on the effects of dietary
fiber on diabetes control, some of which include the use of resistant starches. These studies,
however, are mostly conducted using uncommonly consumed resistant starches, such as green
banana, or utilize adding existing resistant starches to the development of new foods. This
research will explore the known development of resistant starch in cooked and cooled pasta, and
how the consumption of this pasta’s altered state affects postprandial blood glucose response.
This method is more accessible to the general public and fills a gap in the current published
research. This proposed research seeks to compare the difference between fasting and blood
glucose after consuming just-prepared pasta, to the difference in fasting and blood glucose after
consuming pasta that has been cooked, cooled, and reheated. Researchers would perform a
clinical trial, obtaining at least 300 participants to ensure a representative sample size of the
population of Orono, Maine. The participants would be attending two sessions within a 10-day
period, demanding 3 hours of their time in each session, with 6 hours in total. The two blood
glucose measurements would be compared and analyzed for significance using a One-Way
ANOVA. If found to be statistically significant, these results would further solidify the positive
effects of resistant starch on blood glucose response. Significant results would give the public a
simple way to aid in controlling blood glucose. Cooking pasta ahead of time, and reheating it
requires no additional money, and can be a resource for those with diabetes and lower
socioeconomic status.
Needs statement
In the current published research, there are a couple strategies utilized by researchers to

measure the affect of resistant starches on postprandial glucose levels and insulin sensitivity.

Gower et al. (2016) studied the effects of two different concentrations (15g and 30g) of high-

amylose maize resistant starch added to cookies on postprandial insulin response. The results

found that a 30g/day supplementation showed a statistically significant improvement in insulin

sensitivity among participants. In Dainty et al. (2016), researchers also used high-amylose maize

resistant starch, but this time utilizing it by replacing 60% of the wheat flour in a bagel. The

results found that the increased daily intake of high-amylose maize resistant starch significantly

lowered fast, 2-hour postprandial and 3-hour postprandial insulin sensitivity.

In terms of research on retrograde (RS-3) resistant starch’s effect on postprandial blood

glucose, the number of published studies is lacking. In 2021, Robertson et al. (2021) published

an introduction to their new research initiative analyzing the effects of retrograde resistant starch

(RS-3) created in cooked and cooled mashed potatoes on blood glucose. This is an exciting step

in filling the research gap found in RS-3 effects on postprandial blood glucose.

Filling this gap in research is invaluable due to the nature of the development and application

of this dietary fiber-type. Retrograde resistant starch (RS-3) is a much cheaper alternative to

buying high fiber versions of common carbohydrate sources. One pound of spaghetti in

Walmart’s Great Value brand (3g of fiber per serving) is $0.92 (price on 3/20/2022), versus one

pound of Barilla Whole Grain spaghetti (7g of fiber per serving), priced at $1.28 (price on

3/20/2022). This process is also much cheaper for companies to utilize in comparison to

additional ingredients, which drives up cost. Due to this decrease in cost, the availability of this

form of dietary fiber increases, especially to those with lower socioeconomic status.
Additionally, money is not the only savings using this process. Flavor is also maintained, which

is important when offering this tactic to the general population, as many are accustomed to the

flavor of regular white pasta and would likely not enjoy whole wheat pasta.

Research Goals

The goal of this research is to gain knowledge on the effect of retrograde resistant starch

on postprandial blood glucose. Utilizing the retrograde resistant starch (RS-3) found in pasta that

has been cooked and cooled, we hope to find a significant lowering of blood glucose after the

consumption of equal amounts of pasta that has been prepared and eaten hot, and pasta that has

been cooked, cooled, and reheated.

Literature review

Development of Retrograded Starch

As stated in Birt et al. (2013), there are five types of known resistant starches. Retrograded starch

is also known as RS-3 (resistant starch type 3) and occurs when a starch is cooked and cooled.

The cooking and cooling create a certain number of double helices that are less soluble and

cannot be digested, which classifies it as a dietary fiber. (Birt et al, 2013). The amount of these

resistant starches created during the retrogradation process varies from starch to starch.

Known Resistant Starch Effects on Blood Glucose

The creation of dietary fiber through retrogradation of starches slows digestion and increases

insulin sensitivity, which is important in treating and preventing diabetes (Wang et al., 2019).

Researchers in this field have performed tests for these effects in other types of resistant starches,

but very few have specifically used RS-3. One frequently used RS is RS-2, typically in the form
of high-amylose maize. One study found that a supplementation of 30 g/day of high-amylose

maize led to increased insulin sensitivity in their participants (Gower et al., 2016). This study

aims to prove the significant effect of RS-3 on blood glucose levels and fill the gap in research

pertaining to this type of resistant starch.

Research Design

Hypothesis

If participants consume pasta that has been cooked, cooled, and reheated, then they will

have a lower blood glucose than measured after consuming just-prepared pasta.

Methodology

Adults over 18 will be recruited using interest sheets posted at frequently visited areas in

Orono, Maine, as well as boards such as Facebook and Craigslist with the incentive of a $50 gift

card after completion. All responses will be screened for age (over 18), gluten intolerance/

allergy, digestive disorders, and metabolic disorders. A sample of around 300 participants over

the age of 18, with a 50% ± 5% gender split, no gluten intolerance/allergy, no history of

digestive disorders, and no history of metabolic disorder besides Type 2 Diabetes will be chosen

and notified on their participation. The sessions dates would be scheduled at time of notification.

Using the Lengyel gym at the University of Maine, two fasted groups will attend two

three-hour sessions within the three-month research period. During each session, all participants

will be monitored for two hours with a continuous glucose monitoring system (C-GMS). Upon

arriving, checking in and receiving the C-GMS, fasting glucose is measured and participants will

eat a serving (2 oz.) of just-prepared pasta within 20 minutes and are retained for 2 hours to

measure baseline glucose response. The participants then can leave with specific instructions to
not remove their C-GMS device until after their final session. In the next session, the participants

check in and their C-GMS is verified to be in good working order. Researchers again measure

their fasting glucose and participants will eat one serving (2 oz.) cooked, cooled, and reheated

pasta with 20 minutes. Participants are then retained for 2 hours to measure the RS-3 glucose

response. The C-GMS would then be removed, and the participants receive their $50 gift cards.

There will be 24 sessions in total with 25 participants per session (target of 300 participants) and

two sessions per participant.

Method of analysis

Upon completing the clinical research portion of this study, the statistical analysis will

begin. The change in glucose from fasting to the highest blood glucose in the two-hour window

after consuming just-prepared pasta will be compared to the change in glucose from fasting to

the highest blood glucose in the two-hour window after consuming cooked, cooled, and reheated

pasta using One-Way ANOVA. We also expect to complete a secondary analysis comparing this

data between diabetics and non-diabetics, utilizing a Pearson Correlation Coefficient.

Timeline

o September 2022 – Start recruiting

o January 2023 – End recruiting

o January 2023 - Begin testing

o April 2023 – Begin analyzing data

Expected outcome
 The expected outcome of this research is a statistically significant lowering of

postprandial blood glucose after consuming cooked, cooled, and reheated pasta.

Budget

 Indirect costs allotment - $5,000

 Materials and supplies

o Continuous blood glucose monitors (DEXCOM transmitters) - $25,000

- These devices can be purchased at wholesale for much cheaper than retail

value and are much easier to monitor than a classic stick meter.

o 4 tablets for researchers to monitor blood glucose monitors and record data. –

($200)

o 2 scales for measuring pasta servings – $400

o Alcohol Wipes - $100

o Sharps disposal bin and bags - $100

o Pasta - $200

 Salaries/fringe benefits

o $50 incentive for participants - $15,000

o Pay for 4 researchers – $1,000

Total: $50,000

Forms

 IRB approval

 Informed consent from participants

References

Birt, D. F., Boylston, T., Hendrich, S., Jane, J. L., Hollis, J., Li, L., McClelland, J., Moore, S.,

Phillips, G. J., Rowling, M., Schalinske, K., Scott, M. P., & Whitley, E. M. (2013).

Resistant starch: Promise for improving human health. Advances in nutrition (Bethesda,

Md.), 4(6), 587–601. https://doi.org/10.3945/an.113.004325

Dainty, S. A., Klingel, S. L., Pilkey, S. E., McDonald, E., McKeown, B., Emes, M. J., &

Duncan, A. M. (2016). Resistant starch bagels reduce fasting and postprandial insulin in

adults at risk of Type 2 Diabetes. The Journal of nutrition, 146(11), 2252–2259.

https://doi.org/10.3945/jn.116.239418

Gower, B. A., Bergman, R., Stefanovski, D., Darnell, B., Ovalle, F., Fisher, G., Sweatt, S. K.,

Resuehr, H. S., & Pelkman, C. (2016). Baseline insulin sensitivity affects response to

high-amylose maize resistant starch in women: a randomized, controlled trial. Nutrition

& metabolism, 13, 2. https://doi.org/10.1186/s12986-016-0062-5

Robertson, T. M., Brown, J. E., Fielding, B. A., Hovorka, R., & Robertson, M. D. (2021).

Resistant starch production and glucose release from pre-prepared chilled food: The

SPUD project. Nutrition bulletin, 46(1), 52–59. https://doi.org/10.1111/nbu.12476

Wang, Y., Chen, J., Song, Y. H., Zhao, R., Xia, L., Chen, Y., Cui, Y. P., Rao, Z. Y., Zhou, Y.,

Zhuang, W., & Wu, X. T. (2019). Effects of the resistant starch on glucose, insulin,

insulin resistance, and lipid parameters in overweight or obese adults: a systematic

review and meta-analysis. Nutrition & diabetes, 9(1), 19. https://doi.org/10.1038/s41387-

019-0086-9