Diet and the Colonic Exfoliome: A Novel, Non-Invasive Approach to Testing Interventions in Humans
Phase: NA
Conditions: Healthy Subject
Sponsor: Fred Hutchinson Cancer Center

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PROTOCOL 
Diet and the Colonic Exfoliome: A Novel, Non-invasive Approach to  
Testing Interventions in Humans 
R21 CA245456 
MPI: Drs. Johanna Lampe and Meredith Hullar (Fred Hutch) and Robert Chapkin (Texas A&M) 
 
TABLE OF CONTENTS 
1. Protocol Synopsis 
2. Background/Significance 
3. Specific Aims, Objectives/Endpoints 
4. Study Design 
5. Study Population/Recruitment 
6. Eligibility Criteria 
7. Key Measurements and Sample Handling 
8. Laboratory Procedures  
9. Data Analysis Plans 
10. Timeline 
11. Strengths and Limitations 
12. Anticipated Results, Alternative Approaches 
and Future Research  
13. Data Safety and Monitoring Plan 
Appendix 1 – Study supplements – Manufacturer 
Product Information 
Appendix 2 - Adverse Event Relatedness Reference 
Sheet 
                                                                                                                                                                                        
 
1. PROTOCOL SYNOPSIS 
Studies in animals have shown that both long-chain n-3 polyunsaturated fatty acids (PUFA) and a 
bacterial metabolite of dietary fiber (butyrate) may reduce colon tumor formation, and that the two in 
combination are even more effective than either alone. Several different mechanisms of action have 
been suggested. In humans, some epidemiologic studies suggest that people who consume high-fiber 
diets or use n-3 PUFA supplements may have a lower risk of colorectal cancer, but there are currently 
no controlled dietary interventions evaluating the combined effects of these two dietary constituents on 
colorectal cancer risk factors in humans.  We will test the combination of n-3 PUFA and fiber on gene 
expression in colon cells that are part of the pathways important to cancer risk and will seek to 
understand how variation in the gut bacterial community may affect response to intervention. 
 
Design: 
 
A randomized, placebo-controlled, double blinded, crossover dietary intervention 
Sample Size: Recruitment of 40 participants with the goal of having 30 complete study measures. 
Participants: Men and women 
Selection: 
Individuals from the Greater Seattle area who are normal to overweight (BMI of 18-30 
kg/m2) and between the ages of 50-75, who typically consume diets with 20g fiber or less 
per day. We will recruit U.S. racial minorities at an overall level of 32%, which mirrors the 
minority distribution in Seattle/King County 
Exclusion:  
The exclusion criteria will address a mix of medical and practical issues: 1) chronic 
medical illness, history of gastrointestinal disorders (e.g., ulcerative colitis, Crohn 
disease, celiac sprue, HNPCC, familial adenomatous polyposis, pancreatic disease, 
previous gastrointestinal resection, radiation or chemotherapy, and cancer (other than 
non-melanoma skin cancer); 2) weight change greater than 4.5 kg within past year; 3) 
oral or IV antibiotic use within the past 3 months; 4) regular use of aspirin or NSAIDs;  5) 
smoking; 6) known allergy to fish; 7) intention to relocate out of study area within next 4 
months; 8) abnormal liver/kidney function as ascertained at a baseline blood draw. 
Intervention: a) Placebo and b) Supplemental soluble fiber (35 g/d) + supplemental n-3 PUFA (6 g/d    
eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). 
IRO REC'D OCT 04 2022
FHCRC IRB Approval  
10/20/2022 
Document Released Date

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2. BACKGROUND/SIGNIFICANCE 
Diet is an important risk factor for colorectal cancer (CRC) and several dietary constituents implicated in 
CRC are modified by gut microbial metabolism. Microbial fermentation of dietary fiber produces short 
chain fatty acids (SCFA), e.g., acetate, propionate, and butyrate. Dietary fiber has been shown to reduce 
colon tumors in animal models and, in vitro, butyrate influences cellular pathways important to cancer 
risk. Further, work from our group suggests that the combined effects of butyrate and long chain omega-
3 polyunsaturated fatty acids (n-3 PUFA) found in cold-water, oily fish, may enhance the 
chemopreventive potential of these dietary constituents. We postulate that the relatively low intakes of n-
3 PUFA and fiber in Western populations and the failure to address interactions between these dietary 
components may explain why chemoprotective effects of n-3 PUFA and fermentable fibers have not 
been detected consistently in prospective cohort studies. We hypothesize that the combined effects of n-
3 PUFA supplementation and supplemental highly fermentable fiber will alter critical pathways important 
to CRC prevention, particularly intrinsic mitochondrial-mediated programmed cell death resulting from 
the accumulation of lipid reactive oxygen species (ferroptosis), cell proliferation, and the 
eicosanoid/inflammation pathways.  
Gut bacteria ferment dietary fiber to butyrate and other SCFA.1-14 Butyrate is a potent histone 
deacetylase inhibitor15, 16 associated with reduced CRC risk.17-19 Work from our group using preclinical 
models suggests that combined effects of fiber-derived butyrate and n-3 PUFA in fish oils act 
synergistically to enhance the chemopreventive potential above and beyond the contribution of butyrate 
alone, in part, by altering the glutathione peroxidase-4 (GPX-4)-dependent redox environment in the cell 
and increasing programmed cell death (ferroptosis).20-30   These results agree with a recent prospective, 
nested case-control study demonstrating significant CRC risk reduction among pescovegetarians, 
whose diets were high in both dietary fiber and n-3 PUFA-rich fish (HR: 0.57; 95% CI: 0.40, 0.82).31  We 
hypothesize that the combined effects of 35 g supplemental fiber, high in soluble fibers, and 6 g 
supplemental long chain n-3 PUFA (DHA + EPA) will alter pathways important to CRC prevention.  
This is a randomized, controlled crossover pilot study in 40 healthy men and women (50-75 y) to 
compare supplemental soluble fiber (35 g/d) + supplemental n-3 PUFA (6 g/d EPA+DHA), in quantities 
mirroring mean daily intakes associated with lower CRC risk, with a maltodextrin and corn oil control 
(placebo). Stool samples will be collected at the beginning, middle (day 15), and end of each of the two 
30-day intervention periods. Using a novel, cost-effective, non-invasive approach, we will evaluate 
differences in global gene expression signatures in the stool exfoliome (i.e., sloughed colonic epithelial 
cells in stool) using RNA-Seq.32  We will focus on pathways related to colonic cell proliferation and 
apoptosis/ferroptosis, cell phenotype, and inflammatory response. Further, we will evaluate changes in 
gut microbial functional genes involved in butyrate production using droplet digital PCR (ddPCR). The 
collection of multiple samples over the intervention periods will provide a critical measure of longitudinal 
changes in response to the supplemental n-3 PUFA and fermentable fiber.   
This pilot will be the first in humans to integrate and characterize the relationships between n-3 PUFA 
and fermentable fiber exposure that modulate programmed cell death and other CRC-related cell-
signaling pathways through metabolic activities of the gut microbiome. Results of this controlled 
intervention will help to translate the current mechanistic knowledge from preclinical animal models to 
humans and to inform approaches for CRC prevention. Interrogating the stool exfoliome is a novel, cost-
effective, non-invasive approach to studying effects of interventions on the human gut.  
3. SPECIFIC AIMS, OBJECTIVES, ENDPOINTS: 
Aim 1: To test in a pilot feasibility study, in humans, effects of supplemental dietary fiber (35 g) and 
EPA+DHA (6 g) vs placebo on mRNA expression in exfoliated colonic cells. We hypothesize that fiber 
and n-3 PUFA supplementation will uniquely influence colonocyte proliferation and programmed cell 
death-related gene expression and signaling pathways. 
Aim 2: To test whether response to the intervention differs by gut microbial functional capacity related to 
fiber fermentation. We hypothesize that changes in colonocyte gene expression will be modified by the 

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gut microbial capacity to ferment dietary fiber (i.e., SCFA-producing pathways), measured as 
butyrogenic potentialEXW\U\O-CoA:acetate CoA-transferase (but) + butyrate kinase (buk) genes)/(total 
bacterial load 16S rRNA gene) assayed using digital drop PCR. 
Objectives and Endpoints:  
1. To determine whether the combination of supplemental dietary fiber and EPA+DHA vs placebo 
affects mRNA expression in exfoliated colonic cells. The endpoint is mRNA expression profiles in 
the exfoliome. 
2. To determine whether gut microbial functional capacity related to fiber fermentation, as measured 
by presence of fiber-fermenting genes in stool bacteria, alters exfoliome mRNA expression 
response to the intervention on. The endpoint is mRNA expression profiles in the exfoliome. 
Implications: This proposed pilot feasibility study will be the first controlled study in humans to integrate 
and characterize the relationships between n-3 PUFA and prebiotic fiber exposure that modulate 
programmed cell death/CRC-related cell-signaling pathways through changes in the gut microbiome. 
Results of this human mechanistic controlled intervention will help to translate the current mechanistic 
knowledge from preclinical animal models to humans and to inform approaches for CRC prevention. 
Also, importantly, we will further evaluate the utility of interrogating the exfoliome as a novel, non-
invasive approach to studying effects of interventions on the human gut.  
4. STUDY DESIGN 
The study is designed as a randomized, cross-over, placebo-controlled dietary intervention of 
supplemental soluble fiber and supplemental n-3 PUFA compared to corn oil and maltodextrin control.  
We will randomize participants to one of the two treatment orders, either supplemental fiber and fat first 
followed by the placebo in the second period or the placebo first followed by supplemental fiber and fat.  
Each treatment will be 30 days in duration with at least a 60-day washout period between treatments.  
Stool samples will be collected at the beginning, middle (day 15) and end of each of the two 30-day 
intervention periods.  
 
 
 

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It will take participants at least 4 months to complete the trial. Details of the activities and timeline:
4.a. Fiber and Fat Supplements The active treatment period will include supplemental soluble fiber (35 
g/d) + supplemental n-3 PUFA (6 g/d EPA+DHA).  
Supplemental Fiber: The soluble fiber with known fermentation profiles will be provided by Tate & Lyle, a 
well-established food and beverage ingredient company.  For placebo, we will use maltodextrin provided 
in the same packaging and by the same company. 
Supplemental n-3 PUFA: We will use a commercial fish oil supplement (Omega Cure, Omega3 
Innovations), consumed as 2 vials daily. Each vial (11.75 g oil) delivers 3 g EPA +DHA in a 1:1 ratio. 
For placebo, we will use equivalent amounts of corn oil in visually identical packaging provided and by 
the same company.
All study supplements will be in packaged as daily doses.  At the study visits at the beginning and mid-
point of each study period, participants will receive a box with all the supplements needed to cover the 
time in between visits along with instructions regarding proper storage conditions (ie. the fish oil is to be 
refrigerated for better preservation.)  
4.b. Safety and Adverse Effects  Supplemental fiber (30-45 g/d) has been well-tolerated in short and 
long-term interventions (4 wks to 1-10 y).33-36 In a study specifically designed to evaluate tolerance, 
individuals were given 10, 30 and 60 g/d of supplemental fiber for 7 d in addition to their regular diet. All 
doses were well-tolerated.37 Further, the specific product we propose to use, PROMITOR® Soluble Corn 
Fibre (SCF), has been well tolerated by healthy volunteers at 65 g/d.38, 39 SCF, a glucose 
ROLJRVDFFKDULGHFRQWDLQLQJDPL[WXUHRIĮ-6, 1-4, 1-3, and 1-2 glucosidic linkages, has demonstrated 
prebiotic effects.40, 41 Testing of SCF in an in vitro SHIME method that mimics human digestion showed 
that SCF is well fermented in the distal colon leading to positive effects on the GMC, and that these 
effects are translated to immunomodulatory properties and improved gut barrier.42 Long-term trials (6-27 
months) designed to evaluate safety of supplemental n-3 PUFA up to 7 g/d have reported no adverse in 
general, and in bleeding complications specifically, even among patients at higher risk,43, 44  There is 
insufficient data to establish a tolerable upper intake for n-3 PUFA for any population group.45
4.c. Length of intervention periods and washout: Description and rationale.  Intervention Periods:  
Crypt turnover is 3.5 d and 4.2 d for the colon and rectum, respectively46 and dietary n-3 PUFA are 

Fat x Fiber Protocol  Version 1    p. 5 of 18 
incorporated into colonic cell membranes within 7 d.47 1-mo interventions with diet or aspirin result in 
measurable changes in colonocyte gene expression48 and down-stream colon biomarkers.49 Gut 
microbiome changes occur in 2-3 wks.3, 50, 51 Changes in gut mucosal cancer-risk biomarkers, the 
microbiome, and microbial metabolites were also seen in a short-term 2-wk intervention of fat and 
fiber.52 We hypothesize that changes in the gut microbiome influence colonic gene expression, thus 
conservatively we are using a 30-d period to allow sufficient time for both outcomes to change, while 
considering participant burden. Washout: The 60-d washout will ensure that residual intervention effects 
from the previous period are eliminated. As mentioned, the gut epithelium and microbiome change 
rapidly, but blood DHA+EPA clearance requires 45 d.53 
4.d. Monitoring and assessing adherence. Participants will meet with study staff routinely to assess 
any difficulties with study adherence. n-3 PUFA are rapidly incorporated into colonic tissue.54, 55 and 
there is high correlation between membrane PUFA and plasma phospholipid (PLFA) PUFA.56, 57  We will 
assess adherence to n-3 PUFA supplementation using plasma PLFA profiles (Section C.4.a) and by 
monitoring vial counts, calculating adherence % (vials supplied - vials returned/days elapsed).58 
Adherence to supplemental fiber will be monitored by study staff. Participants will be instructed to return 
unconsumed fiber packets for weighing and consideration in intake determination. During the 
intervention, participants will complete daily forms to monitor adherence to the supplements.32   
4.e. Summary of study activities and measures  
5. STUDY POPULATION/RECRUITMENT 
Participants will be healthy, non-smoking men and women, ages 50-75 y, who are normal to overweight 
>ERG\PDVVLQGH[%0,18 DQGNJP2]. Women will be postmenopausal, with no menstrual period 
in 12 months.   We will place ads in newspapers, distribute flyers, and use social media. A screening 
survey will determine eligibility (age, health, medications, habitual fiber intake); individuals will meet with 
the study coordinator for an info session and to obtain consent. We will measure height and weight (for 
BMI) and draw blood (Complete Blood Count (CBC) and Comprehensive Metabolic Panel) analyzed by 
Quest Diagnostics, a CLIA certified lab) will further determine eligibility. 
Study information will be available online. The screening questionnaire will be downloadable. Interested 
persons will fill it out and send it via mail. A contact phone number and email address will also be 
provided for individuals who want to be mailed an information packet. Interested persons will return the 
completed questionnaire to the study staff, who will determine eligibility and will contact eligible persons 
Table 1. Study Activities and Measures 
Reason for Measures 
Screening 
Screening questionnaire 
Exclusion criteria 
Information session 
Informed consent 
Baseline blood sample to Quest Diagnostics 
Exclusion criteria (liver and kidney functions) 
Health and Demographics questionnaire 
Exclusion criteria 
Anthropometrics 
Exclusion criteria 
Two Supplementation Periods 
Each 30 days each 
Cross-over (order randomly assigned) * 
Stool samples (6 total) Beginning, middle and end of each period 
For colonocytes 
Global gene expression signatures in the exfoliome (using RNA-Seq) 
For Microbial  
Microbial DNA extraction, ddPCR and SCFA. 
    Bowel Habits & General Symptoms questionnaire  
Intervention effects 
Blood samples (4 total) Baseline & end of each period 
Plasma phospholipid PUFA 
Weight, Baseline & end of each period 
Monitor biological changes 
Daily calendar 
Compliance 
*  In between feeding periods there will be at least 60 days washout period. 

Fat x Fiber Protocol  Version 1    p. 6 of 18 
by phone or email to schedule a meeting. A packet containing a confirmation letter and the screening 
study consent form will be sent to the person prior to attending the meeting. Participants eligible to enter 
the intervention will meet with our staff to discuss all the details and expectations of the study, review 
informed consent and answer any questions they might have, and if still interested sign the consent form.  
6. ELIGIBILITY CRITERIA 
Inclusion:         Healthy men and women aged 50-75 years who are normal-overweight (BMI of 18-30 
kg/m2), non-smoking and consume fiber intakes of less than <20 g/d. 
Exclusion:   
 
1. medical conditions: hypertension requiring medication; chronic liver disease (cirrhosis, hepatitis); 
kidney dysfunction or disease; previous resection of stomach or intestines; history of 
gastrointestinal disorders: colorectal cancer or other gastrointestinalcancer, polyposis syndromes, 
inflammatory bowel disease (ulcerative colitis, Crohn disease), intestinal blockage requiring 
surgery, pancreatic disease,  radiation treatment to the abdomen within past 5 years;  cancer 
(other than non-melanoma skin cancer);   
2. regular use (more than once per week) of: NSAIDs (aspirin, ibuprofen, naproxen), Coumadin 
(warfarin), Eliquis (apixaban), Plaviz (clopidogrel), Xarelto (rivaroxaban), Lovaza™ (prescription 
fish oil), blood clotting medications; 
3. weight change greater than 4.5 kg within past year;  
4. oral or IV antibiotic use within the past 3 months;  
5. consumption of three or more alcoholic beverages a day; 
6. smoking or use of cannabis products within the past year;  
7. known allergy to fish or fish-containing products; allergy to Vitamin E analogues 
8. intention to relocate out of study area within next 4 months;  
9. abnormal clinical lab tests as ascertained at a baseline blood draw: 
*We will provide participants with a copy of their lab results if these are out of the normal range – 
which will make them ineligible for the study.  We call them with this information prior to sending 
them the results via mail. We will encourage them to contact their physician.   
7. SPECIMEN AND DATA COLLECTION 
7.a. Blood We will obtain 10-hour fasting blood samples at the beginning and at the end of each 
intervention period.  The first sample will be sent to QUEST Diagnostics (a CLIA certified lab) for clinical 
chemistries (to confirm eligibility; see Section 6 for specifics).  Participants will come to the Prevention 
Center at the Fred Hutch where certified personnel will collect four 10-ml vacutainers of blood; for 
plasma and serum.  Specimens will be stored at -80ºC.    
7.b. Stool samples at three timepoints in each intervention arm, beginning, middle and end; for : a) 
exfoliated colonocyte transcriptome (mRNA) analysis, and b) fecal microbiome analyses. Participants 
Ineligible if results fall outside of the following parameters based on Complete Blood 
Count (CBC) and Comprehensive Metabolic Panel performed by Quest Diagnostics 
(a CLIA certified lab).* Blood from the baseline fasting blood draw will be sent to 
Quest with Study IDs not names.  Parameters: 
White blood cell count  3,000-11,000/mm3    
Platelet count  150,000-400,000 mm3     
 
 
Bilirubin 0.2-1.3 mg/dL    
 
 
 
AST  0-35 U/L   
 
 
 
 
 
ALT 0-40 U/L   
 
 
 
 
Alkaline phosphatase 20-125 U/L 
 
 
&UHDWLQLQHPJG/  
 
 
Potassium 3.5-5.0 mmol/L   

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will collect small samples of stool into vials we provide that are prefilled with RNAlater for microbial 
DNA,59 RNAshield for exfoliated colonocyte transcriptomics,60  and in 95% ethanol for SCFA and future 
metabolomics measures.61 Human transcriptome profiles are stable for at least 60 h at 4°C prior to 
addition of RNAshield solution and once RNAshield solution is added, at -80°C for at least 2 yrs.48, 60 
7.c. Questionnaires: Dietary Assessment  We will use an established, validated food frequency 
questionnaire (FFQ) to characterize food and nutrient intakes of the study participants over the 3 months 
prior to baseline to characterize habitual diets (typically low in n-3 PUFA and fiber and high in n-6 
PUFA).    Health and Demographics will also be collected. A General Symptoms and Bowel Habits 
Questionnaire will also be provided every time participants collect stool- this is a useful tool to 
determine intervention effects.  
8. LABORATORY PROCEDURES 
8.a. Plasma n-3 PUFA Assays  To confirm adherence to n-3 PUFA exposure, we will isolate total 
phospholipids from plasma collected on days 0 and 30, trans-esterify, and measure EPA and DHA by 
GC/MS.62 This work will be performed by our Co-Investigator Dr. Robert Chapkin at Texas A&M. 
8.b. Host Gene Expression Using RNA-Seq  Poly A+ RNA will be isolated for exfoliated cells in stool 
as we describe.48, 60, 63 An Agilent 2100 Bioanalyzer will be used to assess integrity of fecal poly A+ RNA  
as we described.64, 65  This work will be performed by MPI, Dr. Robert Chapkin at Texas A&M. 
8.c. Fecal Microbiome Analysis Using ddPCR  We will extract genomic DNA from stool samples and 
use the QX200 ddPCR system (Bio-Rad) to quantify bacterial genes from the two major butyrate 
synthetic pathways, butyryl-CoA:acetate CoA-transferase (but) and butyrate kinase (buk) and total 
bacterial load (16S rRNA genes) according to the manufacturer’s instructions, and primer and probe set 
optimized for the selected genes.66-70 We have chosen this assay because it is more reliable, accurate, 
precise, and less prone to inhibition than qPCR. Butyrogenic potential of the microbiome will be defined 
DVEXW\U\O-CoA:acetate CoA-transferase (but) + butyrate kinase (buk) genes)/(total bacterial load 16S 
rRNA gene). 66-69 This work will be performed in the JLampe lab at Fred Hutch. 
9. DATA ANALYSIS 
9.a. Data entry and quality assurance procedures. We will code and enter data from questionnaires, 
tracking forms, etc., and integrate with laboratory data, and monitor adherence to protocols for data 
entry, sample processing, storage, and analysis.   
9.b. Aims testing, Overview: In Aims 1 and 2 our analyses will investigate the response to the fiber 
and n-3 PUFA intervention at levels of a) individual genes or variables and b) pathways and 
networks.  At the individual marker level (a) we will test genes and the butyrogenic potential (Aim 1 and 
2) for differences due to intervention using univariate models, adjusting for appropriate covariates. At the 
pathway/network level (b), analysis will focus on hypothesized CRC pathways and employ gene-set 
(“global”) enrichment analysis tests to evaluate changes in host gene pathways (Aim 1). When prior 
genetic/metabolic network structure is known within a pathway, the analysis will account for this.  
Models: a) For single-gene/variable analyses (Aims 1 and 2), we use linear mixed models with a 
random effect for participant to account for the repeated measures of marker y during and at the end of 
each intervention. Fixed effects will include an indicator, I, for intervention as well as demographic 
covariates (e.g., age, sex, BMI) which will be tested by randomization and included where appropriate. 
Although randomization to intervention sequence should alleviate the potential for differential carryover 
effects between periods, models will include an indicator S RIUDQGRPL]DWLRQVHTXHQFH(UURUWHUPVࣅik 
are assumed to be correlated within, and independent between, participants. In summary, models take 
the form yik = Di + Iik + Si İik, where Di is a random effect for subject. Robust standard errors will be used 
to test for differential effect of intervention, and all p values will be corrected for multiple testing by 
controlling the FDR using Benjamini-Hochberg71.  b) For pathways, we will apply kernel association 
testing (KAT) procedures.72, 73 These multivariate methods allow for pathway-wide testing and modeling 
in a manner which extends gene-set style tests. They incorporate adjustment for covariates and 
generalize tools such as the globalTest,74 used in genomics studies, and perMANOVA,75-77 common to 
microbiome analyses. Network or structural relationships (e.g., metabolic) between genes in a pathway 

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are easily incorporated into construction of a similarity/kernel matrix on which KATs are based72, 78. 
These known (or estimated) interactions can enhance the KAT testing procedure to increase power.79, 80 
Similar methods analyzing gene-gene networks have been successful in crossover dietary studies.81 We 
will account for the repeated-measures aspect of the crossover design in these models by applying a 
variant of family-based KAT.82, 83 Finally, longitudinal data collected on days 0, 15, and 30 of each period 
will be analyzed using a linear mixed model for each measure, as above.  Terms for time and effect 
modification by intervention (time*intervention) will be added to this model if the data support 
autocorrelation within a measure across time. For longitudinal analyses of multivariate data (i.e., 
pathways/ gene-sets), we will test for longitudinal differences due to intervention by way of a repeated-
measures KAT82, 83 and a variant of GSA.84 We will consider effects of butyrogenic potential (defined as 
above and below the median; Aim 2) on host gene expression responses (Aim 1). 
9.c. Power and sample size. This is a randomized, controlled crossover pilot feasibility study in 30 
healthy men and women, designed to generate preliminary data to inform a larger human mechanistic 
study. Based on our previous experience, we are confident we will detect gene expression changes with 
the fiber + n-3 PUFA intervention. Aim 1: In our FlaxFx study32 with a similar crossover design, we 
detected differential expression of 32 genes between placebo and lignan extract (FDR<0.05) in 
exfoliated cells (n=42 participants). Given the impact of dietary fiber on multiple pathways in animal 
models,85 we anticipate similar or better power in the current study. In addition, we also considered 
power for single-gene analysis in this crossover study (n=30). Using a 2-sided paired t-test at 
significance level alpha=0.05 and a range of effect sizes, conservative estimates are: power of at least 
80% for effect sizes larger than 0.5.  Aim 2: a power estimate for ddPCR data, based on differences in 
but gene abundance with low- (4.96*1011 ± 3.22*1010 copies/g feces) and high-fiber diets (1.37*1012 
±1.47*1011) in healthy individuals (n=30),86 at significance level alpha=0.05 and effect sizes larger than 
0.8 , we estimate power at least 95 % to detect diet-related differences in this gene. We anticipate a 
similar distribution for buk.  
 
10. PROJECT TIMELINE 
 
 
11. STRENGTHS AND LIMITATIONS OF PROJECT   
The strengths are: (i) translation and testing of a hypothesis in humans that has been well characterized 
in animal models and is supported by cogent mechanistic evidence; (ii) a robust, randomized crossover 
study design with within-person pairwise comparison; (iii) feeding of n-3 PUFA and prebiotic dietary 
fiber40, 41 at levels relevant to CRC prevention; (iv) pathway-based evaluation; (v) novel, non-invasive 
approach to measure gene expression using the stool exfoliome; (vi) targeted measures of butyrogenic 
potential; and (vii) extensive experience and expertise of our research team in all aspects of the 
proposed work.  Limitations relate to the inherent challenges of short-term human dietary interventions 
without disease as an outcome and the R21 budget constraints. The latter limits the sample size, 
number of treatments (e.g., no test of n-3 PUFA and fiber individually), and comparison of exfoliome and 
biopsy gene expression. However, our cost-effective, robust study design, relevant dosing, and 
pathway-related outcomes will inform the utility of further work in this area. Further, we focus on 

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pathways that show, in animal models, that fish oil/fermentable fiber-containing diets protect against 
colon cancer by up-regulating ferroptosis/apoptosis and suppressing proliferation and inflammatory 
mediators.21, 23, 24, 87-89 
 
12. ANTICIPATED RESULTS, ALTERNATIVE APPROACHES AND FUTURE RESEARCH  
We anticipate generating strong pilot data on the utility of n-3 PUFA + fermentable fiber to affect colonic 
pathways relevant to CRC prevention. With our experience in conducting human intervention studies 
and analyzing the proposed specimens, we do not anticipate any difficulties completing this small, pilot 
intervention.  Furthermore, an understanding of the inter-individual differences in butyrate and n-3 PUFA 
disposition may inform development of tailored diet strategies for CRC-risk reduction. Specimens 
collected as part of this study will also be stored for additional microbiome and metabolomic analysis as 
future funding permits. 
 
13. DATA SAFETY AND MONITORING PLAN 
Oversight for this study will be provided by the Principal Investigators, Drs. Johanna Lampe and 
Meredith Hullar (Fred Hutch) and Robert Chapkin (Texas A&M), with delegation of responsibilities to 
designated study personnel. They will ensure all entry criteria are met prior to the initiation of the 
protocol, and all study procedures and reporting of adverse events are performed according to the IRB-
approved protocol.    
Dr. Rachel Issaka, a board certified gastroenterologist at SCCA and UW Medical Center and a health 
services researcher with the Fred Hutchinson Institute for Cancer Outcomes Research will serve as 
medical advisor for the study.  She will be available for consult in adverse events cases.        
All adverse events related to the study procedures will be fully documented on the appropriate case 
report form(s) and entered in a study database.  For each adverse event, the investigator will provide the 
onset, duration, intensity, treatment required, and outcome, including documentation of need for 
premature termination of any study procedures. 
Anticipated adverse events related to this study are: 
Blood Draws Participants may experience a little discomfort or have a temporary bruise from having 
blood drawn.  Occasionally a participant may feel lightheaded or feel faint when having blood drawn.  
Blood will be taken only by trained personnel in the Fred Hutch Prevention Center Shared Resource 
Research Clinic using established procedures.  The participants will be sitting down when the blood is 
drawn.  If a participant feels faint, they will be instructed to lie down until the feeling goes away.   
Dietary Supplements We will provide two dietary supplements during the active intervention and similar 
controls during the placebo periods.  All participants will go through both intervention periods.  Fiber:  
~35 g highly-soluble, well-tolerated commercial fibers. Supplemental fiber (30-45 g/d) has been well-
tolerated in short- and long-term interventions (4 wks to 1-10 y).  Control:  We will use maltodextrin (a 
food-ingredient polysaccharide made from partially hydrolysed starch) provided in the same packaging.  
Mild bloating and abdominal distension might occur while on the fiber treatment since the fiber content 
will be higher than their habitual diet.  Participants will be monitored by the study staff.  Most people 
adapt to the higher fiber intake within 3-5 days.  Fish Oil: n-3 PUFA: We will provide a daily dose of 6 g 
EPA+DHA, consumed as 2 vials daily for 30 d. Control: We will use equivalent amounts of corn oil in 
visually identical packaging. Although generally safe, high amounts of EPA and DHA may thin the blood 
(increase clotting time) and slightly lower blood pressure. Fish oil may cause belching, bad breath, 
heartburn, nausea, loose stools.  Participant will be instructed to report all symptoms.  Study staff will 
contact participants after the first week on each study period to ascertain compliance and ask about any 
concerns they may have.  Study staff will see the participants after 2 weeks provide them the needed 
supplements and will, again, ascertain compliance and ask about any concerns they may have.  We 
also will maintain text and email contact with them.  They fill out a daily form that includes space for 
commenting about symptoms or issues; thus, we are able to address the issues as they present 
themselves and are able to suggest ways to ways to minimize these effects.  Fish Oil - Risk of allergic 

Fat x Fiber Protocol  Version 1    p. 10 of 18 
reaction will be minimized through eligibility questions addressing history of medication, food allergies / 
intolerances. Taking fish oil supplements with meals can often decrease gastrointestinal side effects. 
Participants will keep a daily study log which will include questions on health status and possible 
adverse experiences. A general symptom questionnaire will be administered at baseline and each 
subsequent visit.  If there are any indications that a participant has a food allergy that was not previously 
known, we immediately have them stop the intervention.  The study coordinator has an emergency cell 
phone and can be reached 24/7 by study participants.  
Adverse event grading scale, based on Common Terminology Criteria for Adverse Events (CTCAE) v5.0 
(Publish Date: November 27, 2017):  
Grades Grade refers to the severity of the AE. The CTCAE displays Grades 1 through 5 with unique 
clinical descriptions of severity for each AE based on this general guideline:  
Grade 1 Mild; asymptomatic or mild symptoms; clinical or diagnostic observations only; intervention 
not indicated.  
Grade 2 Moderate; minimal, local or noninvasive intervention indicated; limiting age appropriate 
instrumental ADL*.  
Grade 3 Severe or medically significant but not immediately life-threatening; hospitalization or 
prolongation of hospitalization indicated; disabling; limiting self care ADL**.  
Grade 4 Life-threatening consequences; urgent intervention indicated.  
Grade 5 Death related to AE. 
Activities of Daily Living (ADL) *Instrumental ADL refer to preparing meals, shopping for groceries 
or clothes, using the telephone, managing money, etc. **Self care ADL refer to bathing, dressing 
and undressing, feeding self, using the toilet, taking medications, and not bedridden. 
Plan for unanticipated AE reporting: All unanticipated AEs related to the study procedures that are 
severe or serious will be reported by Dr. Johanna Lampe the IRB within 10 days of notification of the 
investigator.  We will use as a guide the Adverse Events Relatedness Reference Sheet (Cancer 
Consortium Clinical Research Support) see Appendix 2.  And we will follow:   
Hutch IRB Policy 2.6 
Adverse Biomedical Events 
When, in the opinion of the principal investigator, or PI, any adverse event meets all three of the 
following criteria, whether they occur on- or off-site, they must be submitted to the IRB within 10 
calendar days. 
x Unexpected: An adverse event is unexpected when its nature (specificity), severity or frequency 
are not consistent with (a) the known or foreseeable risk of adverse events associated with the 
research procedures described in the protocol-related documents, such as the IRB-approved 
research protocol, informed consent document and other relevant sources of information such 
as product labeling and package inserts; and are also not consistent with (b) the characteristics 
of the subject population being studied, including the expected natural progression of any 
underlying disease, disorder or condition, or any predisposing risk-factor profile for the adverse 
event. 
x Related or possibly related to the research: An adverse event is related or possibly related to 
research procedures if, in the opinion of the PI, it was more likely than not caused by the 
research procedures. Adverse events that are solely caused by an underlying disease, disorder 
or condition or by other circumstances unrelated to the research or any underlying disease, 
disorder or condition are not "related or possibly related." If there is any question whether an 
adverse event is related or possibly related to the research, report the adverse event. 
Serious: An adverse event is serious when it results in any of the following outcomes: death, risk of 
death, inpatient hospitalization or prolongation of existing hospitalization, a persistent or significant 
disability/incapacity/or change in psychosocial status, a congenital anomaly or the requirement of an 

Fat x Fiber Protocol  Version 1    p. 11 of 18 
intervention to prevent permanent impairment or damage.  Plan for anticipated AE reporting: All 
serious anticipated AEs related to the study procedures will be reported by Dr. Johanna Lampe to the 
IRB within 10 days of notification of the investigator.  
Plan for ongoing review of results: The PIs will be notified within 24 hours by the research manager 
of any early terminations due to an adverse event.  
Plan for safety review: the PI will perform a cumulative review of all adverse events and premature 
terminations review every 6 months after study initiation or after completion of 50% of participant visits, 
whichever occurs first.  
Plan for annual reporting: A summary of the investigation including all adverse events and how they 
were handled, enrollment, drop-outs and reason for discontinuation and any protocol modifications will 
be provided to the IRB on an annual basis.  
Annual Reports will contain:  
x 
The number of adverse events and an explanation of how each event was handled  
x 
The number of complaints and how each complaint was handled  
x 
The number of subject withdrawals and an explanation of why the subject withdrew or was 
withdrawn  
x 
The number of protocol deviations and how each was handled  
The occurrence of any serious and unexpected event may prompt changes in study protocol. Any such 
change will be approved by the IRB before implementation.   
Monitoring for data integrity and safety will be the responsibility of all the investigators. The following 
will be included:  
Validity and integrity of data: Data are checked for missing, unusual, or impossible records when 
they are entered into the study’s computer database.  The investigators may consider modifications 
to the data-collection protocol with permission from IRB if they deem the change(s) is necessary to 
ensure the validity and integrity of data collected. 
Enrollment rate relative to expectation: Early lags in recruitment will be rectified with increased 
recruitment efforts so that recruitment will be completed on time.  The investigators will monitor this 
closely to ensure full enrollment of appropriate participants.   
Retention of participants and adherence to protocol: The investigators will monitor retention and 
adherence to study protocol. Adherence to the study supplements will be monitored via the Daily 
Record that includes a calendar where participants mark daily capsules consumed.  
Oversight for this study will be provided by Dr. Johanna Lampe (corresponding MPI on this grant) with 
delegation of responsibilities to the Study Manager, Lisa Levy. They will ensure all entry criteria are met 
prior to the initiation of the protocol, and all study procedures and reporting of adverse events are 
performed according to the IRB-approved protocol.  All adverse events related to the study procedures 
will be fully documented on the appropriate case report form(s) and entered in a study database.  For 
each adverse event, the investigator will provide the onset, duration, intensity, treatment required, and 
outcome, including documentation of need for premature termination of any study procedures. 
 
 
 

Fat x Fiber Protocol  Version 1    p. 12 of 18 
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Hippe B, Zwielehner J, Liszt K, Lassl C, Unger F, Haslberger AG. Quantification of butyryl CoA:acetate 
CoA-transferase genes reveals different butyrate production capacity in individuals according to diet and 
age. FEMS Microbiol Lett 2011;316:130-5. 
87. 
Coussens LM, Zitvogel L, Palucka AK. Neutralizing tumor-promoting chronic inflammation: a magic bullet? 
Science 2013;339:286-91. 
88. 
Tabbaa M, Golubic M, Roizen MF, Bernstein AM. Docosahexaenoic acid, inflammation, and bacterial 
dysbiosis in relation to periodontal disease, inflammatory bowel disease, and the metabolic syndrome. 
Nutrients 2013;5:3299-310. 

Fat x Fiber Protocol  Version 1    p. 16 of 18 
89. 
Monk JM, Turk HF, Fan YY, Callaway E, Weeks B, Yang P, McMurray DN, Chapkin RS. Antagonizing 
arachidonic acid-derived eicosanoids reduces inflammatory Th17 and Th1 cell-mediated inflammation and 
colitis severity. Mediators Inflamm 2014;2014:917149. PMC4127240. 
 
 
 

Fat x Fiber Protocol  Version 1    p. 17 of 18 
APPENDIX 1   
Study supplements – Manufacturer Product Information  

 
 
 
 
 
PROMITOR™ Soluble Corn Fiber 85A 
Specification Sheet 
Page 1 of 6 
 
Version 1.8 (Date of Last Change: 09.06.2016) 
Date Printed: 12/21/2016 
 
 
Specification Sheet 
300000001508 
    
 
PROMITOR™ Soluble Corn Fiber 85A 
1810012115021, 1810012115003, 18100121 
 
    
 
 
 
Product Description : PROMITOR™ Soluble Corn Fiber is a versatile soluble fiber that imparts no 
color or off flavor in a wide variety of food and beverage applications. 
 
Appearance: Powder 
Color: Off-White 
Odor: Starch 
 
Solubility Statement: Soluble In Water 
 
Quality Inspection Plan (Official Specification Values) 
 
Parameter 
Target 
Range/ 
Attribute 
Unit 
Method 
SEDIMENT 
 
<= 16 
ppm 
Sediment - TN70570 
COLOR 
 
<= 2.5 
 
Color - TN22505 
MOISTURE 
 
<= 6.00 
% 
Moisture - TN46015 
pH 
 
3.5 - 5.5 
 
pH - TN60520 
FIBER 
 
>= 85.0 
%ds 
Repolymerized Compounds - 
TN65645 
SCREEN ON US#20 
 
<= 10.00 
% 
Screen Size - TN69700 
SCREEN ON US#100 
 
>= 60.00 
% 
Screen Size - TN69700 
SUGARS, AOAC 
 
<= 2.0 
% 
Sugars, AOAC - TN67475 
DENSITY 
 
20.00 - 24.00 
lb/scf 
Bulk Density - TN24551 
SOLUBILITY 
 
<= 30.0 
s 
Solubility - TN76000 
BACTER TOTAL 
 
<= 1000 
n/g 
Total Plate Count - TN10560 
OSMOPHYLIC YEAST 
 
<= 50 
n/g 
OSMOPHILIC MOLD - 
TN10525 
OSMOPHYLIC MOLD 
 
<= 50 
n/g 
OSMOPHILIC MOLD - 
TN10525 
COLIFORMS 
 
<= 3 
n/g 
Coliforms - TN10510 
E. COLI 
 
None Detected 
 
Coliforms - TN10510 
SALMONELLA 
 
Negative 
 
Salmonella - TN10547 
 
 
Product Composition / Labeling 
Labelling regulations may vary from one country to another, please consult with your own regulatory personnel 
when determining how to label your finished product. 

 
 
 
 
 
PROMITOR™ Soluble Corn Fiber 85A 
Specification Sheet 
Page 2 of 6 
 
Version 1.8 (Date of Last Change: 09.06.2016) 
Date Printed: 12/21/2016 
 
 
If you require any further information on our ingredient, please contact your local Tate & Lyle representative. 
 
Regulatory Status 
Compliance Statement: 
x 
Complies with US FDA Regulation 21CFR184.1444 - generally 
recognized as safe. 
 
21 CFR References: 
21CFR184.1444 
 
Country of Origin 
 
Country of Manufacture: United States 
 
Kosher Status 
 
Certified Kosher: Yes 
Certified Kosher for Passover: No 
 
The above-mentioned ingredient or its components is certified Kosher by: 
x 
Orthodox Union (OU) 
 
Halal Status 
 
Certified Halal: Yes  
 
Vegetarian Status 
 
Vegetarian: Suitable  
Vegan: Suitable  
Ovo-Vegetarian: Suitable  
Lacto-Vegetarian: Suitable  
Lacto-Ovo-Vegetarian: Suitable  
 
Presence of Allergens 
 
If present in this Tate & Lyle ingredient, allergens will be denoted as “Present” in the table below. 
Allergen 
Status 
Crustacean & Shellfish 
Absent 
Mollusk 
Absent 
Fish 
Absent 
Egg 
Absent 
Milk (including Lactose) 
Absent 
Peanut & Oils 
Absent 
Soybean & Oils 
Absent 
Gluten Containing: Wheat, Rye, Barley, Oats, Spelt, 
Absent 

 
 
 
 
 
PROMITOR™ Soluble Corn Fiber 85A 
Specification Sheet 
Page 3 of 6 
 
Version 1.8 (Date of Last Change: 09.06.2016) 
Date Printed: 12/21/2016 
 
 
Kamut, Triticale 
Seeds, Sesame & Oils 
Absent 
Celery/Celeriac 
Absent 
Mustard 
Absent 
Lupin 
Absent 
Tree Nuts, Almond 
Absent 
Tree Nuts, Brazil 
Absent 
Tree Nuts, Cashew 
Absent 
Tree Nuts, Chestnut 
Absent 
Tree Nuts, Coconut and Oils 
Absent 
Tree Nuts, Hazelnut / Filbert 
Absent 
Tree Nuts, Hickory 
Absent 
Tree Nuts, Macadamia 
Absent 
Tree Nuts, Pine 
Absent 
Tree Nuts, Pistachio 
Absent 
Tree Nuts, Queensland 
Absent 
Tree Nuts, Walnut 
Absent 
Buckwheat 
Absent 
Bee Pollen/ Propolis 
Absent 
Royal Jelly 
Absent 
Pork 
Absent 
Peach 
Absent 
Tomato 
Absent 
Sulfur Dioxide and Sulfite > 10 ppm 
Absent 
 
Meaning of Present/Absent/Present-Exempt 
Present - Intentionally added during the production process. 
Absent - Not intentionally added during the production   process. 
Present-Exempt - Intentionally added during the production process, however have been given exemption from 
being required to be labeled on the final package. 
 
The above list of allergens is in accordance with all applicable EU and US legal requirements. 
 
Additional allergen information is available in the Product Information Sheet for  
PROMITOR™ Soluble Corn Fiber 85A 
 
GMO Status 
 
 
 
For additional information regarding the GM status of the above-mentioned ingredient, please refer to the 
Product Information Sheet. 
 
Storage Conditions 

 
 
 
 
 
PROMITOR™ Soluble Corn Fiber 85A 
Specification Sheet 
Page 4 of 6 
 
Version 1.8 (Date of Last Change: 09.06.2016) 
Date Printed: 12/21/2016 
 
 
 
Container 
Storage Temperature 
Storage conditions to achieve maximum 
shelf life 
Multiwall Bag 
 
Store at ambient temperature and humidity. 
 
Shelf Life 
 
Package 
Shelf Life 
Sulfur Dioxide and Sulfite Level 
Multiwall Bag 
730 Days 
< 10 ppm 
 
Organic Statement 
 
Certified for Organic Labeling: No 
 
For more information please consult the Product Information Sheet for PROMITOR™ Soluble Corn Fiber 85A 
 
Nutritional Information 
 

 
 
 
 
 
PROMITOR™ Soluble Corn Fiber 85A 
Specification Sheet 
Page 5 of 6 
 
Version 1.8 (Date of Last Change: 09.06.2016) 
Date Printed: 12/21/2016 
 
 
 
 
 
 
Supplier Information 
 
Name:  
 
 
Tate & Lyle 
Address: 
 
 
2200 E Eldorado 
 
 
 
 
Decatur, IL 62525 
 
 
 
 
USA 
Main Phone:  
 
1-217-423-4411 
Main Fax: 
 
 
1-217-421-2628 
E-mail:  
 
 
orders@tateandlyle.com 
 
 
Disclaimer 
 

 
 
 
 
 
PROMITOR™ Soluble Corn Fiber 85A 
Specification Sheet 
Page 6 of 6 
 
Version 1.8 (Date of Last Change: 09.06.2016) 
Date Printed: 12/21/2016 
 
 
THIS INFORMATION IS PROVIDED IN GOOD FAITH AND IS BELIEVED TO BE ACCURATE, BUT IS 
INTENDED FOR INFORMATIONAL PURPOSES ONLY AND NOT AS A RECOMMENDATION OR 
ENDORSEMENT. PRODUCT NAMES ARE NOT NECESSARILY DESCRIPTIVE OR INDICATIVE OF 
COMPOSITION OR FUNCTION. TATE & LYLE CANNOT ANTICIPATE OR CONTROL HOW THIS 
INFORMATION COULD BE USED. THEREFORE, YOU SHOULD NOT RELY ON IT FOR ANY LEGAL 
POSITION OR REGULATORY ASPECT, BUT YOU SHOULD CONDUCT INDEPENDENT INQUIRY, 
RESEARCH, AND TESTING BEFORE USING THE INFORMATION, INCLUDING, WITHOUT LIMITATION, 
THE DESIGNATION OF PRODUCT CLAIMS. TATE & LYLE DISCLAIMS ALL LIABILITY FOR ANY LOSS OR 
DAMAGE, TO THE FULLEST EXTENT PERMITTED BY LAW, INCLUDING LOSSES OR DAMAGES 
CAUSED BY CLAIMS OF INTELLECTUAL PROPERTY INFRINGEMENT. ALL WARRANTIES ARE HEREBY 
DISCLAIMED, INCLUDING THE IMPLIED WARRANTIES OF FITNESS FOR A PARTICULAR PURPOSE 
AND OF MERCHANTABILITY. THIS INFORMATION IS CONFIDENTIAL AND DISCLOSURE OR 
DISTRIBUTION IS PROHIBITED. 
 

 
 
 
 
 
STAR-DRI® 1015 A Maltodextrin 
Specification Sheet 
Page 1 of 6 
 
Version 1.3 (Date of Last Change: 07.07.2015) 
Date Printed: 12/16/2016 
 
 
Specification Sheet 
300000001486 
    
 
STAR-DRI® 1015 A Maltodextrin 
1630011115800, 1630011115021, 16300111 
 
    
 
 
 
 
 
Appearance: Powder 
Color: White 
Odor:  
 
Solubility Statement: Soluble In Water 
 
Quality Inspection Plan (Official Specification Values) 
 
Parameter 
Target 
Range/ 
Attribute 
Unit 
Method 
MOISTURE 
 
<= 6.00 
% 
Moisture - TN46020 
pH 
 
4.0 - 6.0 
 
pH - TN60550 
FOREIGN MATTER 
 
<= 4 
ppm 
Foreign Matter - TN32545 
DENSITY 
 
15.00 - 18.00 
lb/scf 
Bulk Density - TN24551 
LOOSE BULK DENSITY 
 
240 - 289 
g/dm^3 
BULK DENSITY - TN24505 
PACKED BULK 
DENSITY 
 
288 - 385 
g/dm^3 
BULK DENSITY - TN24505 
PACKED BULK 
DENSITY 
 
18.0 - 24.0 
lb/scf 
BULK DENSITY - TN24505 
FLOW RATE 
 
5.0 - 10.0 
 
Flow Rate - TN31340 
SOLUBILITY 
 
<= 30.0 
s 
Solubility - TN76000 
DE 
 
9.00 - 12.00 
% 
Dextrose Equivalent - 
TN25617 
SCREEN THRU US#20 
 
>= 97.00 
% 
Screen Size - TN69700 
SCREEN ON US#100 
 
>= 65.00 
% 
Screen Size - TN69700 
SCREEN THRU US#200 
 
<= 10.00 
% 
Screen Size - TN69700 
BACTER TOTAL 
 
<= 2000 
n/g 
Total Plate Count - TN10560 
YEAST SPORE 
 
<= 50 
n/g 
Mold & Yeast - TN47010 
MOLD 
 
<= 50 
n/g 
Mold & Yeast - TN47010 
THERMOPHILES, TOT 
 
<= 150 
n/10g 
Thermophilic Spores (NFPA) - 
TN16010 
FLAT SOURS 
 
<= 75 
n/10g 
Thermophilic Spores (NFPA) - 
TN16010 
SULFIDE SPOR 
 
<= 5 
Mold/10gr 
Thermophilic Spores (NFPA) - 
TN16010 

 
 
 
 
 
STAR-DRI® 1015 A Maltodextrin 
Specification Sheet 
Page 2 of 6 
 
Version 1.3 (Date of Last Change: 07.07.2015) 
Date Printed: 12/16/2016 
 
 
GAS FORMERS 
 
<= 4 
per 6 tube 
Thermophilic Spores (NFPA) - 
TN16010 
SALMONELLA 
 
Negative 
 
SAL B/HW - TN10545 
E. COLI 
 
None Detected 
 
Coliforms - TN10510 
COLIFORMS 
 
<= 10 
n/g 
Coliforms - TN10510 
 
 
Product Composition / Labeling 
Maltodextrin 
 
Regulatory Status 
Compliance Statement: 
x 
Complies with FDA Regulation 21CFR184.1444 as Maltodextrin - 
generally recognized as safe. 
 
21 CFR References: 
GRAS, 21CFR184.1444 
 
Country of Origin 
 
Country of Manufacture: United States 
 
Kosher Status 
 
Certified Kosher: Yes 
Certified Kosher for Passover: No 
 
The above-mentioned ingredient or its components is certified Kosher by: 
x 
Orthodox Union (OU) 
 
Halal Status 
 
Certified Halal: Yes  
 
Vegetarian Status 
 
Vegetarian: Suitable  
Vegan: Suitable  
Ovo-Vegetarian: Suitable  
Lacto-Vegetarian: Suitable  
Lacto-Ovo-Vegetarian: Suitable  
 
Presence of Allergens 
 
If present in this Tate & Lyle ingredient, allergens will be denoted as “Present” in the table below. 
Allergen 
Status 

 
 
 
 
 
STAR-DRI® 1015 A Maltodextrin 
Specification Sheet 
Page 3 of 6 
 
Version 1.3 (Date of Last Change: 07.07.2015) 
Date Printed: 12/16/2016 
 
 
Crustacean & Shellfish 
Absent 
Mollusk 
Absent 
Fish 
Absent 
Egg 
Absent 
Milk (including Lactose) 
Absent 
Peanut & Oils 
Absent 
Soybean & Oils 
Absent 
Gluten Containing: Wheat, Rye, Barley, Oats, Spelt, 
Kamut, Triticale 
Absent 
Seeds, Sesame & Oils 
Absent 
Celery/Celeriac 
Absent 
Mustard 
Absent 
Lupin 
Absent 
Tree Nuts, Almond 
Absent 
Tree Nuts, Brazil 
Absent 
Tree Nuts, Cashew 
Absent 
Tree Nuts, Chestnut 
Absent 
Tree Nuts, Coconut and Oils 
Absent 
Tree Nuts, Hazelnut / Filbert 
Absent 
Tree Nuts, Hickory 
Absent 
Tree Nuts, Macadamia 
Absent 
Tree Nuts, Pine 
Absent 
Tree Nuts, Pistachio 
Absent 
Tree Nuts, Queensland 
Absent 
Tree Nuts, Walnut 
Absent 
Buckwheat 
Absent 
Bee Pollen/ Propolis 
Absent 
Royal Jelly 
Absent 
Pork 
Absent 
Peach 
Absent 
Tomato 
Absent 
Sulfur Dioxide and Sulfite > 10 ppm 
Absent 
 
Meaning of Present/Absent/Present-Exempt 
Present - Intentionally added during the production process. 
Absent - Not intentionally added during the production   process. 
Present-Exempt - Intentionally added during the production process, however have been given exemption from 
being required to be labeled on the final package. 
 
The above list of allergens is in accordance with all applicable EU and US legal requirements. 
 
Additional allergen information is available in the Product Information Sheet for  
STAR-DRI® 1015 A Maltodextrin 
 

 
 
 
 
 
STAR-DRI® 1015 A Maltodextrin 
Specification Sheet 
Page 4 of 6 
 
Version 1.3 (Date of Last Change: 07.07.2015) 
Date Printed: 12/16/2016 
 
 
GMO Status 
 
The above-mentioned ingredient is not considered Non-GMO 
 
For additional information regarding the GM status of the above-mentioned ingredient, please refer to the 
Product Information Sheet. 
 
Storage Conditions 
 
Container 
Storage Temperature 
Storage conditions to achieve maximum 
shelf life 
Multiwall Bag 
 
Cool dry environment. 
 
Shelf Life 
 
Package 
Shelf Life 
Sulfur Dioxide and Sulfite Level 
Multiwall Bag 
730 Days 
< 10 ppm 
 
Organic Statement 
 
Certified for Organic Labeling: No 
 
For more information please consult the Product Information Sheet for STAR-DRI® 1015 A Maltodextrin 
 
Nutritional Information 
 

 
 
 
 
 
STAR-DRI® 1015 A Maltodextrin 
Specification Sheet 
Page 5 of 6 
 
Version 1.3 (Date of Last Change: 07.07.2015) 
Date Printed: 12/16/2016 
 
 
 
 
 
 
 
Supplier Information 
 
Name:  
 
 
Tate & Lyle 
Address: 
 
 
2200 E Eldorado 
 
 
 
 
Decatur, IL 62525 
 
 
 
 
USA 
Main Phone:  
 
1-217-423-4411 
Main Fax: 
 
 
1-217-421-2628 
E-mail:  
 
 
orders@tateandlyle.com 
 
 
Disclaimer 
 
THIS INFORMATION IS PROVIDED IN GOOD FAITH AND IS BELIEVED TO BE ACCURATE, BUT IS 
INTENDED FOR INFORMATIONAL PURPOSES ONLY AND NOT AS A RECOMMENDATION OR 
ENDORSEMENT. PRODUCT NAMES ARE NOT NECESSARILY DESCRIPTIVE OR INDICATIVE OF 

 
 
 
 
 
STAR-DRI® 1015 A Maltodextrin 
Specification Sheet 
Page 6 of 6 
 
Version 1.3 (Date of Last Change: 07.07.2015) 
Date Printed: 12/16/2016 
 
 
COMPOSITION OR FUNCTION. TATE & LYLE CANNOT ANTICIPATE OR CONTROL HOW THIS 
INFORMATION COULD BE USED. THEREFORE, YOU SHOULD NOT RELY ON IT FOR ANY LEGAL 
POSITION OR REGULATORY ASPECT, BUT YOU SHOULD CONDUCT INDEPENDENT INQUIRY, 
RESEARCH, AND TESTING BEFORE USING THE INFORMATION, INCLUDING, WITHOUT LIMITATION, 
THE DESIGNATION OF PRODUCT CLAIMS. TATE & LYLE DISCLAIMS ALL LIABILITY FOR ANY LOSS OR 
DAMAGE, TO THE FULLEST EXTENT PERMITTED BY LAW, INCLUDING LOSSES OR DAMAGES 
CAUSED BY CLAIMS OF INTELLECTUAL PROPERTY INFRINGEMENT. ALL WARRANTIES ARE HEREBY 
DISCLAIMED, INCLUDING THE IMPLIED WARRANTIES OF FITNESS FOR A PARTICULAR PURPOSE 
AND OF MERCHANTABILITY. THIS INFORMATION IS CONFIDENTIAL AND DISCLOSURE OR 
DISTRIBUTION IS PROHIBITED. 
 

Fat x Fiber Protocol  Version 1    p. 18 of 18 
APPENDIX 2   
Adverse Events Relatedness Reference Sheet (Cancer Consortium Clinical Research Support) 
 

Clinical Research Support
Adverse Event Relatedness Reference Sheet
06045 AE Relatedness Reference Sheet
Page 1 of 1
Version: 02/10/2016
Relatedness (attribution) of adverse events to a study intervention must be assessed in accordance with the study 
protocol. In the event that a protocol does not provide assessment criteria, the descriptions below may be used. 
Relatedness of Adverse Events to an Intervention
Definite
(must have all 4)
x
Has a reasonable temporal relationship to the intervention
x
Is not readily associated with the subject’s clinical state, concomitant 
medications/interventions, or other identifiable factors
x
Follows a known pattern of response related to the specific intervention
x
Disappears or decreases with reduction in dose or cessation of intervention and recurs 
with re-exposure (if applicable)
Probable
(must have 3)
x
Has a reasonable temporal relationship to the intervention
x
Is not readily associated with the subject’s clinical state, concomitant 
medications/interventions, or other identifiable factors
x
Follows a known pattern of response to intervention
x
Disappears or decreases with reduction in dose or cessation of intervention
Possible
(must have 2)
x
Has a reasonable temporal relationship to the intervention
x
Has unknown association with the subject’s clinical state, concomitant 
medications/interventions, or other identifiable factors
x
Follows a known pattern of response to intervention
Unlikely
(must have 2)
x
Does not have a temporal relationship to the intervention
x
May be associated with the subject’s clinical state, concomitant 
medications/interventions, or other identifiable factors
x
Does not follow a known pattern of response to intervention
x
Does not change with reduction in dose or cessation of intervention, and/or does not 
reappear or worsen with reintroduction of intervention (if applicable)
Unrelated
(must have all 4)
x
Does not have a temporal relationship to the intervention
x
Is associated with the subject’s clinical state, concomitant medications/interventions, or 
other identifiable factors
x
Does not follow a known pattern of response to intervention
x
Does not change with reduction in dose or cessation of intervention, and/or does not 
reappear or worsen with reintroduction of intervention (if applicable)
**This content is based in part on the National Institute of Allergy and Infectious Disease (NIAID) definitions of
relatedness to adverse events.**