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Confidentiality Note
Dole Food Company at the NCRC Nicholas D. Gillitt, Ph.D. Director Dole Nutrition Research Laboratory (DNRL)
2 March 18, 2013 Confidentiality Note
Fresh Fruit $4.7 Billion (69% of Sales)
Fresh Vegetables $1.0 Billion (15% of Sales)
Packaged Foods $1.1 Billion (16% of Sales)
Dole Food Company, Inc. $6.8 Billion Sales
(LTM as of 6/19/2010)
Generates net revenues of $6.8 billion
Dole Food Company Inc.
3 March 18, 2013 Confidentiality Note
Operates in over 90 countries worldwide
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Dolefil Pineapple Plantation - Philippines
Employs 76,000 people
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Murdock’s Vision
Believes in promoting the health benefits of a diet rich in fruits and vegetables – as part of a business strategy and a personal mission.
6 March 18, 2013 Confidentiality Note
• Inspiration was a life-change loss: death of his wife from cancer when she was 43 years of age – the same age his mother also died of cancer. “We used to eat steak and fried foods and sweets…we didn’t consider the impact until Gabriele was diagnosed with cancer. We tried everything to save her…but to no avail. That has made me all the more determined to try and save others.”
A Very Personal Mission…
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Advance scientific discovery • Bringing together a variety of instruments and disciplines under one roof. • Collaboration among top scientists, researchers and universities. • Areas of study will be biotechnology, obesity, cancer, agriculture and medical devices. • Collective power of shared knowledge. • New breakthroughs to market faster than ever before.
David H. Murdock & the NCRC
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The NCRC - Where Dole Does Science
Kannapolis, North Carolina
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North Carolina Research Campus Historic collaboration of eight top research universities, including Duke, NC State and UNC.
A place where life meets science
The NCRC – The Dream…
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300 acres - site of the old Cannon Mills textile operation – 27,000 people worked on this site alone…
The NCRC – The Reality…
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The NCRC – The Reality… David H. Murdock Core Laboratory
300,000 sq.ft. lab space, houses all the whistles and bells an lab could need!
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The NCRC – The Reality…
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Universities
The Model: Academia & Industry
Businesses
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Cannon Village
600 Laureate Way
DNRL: 10,000 sq ft lab/office space on 2nd floor dedicated to analytical chemistry and featuring HPLC, GC-MS, LC-MS/MS & NMR capability
DNRL – Site Geography
15 March 18, 2013 Confidentiality Note
Why is Dole at the NCRC?
• "Diets rich in fruits and vegetables may reduce the risk of some types of cancer and other chronic diseases”
SOURCE: Authorized by the National Cancer Institute and endorsed by the FDA
• "Diets low in saturated fat and cholesterol and rich in fruits, vegetables, and grain products that contain some types of dietary fiber, particularly soluble fiber, may reduce the risk of heart disease, a disease associated with many factors. .“
SOURCE: FDA Approved Health Claim
• “Second, consumption of fruits and vegetables is associated with reduced risk of many chronic diseases. Specifically, moderate evidence indicates that intake of at least 2-1/2 cups of vegetables and fruits per day is associated with a reduced risk of cardiovascular disease, including heart attack and stroke. Some vegetables and fruits may be protective against certain types of cancer.”
SOURCE: 2010 Dietary Guidelines for Americans
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Is F&V that important to our diet?
• What we should be eating…
7-13 servings F&V / day 2010 Dietary Guidelines for Americans
• What we are eating…
Only 30% consume 5 or more CDC 2005 Behavioral Risk Factor Surveillance System (BRFSS)
• According to the CDC about 2.4 million Americans die each year from any cause… - Heart Disease: 500,000 (American Heart Association) - Cancer: 500,000 (National Cancer Institute) - Diabetes 250,000 (American Diabetes Association)
50% of all deaths occur due to diseases that have a lifestyle/dietary dietary component
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What is Dole doing Differently?
• EDUCATE - Packaging claims, facts panels etc. - DNI, Dole.com, newsletters, books
brochures etc.
We NEED to Convince the 200 million people that are not eating enough F&V to eat more…
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SCIENTIFIC RESEARCH KEY: Produce exciting science that brings attention to our products and supports our ongoing educational efforts Through our work at the NCRC Dole aims to help provide a solid scientific foundation for the benefits of a diet rich in fruits and vegetables as well as providing both more nutritious products and better education to our consumers.
What is Dole doing Differently?
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We have our own cutting edge analytical instrumentation at our disposal
Nutrition Research Laboratory
NMR
LC-MS/MS
HPLC
GC-MS
UV/Vis
The Dole Lab…
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0
200
400
600
800
0 10 20 30 40Time, min
[p-N
itrop
heno
l], µ
mol
ml-1
Chemical Analysis…
0
50
100
150
200
250
300
350
400
[Phe
nolic
com
poun
ds],
GA
E m
g/10
0g
Stage 2Flesh
Stage 2Peel
Stage 4Flesh
Stage 4Peel
Stage 7Flesh
Stage 7Peel
PostStage 7Flesh
PostStage 7
Peel
Non-Water Soluble Phenolics Water Soluble Phenolics
Nutrient Content Variation
Enzymatic Activity Determination
Structural Elucidation
Dole scientists are: • Measuring the phytochemical content of fruits and
vegetables (inc. peels) as a function of variety, ripening stage, farming technique etc.
• Working with our agronomists to create more nutritious products
• Creating new product formulations based on nutrient density
• Providing information for consumer education
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In addition Dole is actively funding studies that investigate how foods can directly affect our health…
Obesity and overweight
Blood pressure, heart disease and stroke
Athletic performance and recovery time
Diabetes
Clinical Studies
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The Practicality of Science to Dole
Idea/Need/ Opportunity
Research & Validate
Results
Create & Innovate
Test
Sell
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Study 1: Chia Seed and Inflammation
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RESULTS
0
5
10
15
20
25
30
35
40
Whole Chia Seed(N=16)
Milled Chia Seed(N=14)
Poppy Seed (N=26)
EPA
(µg/
ml)
Pre-Study 10-weeks
3 x 2 interaction effect, P=0.016 *
0
10
20
30
40
50
60
70
Whole Chia Seed(N=16)
Milled Chia Seed(N=14)
Poppy Seed (N=26)
ALA
(µg/
ml)
Pre-Study 10-weeks
3 x 2 interaction effect, P=0.002 *
01
234
56
78
Whole Chia Seed(N=16)
Milled Chia Seed(N=14)
Poppy Seed (N=26)
CRP
(mg/
L)
Pre-Study 10-weeks
00.5
11.5
2
2.53
3.54
Whole Chia Seed(N=16)
Milled Chia Seed(N=14)
Poppy Seed (N=26)
IL-6
(pg/
ml)
Pre-Study 10-weeks
25 March 18, 2013 Confidentiality Note
Dole & ASU Collaboration
• Published July 2012
• Journal of Alternative and Complimentary Medicine is a well respected journal with a reasonable impact factor (1.585 in 2011)
• Press surrounding publication has been VERY positive
• Study PI David Nieman has been contacted numerous times for interviews by websites magazines etc.
PUBLICATION
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Science = Products…
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The Practicality of Science
Idea/Need/
Opportunity Resea
rch & Validat
e
Results Creat
e & Innovate
Test
Sell
NEXT STEPS
• Since chronic chia seed ingestion did not show any inflammation marker changes we propose investigating acute ingestion effects with a pharmacokinetic study…
STUDY CONCLUSION
• Milled but NOT whole chia seed increased plasma ALA (58%) and EPA (39%)
ACTION
• Reformulation of product to include milled chia seed making ALA bio-accessible.
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Hot off the Bench…
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Study 2: Bananas as an Energy Source
One medium banana (~118 g) contains: • 105 kcal • 27 g carbohydrate • 14.4 g sugars = glucose (5.9 grams), fructose
(5.7 grams), and sucrose (2.8 grams) • 3.1 g fiber (12% DV) • GI = 52 (low-to-medium rating) • 422 mg potassium (12% DV) • 15mg vitamin C (25% DV) • 1037 ORAC units (μmol Trolox Equivalents)
So far there are no studies looking at: 1) The effects of banana consumption as an
energy source for physical exertion 2) The countermeasure effects to oxidative stress
and inflammation induced by strenuous exercise.
30 March 18, 2013 Confidentiality Note
• Hypothesis: The antioxidant compounds in bananas would reduce inflammation in cyclists compared to a non antioxidant containing calorie matched control.
• Subjects: 15 male competitive cyclists aged 22 to 45 years. • Design: Cyclists to receive both banana or calorie matched placebo during a 75
km ride over about 2 hrs in a crossover design with washout period.
• Outcome Measures: - Performance & energy utilization (blood lactate and blood glucose) - Traditional biomarkers of inflammation/oxidative stress and immune function:
plasma cytokines e.g., IL-6, IL-10 etc. TNF-α, blood total leukocytes (neutrophils and monocytes) and oxidative burst, blood FRAP and ORAC.
- Metabolomics (measure all metabolites in the blood)
Total Phenolic Comparison of Bananas and a Calorie matched Control
0
20
40
60
80
Banana Control
Tota
l Phe
nolic
s, m
g (G
AE
) 100
cal
orie
s-1Research Design
31 March 18, 2013 Confidentiality Note
interaction effect, p=0.258 interaction effect, p=0.178
Variable Banana Control Mean HR 166±3.3 (91% HR max) 163±2.5 (89% HR max) Mean RPE 14.6±0.4 14.4±0.3 Weight change 3.3±0.4* 2.4±0.7 Watts (% maximal) 59.4% 61.5%
Control Control Banana Banana
RESULTS
32 March 18, 2013 Confidentiality Note
RESULTS Effect of Supported and Non-Supported Exercise on Plasma IL-10
0
10
20
30
40
50
60
70
80
90
Pre Exercise H2O vs 6% CHO
Nieman et al 2005
Post Exercise H2O vs 6% CHO
Nieman et al 2005
Pre Exercise BAN vs 6% CHO
Current Data
Post Exercise BAN vs 6% CHO
Current data
Plas
ma
IL-1
0 (p
g/m
l) This study
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460 442
604
521
575
510
0
100
200
300
400
500
600
700
Banana Gatorade
FRAP
(asc
orbi
c ac
id e
quiv
alen
ts, µ
mol
/L)
Pre-Exerc 1-h Exerc Post-Exerc
Interaction effect, p=0.012 *
*
Control Banana
RESULTS
34 March 18, 2013 Confidentiality Note
RESULTS - METABOLOMICS
“Metabolomics” is the measure of all the metabolites found in a body fluid (e.g., whole blood, plasma, urine etc.). A metabolite is any small molecule that is present in the sample which is not a protein (proteomics). Each peak on a chromatogram represents a different compound. There are three basic steps to a metabolomics interpretation… STEP 1: Compare the average metabolic profile from a number of subjects “pre” and “post” intervention to see if the identified metabolites increase or decrease as a function of that intervention (see blue arrows in above figure). STEP 2: Assign statistical significance to the changes and list the metabolites that change by more than a certain amount. STEP3: Identify which metabolic pathways these compounds are involved in and construct a discussion of how the intervention affected them.
“Pre”
“Post”
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RESULTS - METABOLOMICS
Intervention (banana vs sports drink): The only identified metabolite that was statistically significantly different between the banana and sports drink groups was dopamine which is known to occur naturally in bananas. Dopamine has two known roles in the body:
Neuronal: Dopamine is a neurotransmitter involved in the sensations relating to motivation, mood, pleasure and reward. However, its neuronal actions are limited to the brain and central nervous system. Since dopamine cannot cross the blood brain barrier dietary dopamine can not influence this metabolism. Hormonal: Hormones travel in the circulatory system and as such can have a wide reaching impact on human metabolism. Dopamine is linked to two potential hormonal effects: Cardiac Function: IV administered dopamine is used as a heart stimulant. It causes the heart to beat more powerfully and at a faster rate. Immunoregulation: White blood cells have dopamine receptors and in the presence of dopamine can be either activated or deactivated.
36 March 18, 2013 Confidentiality Note
Exercise (“pre” vs “post”): If we remove dopamine from the statistical analysis there is no metabolic difference between the banana and sports drinks groups. This allows us to condense the two data sets into one and look at an overall exercise effect. The following conclusions were reached and are to be presented at the 2012 annual meeting of the American College of Sports Medicine in San Francisco.
• 103 metabolites identified (56 significant). Score plots from the partial least squares discriminant analysis (PLS-DA) model visualized the global metabolic differences between pre-exercise, immediately post-exercise, and 1-h post-exercise, indicating a distinct separation between time points.
• The shifts in the 15 most significantly different metabolites following supported exercise with bananas or sports drinks over a 75-km cycling time trial indicated a similar pattern of heightened production of glutathione and utilization of fuel substrates in several pathways including glycolysis, lipolysis, and amino acid catabolism.
RESULTS - METABOLOMICS
Blue = “pre” Red = “post” Black = “post 1h”
37 March 18, 2013 Confidentiality Note
Dole & ASU Collaboration
• Published June 2012
• PLoS one is a well respected journal with a reasonable impact factor (4.092 in 2011)
• Press surrounding publication has been VERY positive
• Dole has built a summer marketing campaign around this study positioning bananas as natures original energy bar
• Nieman has been contacted numerous times for interviews by websites magazines etc.
PUBLICATION
38 March 18, 2013 Confidentiality Note
http://www.dole.com/bananas/energy.aspx
SUMMER CAMPAIGN
39 March 18, 2013 Confidentiality Note
Press from worldwide media coverage
40 March 18, 2013 Confidentiality Note
BANANAS vs. SPORTS DRINKS, Equal Performance, More Nutrients, 1/3 Price!
Clipart from worldwide media coverage
41 March 18, 2013 Confidentiality Note
NEXT on Doles “Science To Do” List…
• Bananas & Heart Health Dole/ASU Effect of 2 bananas a day for 5 weeks on plasma nutrient levels important to heart health, e.g., potassium and vitamins B6 and C as well as common heart disease markers, e.g., cholesterol levels CRP, blood lipids, weight and waist circumference etc.
• Pineapples and Inflammation Dole/DHMRI Effect of fresh pineapple on COX-2 mediated inflammation cascades in human cells. Bromelain is an enzyme found in pineapple which is believed to reduce inflammation. This study will look at the COX- 2 expression in synoviocytes as a function of bromelin introduction. Dole is developing a high bromelain pineapple which would allow market differentiation if a scientifically demonstrable human health benefit could be found.
• Broccoli & Human Health Dole/DHMRI/NC State Campus wide effort to investigate the effects of broccoli on human health. This will be the first time a collective collaborative effort involving Universities, seed producers, growers, marketers and sellers will be directed at discovering the health benefits of such a commonly eaten commodity vegetable.
• Dole Portabella Vitamin D Powder & Muscle Function in High School Athletes Dole/ASU Effect of Dole vitamin D capsules delivering 1x RDA of vitamin D2 made from portabella mushrooms on vitamin D status and muscle function and recovery in high school athletes.
42 March 18, 2013 Confidentiality Note
Cont’d…
• Effect of a F&V Juice on Cyclists Undergoing Strenuous Exercise Dole/ASU
Effect of 16 fl oz of a freeze dried reconstituted fruit and vegetable cocktail on conventional oxidative stress and inflammation markers as well as the small molecule blood plasma metabolome of cyclists under extreme exercise conditions.
• Nutrasorb, A Blueberry/Green Tea Polyphenol Laden Protein Supplement: Effects on Runners Undergoing Strenuous Exercise Dole/ASU/NC State/Rutgers
Effect of a polyphenolic rich blueberry/green tea sorbed protein supplement on conventional oxidative stress and inflammation markers as well as the small molecule blood plasma metabolome of cyclists under extreme exercise conditions.
• Dole Portabella Vitamin D Powder & Muscle Function in a NASCAR pit crew Dole/ASU Effect of Dole vitamin D capsules delivering 10x the RDA for vitamin D2 made from portabella mushrooms on vitamin D status and muscle function and recovery in high school athletes.
43 March 18, 2013 Confidentiality Note