SHORT TITLE: IRON BIOFORTIFICATION FOR COGNITIVE BENEFIT

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NFS 430 Consumption of Iron-Biofortified Beans Positively Affects Cognitive Performance in 18- to 27-Year- Old Rwandan Female College Students in an 18-Week Randomized Controlled Efficacy Trial SHORT TITLE: IRON BIOFORTIFICATION FOR COGNITIVE BENEFIT STUDY ANALYSIS BY: JESSICA M. PAGE

DISCUSSION OUTLINE AUTHORS BACKGROUND PURPOSE & HYPOTHESIS RESEARCH METHODS RESULTS LIMITATIONS IMPLICATIONS REFERENCES

AUTHORS CREDENTIALS

MURRAY-KOLB WENGER LUNG'AHO HAAS SCOTT* RHOTEN* PH.D., THE PENNSYLVANIA STATE UNIVERSITY, 03. MS, THE PENNSYLVANIA STATE UNIVERSITY, 98. PH.D., EXPERIMENTAL PSYCHOLOGY, STATE UNIVERSITY OF NEW YORK AT BINGHAMTON, 94. PH.D., DOCTOR OF PHILOSOPHY, FOOD SCIENCE AND INTERNATIONAL NUTRITION, CORNELL. PH.D., BIOLOGICAL ANTHROPOLOGY, PENNSYLVANIA STATE UNIVERSITY. PH.D. IN PROGRESS, DOCTOR OF LAW, PENN LAW 18. MS, ENV STUDIES, UNIVERSITY OF PENNSYLVANIA, 15. MS, PH.D. CANDIDATE, UNIVERSITY OF OKLAHOMA. (SENIOR OUTCOMES RESEARCHER AT ICON)

IRON DEFICIENCY ANEMIA & BIOFORTIFICATION BACKGROUND

IRON OXYGEN TRANSPORT 2/3: Hemoglobin of RBCs ¼: Stores of Ferritin + Hemosiderin Remaining: Myoglobin & Enzymes (RedOx) WRA 18 mg/d 10% ABS: 1.8 mg/d Absorbed Not All Sources Equal Meat vs. Plants: 50% vs. 10%

IRON DEFICIENCY ANEMIA PREVALENCE #1 Most Common Nutrient Deficiency IDA 34% of Population, 2 Billion People High in Rwanda Highest in WRA and Pregnant

IRON DEFICIENCY ANEMIA CAUSES CAUSES: Low Iron Intake Meat, Beans, Greens, Nuts Poor Absorption Heme vs. Non-heme Iron Increased Requirements Chronic Blood Loss

IRON DEFICIENCY ANEMIA DEFINITION IRON STORES ARE USED UP SUPPLY OF IRON TO THE TISSUES IS DECREASED DECREASED ABILITY OF CELLS TO GET ENERGY HGB <120 G/L

IRON DEFICIENCY ANEMIA EFFECTS Fatigue & Dizziness Palpitations Brittle Nails/Koilonychia Pallor Increased Maternal Mortality Reduced Physical Work Capacity Reduced Cognition Learning Memory Attention

IRON DEFICIENCY ANEMIA TREATMENTS Diet Change Fortified Foods Supplementation IV Iron Therapy Blood Transfusions Iron Biofortified Crops

BIOFORTIFICATION HARVESTPLUS Identify High Micronutrient Crops Conventional Breeding Variety Blending Increased Target Nutrients High Yield Climate & Pest Guards 290 Varieties of 12 Crops Iron, Zinc, Vitamin A Beans, Sweet Potato, Rice, Wheat, Corn, Pearl Millet

PURPOSE & HYPOTHESIS

STUDY PURPOSE & HYPOTHESIS PURPOSE: "...to determine the efficacy of iron-biofortified beans in improving cognition in WRA compared with control beans." HYPOTHESIS: "...iron repletion by way of a biofortified staple food (beans) would produce improvements in cognitive performance in WRA."

RESEARCH METHODS

RESEARCH METHODS DESIGN DESIGN Randomized Controlled Efficacy Trial Double Blinding 18-Week Intervention: January 7-May 23, 2013 LOCATION University of Rwanda Huye Campus Butare, Rwanda SAMPLING Parent Study Recruitment: "Healthy" & Iron Depleted (SF<20 μg/l) 2-Tailed 5% Type I Error rate & 90% Power = 60 subjects/group

RESEARCH METHODS SCREENING INCLUSIONS Females Attending UR Serum Ferritin <20 μg/l EXCLUSONS Males Age <18 & >27 BMI <16 Hgb <90 g/l SF >20 μg/l Major Medical Conditions Iron Supplements Interfering Medications Pregnant/Lactating STARTING SAMPLE: n=239 120 Biofortified + 119 Control

S A M P L E ANALYSIS SAMPLE: n=150 72 Biofortified + 78 Control

RESEARCH METHODS INTERVENTION GROUPS Randomly Assigned 1 of 4 Color Groups 2 Intervention, 2 Control Intervention: 86.1 mg/kg Iron Beans Control: 50.1 mg/kg Iron Beans

RESEARCH METHODS INTERVENTION 2x / Day Buffet-Style Campus Dining 2-Week Rotating Menu Served 175g Beans by Plate Color MEALS Ad Libitum Weighed Pre & Post Breakfast/Snack 24-Hour Recalls 3-Day Nutrient Intakes

RESEARCH METHODS INTERVENTION STARCHES White Potatoes Sweet Potatoes Rice Pasta Cassava Maize Porridge Plantains MEAL CONTENTS VEGETABLES NO MEAT Cabbage Cassava Greens DRINK Water Only SUNDAYS Ice Cream Fruit

RESEARCH METHODS BLOOD TESTS BASELINE & ENDLINE with Random Serial Sampling Whole Blood Samples & CBC Hct MCV MCHC RDW Serum Ferritin (SF) Total Body Iron (BI) Ratio TfR : SF C-Reactive Protein(CRP) α-1-acid Glycoprotein(AGP)

RESEARCH METHODS BLOOD TESTS DEFINITIONS Anemia: Hgb <120 g/l Iron Deficiency: SF <15 μg/l, TfR >8.3 mg/l or BI <0 mg/kg

RESEARCH METHODS COGNITIVE TESTS Computer-Based Tests 60-90 minutes Baseline 3 Weeks In Endline 3 Weeks Before Trial End Tasks: Taps Iron-Dependent Cortical Systems Used On Humans in Research External Validity Check

T E SRT: Speed of simplest reaction time. GNG: Assesses inhibitory control and sustained attention. Press button with proper stimulus. S ANT: Tests alerting, orienting, and executive control. T S SMS: Tests short-term memory; 1, 3, or 6 symbols and recalling those previously seen. CRT: Short-term visual memory, shown common objects. Speed, accuracy, and efficiency of recognition.

RESEARCH RESULTS

STATISTICS BLOOD Chi-Square: Group Differences at Baseline 44% ID Anemia 92% Iron Deficient No differences between: Groups Sub-sample & Larger Sample

STATISTICS BLOOD Group Comparison: Raw Means Hemoglobin Sig Increase Biofortified Serum Ferritin Sig Increase in Both, Higher in Biofortified Body Iron Sig Increase Biofortified

ANOVA Confirmed Effect: Hgb Ferritin Body Iron None for: Transferrin Receptor

STATISTICS TESTS SRT, GNG, ANT Accuracy VERY High (98-99%) Non-Sig Effects for SRT & GNG 1 ANT (RT, orienting) Variable with Group Effect

STATISTICS TESTS CRT: Biofortified Sig Improvement Faster Reactions & Remembered Old Images Better PCC: More Adaptable SMS: Biofortified Sig Improvement Decreased Slope & Intercept =Memory Speed & Efficiency Improved

STATISTICS Z-SCORES Per Protocol Analysis Significant Results Only Z-Scores Shows Grand Mean Change

RESULTS ATTRITION ANALYSIS EXCLUSION 160: Intervention & Cognitive Tests 150: Complete Data 10 Excluded 6.25% Attrition 6.49% Biofortified & 6.02% Control

RESULTS SUMMARY SELECTIVE ATTENTION SPEED 17% Larger Improvement in Speed P < 0.01 MEMORY RETRIEVAL 7-Fold: Memory Speed 68% Greater Improvement Memory Efficacy 2-Fold: Specificity MEMORY SEARCH 2-Fold: Speed 3-Fold: Memory Search Efficiency

RESEARCH LIMITATIONS

LIMITATIONS HOMOGENOUS SAMPLE All WRA All Medical Students Not General Population Generalizability Limited NORMAL BEAN INTAKE May Consume More in Trial Not Representative of Population's Average Intake

CLINICAL & PUBLIC IMPLICATIONS

IMPLICATIONS CLINICAL Hospital Diets for ID Anemia Biofortified Beans Vitamin C Prescribing Biofortified Beans for IDA Improvements PUBLIC Developing Countries Cost Effective Nutritionally Dense Malnutrition Nutrient Deficiencies HarvestPlus: Reach 1 Billion People, 2030

THANK YOU.

References for Research Paper Murray-Kolb LE, Wenger MJ, Scott SP, Rhoten SE, Lung'aho MG, Haas JD. Consumption of Iron-Biofortified Beans Positively Affects Cognitive Performance in 18- to 27- Year-Old Rwandan Female College Students in an 18-Week Randomized Controlled Efficacy Trial. J Nutr. 2017;147(11):2109-2117. Doi:10.3945/jn.117.255356.

References for Information Crops. Biofortification: The Nutrition Revolution Is Now. https://www.harvestplus.org/what-wedo/crops. Published November 8, 2018. Accessed November 10, 2018. Haas JD, Luna SV, Lungaho MG, et al. Consuming Iron Biofortified Beans Increases Iron Status in Rwandan Women after 128 Days in a Randomized Controlled Feeding Trial. JNutr. 2016;146(8):1586-1592. doi:10.3945/jn.115.224741. Iron. Nutrient Reference Values. https://www.nrv.gov.au/nutrients/iron. Published September 4, 2014. Accessed November 10, 2018. More S, Shivkumar VB, Gangane N, Shende S. Effects of iron deficiency on cognitive function in school going adolescent females in rural area of Central India. Anemia. 2013;2013:1-5. doi:10.1155/2013/819136. Stoltzfus RJ, Dreyfuss ML. Guidelines for the use of iron supplements to prevent and treat iron deficiency anemia. World Health Organization. http://www.who.int/nutrition/publications/micronutrients/guidelines_for_iron_supp lementation.pdf?ua=1&ua=1. Published 1998. Accessed October 24, 2018.

References for Pictures https://www.walmart.com/ip/nature-s-blend-ferrous-gluconate-5-gr-elemental-iron-324-mg- 100-Tablets/156396557 http://www.haimanotgb.com/pinto-beans/ https://www.yourgenome.org/facts/what-is-selective-breeding https://www.harvestplus.org/content/iron https://www.kisspng.com/png-iron-deficiency-anemia-iron-deficiency-human-iron- 273852/preview.html