Effect of Iron- and Zinc-Biofortified Pearl Millet (ICTP8203-Fe) Consumption on Growth and Immunity in Children Aged 12-18 Months in India
Phase: NA
Conditions: Iron Deficiency, Zinc Deficiency
Sponsor: Cornell University

Effect of Iron- and Zinc-Biofortified Pearl Millet Consumption on Growth and 
1 
Immune Competence in Children Aged 12-18 Months in India—Study Protocol 
2 
for a Randomised Controlled Trial 
3 
 
4 
Clinical Trials ID: NCT02233764 
5 
 
6 
September 6, 2017 
 
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2 
Effect of Iron- and Zinc-Biofortified Pearl Millet Consumption on Growth and 
1 
Immune Competence in Children Aged 12-18 Months in India—Study Protocol 
2 
for a Randomised Controlled Trial  
3 
 
4 
Saurabh Mehta1,2, Julia L. Finkelstein1,3, Sudha Venkatramanan1, Samantha L. 
5 
Huey1, Shobha A. Udipi4, Padmini Ghugre5, Caleb Ruth 6, Richard L. Canfield1, 
6 
Anura V. Kurpad3, Ramesh D. Potdar7, Jere D. Haas1  
7 
 
8 
1Division of Nutritional Sciences, Cornell University, Ithaca, New York, US  
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2Institute for Nutritional Sciences, Global Health, and Technology, Cornell University, 
10 
Ithaca, New York, US  
11 
3St. John’s Research Institute, Bangalore, India (SJRI) 
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4Kasturba Health Society Medical Research Centre (KHS-MRC), Mumbai, India 
13 
5Shreemati Nathibai Damodar Thackersey, Women’s University (SNDT), Mumbai, 
14 
India 
15 
6 Data Performance LLC, Ithaca, New York, US 
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7Center for the Study of Social Change, Mumbai, India (CSSC)  
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18 
Corresponding Author (Principal Investigator):  
19 
Saurabh Mehta, MBBS, ScD  
20 
314 Savage Hall, Ithaca, New York 14853, US  
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Phone: +1-607-255-2640; Fax: +1-607-255-1033  
22 
E-mail: smehta@cornell.edu  
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Tables and Figures: 2 
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Keywords: Biofortification, iron, zinc, pearl millet, children, growth  
1 
ABSTRACT 
2 
 
3 
Introduction: Biofortified crops represent a sustainable agricultural solution for the 
4 
widespread micronutrient malnutrition in India and other resource-limited settings. 
5 
This study aims to investigate the effect of the consumption of iron and zinc-
6 
biofortified pearl millet by children on biomarkers of iron and zinc status and growth 
7 
outcomes. Additionally, we will assess immune function in the participants. 
8 
 
9 
Methods and Analysis: We will conduct a randomised controlled feeding trial in 
10 
identified slums of Mumbai, India among 250 children between 12-18 months of age 
11 
for 9 months. Children will be randomised to receive the biofortified pearl millet 
12 
(FeZn-PM, ICTP8203-Fe) or non-biofortified pearl millet. Anthropometric and 
13 
morbidity data will be gathered every month for 9 months. Biological samples will be 
14 
collected at baseline, midline, and endline, to assess iron and zinc status, including 
15 
haemoglobin, serum ferritin, serum transferrin receptor, serum zinc, C-reactive 
16 
protein, and alpha-1 acid glycoprotein. Biological samples will be archived for future 
17 
analyses. The midline measurement will be a random serial sample between 
18 
baseline and endline. Immune function will be assessed at each time point by the 
19 
measurement of T cell counts in a subset, respectively. 
20 
 
21 
Ethics and Dissemination: This study has obtained clearance from the Health 
22 
Ministry Screening Committee (HMSC) of the Indian Council of Medical Research 
23 
(ICMR). Ethical clearance has been obtained from Cornell University’s Institutional 
24 
Review Board, the Inter System Biomedica Ethics Committee (ISBEC), and St. 
25 
John’s Research Institute Institutional Ethics Review Board. The results of this study 
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4 
will be disseminated at several research conferences and as published articles in 
1 
peer-reviewed journals.  
2 
 
3 
Registration Details: Clinicaltrials.gov registration number: NCT02233764 
4 
(registered on September 4, 2014). Clinical Trials Registry of India (CTRI), reference 
5 
number REF/2014/10/007731, CTRI number CTRI/2015/11/006376. 
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Strengths and limitations of this study 
1 
• This is the first longitudinal randomised controlled trial to determine the 
2 
efficacy of consuming complementary foods prepared using iron- and zinc-
3 
biofortified pearl millet on both nutritional status and functional outcomes, 
4 
among children 12-18 months of age. 
5 
• The longitudinal random midline serial sampling strategy both increases 
6 
sensitivity and the power of the proposed study, while reducing cost and 
7 
invasiveness (by decreasing the number of biological samples from each 
8 
participant). 
9 
• Data will be collected on iPads or laptops, using a mobile electronic data 
10 
capture system framework on the iOS platform. This will decrease the 
11 
potential for error in data entry compared to standard written hard copies of 
12 
forms, and allow direct uploading of data using a secure server. 
13 
• One limitation of this study is that blinding may not be possible due to 
14 
potential sensory differences between the crops used in the two arms of the 
15 
study. 
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INTRODUCTION 
1 
The burden of iron and zinc deficiency 
2 
Deficiencies in iron and zinc are two of the most important public health problems 
3 
globally1 2. The role of iron deficiency in growth and development of children has 
4 
been demonstrated in several studies3-5. Similarly, it appears that a major fraction of 
5 
stunting can be explained by zinc deficiency6. Suboptimal iron and zinc status also 
6 
impairs immune functioning through a number of mechanisms, including a reduction 
7 
in the proportion of circulating T-lymphocytes and lymphocyte proliferative 
8 
responses7. Cognitive deficits are also observed with iron and zinc deficiency, 
9 
including irreversible impairments in neurological and psychomotor development of 
10 
children8.  
11 
 
12 
Iron and zinc biofortification  
13 
Biofortification has the potential to be a more sustainable and cost-effective 
14 
approach compared to other strategies such as diet diversification, fortification, and 
15 
supplementation, to address micronutrient malnutrition among vulnerable 
16 
populations9. In India, staple crops such as pearl millet (Pennisetum glaucum) are 
17 
consumed as part of the daily diet, particularly in Maharashtra, Gujarat, Rajasthan, 
18 
and Karnataka. The iron and zinc concentration in biofortified PM is reported to be 
19 
70-85 and 35-40 parts per million (ppm), respectively10. Previous research from 
20 
southern India and preliminary data from our acceptability study indicate that the 
21 
mean consumption of pearl millet (PM) flour in children is 61-80 g/day among Indian 
22 
children11 12, showing the suitability of this crop as a target for biofortification. Thus, 
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for children, iron and zinc intakes from PM would be 7-8 mg/100g and 3-4 mg/100g, 
24 
respectively10, and would help meet 50 to 70% of the recommended dietary 
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allowances (RDA) for children between 1-3 years of age. These estimates are based 
1 
on the RDA recently recommended for Indians by the Indian Council of Medical 
2 
Research13.  
3 
 
4 
The primary objectives of this randomised controlled trial are to study the effect of 
5 
the daily consumption of high iron- and zinc-biofortified pearl millet on biomarkers of 
6 
iron and zinc status, as defined by haemoglobin, serum ferritin, serum transferrin 
7 
receptor, serum zinc, and C-reactive protein; and growth, as defined by length-for-
8 
age, weight-for-age and weight-for-length, among 12-18-month-old children, as 
9 
compared to children receiving conventional pearl millet. Additionally, we will assess 
10 
the effect of high iron- and zinc-biofortified pearl millet by 12-18-month-old children 
11 
on immune outcomes.  
12 
 
13 
METHODS AND ANALYSIS 
14 
Study setting  
15 
This study will take place in identified slums of urban Mumbai, Maharashtra, India. 
16 
Mumbai is the commercial capital of India, and about 41.3% of city’s populace 
17 
resides in urban slums14.  
18 
 
19 
Study design 
20 
We will conduct a randomised controlled trial in which children (12 to 18 months old) 
21 
will be fed complementary foods prepared using iron- and zinc-biofortified pearl 
22 
millet (FeZn-PM) or the comparator conventional pearl millet for nine months. 
23 
Caregivers will be requested to provide three (3) informed consent forms for 1) 
24 
census; 2) screening; and 3) enrolment into the trial. Caregivers who provide all 
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three consents are considered eligible to be screened. Details of the informed 
1 
consent process can be found the section “Informed consent process.”  
2 
 
3 
1. Census: Before the study begins, a census will be conducted in the identified 
4 
slums to gather information on the age of the child, sex, and location. 
5 
 
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2. Screening: Consent will be obtained for screening and inclusion in the trial. Non-
7 
invasive data will be collected on: dietary allergies, use of iron or zinc dietary 
8 
supplements, availability of a caregiver, and if the caregivers are planning to stay in 
9 
the slum for the duration of the study. Anthropometry and dietary intake will also be 
10 
measured. If the children were not dewormed recently, they will be provided liquid 
11 
albendazole by the study physician at the study clinic (5 mL of syrup equivalent to 
12 
200 mg per dose). If still eligible for the study, blood will be collected to measure 
13 
complete blood counts including haemoglobin for further determination of eligibility.  
14 
 
15 
3. Randomization: A statistician from the Cornell Statistical Consulting Unit will 
16 
generate the random allocation sequence using a statistical software package (SAS 
17 
version 9.4); the randomization sequence key will be blinded to all study personnel 
18 
except the study statistician and the Filemaker database developer until follow-up is 
19 
over. Randomization will be allocated at the individual level. Individuals will be 
20 
randomized in blocks of 60. Pearl millet food products will coded by study arm (Arm 
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1: iron- and zinc-biofortified pearl millet; Arm 2: conventional pearl millet). Children 
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will be randomized to either study arm and will be monitored to ascertain they 
23 
consume the assigned food product throughout the study duration. The midline 
24 
measurement (including biological sample collection) will be a random serial sample 
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taken at any month (months 2-7) between the baseline and endline measurements15. 
1 
Longitudinal midline random serial sampling is often used in population 
2 
pharmacokinetic research and has been shown to be a useful strategy for iron 
3 
fortification efficacy studies, by describing the pattern of iron repletion15.  
4 
 
5 
Intervention and comparator 
6 
The intervention is iron-and-zinc-biofortified pearl millet (ICTP-8203) developed by 
7 
the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), and 
8 
the comparator is a conventional pearl millet that is commercially available on the 
9 
market. The comparator was chosen because it is similar to the intervention in all 
10 
aspects, except for iron and zinc content, which allows direct analysis of the impact 
11 
of the iron-and-zinc-biofortified variety on our outcome measurements. The nutrient 
12 
composition of biofortified and the conventional pearl millet flour are presented in 
13 
Table 1. We expect a child to consume an average of 60 g of pearl millet per day, 
14 
depending on the age of the child11.  
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18 
 
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24 
 
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Table 1: Nutrient composition of pearl millet 
varieties 
 
Biofortified 
(per 100 g 
flour) 
Control 
(per 100 g 
flour) 
Moisture (g) 
6.89 
7.16 
Fat (g) 
7.88 
8.74 
Protein (g) 
13.46 
13.46 
Carbohydrate 
(g) 
30.22 
30.98 
Energy (Kcal) 
246 
256 
Ash (g) 
1.99 
1.63 
Phytate (mg) 
876.59 
998.44 
Iron (mg) 
6.64 
2.56 
Zinc (mg) 
4.43 
1.24 

 
 
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Inclusion and exclusion criteria  
1 
Inclusion criteria  
2 
Participants included in this study will be 12-18-month-old male and female children 
3 
with haemoglobin concentration greater than or equal to 9 g/dL, living in urban slums 
4 
of Mumbai.  
5 
Exclusion criteria  
6 
Children will be excluded if: (1) they are younger than 12 months, 0 days old or older 
7 
than 18 months, 30 days at enrolment; (2) their haemoglobin concentration is less 
8 
than 9 g/dL or haemoglobinopathy is present (as indicated via abnormal peak via 
9 
hemoglobin variant analysis and confirmed by CSSC physicians); (3) they show 
10 
signs of severe malnutrition (a weight-for-length -3)16; (4) their caregiver reports prior 
11 
known diagnosis of HIV, tuberculosis, or current diagnoses of HIV, malaria, 
12 
tuberculosis, and/or dengue fever; (5) their caregiver is unavailable to bring the child 
13 
to the feeding centre during follow-up; (6) the child has any known dietary allergies; 
14 
and (7) their caregivers will leave the study site for greater than 4 weeks during the 
15 
follow-up period; (8) prior (within the past one year) or current consumption of iron or 
16 
zinc supplements.  Children who are severely anemic will be referred to physicians 
17 
at the Center for the Study of Social Change (CSSC).  
18 
 
19 
Informed consent process 
20 
Research assistants will obtain informed consent from caregivers. Three consent 
21 
forms will be collected at the Screening Visit: 1) Pre-Screening Consent (non-
22 
invasive questionnaires to determine age eligibility, use of dietary supplements, 
23 
availability of caregiver, etc. as described in Exclusion Criteria above; 2) Screening 
24 
Consent (invasive procedures including blood collection and anthropometric 
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measurements to assess malnutrition); and 3) Intervention Consent (including 
1 
baseline procedures such as a full doctor’s exam and detailed background 
2 
questionnaires).  
3 
 
4 
Feeding Centre 
5 
Feeding centres will be located near to where children and their caregivers’ 
6 
dwellings are clustered, to ensure that travel time from their home to the feeding 
7 
centre is within walking distance.  
8 
 
9 
Sample size estimation 
10 
Our estimates of sample size are based on assumptions about mean values and 
11 
associated variation in haemoglobin and serum ferritin. We expect 50–75% of these 
12 
children to have iron and zinc deficiency based on published literature17 and 
13 
preliminary results (unpublished). We assume that absorbed iron will be transferred 
14 
to body stores in the liver, and that the change in liver stores is reflected in changes 
15 
in serum ferritin at a rate of 8 µg/L of serum ferritin per mg liver iron in non-anaemic, 
16 
iron-depleted children. Iron-deficient, anaemic (haemoglobin < 11 g/dL18) children 
17 
will have their absorbable iron directed to haemoglobin synthesis in the early months 
18 
of the feeding trial. We estimate that the children consuming biofortified FeZn-PM 
19 
will demonstrate a gain of 1 g/dL in haemoglobin concentrations in 2.4 months, 
20 
whereas children consuming control pearl millet will need more than 9 months to 
21 
demonstrate the same increase. To detect a significant increase in ferritin from the 
22 
baseline value of 1.79 (6 mg/L) with standard deviation of 1.2, in the experimental 
23 
group, compared to the control group, at a power of 80% and 5% significance level, 
24 
96 participants will be required.  
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1 
Follow-up 
2 
Assessments 
3 
At baseline, we will assess anthropometry; collect blood for analysis of hemoglobin, 
4 
iron, zinc, CRP, and AGP biomarkers in the blood (all blood analyses other than 
5 
complete blood counts and T cell counts will be stored at -80 and then performed in 
6 
batch at the end of the trial); nutrient intake measures using multiple-pass 24-h 
7 
dietary recall19 for children and mothers; socio-economic and demographic 
8 
information; and morbidity history. Follow-up will continue for 9 months and will 
9 
include monthly anthropometric measurements, morbidity assessments, Infant and 
10 
Young Child Feeding Questionnaire20, and 24-hour dietary recall19. Both maternal 
11 
and infant dietary data will be collected via paper forms and entered into the CS 
12 
Dietary System Rel. 1.10, to calculate energy and nutrient intakes. Additionally, we 
13 
will collect rectal swab or stool samples to determine microbiome composition in 
14 
potential future ancillary analyses. The midline time point will be a random serial 
15 
sample that occurs between the baseline and endline measures. Children may visit 
16 
the CSSC clinic at any time during the trial for healthcare treatment. All blood 
17 
collection procedures will include the use a local anaesthetic (Prilox- Lidocaine and 
18 
Prilocaine cream) to decrease pain, and a Vein Finder device to illuminate the veins 
19 
to better identify the injection site. 
20 
 
21 
Administration of intervention 
22 
Both arms will receive complementary foods prepared with pearl millet three times 
23 
per day, 6 days per week, for 9 months. Culturally acceptable pearl millet based 
24 
complementary foods were developed by SNDT University and the acceptability of 
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these food products was tested on the caregivers and the children from the slums of 
1 
Mumbai10. We will conduct a run-in/pilot phase of the study with 1-3 selected feeding 
2 
centres. 
3 
 
4 
Each day, the child’s caregiver will bring the child to their feeding centre. Two meals 
5 
will be consumed at the feeding centre; the third meal may be consumed at home. 
6 
The food intake of the two meals at the feeding centre will be measured directly 
7 
before and after consumption. To measure the intake of the third meal, the weight of 
8 
unconsumed food from the third meal will be measured the next morning at the 
9 
feeding centre. To assure adherence, healthcare workers will follow up with the 
10 
participants’ caregivers and record reasons for non-adherence. Throughout the 
11 
study, we will conduct periodic analysis of random samples of both grain varieties 
12 
and prepared food products to ensure food safety and quality of the intervention. 
13 
 
14 
Primary outcome measurements 
15 
Biomarkers of iron and zinc status 
16 
We will determine if biofortified pearl millet improves iron and zinc status compared 
17 
to children who consume non-biofortified pearl millet. Specifically, iron and zinc 
18 
status will be assessed by measuring concentrations of haemoglobin (Hb), serum 
19 
ferritin, serum transferrin receptor (sTfR) and plasma zinc at enrolment (baseline), 
20 
midline, and endline. Additionally, we will measure concentrations of inflammatory 
21 
biomarkers C-reactive protein (CRP) and alpha 1-acid glycoprotein (AGP), as iron 
22 
and zinc biomarkers can be influenced by inflammation. 
23 
  
24 
Growth 
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We will assess change in a child’s growth as another outcome variable to determine 
1 
if iron-biofortified pearl millet reduces the risk of underweight (weight-for-age z score 
2 
< -2), wasting (weight-for-height z-score < -2) and stunting (length-for-age z score < -
3 
2) during the 9-month intervention period, compared to those who receive non-
4 
biofortified pearl millet. We will measure weight (kg) and length (cm) at baseline and 
5 
monthly throughout follow-up. The weight and length data will be converted to 
6 
weight-for-age, weight-for-length z-score, and length-for-age z-score. The growth 
7 
rates (as kg/month, and cm/month) will also be compared from the absolute 
8 
measurements after controlling for age.  
9 
 
10 
Additional outcomes 
11 
Immune function 
12 
We will assess immune function by measuring T cell counts and vaccine responses. 
13 
Additionally, we will collect morbidity data, including changes in types and 
14 
frequencies of morbidities such as diarrhoeal illness, pneumonia, and any chronic 
15 
disease throughout follow-up during each clinic visit. Caregivers will also be 
16 
encouraged to come to the clinic at any time for any health-related reasons, which 
17 
will also be recorded. This will provide an accurate representation of both innate and 
18 
adaptive immunity in participants. 
19 
 
20 
Cognitive function 
21 
To determine if consumption of biofortified pearl millet improves child cognitive 
22 
function, compared to consumption of non-biofortified pearl millet, we will assess in a 
23 
subset of children aspects of cognition that (1) previously have been shown to be 
24 
sensitive to the effects of early iron and/or zinc deficiency, or (2) draw heavily upon 
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brain structures or processes thought to be vulnerable to early iron and/or zinc 
1 
deficiency, based on studies of animal models and humans. Thus, we will assess 
2 
multiple specific aspects of memory, attention and processing speed using 
3 
automated eye trackers21. In addition, we will also assess higher-level, integrative 
4 
cognitive abilities, including problem-solving and exploratory behavior and global 
5 
aspects of attention and inattention during free play with toys22.  
6 
 
7 
Data collection and storage  
8 
Prior to data collection, the field staff will be trained on ethical and data collection 
9 
procedures. The protocol of data collection forms to be used is shown in Figure 1. All 
10 
data except dietary data will be collected on iPads or laptops for the proposed 
11 
project. We will use a mobile electronic data capture (mEDC) framework on the iOS 
12 
platform, Connedct, specifically designed for this project23. A secure server will be 
13 
utilized for uploading, storing, and accessing the data. This will enable real-time 
14 
feedback and error checking, as well as eliminate errors associated with data entry, 
15 
facilitating faster data analysis and rapid dissemination of results. All information will 
16 
be kept confidential. All names will be removed from the data for analysis. The 
17 
identifying codes and linked names will be securely stored in password-protected 
18 
computers. All data will be securely stored on a third-party server, which will have 
19 
limited access by study team members. All biological specimens will be collected by 
20 
trained medical professionals and evaluated by certified laboratories within India. 
21 
Biological specimens will be stored appropriately throughout the duration of the 
22 
study and after the study for future analysis.  
23 
 
24 
Data analysis  
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16 
We will use an intention-to-treat approach to determine the effect of biofortified pearl 
1 
millet on the outcomes described above. Advanced analysis will use mixed models 
2 
to account for pearl millet-based complementary foods consumed and use each 
3 
block of 60 as a random effect. We will also plot dose-response curves using 
4 
restricted cubic splines that will help us detect any threshold effects, as well as non-
5 
linear associations. Nonparametric tests, such as the Hodges-Lehmann test, will be 
6 
used where relevant, as in the case of non-normally distributed variables including 
7 
serum ferritin. 
8 
  
9 
Study monitoring board 
10 
A Data Safety Monitoring Board (DSMB) will be established for this study and will 
11 
include experts representing Cornell University and SNDT University who are not 
12 
directly involved in the trial. The DSMB members will oversee the study and will 
13 
periodically monitor the progress and outcomes of the intervention.  
14 
 
15 
Reporting of adverse events  
16 
All adverse events will be reported by the study physician via an Adverse 
17 
Event/Serious Adverse Event Report (AE/SAE) form. When an adverse event 
18 
occurs, study physicians, research assistants and/or other study personnel will 
19 
undertake all necessary precautions to ensure the safety and well-being of the 
20 
participant.  
21 
 
22 
The following study protocol endpoints will be considered to define safety and 
23 
efficacy outcomes, and establish un-blinding and stopping guidelines in this trial, as 
24 
per the discretion of the DSMB: 1. Diagnosis of the development of severe acute 
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malnutrition such as Kwashiorkor; 2. Occurrence of all-cause death. Demonstration 
1 
of efficacy, namely a significant beneficial effect on mortality and other adverse 
2 
outcomes will be used as a guideline to determine if the study should be un-blinded, 
3 
stopped, or terminated. 
4 
 
5 
 
6 
ETHICS AND DISSEMINATION 
7 
Before conducting interviews or allowing participation in the study, written informed 
8 
consent will be obtained from each participant’s guardians/primary caregivers, and 
9 
research assistants will record each granting of informed consent using audio and 
10 
video technology. Severely anaemic (<7 g/dL) and malnourished children will be 
11 
provided with appropriate medical care and referred to CSSC. The caregivers will be 
12 
assured of the confidentiality and anonymity of reports and publications generated 
13 
from this study. Participation will be voluntary, and participants will be assured that 
14 
refusal to participate in the study will not impact their access to care. The ethical 
15 
clearance for this study has been obtained from the Institutional Review Board of 
16 
Cornell University; Intersystem Biomedical Ethics Committee (ISBEC) of SNDT 
17 
University, Mumbai, India; and the Institutional Review Board of St. John’s Research 
18 
Institute, Bangalore, India. This study has received clearance from Health Ministry’s 
19 
Screening Committee (HMSC) of the ICMR. The results of this study will be 
20 
disseminated at several research conferences and as published articles in peer-
21 
reviewed journals. The present study protocol was prepared in accordance to the 
22 
SPIRIT statement24. This trial has been registered at Clinicaltrials.gov (registration 
23 
number NCT02233764) and the Clinical Trials Registry of India (CTRI) (reference 
24 
number REF/2014/10/007731, CTRI number CTRI/2015/11/006376).  
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1 
 
2 
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AUTHORS’ CONTRIBUTIONS: Saurabh Mehta, Julia L. Finkelstein and Jere D. 
30 
Haas designed the study, conceived the research questions and prepared the study 
31 
protocol. Saurabh Mehta is the principal investigator of the study and Julia L. 
32 
Finkelstein, Jere D. Haas, Shobha A. Udipi, Padmini Ghugre, Richard L. Canfield, 
33 
Anura V. Kurpad, and Ramesh D. Potdar are co-investigators. Caleb Ruth 
34 
developed the database used in this trial and provided feedback on the protocol. 
35 
Samantha L. Huey and Sudha Venkatramanan contributed to the editing of the 
36 
protocol and will supervise the data collection under the guidance of the 
37 
investigators. All authors read and approved the final protocol. 
38 
 
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ABBREVIATIONS: AGP: Alpha 1-acid glycoprotein; CRP: C-reactive protein; Fe: 
1 
Iron; FeZn-PM: iron- and zinc-enhanced variety of pearl millet; ICRISAT: 
2 
International Crops Research Institute for the Semi-Arid Tropics; ICTP8203-Fe: high 
3 
iron and zinc-biofortified pearl millet variety; ICMR: Indian Council of Medical 
4 
Research; ISBEC: Intersystem Biomedical Ethics Committee; IRB: Institutional 
5 
Review Board; KHS: Kasturba Health Society; PM: Pearl Millet; RDA: 
6 
Recommended Daily Allowance; SJRI: St. John’s Research Institute; SNDT: 
7 
Shreemati Nathibai Damodar Thackersey; sTfR: Serum transferrin receptor; Zn: Zinc 
8 
 
9 
FUNDING: This work was supported by HarvestPlus, grant number 2014H8302.  
10 
Address: 
11 
HarvestPlus 
12 
2033 K St NW 
13 
Washington, DC 20006 
14 
 
15 
CONFLICTS OF INTEREST: SM is an unpaid board member for a diagnostic start 
16 
up focused on developing point-of-care assays for nutritional status informed by his 
17 
research as a faculty member at Cornell University. All other authors report no 
18 
conflict of interest.  
19 
 
20 
Figure Legends: 
21 
Figure 1. Form collection protocol. 
22