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    Setareh Shahab-Ferdows

    Background: Few data are available on the effectiveness of large-scale food fortification programs.Objective: We assessed the impact of mandatory wheat flour fortification on micronutrient status in Yaoundé and Douala, Cameroon.Methods:... more
    Background: Few data are available on the effectiveness of large-scale food fortification programs.Objective: We assessed the impact of mandatory wheat flour fortification on micronutrient status in Yaoundé and Douala, Cameroon.Methods: We conducted representative surveys 2 y before and 1 y after the introduction of fortified wheat flour. In each survey, 10 households were selected within each of the same 30 clusters (n = ∼300 households). Indicators of inflammation, malaria, anemia, and micronutrient status [plasma ferritin, soluble transferrin receptor (sTfR), zinc, folate, and vitamin B-12] were assessed among women aged 15-49 y and children 12-59 mo of age.Results: Wheat flour was consumed in the past 7 d by ≥90% of participants. Postfortification, mean total iron and zinc concentrations of flour samples were 46.2 and 73.6 mg/kg (target added amounts were 60 and 95 mg/kg, respectively). Maternal anemia prevalence was significantly lower postfortification (46.7% compared with 39....
    Human milk is recommended as the sole food source for infants 0-6 mo. Its micronutrient (MN) content is used as basis for making intake recommendations for breast-fed infants and lactating women. P...
    Anaemia has serious effects on human health and has multifactorial aetiologies. This study aimed to determine putative risk factors for anaemia in children 6–59 months and 15‐ to 49‐year‐old non‐pregnant women living in Ghana. Data from a... more
    Anaemia has serious effects on human health and has multifactorial aetiologies. This study aimed to determine putative risk factors for anaemia in children 6–59 months and 15‐ to 49‐year‐old non‐pregnant women living in Ghana. Data from a nationally representative cross‐sectional survey were analysed for associations between anaemia and various anaemia risk factors. National and stratum‐specific multivariable regressions were constructed separately for children and women to calculate the adjusted prevalence ratio (aPR) for anaemia of variables found to be statistically significantly associated with anaemia in bivariate analysis. Nationally, the aPR for anaemia was greater in children with iron deficiency (ID; aPR 2.20; 95% confidence interval [CI]: 1.88, 2.59), malaria parasitaemia (aPR 1.96; 95% CI: 1.65, 2.32), inflammation (aPR 1.26; 95% CI: 1.08, 1.46), vitamin A deficiency (VAD; aPR 1.38; 95% CI: 1.19, 1.60) and stunting (aPR 1.26; 95% CI: 1.09, 1.46). In women, ID (aPR 4.33; 9...
    Increased first-trimester low-density lipoprotein (LDL-C) concentration has been associated with adverse pregnancy outcomes, such as gestational diabetes. The B vitamins folate, B-6, and total B-12 are key for the methyl group-dependent... more
    Increased first-trimester low-density lipoprotein (LDL-C) concentration has been associated with adverse pregnancy outcomes, such as gestational diabetes. The B vitamins folate, B-6, and total B-12 are key for the methyl group-dependent endogenous synthesis of phosphatidylcholine, which is needed for lipoprotein synthesis, e.g., very low-density lipoprotein (VLDL), the precursor of circulating LDL-C. Maternal B-vitamin concentration usually declines across trimesters. Whether changes in maternal B-vitamin concentrations are associated with total cholesterol (TC), triglycerides (TG), and lipoprotein concentrations is unknown. Therefore, we explored the association between plasma folate, vitamin B-6 in the form of pyridoxal 5′-phosphate (PLP), and total B-12 with serum TC, LDL-C, HDL-C, and TG concentrations across trimesters. This secondary analysis used data of a prospective pregnancy cohort study included apparently healthy adult women (n = 179) from Rio de Janeiro, Brazil. The bio...
    Choline and related metabolites are key factors in many metabolic processes, and insufficient supply can adversely affect reproduction and fetal development. Choline status is mainly regulated by intake, and human milk is the only choline... more
    Choline and related metabolites are key factors in many metabolic processes, and insufficient supply can adversely affect reproduction and fetal development. Choline status is mainly regulated by intake, and human milk is the only choline source for exclusively breastfed infants. Further, maternal status, genotype, and phenotype, as well as infant outcomes, have been related to milk choline concentrations. In order to enable the rapid assessment of choline intake for exclusively breastfed infants and to further investigate the associations between milk choline and maternal and infant status and other outcomes, we have developed a simplified method for the simultaneous analysis of human milk choline, glycerophosphocholine, phosphocholine, and the less abundant related metabolites betaine, carnitine, creatinine, dimethylglycine (DMG), methionine, and trimethylamine N-oxide (TMAO) using ultraperformance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS). These analytes have mi...
    Our previously reported, first validated, UPLC-MS/MS-based simultaneous analysis of five human milk B-vitamins revealed severe matrix effects. High levels of endogenous lactose fouled the electrospray ionization source affecting the... more
    Our previously reported, first validated, UPLC-MS/MS-based simultaneous analysis of five human milk B-vitamins revealed severe matrix effects. High levels of endogenous lactose fouled the electrospray ionization source affecting the analysis. We evaluated solid-phase extraction (SPE), liquid-solid extraction (LSE), protein precipitation (PPT), and liquid chromatography effluent diversion for lactose-removal. SPE failed to separate lactose from vitamins; LSE using 2-propanol reduced lactose and vitamin recoveries. PPT-solvent, milk volume, and reconstitution solvent influenced flavin adenine dinucleotide, pyridoxal and nicotinamide recoveries. Using an optimized LC-gradient enabled chromatographic separation of lactose from vitamins and its removal using a post-column switch-valve. Only 40 µL milk was subjected to methanol-PPT and non-polar matrix removal by methyl tert-butyl ether. B-vitamin recoveries were established (81.9-118.6%; CV ≤ 11.9%; precision: 4.9-13.7%) with greatly reduced matrix effects, and improved process efficiency, and recovery.
    Background Infantile beriberi–related mortality is still common in South and Southeast Asia. Interventions to increase maternal thiamine intakes, and thus human milk thiamine, are warranted; however, the required dose remains unknown.... more
    Background Infantile beriberi–related mortality is still common in South and Southeast Asia. Interventions to increase maternal thiamine intakes, and thus human milk thiamine, are warranted; however, the required dose remains unknown. Objectives We sought to estimate the dose at which additional maternal intake of oral thiamine no longer meaningfully increased milk thiamine concentrations in infants at 24 wk postpartum, and to investigate the impact of 4 thiamine supplementation doses on milk and blood thiamine status biomarkers. Methods In this double-blind, 4–parallel arm randomized controlled dose–response trial, healthy mothers were recruited in Kampong Thom, Cambodia. At 2 wk postpartum, women were randomly assigned to consume 1 capsule, containing 0, 1.2 (estimated average requirement), 2.4, or 10 mg of thiamine daily from 2 through 24 weeks postpartum. Human milk total thiamine concentrations were measured using HPLC. An Emax curve was plotted, which was estimated using a non...
    Background Little is known regarding the associations between maternal factors and B-vitamin and choline concentrations in early milk and the trajectories of these vitamins during lactation. Objectives In this hypothesis-generating study,... more
    Background Little is known regarding the associations between maternal factors and B-vitamin and choline concentrations in early milk and the trajectories of these vitamins during lactation. Objectives In this hypothesis-generating study, we modeled the association between maternal and offspring factors and longitudinal changes in milk B-vitamin and choline concentrations throughout lactation. Methods A hundred women were studied in a prospective birth cohort and milk samples from 52 women were collected at 2–8 d, 76 women at 28–50 d, and 42 women at 88–119 d postpartum. Maternal dietary intake during pregnancy and lactation was assessed by an FFQ. Linear mixed-effects models with interaction terms were used to evaluate changes in milk B-vitamin and choline concentrations over time based on maternal factors and the early postpartum concentrations of these micronutrients. Results The women with higher early postpartum milk concentrations of niacin (βinteraction = −0.02; SE = 0.00; P ...
    Background Maternal micronutrient deficits during preconception and pregnancy may persist during lactation and compromise human milk composition. Objective We measured micronutrient concentrations in human milk and investigated their... more
    Background Maternal micronutrient deficits during preconception and pregnancy may persist during lactation and compromise human milk composition. Objective We measured micronutrient concentrations in human milk and investigated their association with maternal micronutrient intakes, status, and milk volume. Methods Infant milk intake (measured via a deuterium dose-to-mother technique), milk micronutrient and fat concentrations, and maternal micronutrient intakes were assessed at 2 and 5 mo postpartum in 212 Indonesian lactating mother–infant pairs. Maternal hemoglobin, ferritin, transferrin receptors, retinol binding protein (RBP), zinc, selenium, and vitamin B-12 were measured at 5 mo (n = 163). Multivariate or mixed effects regression examined associations of milk micronutrient concentrations with maternal micronutrient intakes, status, and milk volume. Results Prevalence of anemia (15%), and iron (15% based on body iron), selenium (2.5%), and vitamin B-12 deficiency (0%) were low ...
    Background Maternal supplementation during lactation could increase milk B-vitamin concentrations, but little is known about the kinetics of milk vitamin responses. Objectives We compared acute effects of maternal lipid-based nutrient... more
    Background Maternal supplementation during lactation could increase milk B-vitamin concentrations, but little is known about the kinetics of milk vitamin responses. Objectives We compared acute effects of maternal lipid-based nutrient supplement (LNS) consumption (n = 22 nutrients, 175%–212% of the RDA intake for the nutrients examined), as a single dose or at spaced intervals during 8 h, on milk concentrations and infant intake from milk of B-vitamins. Methods This randomized crossover trial in Quetzaltenango, Guatemala included 26 mother–infant dyads 4–6 mo postpartum who were randomly assigned to receive 3 treatments in a random order: bolus 30-g dose of LNS (Bolus); 3 × 10-g doses of LNS (Divided); and no LNS (Control), with control meals. Mothers attended three 8-h visits during which infant milk consumption was measured and milk samples were collected at every feed. Infant intake was assessed as $\mathop \sum \nolimits_{i\ = \ 1}^n ( {{\rm{milk\ volum}}{{\rm{e}}_{{\rm{feed\ }}...
    Objectives Choline and related metabolites play important roles in metabolic processes. Inadequate provision of these nutrients to the exclusively breast-fed infant can negatively impact its healthy growth and development. Methods We... more
    Objectives Choline and related metabolites play important roles in metabolic processes. Inadequate provision of these nutrients to the exclusively breast-fed infant can negatively impact its healthy growth and development. Methods We developed an UPLC-MS/MS method for analyzing choline (Cho), phospho-choline (PCho), glycerophospho-choline (GPCho), total choline (tCho = Cho + PCho + GPCho), betaine, carnitine, creatinine, dimethyl glycine (DMG), methionine, and trimethylamine N-oxide (TMAO) in human milk. Results Optimized results were obtained using a Phenomenex Luna Silica (2) column, 100 × 2 mm, 3 µm, and a gradient of 0.1% aqueous propionic acid (A) and acetonitrile (B) from 60% to 90% A over 2 min (Waters ACQUITY UPLC I-Class - SCIEX 4500TQ mass spectrometer). Sample preparation required only 5–10µL of milk, diluted 1:80 in methanol/water, 4:1, v/v, prior to analysis. Quantification was done using isotopically labeled internal standards and an external standard curve. Pooled hum...
    Background Except for low thiamin content, little is known about vitamins or macronutrients in milk from Cambodian mothers, and associations among milk nutrients. Objectives We measured fat-soluble vitamins (FSVs) and water-soluble... more
    Background Except for low thiamin content, little is known about vitamins or macronutrients in milk from Cambodian mothers, and associations among milk nutrients. Objectives We measured fat-soluble vitamins (FSVs) and water-soluble vitamins (WSVs), and macronutrients, and explored internutrient associations in milk from Cambodian mothers. Methods Milk from women (aged 18–45 y, 3–27 wk postpartum, n = 68) who participated in a thiamin-fortification trial were analyzed for vitamins B-2 (riboflavin, FAD), B-3 (nicotinamide), B-5, B-6 (pyridoxal, pyridoxine), B-7, B-12, A, E [α-tocopherol and γ-tocopherol (γ-TPH)], carotenoids, carbohydrate (CHO), fat, and protein. Milk vitamin B-1 [thiamin, thiamin monophosphate (TMP), thiamin pyrophosphate (TPP)] was previously assessed for fortification effects. Milk nutrient concentrations were compared with the Adequate Intake (AI) values for infants aged 0–6 mo. Pearson correlation was used to examine internutrient associations after excluding nut...
    Objectives Little is known about the influence of maternal micronutrient (MN) supplementation during pregnancy/lactation on human milk (HM) MN concentrations. We evaluated B-vitamin (BV) concentrations in milk from mothers in Ghana and... more
    Objectives Little is known about the influence of maternal micronutrient (MN) supplementation during pregnancy/lactation on human milk (HM) MN concentrations. We evaluated B-vitamin (BV) concentrations in milk from mothers in Ghana and Malawi who participated in the International Lipid-Based Nutrient Supplements (iLiNS) Project. Methods Women (<20 wk gestation) were randomized to receive: 1 x daily iron/folic acid (IFA) during pregnancy only, or a multiple micronutrient (MMN) tablet or LNS containing equal amounts of vitamins during pregnancy and 6 mo of lactation. HM at 6mo postpartum was analyzed for vitamins B1, B2, B3 (nicotinamide only), B5, B6, and B12 and compared to the recommended Adequate Intakes (AI) for infants 0–6 mo, which are based on values from milk from well-nourished mothers, assuming 780 mL/d milk intake. Results Concentrations and responses to supplementation differed greatly between countries. Comparing the IFA groups’ concentrations (geom. mean), women in G...
    Objectives Targeted metabolomics are commercially available for human plasma, but not for human milk. However, metabolite analyses could provide a novel and efficient approach to understanding human milk composition and relationships to... more
    Objectives Targeted metabolomics are commercially available for human plasma, but not for human milk. However, metabolite analyses could provide a novel and efficient approach to understanding human milk composition and relationships to maternal and infant status. Methods Pooled human milk was used to evaluate and validate the Biocrates AbsoluteIDQ® p180 kit for human milk metabolomics (40 acylcarnitines, 42 acids/biogenic amines, 91 phospholipids, 15 sphingolipids, sum of hexoses) using an ABSciex 5500QTRAP mass spectrometer in LC-MS/MS and flow injection analysis (FIA) mode. In a feasibility study, milk collected <6 mo lactation from A) Bangladeshi healthy mothers (BMI >18.5, n = 12) and from B) mothers with stunted infants (HAZ-score 
    Human-milk-targeted metabolomics analysis offers novel insights into milk composition and relationships with maternal and infant phenotypes and nutritional status. The Biocrates AbsoluteIDQ® p180 kit, targeting 40 acylcarnitines, 42 amino... more
    Human-milk-targeted metabolomics analysis offers novel insights into milk composition and relationships with maternal and infant phenotypes and nutritional status. The Biocrates AbsoluteIDQ® p180 kit, targeting 40 acylcarnitines, 42 amino acids/biogenic amines, 91 phospholipids, 15 sphingolipids, and sum of hexoses, was evaluated for human milk using the AB Sciex 5500 QTRAP mass-spectrometer in liquid chromatography-tandem mass-spectrometry (LC-MS/MS) and flow-injection analysis (FIA) mode. Milk (<6 months lactation) from (A) Bangladeshi apparently healthy mothers (body mass index (BMI) > 18.5; n = 12) and (B) Bangladeshi mothers of stunted infants (height-for-age Z (HAZ)-score 18.5 was lower in 6 amino acids/biogenic amines but higher in isovalerylcarnitine, two phospholipids, and one sphingomyelin (p < 0.05 for all). Associations between milk metabolites differed between groups. The AbsoluteIDQ® p180 kit is a rapid analysis tool suitable for human milk analysis and reduce...
    Background Anemia in young children is a global health problem. Risk factors include poor nutrient intake and poor water quality, sanitation, or hygiene. Objective We evaluated the effects of water quality, sanitation, handwashing, and... more
    Background Anemia in young children is a global health problem. Risk factors include poor nutrient intake and poor water quality, sanitation, or hygiene. Objective We evaluated the effects of water quality, sanitation, handwashing, and nutrition interventions on micronutrient status and anemia among children in rural Kenya and Bangladesh. Design We nested substudies within 2 cluster-randomized controlled trials enrolling pregnant women and following their children for 2 y. These substudies included 4 groups: water, sanitation, and handwashing (WSH); nutrition (N), including lipid-based nutrient supplements (LNSs; ages 6–24 mo) and infant and young child feeding (IYCF) counseling; WSH+N; and control. Hemoglobin and micronutrient biomarkers were measured after 2 y of intervention and compared between groups using generalized linear models with robust SEs. Results In Kenya, 699 children were assessed at a mean ± SD age of 22.1 ± 1.8 mo, and in Bangladesh 1470 participants were measured...
    Little valid information is available on human milk nutrient concentrations, especially for micronutrients (MN), and there are no valid Reference Values (RVs) across lactation. In this multi-center collaborative study, RVs will be... more
    Little valid information is available on human milk nutrient concentrations, especially for micronutrients (MN), and there are no valid Reference Values (RVs) across lactation. In this multi-center collaborative study, RVs will be established for human milk nutrients across the first 8.5 months postpartum. Well-nourished, unsupplemented women in Bangladesh, Brazil, Denmark, and The Gambia (n = 250/site) were recruited during the third trimester of pregnancy. Milk, blood, saliva, urine, and stool samples from mothers and their infants are collected identically at three visits (1–3.49, 3.5–5.99, 6.0–8.49 months postpartum). Milk analyses include macronutrients, selected vitamins, trace- elements and minerals, iodine, metabolomics, amino acids, human milk oligosaccharides, and bioactive peptides. We measure milk volume; maternal and infant diets, anthropometry, and morbidity; infant development, maternal genome, and the infant and maternal microbiome. RVs will be constructed based on m...
    To utilize complementary biochemical and dietary data collected before the initiation of national flour fortification to (i) identify micronutrient insufficiencies or deficiencies and dietary inadequacies in Palestinian women and children... more
    To utilize complementary biochemical and dietary data collected before the initiation of national flour fortification to (i) identify micronutrient insufficiencies or deficiencies and dietary inadequacies in Palestinian women and children in vulnerable communities and (ii) assess the suitability of the current wheat flour fortification formula. Quantitative dietary intake questionnaires were administered and fasting venous blood samples collected in randomly selected households in Gaza City and Hebron. The impact of fortification was simulated by estimating the additional micronutrient content of fortified wheat flour. Households in Gaza City and Hebron that were not receiving food aid from social programmes. Non-pregnant women (18-49 years) and children aged 36-83 months. The micronutrients with highest prevalence of insufficiency were vitamin D in women (84-97 % with serum 25-hydroxyvitamin D <50 nmol/l) and vitamin B12 in women and children (43-82 % with serum B12 <221 pmol/l). Deficiencies of vitamin A, Fe and Zn were also of public health concern. Current levels of wheat flour fortificants were predicted to improve, but not eliminate, micronutrient intake inadequacies. Modification of fortificant concentrations of vitamin D, thiamin, vitamin B12, Zn and folic acid may be indicated. Micronutrient insufficiencies or deficiencies and intake inadequacies were prevalent based on either biochemical or dietary intake criteria. Adjustments to the current fortification formula for wheat flour are necessary to better meet the nutrient needs of Palestinian women and children.
    It is uncertain whether vitamin B-12 supplementation can improve neurophysiologic function in asymptomatic elderly with low vitamin B-12 status or whether folate status affects responses to vitamin B-12 supplementation. We assessed the... more
    It is uncertain whether vitamin B-12 supplementation can improve neurophysiologic function in asymptomatic elderly with low vitamin B-12 status or whether folate status affects responses to vitamin B-12 supplementation. We assessed the effects of a single intramuscular injection of 10 mg vitamin B-12 (which also contained 100 mg vitamin B-6 and 100 mg vitamin B-1) on vitamin B-12 status and neurophysiologic function in elderly community-dwelling Chileans with low serum vitamin B-12 concentrations who were consuming bread fortified with folic acid. A pretreatment and posttreatment study was conducted in 51 participants (median ± SD age: 73 ± 3 y; women: 47%) with serum vitamin B-12 concentrations <120 pmol/L at screening. Vitamin B-12 status was defined by combining vitamin B-12, plasma total homocysteine (tHcy), methylmalonic acid (MMA), and holotranscobalamin into one variable [i.e., a combined indicator of vitamin B-12 status (cB-12)]. The response to treatment was assessed by ...
    Little information is available on B vitamin concentrations in human milk or on how they are affected by maternal B vitamin deficiencies, antiretroviral therapy, or maternal supplementation. The objective was to evaluate the effects of... more
    Little information is available on B vitamin concentrations in human milk or on how they are affected by maternal B vitamin deficiencies, antiretroviral therapy, or maternal supplementation. The objective was to evaluate the effects of antiretroviral therapy and/or lipid-based nutrient supplements (LNSs) on B vitamin concentrations in breast milk from HIV-infected women in Malawi. Breast milk was collected from 537 women recruited within the Breastfeeding, Antiretrovirals, and Nutrition study at 2 or 6 wk and 24 wk postpartum. Women were assigned to receive antiretrovirals and LNSs, antiretrovirals only, LNSs only, or a control. Antiretrovirals and LNSs were given to the mothers from weeks 0 to 28. The antiretrovirals were zidovudine/lamivudine and nelfinavir or lopinavir/ritonavir. LNSs provided 93-118% of the Recommended Dietary Allowances of thiamin, riboflavin, niacin, pyridoxine, and vitamin B-12. Infants were exclusively breastfed. LNSs increased milk concentrations of all vit...
    Representative data on folate and vitamin B-12 dietary intake and status in low-income countries are rare, despite the widespread adoption of folic acid fortification. The purpose was to evaluate folate and vitamin B-12 intake, status,... more
    Representative data on folate and vitamin B-12 dietary intake and status in low-income countries are rare, despite the widespread adoption of folic acid fortification. The purpose was to evaluate folate and vitamin B-12 intake, status, and risk factors for deficiency before a national fortification program in Cameroon. A nationally representative cross-sectional cluster survey was conducted in 3 ecologic zones of Cameroon (South, North, and the 2 largest cities, Yaoundé/Douala), and information on dietary intake was collected from 10 households in each of 30 randomly selected clusters per zone. In a subset of women and their 12- to 59-mo-old children (n = 396 pairs), plasma folate and vitamin B-12, as well as breast milk vitamin B-12, were analyzed. Vitamin B-12 and folate dietary intake patterns and plasma concentrations were similar for women and children. In the subsample, 18% and 29% of women and 8% and 30% of children were vitamin B-12 (≤221 pmol/L) and folate (<10 nmol/L) d...
    Vitamin B12 deficiency is highly prevalent in Bangladesh and other countries with low animal source food intake. Poor B12 status is associated with several adverse outcomes in pregnancy and infancy, and low breast milk B12; however no B12... more
    Vitamin B12 deficiency is highly prevalent in Bangladesh and other countries with low animal source food intake. Poor B12 status is associated with several adverse outcomes in pregnancy and infancy, and low breast milk B12; however no B12 supplementation trials have been carried out in pregnant women. In a pilot study, women (n=69, 18–35 y, Hb <115 g/L, 11–14 wk pregnant) were randomized to 250 ug B12/d or placebo through 4 mo postpartum. Both groups also received 400 ug folic acid + 60 mg Fe. Women were immunized with HINI vaccine at 26–28 wk gestation. There were no sig. differences between groups at baseline, when 26% women were B12 deficient (<150 pM), 41% had marginal status (150–221 pM), 26% elevated MMA and 61% tHcy (>8uM). Supplementation increased plasma B12 and lowered MMA in cord blood and in mothers and infants at 4 mo; it also reduced tHcy in cord blood and infants (all P<0.001) but not in mothers of whom 12% remained elevated (>12 uM) after supplementati...
    Given the importance of breast milk for providing micronutrients (MN) in the first 624 mo our objective is to determine if its MN content is adequate in populations with poor dietary quality and how it is affected by maternal... more
    Given the importance of breast milk for providing micronutrients (MN) in the first 624 mo our objective is to determine if its MN content is adequate in populations with poor dietary quality and how it is affected by maternal supplementation. We developed methods, including HPLC-MS/MS, to analyze multiple MN simultaneously in the milk matrix improving accuracy and efficiency of older methods. Available samples collected at 24 mo lactation were identified from multiple countries. Average population concentrations were expressed as % of those accepted by the IOM when setting DRIs for infants and lactation. Examples of the large differences in composition include: B1, 60% in India; riboflavin, 70% India, 30% Malawi, 20% Philippines and 10% in Bangladesh; niacin, 1020% (including California); B6, 220% California, 80% India, Malawi and Bangladesh, and 20% Philippines; B12, 60% Malawi, 30% India, and a very low 20% in Guatemala. Supplementation of HIV+ women in Malawi with 1X RDA from 06 ...
    The purpose of this research was to compare and validate methods for the analysis of vitamin B12 in human milk. The Adequate Intake (AI) for infants is 0.4 μg based on an assumed milk concentration of 378pM. However, some older... more
    The purpose of this research was to compare and validate methods for the analysis of vitamin B12 in human milk. The Adequate Intake (AI) for infants is 0.4 μg based on an assumed milk concentration of 378pM. However, some older concentration values were determined with inaccurate and non-validated methods. Poor maternal B12 intake or status causes low milk B12 and subsequent challenges for detection. Also, haptocorrin (HC) in human milk can produce falsely elevated or decreased values. We evaluated two assays (SimulTRAC-SNC and IMMULITE) for their suitability for analyzing B12 in human milk. SimulTRAC-SNB gave poor results and was highly influenced by HC, while IMMULITE can be applied without HC removal with an overall recovery rate of 78.9 % ± 9.1 (standard addition: 100 – 900 pg/mL). Diluted milk samples (24 – 193pM) were linearly correlated (r > 0.992). Analysis of milk samples from supplemented vs. unsupplemented women respectively in Bangladesh (± 250μg/d; milk concentration...
    Risk factors for folate deficiency differ from those for vitamin B12 deficiency in Cameroonian women and children (119.7) Abstract To assess nutritional status and risk factors for deficiency prior to a national fortification program, a... more
    Risk factors for folate deficiency differ from those for vitamin B12 deficiency in Cameroonian women and children (119.7) Abstract To assess nutritional status and risk factors for deficiency prior to a national fortification program, a nationally representative survey was conducted in 3 ecologic zones of Cameroon (South, North, and Cities). 24h dietary recall data were collected from 10 households (HH) in each of 30 randomly selected clusters/zone.. In a subset of women of reproductive age and their 12-59 mo old children (n= 396 pairs), plasma B12 & folate were analyzed by radioassay. Based on plasma values, 17% and 29% of women and 8% and 30% of children were folate (<10nm/L) and B12 (<221pm/L) deficient respectively. Folate deficiency did not differ among zones, but B12 deficiency was higher in the North/ vs. Cities. Folate and B12 dietary intake patterns were similar for women and children. Risk factors for folate and B12 deficiency differed: plasma folate was higher in th...
    Maternal supplementation in lactation could be an effective approach for improving breast milk quality in populations consuming poor quality diets. To determine acute effects of micronutrient (MN) supplementation on concentrations in milk... more
    Maternal supplementation in lactation could be an effective approach for improving breast milk quality in populations consuming poor quality diets. To determine acute effects of micronutrient (MN) supplementation on concentrations in milk during a 24h period, we recruited 18 healthy Bangladeshi women at 2-4 mo lactation for a 3d supplementation study. On d 1, no supplements were given, while in the morning of d 2 and 3 the participants received a dose of ~1x and 2x RDA respectively. Aliquots of a full breast milk expression were collected during every feed from the same breast on each study day and analyzed for vitamins A, E, B1, B2, B3, B6, and B12. All but vitamin A showed differences in mean concentrations based on collection day (P < 0.05, n = 448). To explore kinetics of vitamin secretion into milk, samples were grouped into 2h time intervals/d and mean values of the same intervals across days were compared. B2 and B6 concentrations both spiked after supplement consumption. ...
    Vitamin B12 is important for infant growth and supplementation to lactating mothers could increase intakes to exclusively breastfed (EBF) infants in areas where maternal diet is poor in B12. HIV+ mothers in the Breastfeeding,... more
    Vitamin B12 is important for infant growth and supplementation to lactating mothers could increase intakes to exclusively breastfed (EBF) infants in areas where maternal diet is poor in B12. HIV+ mothers in the Breastfeeding, Antiretrovirals, and Nutrition study were randomized to receive a lipid-based nutrient supplement (LNS), containing the RDA for B12 (2.6μg), or no LNS. We examined the relationships among maternal and infant plasma (MP, IP) and breastmilk (BM) concentrations of B12 in a sub-sample (n=521) at 2 or 6 wk (baseline) and 24 wk. At baseline MP B12 was positively correlated with BM (r=0.36) and IP (r=0.42), and IP and BM B12 were correlated (r=0.32) (all p<0.001). At 24 wk the relationships remained significant: MP B12 with BM (r=0.42) and IP (r=0.32), and IP with BM (r=0.29). At baseline there were no differences between LNS and no LNS in MP, BM or IP B12 concentrations. At 24 wk MP and BM B12 were higher in the LNS vs. no LNS group (p<0.005).However IP B12 was...
    In resource-limited settings without safe alternatives to breastfeeding, the WHO recommends exclusive breastfeeding and antiretroviral (ARV) prophylaxis. Given the high prevalence of anemia among HIV-infected women, mothers and their... more
    In resource-limited settings without safe alternatives to breastfeeding, the WHO recommends exclusive breastfeeding and antiretroviral (ARV) prophylaxis. Given the high prevalence of anemia among HIV-infected women, mothers and their infants (via fetal iron accretion) may be at risk of iron deficiency. We assessed the effects of maternal micronutrient-fortified lipid-based nutrient supplements (LNS) and maternal ARV treatment or infant ARV prophylaxis on maternal and infant iron status during exclusive breastfeeding from birth to 24 weeks. The Breastfeeding, Antiretrovirals, and Nutrition Study was a randomized controlled trial conducted in Lilongwe, Malawi from 2004-2010. HIV-infected mothers (CD4>200 cells/ul) and their infants were randomly assigned to 28-week interventions: maternal-LNS/maternal-ARV (n=424), maternal-LNS/infant-ARV (n=426), maternal-LNS (n=334), maternal-ARV (n=425), infant-ARV (n=426), or control (n=334). Longitudinal models tested intervention effects on hemoglobin (Hb). In a subsample (n=537) with multiple iron indicators, intervention effects on Hb, transferrin receptors (TfR) and ferritin were tested with linear and Poisson regression. In longitudinal models, LNS effects on maternal and infant Hb were minimal. In subsample mothers, maternal ARVs were associated with tissue iron depletion (TfR>8.3 mg/L) (Risk ratio (RR): 3.1, p<0.01), but not in ARV-treated mothers receiving LNS (p=0.17). LNS without ARVs, was not associated with iron deficiency or anemia (p>0.1). In subsample infants, interventions were not associated with impaired iron status (all p-values>0.1). Maternal ARV treatment with protease inhibitors is associated with maternal tissue iron depletion; but LNS mitigates adverse effects. ARVs do not appear to influence infant iron status; however, extended use needs to be evaluated.
    To utilize complementary biochemical and dietary data collected before the initiation of national flour fortification to (i) identify micronutrient insufficiencies or deficiencies and dietary inadequacies in Palestinian women and children... more
    To utilize complementary biochemical and dietary data collected before the initiation of national flour fortification to (i) identify micronutrient insufficiencies or deficiencies and dietary inadequacies in Palestinian women and children in vulnerable communities and (ii) assess the suitability of the current wheat flour fortification formula. Quantitative dietary intake questionnaires were administered and fasting venous blood samples collected in randomly selected households in Gaza City and Hebron. The impact of fortification was simulated by estimating the additional micronutrient content of fortified wheat flour. Households in Gaza City and Hebron that were not receiving food aid from social programmes. Non-pregnant women (18-49 years) and children aged 36-83 months. The micronutrients with highest prevalence of insufficiency were vitamin D in women (84-97 % with serum 25-hydroxyvitamin D <50 nmol/l) and vitamin B12 in women and children (43-82 % with serum B12 <221 pmol/l). Deficiencies of vitamin A, Fe and Zn were also of public health concern. Current levels of wheat flour fortificants were predicted to improve, but not eliminate, micronutrient intake inadequacies. Modification of fortificant concentrations of vitamin D, thiamin, vitamin B12, Zn and folic acid may be indicated. Micronutrient insufficiencies or deficiencies and intake inadequacies were prevalent based on either biochemical or dietary intake criteria. Adjustments to the current fortification formula for wheat flour are necessary to better meet the nutrient needs of Palestinian women and children.
    A high prevalence of low serum vitamin B-12 concentrations has been reported in studies and surveys in Latin America including Mexico, but the functional consequences are unknown. This randomized controlled trial assessed the response to... more
    A high prevalence of low serum vitamin B-12 concentrations has been reported in studies and surveys in Latin America including Mexico, but the functional consequences are unknown. This randomized controlled trial assessed the response to a high-dose vitamin B-12 supplementation of women in rural Querétaro, Mexico. Participants aged 20-59 y were stratified at baseline to deficient, marginal, and adequate status groups (serum vitamin B-12, 75-148, 149-220, and >220 pmol/L, respectively), and each group was randomized to vitamin B-12 treatment (single dose of 1 mg i.m. then 500 μg/d orally for 3 mo, n = 70) or placebo (n = 62). Measures at baseline and 3 mo included: complete blood count, serum vitamin B-12, holotranscobalamin (holoTC), folate, ferritin, C-reactive protein (CRP), bone alkaline phosphatase, and methylmalonic acid (MMA) and plasma total homocysteine (tHcy). At baseline, 11% of the women were vitamin B-12 deficient and 22% had marginal status. HoloTC was low (<35 pm...
    Pregnant women in resource-poor areas are at risk of multiple micronutrient deficiencies, and indicators of low vitamin B-12 status have been associated with adverse pregnancy outcomes, including anemia, low birth weight, and intrauterine... more
    Pregnant women in resource-poor areas are at risk of multiple micronutrient deficiencies, and indicators of low vitamin B-12 status have been associated with adverse pregnancy outcomes, including anemia, low birth weight, and intrauterine growth retardation. To evaluate whether daily oral vitamin B-12 supplementation during pregnancy increases maternal and infant measures of vitamin B-12 status, we performed a randomized, placebo-controlled clinical trial. Pregnant women <14 wk of gestation in Bangalore, India, were randomly assigned to receive daily oral supplementation with vitamin B-12 (50 μg) or placebo through 6 wk postpartum. All women were administered iron and folic acid supplements throughout pregnancy. One hundred eighty-three women were randomly assigned to receive vitamin B-12 and 183 to receive placebo. Compared with placebo recipients, vitamin B-12-supplemented women had significantly higher plasma vitamin B-12 concentrations at both the second (median vitamin B-12 concentration: 216 vs. 111 pmol/L, P < 0.001) and third (median: 184 vs. 105 pmol/L, P < 0.001) trimesters. At 6 wk postpartum, median breast milk vitamin B-12 concentration was 136 pmol/L in vitamin B-12-supplemented women vs. 87 pmol/L in the placebo group (P < 0.0005). Among vitamin B-12-supplemented women, the incidence of delivering an infant with intrauterine growth retardation was 33 of 131 (25%) vs. 43 of 125 (34%) in those administered placebo (P = 0.11). In a subset of infants tested at 6 wk of age, median plasma vitamin B-12 concentration was 199 pmol/L in those born to supplemented women vs. 139 pmol/L in the placebo group (P = 0.01). Infant plasma methylmalonic acid and homocysteine concentrations were significantly lower in the vitamin B-12 group as well. Oral supplementation of urban Indian women with vitamin B-12 throughout pregnancy and early lactation significantly increases vitamin B-12 status of mothers and infants. It is important to determine whether there are correlations between these findings and neurologic and metabolic functions. This trial was registered at clinicaltrials.gov as NCT00641862.
    Lipid-based nutrient supplements (LNS) have been effective in the treatment of acute malnutrition among children. We evaluated the use of LNS supplementation for improving the micronutrient status of young children. A 12-month randomised... more
    Lipid-based nutrient supplements (LNS) have been effective in the treatment of acute malnutrition among children. We evaluated the use of LNS supplementation for improving the micronutrient status of young children. A 12-month randomised controlled trial was conducted among children aged 6-18 months living in Intibucá, Honduras. Communities (n = 18) were randomised into clusters matched by poverty indicators (9 intervention, n = 160 and 9 controls, n = 140). Intervention participants received LNS. All children received food vouchers and nutrition education. Primary outcomes included measures of micronutrient status: at baseline, 6 and 12 months' blood were collected for assessment of folate, iron, zinc, riboflavin, and vitamin B12 status; haemoglobin was measured every 3 months; and dietary and anthropometry collected monthly. Longitudinal analyses were based on intent to treat and LNS adherence. Generalised estimating equations were used in the estimation of generalised linear regression models specified for the data. At 6-month follow-up, children in the intervention group had a lower proportion classified as deficient for B12 (43.6%) compared with the control (67.7%; P = 0.03). The intervention group had a higher mean concentration for folate at 6 months (P = 0.06), and improvements continued through 12 months for folate (P = 0.002) and vitamin A deficiency (P = 0.03). This pattern of results, with improved significance, remained in subanalysis based on LNS adherence. These data demonstrate that LNS improved select micronutrient status in young non-malnourished Honduran children.
    Methylmalonic acid (MMA) is a metabolic intermediate transformed to succinic acid (SA) by a vitamin B(12)-dependent catalytic step, and is broadly used as a clinical biomarker of functional vitamin B12 status. However, reported methods... more
    Methylmalonic acid (MMA) is a metabolic intermediate transformed to succinic acid (SA) by a vitamin B(12)-dependent catalytic step, and is broadly used as a clinical biomarker of functional vitamin B12 status. However, reported methods use between 100 and 1000 μL of serum or plasma making them sub-optimal for sample-limited studies, including those with neonates and infants. LC-MS/MS based protocols to measure MMA as n-butyl esters in the presence of tri-deuterated MMA (MMA-d(3)) were modified for use with 25 μL of human serum by scaling down sample processing volumes and analysis by UPLC-MS/MS. Plasma-based calibration solutions were found to be unnecessary, and chromatographic resolution and peak shape of SA and MMA was optimized in <4 min with isocratic 53:47 methanol/1.67 mM (pH 6.5) ammonium formate. Additionally, 1-cyclohexyl-urido-3-dodecanoic acid (CUDA) was included as internal standard allowing direct assessment of MMA recovery. Sample concentrations in the low normal range produced a signal:noise of >100:1. MMA intra- and inter-assay variability was under 10%. MMA-d(3) surrogate recovery averaged 93±14%. MMA stability exceeded three years in frozen samples and was unaffected by up to five freeze/thaw cycles. In conclusion, we report that methylmalonic acid can be measured with 25 μL of serum using water based standards. The assay signal:noise per concentration indicates that the method could perform as implemented with as little as 5 μL of serum. The reported method is applicable for studies of functional B12 status in sample limited experiments including investigations of nutritional status in neonates and in studies where low normal MMA levels are expected.
    Contribution of thiamin and riboflavin vitamers to total breast milk content (623.13) Abstract Thiamin (T) and riboflavin (R) are part of the B-vitamin complex necessary for many metabolic processes. During lactation, nutrient needs... more
    Contribution of thiamin and riboflavin vitamers to total breast milk content (623.13) Abstract Thiamin (T) and riboflavin (R) are part of the B-vitamin complex necessary for many metabolic processes. During lactation, nutrient needs increase to support maternal and infant requirements and inadequate supply has been described even in well-nourished mothers. Both vitamins occur in several forms in breast milk (BM) contributing to its total content. We analyzed T, thiamin-monophosphate (TMP), thiamin-pyrophosphate (TPP), R, and flavin adenine dinucleotide (FAD) in existing BM samples from Malawian mothers collected at 2 and 24 wk. (n = 118). Concentrations were log-transformed and means at 2/24wk were compared by paired t-test (2wk: total T median: 154µg/L, TPP/TMP/T 5/88/7%; total R: 210µg/L, R/FAD 6/94%; 24wk: total T: 196µg/L, 2/77/21%; total R: 173µg/L, 8/92%). Free and total T sig. increased over time (p < 0.05), while TMP showed no change. TPP, a form previously not reported i...