Adequate vitamin A liver stores estimated by the modified relative dose response test are positively associated with breastfeeding but not vitamin A supplementation in Senegalese urban children 9–23 months old: A comparative cross-sectional study

Vitamin A supplementation (VAS) in 6-59-month-old children is recommended but its sustainability is currently questioned. In Senegal, available data suggest that VAS should be maintained, but geographic and age-related specificities need to be addressed to better implement and target VAS programming. The objective of this comparative cross-sectional study, conducted in urban settings of Dakar, was to compare the vitamin A liver stores (VALS) assessed using the modified-relative dose response (MRDR) test between supplemented and non-supplemented 9–23 month-old children and to study their relationship with VAS. The supplemented group (n = 119) received VAS (either 100 000 UI or 200 000 UI) 2 to 6 months before evaluation while the non-supplemented group (n = 110) had not received VAS during the past 6 months. In addition to MRDR, serum retinol concentrations (SR), and biomarkers of subclinical inflammation were measured. Children’s health-related data and feeding patterns were collected. Mean MRDR values (VAS: 0.030 ± 0.017, non-VAS: 0.028 ± 0.016, P = 0.389) and inflammation-adjusted SR (VAS: 1.34 ± 0.37, non-VAS: 1.3 ± 0.35, P = 0.515) of children were adequate. Low prevalence of VALS (VAS: 5.2%, non-VAS: 5.4%) and inflammation-adjusted VAD (VAS: 2.6%, non-VAS: 0.9%) were detected despite high presence of infections and inflammation. Children were mostly still being breastfed (VAS: 85.7%, non-VAS: 77.3%) and complementary feeding indicators were similar in both groups. Only breastfeeding was associated with VALS and was found to reduce by 76% at least, the odds of VAD (adjusted OR = 0.24, 95% CI: 0.07–0.8, P = 0.020). Based on MRDR values, VAS was not related to improved VALS and SR as well as VAD reduction among these children with adequate VALS. Reinforcing breastfeeding advocacy and morbidity prevention/control are essential in this setting. Scaling-back VAS in this subpopulation should be examined regarding the risk of hypervitaminosis A after an evaluation of dietary vitamin A intake sufficiency and a more quantitative assessment of VALS.

Retinol-binding protein, retinol, and modified-relative-dose response in Ugandan children aged 12–23 months and their non-pregnant caregivers

Retinol-binding protein (RBP), retinol, and modified-relative-dose response (MRDR) are used to assess vitamin A status. We describe vitamin A status in Ugandan children and women using dried blood spot (DBS) RBP, serum RBP, plasma retinol, and MRDR and compare DBS-RBP, serum RBP, and plasma retinol. Blood was collected from 39 children aged 12–23 months and 28 non-pregnant mothers aged 15–49 years as a subsample from a survey in Amuria district, Uganda, in 2016. DBS RBP was assessed using a commercial enzyme immunoassay kit, serum RBP using an in-house sandwich enzyme-linked immunosorbent assay, and plasma retinol/MRDR test using high-performance liquid chromatography. We examined (a) median concentration or value (Q1, Q3); (b) R2 between DBS-RBP, serum RBP, and plasma retinol; and (c) Bland-Altman plots. Median (Q1, Q3) for children and mothers, respectively, were as follows: DBS-RBP 1.15 µmol/L (0.97, 1.42) and 1.73 (1.52, 1.96), serum RBP 0.95 µmol/L (0.78, 1.18) and 1.47 µmol/L (1.30, 1.79), plasma retinol 0.82 µmol/L (0.67, 0.99) and 1.33 µmol/L (1.22, 1.58), and MRDR 0.025 (0.014, 0.042) and 0.014 (0.009, 0.019). DBS RBP-serum RBP R2 was 0.09 for both children and mothers. The mean biases were −0.19 µmol/L (95% limits of agreement [LOA] 0.62, −0.99) for children and −0.01 µmol/L (95% LOA −1.11, −1.31) for mothers. DBS RBP-plasma retinol R2 was 0.11 for children and 0.13 for mothers. Mean biases were 0.33 µmol/L (95% LOA −0.37, 1.03) for children, and 0.29 µmol/L (95% LOA −0.69, 1.27) for mothers. Serum RBP-plasma retinol R2 was 0.75 for children and 0.55 for mothers, with mean biases of 0.13 µmol/L (95% LOA −0.23, 0.49) for children and 0.18 µmol/L (95% LOA −0.61, 0.96) for mothers. Results varied by indicator and matrix. The serum RBP-retinol R2 for children was moderate (0.75), but poor for other comparisons. Understanding the relationships among vitamin A indicators across contexts and population groups is needed.

Comparison of Serum Retinol and Retinol-Binding Protein with the Modified Relative Dose-Response Test in Estimating Vitamin A Deficiency Among Ghanaian Women and Children

Objectives To evaluate the sensitivity and specificity of serum retinol (SR) and retinol-binding protein (RBP) in determining vitamin A deficiency (VAD) using the modified relative dose-response (MRDR) test as the reference. Methods Subjects included a subset of women and children participating in the Ghana Micronutrient Survey 2017. VAD was determined by the following cut-offs: SR or RBP < 0.7 mmol/L; MRDR ratio of 3,4-didehydroretinol to SR ³0.060. Sensitivity, specificity and the area-under-the-receiver operating characteristic (ROC) curve were calculated for unadjusted and inflammation-adjusted VAD cut-offs (based on C-reactive protein (CRP) and a1-acid-glycoprotein (AGP) for SR and RBP using the MRDR test as the reference. Results In 167 children and 178 women, inflammation (elevated CRP and/or elevated AGP) was present in 41% and 16%, respectively. Prevalence of VAD ranged, depending on the indicator used, from 7% (MRDR) to 40% (unadjusted SR) in children and 1% (RBP) to 4% (SR and MRDR) in women. Among children, sensitivity and specificity of unadjusted and adjusted SR and RBP were highly variable among the children. Highest sensitivity was achieved by unadjusted SR (80% of children with VAD correctly identified), while highest specificity was achieved by adjusted RBP (86% of children without VAD correctly identified). The best predictor of VAD in children compared with MRDR was adjusted SR, with a sensitivity of 78%, specificity of 73%, and an area under the ROC curve of 0.76. Among women, specificity was 97% for unadjusted and adjusted SR with an area under the ROC curve of 0.48; additional values could not be calculated due to lack of VAD cases. Conclusions SR and RBP were only moderately sensitive and specific for identifying VAD in children in Ghana. Low specificity—falsely identifying VAD—is especially problematic when populations are covered by one or more vitamin A interventions. Overlapping sources of preformed vitamin A can lead to hypervitaminosis, which may affect bone metabolism and growth. More accurate measures should be used in conjunction with SR and RBP for evaluating VAD especially in vulnerable populations. Funding Sources UNICEF, Canada's Ministry of Foreign Affairs, Trade and Development.

Modified relative dose response values differ between lactating women in the United States and Indonesia

The modified relative dose response (MRDR) test distinguishes between vitamin A (VA) deficiency and sufficiency using the molar ratio of 3,4-didehydroretinol to retinol (DR:R) in serum 4–6 h after administering 3,4-didehydroretinyl acetate (DRA). Research in sows demonstrated that serum and milk DR:R are correlated. Two studies determined VA status in lactating women and investigated breast milk as a surrogate for serum in the MRDR test in VA-adequate women. A secondary outcome compared serum with milk carotenoids in US women. Lactating US (24–40 years old, n = 25) and Indonesian (22–40 years old, n = 18) women were given 8.8 µmol DRA. Subjects were allocated to three collections (3–48 h post-dose) for blood and milk ( n = 4–10/time point). DR, retinol, and carotenoids were determined by HPLC. Serum DR:R time-courses were evaluated in all women and DR kinetics analyzed by noncompartmental modeling in the US and VA-deficient Indonesian women. Indonesian women had a higher proportion of VA deficiency by MRDR (61%) than US women (0%). Milk DR concentration was higher than serum. In US women, serum and milk DR:R were correlated at 5 h ( r = 0.86, P < 0.01) but not at 6 or 7 h. Serum DR kinetics ( tmax = 8, t1/2 = 15.3 h) were similar to those in VA-adequate lactating sows ( tmax = 7.5 ± 1.9, t1/2 = 17.4 ± 4.5 h) but varied in milk (human: tmax = 12, t1/2 = 22.4 h; sow: tmax = 14.4 ± 9.8, t1/2 = 71.8 ± 51.2 h). Carotenoids in milk differed from serum ( P < 0.001), with β-cryptoxanthin enrichment. Serum but not milk VA kinetics were similar between women and sows. Milk DR:R may represent a simpler VA biomarker in lactating women vulnerable to VA deficiency, but requires further validation. Impact statement Vitamin A (VA) deficiency is a major health issue globally, and lactating women are particularly vulnerable due to increased needs for milk production. Accurate detection of VA deficiency is important; however, most population surveys measure VA status using serum retinol, which is affected by inflammation and lacks sensitivity. The modified relative dose response (MRDR) test qualitatively distinguishes between VA deficiency and sufficiency and could improve population surveys if completed in a randomly selected subsample of individuals in surveys. The original relative dose response test required two blood samples, while MRDR requires only one, a significant improvement in accessibility of the technique by decreasing burden on subjects and investigators. This work demonstrates significant deficiency in Indonesian women compared with US women. In combination with previous research using lactating sows, these human data support milk as a surrogate for blood in the MRDR, which may be less invasive, but requires further validation.

Vitamin A intake and infection are associated with plasma retinol among pre-school children in rural Zambia

ObjectiveTo determine the prevalence of vitamin A deficiency, infection and adequacy of vitamin A intakes among Zambian children, and the contribution of dietary vitamin A and infection to vitamin A status.DesignA cross-sectional survey of vitamin A intakes by the 24 h recall method, vitamin A status by plasma retinol and the modified relative dose-response test, and infection by acute-phase proteins.SettingRural communities in Central and Eastern Provinces of Zambia.SubjectsChildren 2–5 years of age.ResultsThe prevalence of vitamin A deficiency was 56 % by plasma retinol, 48 % with infection-adjusted plasma retinol and 22 % by the modified relative dose-response test. The majority of children (61 %) had a current infection. Vitamin A intakes were relatively high (331 to 585 μg retinol activity equivalents/d in the harvest/early post-harvest and late post-harvest seasons, respectively) and the prevalence of inadequate intakes was <1 % when compared with the Estimated Average Requirement (210 and 275 μg retinol activity equivalents/d for children aged 1–3 and 4–8 years, respectively). Elevated α-1-acid glycoprotein was negatively associated with plasma retinol (P< 0·0 0 1) and vitamin A intake was positively associated with plasma retinol (P< 0·05), but only when estimated assuming a 26:1 retinol equivalence for provitamin A from green and yellow vegetables.ConclusionsInfection and vitamin A intakes were significant determinants of plasma retinol. We cannot conclude which indicator more accurately represents the true vitamin A status of the population. Reasons for the persistent high prevalence of vitamin A deficiency in the presence of adequate vitamin A intakes are unclear, but the high rates of infection may play a role.

The Vitamin A Status of Zambian Children in a Community of Vitamin A Supplementation and Sugar Fortification Strategies as measured by the Modified Relative Dose Response (MRDR) Test

The main objective of the study was to determine the vitamin A status of Zambian children less than five years of age in a community where strategies of vitamin A supplementation and consumption of vitamin A fortified sugar have been introduced. In a cross-sectional study, a total of 537 children were enrolled. Their vitamin A status was measured using the modified relative dose response (MRDR) test. Their vitamin A status was compared to the status measured using a similar method in 1996, before vitamin A supplementation through capsule distribution and fortification of sugar was implemented as strategies to reduce vitamin A deficiency in the country. Results showed that the vitamin A status of children improved markedly as a result of these strategies.