Use of Model-Based Compartmental Analysis and a Super-Child Design to Study Whole-Body Retinol Kinetics and Vitamin A Total Body Stores in Children from 3 Lower-Income Countries.

BACKGROUND: Model-based compartmental analysis has been used to describe and quantify whole-body vitamin A metabolism and estimate total body stores (TBS) in animals and humans. OBJECTIVES: We applied compartmental modeling and a super-child design to estimate retinol kinetic parameters and TBS for young children in Bangladesh, Guatemala, and the Philippines. METHODS: Children ingested [13C10]retinyl acetate and 1 or 2 blood samples were collected from each child from 6 h to 28 d after dosing. Temporal data for fraction of dose in plasma [13C10]retinol were modeled using WinSAAM software and a 6-component model with vitamin A intake included as weighted data. RESULTS: Model-predicted TBS was 198, 533, and 1062 µmol for the Bangladeshi (age, 9-17 mo), Filipino (12-18 mo), and Guatemalan children (35-65 mo). Retinol kinetics were similar for Filipino and Guatemalan groups and generally faster for Bangladeshi children, although fractional transfer of plasma retinol to a larger exchangeable storage pool was the same for the 3 groups. Recycling to plasma from that pool was ∼2.5 times faster in the Bangladeshi children compared with the other groups and the recycling number was 2-3 times greater. Differences in kinetics between groups are likely related to differences in vitamin A stores and intakes (geometric means: 352, 727, and 764 µg retinol activity equivalents/d for the Bangladeshi, Filipino, and Guatemalan children, respectively). CONCLUSIONS: By collecting 1 or 2 blood samples from each child to generate a composite plasma tracer data set with a minimum of 5 children/time, group TBS and retinol kinetics can be estimated in children by compartmental analysis; inclusion of vitamin A intake data increases confidence in model predictions. The super-child modeling approach is an effective technique for comparing vitamin A status among children from different populations. These trials were registered at www.clinicaltrials.gov as NCT03000543 (Bangladesh), NCT03345147 (Guatemala), and NCT03030339 (Philippines).

Inclusion of Vitamin A Intake Data Provides Improved Compartmental Model-Derived Estimates of Vitamin A Total Body Stores and Disposal Rate in Older Adults.

BACKGROUND: Sampling times and study duration impact estimates of kinetic parameters and variables including total body stores (TBS) and disposal rate (DR) when compartmental analysis is used to analyze vitamin A kinetic data. OBJECTIVE: We hypothesized that inclusion of dietary intake (DI) of vitamin A as an additional input would improve confidence in predictions of TBS and DR when modeling results appear to indicate that studies are not long enough to accurately define the terminal slope of the plasma retinol isotope response curve. METHODS: We reanalyzed previously published data on vitamin A kinetics monitored over 52 d in 7 US and 6 Chinese adults (means: 56 y, BMI 26.6 kg/m2, 38% males), adding an estimate for vitamin A intake [2.8 µmol/d (mean RDA)] as an input during application of the Simulation, Analysis and Modeling software. RESULTS: Use of a model with 1 extravascular compartment (1 EV), as in the original analysis, resulted in predictions of vitamin A intake that were higher than physiologically reasonable; inclusion of intake data in a model with 2 extravascular compartments (2 EV DI) resulted in more realistic estimates of intake and DR. Specifically, predictions of DR by the 2 EV DI (versus 1 EV) model were 2.10 compared with 12.2 µmol/d (US) and 2.21 compared with 5.13 µmol/d (Chinese). Predictions of both TBS [2056 compared with 783 µmol (US) and 594 compared with 219 µmol (Chinese)] and days of vitamin A stores [981 compared with 64 d (US) and 269 compared with 43 d (Chinese)] were higher using the new approach. CONCLUSIONS: Inclusion of vitamin A intake as additional data input when modeling vitamin A kinetics can compensate for less-than-optimal study duration, providing more realistic predictions of vitamin A TBS and DR. This approach advances the application of compartmental analysis to the study of vitamin A and, potentially, other nutrients.

A Population-Based (Super-Child) Approach for Predicting Vitamin A Total Body Stores and Retinol Kinetics in Children Is Validated by the Application of Model-Based Compartmental Analysis to Theoretical Data.

BACKGROUND: Public health nutritionists need accurate and feasible methods to assess vitamin A status and to evaluate efficacy of interventions, especially in children. The application of population-based designs to tracer kinetic data is an effective approach that reduces sample burden for each child. OBJECTIVES: Objectives of the study were to use theoretical data to validate a population-based (super-child) approach for estimating group mean vitamin A total body stores (TBS) and retinol kinetics in children and to use population-based data to improve individual TBS predictions using retinol isotope dilution (RID). METHODS: We generated plasma retinol kinetic data from 6 h to 56 d for 50 theoretical children with high vitamin A intakes, assigning values within physiologically reasonable ranges for state variables and kinetic parameters ("known values"). Mean data sets for all subjects at extensive (n = 36) and reduced (n = 11) sampling times, plus 5 data sets for reduced numbers (5/time, except all at 4 d) and times, were analyzed using Simulation, Analysis and Modeling software. Results were compared with known values; population RID coefficients were used to calculate TBS for individuals. RESULTS: For extensive and reduced data sets including all subjects, population TBS predictions were within 1% of the known value. For 5 data sets reflecting numbers and times being used in ongoing super-child studies, predictions were within 1-17% of the known group value. Using RID equation coefficients from population modeling, TBS predictions at 4 d were within 25% of the known value for 66-80% of subjects and reflected the range of assigned values; when ranked, predicted and assigned values were significantly correlated (Rs  = 0.93, P < 0.0001). Results indicate that 7 d may be better than 4 d for applying RID in children. For all data sets, predictions for kinetic parameters reflected the range of known values. CONCLUSION: The population-based (super-child) approach provides a feasible experimental design for quantifying retinol kinetics, accurately estimating group mean TBS, and predicting TBS for individuals reasonably well.

Use of a "Super-child" Approach to Assess the Vitamin A Equivalence of Moringa oleifera Leaves, Develop a Compartmental Model for Vitamin A Kinetics, and Estimate Vitamin A Total Body Stores in Young Mexican Children.

Background: Worldwide, an estimated 250 million children <5 y old are vitamin A (VA) deficient. In Mexico, despite ongoing efforts to reduce VA deficiency, it remains an important public health problem; thus, food-based interventions that increase the availability and consumption of provitamin A-rich foods should be considered.Objective: The objectives were to assess the VA equivalence of 2H-labeled Moringa oleifera (MO) leaves and to estimate both total body stores (TBS) of VA and plasma retinol kinetics in young Mexican children.Methods: ß-Carotene was intrinsically labeled by growing MO plants in a 2H2O nutrient solution. Fifteen well-nourished children (17-35 mo old) consumed puréed MO leaves (1 mg ß-carotene) and a reference dose of [13C10]retinyl acetate (1 mg) in oil. Blood (2 samples/child) was collected 10 times (2 or 3 children each time) over 35 d. The bioefficacy of MO leaves was calculated from areas under the composite "super-child" plasma isotope response curves, and MO VA equivalence was estimated through the use of these values; a compartmental model was developed to predict VA TBS and retinol kinetics through the use of composite plasma [13C10]retinol data. TBS were also estimated with isotope dilution.Results: The relative bioefficacy of ß-carotene retinol activity equivalents from MO was 28%; VA equivalence was 3.3:1 by weight (0.56 µmol retinol:1 µmol ß-carotene). Kinetics of plasma retinol indicate more rapid plasma appearance and turnover and more extensive recycling in these children than are observed in adults. Model-predicted mean TBS (823 µmol) was similar to values predicted using a retinol isotope dilution equation applied to data from 3 to 6 d after dosing (mean ± SD: 832 ± 176 µmol; n = 7).Conclusions: The super-child approach can be used to estimate population carotenoid bioefficacy and VA equivalence, VA status, and parameters of retinol metabolism from a composite data set. Our results provide initial estimates of retinol kinetics in well-nourished young children with adequate VA stores and demonstrate that MO leaves may be an important source of VA.

Should We Restrict Vitamin A Intake, a Minor Contributor to Plasma Retinol Turnover, When Using Retinol Isotope Dilution Equations to Estimate an Individual's Vitamin A Status, or Should Vitamin A Balance Be Maintained?

We discuss whether dietary vitamin A intake should be restricted or maintained at balance when retinol isotope dilution equations are applied to estimate an individual's vitamin A total body stores (TBS) after oral administration of a labeled dose of vitamin A. Although, at first glance, restriction makes sense as a way to prevent dilution of tracer in plasma, further investigation of the assumptions underlying the widely used isotope dilution equation presented by Olson's laboratory in 1989, as well as the compartmental modeling results presented in this article, indicate that, in fact, restriction leads to an incorrect prediction of TBS if steady state retinol isotope dilution equations are applied at the traditional time (21 d). Our results show that newly ingested vitamin A is a minor contributor to total plasma retinol turnover and that restriction of vitamin A intake leads to a higher plasma retinol specific activity than the value obtained when vitamin A input equals output (balance). When that higher specific activity is used in the traditional retinol isotope dilution equation, it results in a small but notable underestimation of vitamin A TBS. We conclude that, especially if blood is sampled at the traditional time, the most accurate results will be obtained when vitamin A balance is maintained. If sampling is done soon after dosing (e.g., 4 d), dietary intake has less effect on plasma retinol specific activity and thus on predictions of vitamin A status. Vitamin A status can also be estimated if intake is completely restricted and a different (non-steady state) equation is applied at an appropriate time after isotopic equilibrium has been reached.

Retinol Isotope Dilution Is Applied during Restriction of Vitamin A Intake to Predict Individual Subject Total Body Vitamin A Stores at Isotopic Equilibrium.

BACKGROUND: Retinol isotope dilution (RID) equations are used to determine vitamin A status and the efficacy of vitamin A intervention programs. Recent work related to RID methods has focused on modifying the "Olson equation" to improve the accuracy of predictions of vitamin A total body stores (TBS) in individual subjects. OBJECTIVE: We investigated the hypothesis that short-term restriction of vitamin A intake would result in accurate RID prediction of vitamin A TBS in individuals. METHODS: We applied model-based compartmental analysis to a 6-component model derived from published retinol kinetic studies on 12 individuals with a wide range of vitamin A stores and determined vitamin A TBS in the steady state. Then we simulated the impact of eliminating or strictly limiting vitamin A intake at the time of isotope administration, while maintaining plasma retinol homeostasis, on retinol specific activity in plasma (SAp; fraction of dose/µmol retinol) and stores, and we calculated TBS using the simplified RID equation TBS = 0.75 × 1/SAp, where the fractional absorption of tracer was set at 0.75 and SAp was simulated 5 d after dosing. RESULTS: When vitamin A intake was zero or strictly limited (0.25 µmol/d), mean TBS predicted by the equation at 5 d after dose administration divided by TBS determined by using the model was 1.00 (range: 0.959-1.04) or 1.02 (range: 0.983 - 1.06), respectively. CONCLUSIONS: By eliminating or strictly limiting vitamin A input, isotopic equilibrium was reached by 5 d. At isotopic equilibrium, SAp is the same as that in the body's exchangeable vitamin A pools; under these conditions, SAp may be measured at any time from 5 d on and used to calculate TBS.