Consuming fruit juice/smoothies could help overcome barriers to fruit and vegetable (F&V) intake. However, their contribution towards F&V intake within a healthy diet is contentious. We investigated how F&V intake is affected by UK 5-a-day advice, with and without one portion/d of fruit juice/smoothies and explored how these interventions impacted markers of health. Healthy individuals (n 42) with low F&V intake (≤ 2 servings/d) completed a 4-week, parallel-group randomised controlled trial (ClinicalTrials.gov ID: NCT06628401). Participants were randomised to (1) control, (2) whole F&V (FV) or (3) whole F&V plus fruit juice/smoothies (FV + FJ). All groups received weekly financial support (to remove financial barriers to F&V purchase in the intervention groups). FV and FV + FJ also received a co-designed educational booklet. We investigated the intervention effects on self-reported F&V intake (primary outcome), biomarkers of intake and metabolism, mood, gut symptoms and acceptability. Post-intervention F&V intake differed between groups (P < 0·001; η p 2 = 0·62). It was significantly higher in FV ((estimated marginal means (se)); 8·9 (0·64) portions/d, P < 0·001) and FV + FJ (6·6 (0·64), P < 0·001) v. control (2·45 (0·64)), but there was no difference between FV and FV + FJ (P = 0·051). Both interventions showed good acceptability. Depression symptoms differed between groups (P = 0·01; η p 2 = 0·21); they were significantly lower in FV + FJ than control. There were no differences in anxiety or gut symptoms, nor in intake or metabolic biomarkers. A financial and educational intervention based on UK 5-a-day recommendations, with or without fruit juice/smoothies, significantly increased short-term F&V intake and benefited mood without adversely impacting health markers in the short-term. Funder: Fruit Juice Science Centre.

Vitamin A (VitA) is a fat-soluble micronutrient involved in various physiological processes. Its sustained intake exceeding physiological needs leads to increased hepatic accumulation warranting careful monitoring for potential health implications. Despite being widely studied in animal models, there are limited data on histopathology associated with VitA toxicity in human liver that are essential to understand organ response and its pathogenesis to guide clinical management. A literature search was conducted, focusing on peer-reviewed articles that explored the histological effects of excessive liver VitA accumulation in humans, excluding veterinary studies, reviews and articles without histological findings. Data on demographic, clinical, laboratory and histological information were extracted. Thirty-nine studies, including results from 84 liver tissue samples, were reviewed (1960s-2025). Patients' mean age was 40 (3-78) years, with a balanced gender distribution. The majority of cases were due to medical therapy (67.9%) or dietary supplements (27.4%). The mean VitA intake was 112,006 international units (IU)/day (equivalent to 33,600 μg/day) over 6.8 years. The most common histological findings were hepatic stellate cell (HSC) hypertrophy/hyperplasia (90.1%), sinusoidal fibrosis (44.1%), cirrhosis (25.0%), chronic hepatitis (23.8%) and variable steatosis, fibrosis and cholestasis. In humans, the hepatic response to excess VitA begins with HSC hypertrophy and hyperplasia, which represent the main adaptive storage mechanism; hepatotoxicity subsequently manifests through sinusoidal dilatation, chronic inflammation and fibrosis, occasionally progressing to cirrhosis. The severity of liver injury correlates with the duration and dose of VitA intake, with genetic and environmental factors potentially influencing clinical outcomes.

The retinol isotope dilution (RID) technique relies on the administration of isotopically labeled vitamin A tracers to human subjects. Here, we present a method to guide research teams on how to prepare vitamin A stable isotope tracers for both research studies and population surveys. The article provides an overview of the preparation, sourcing of materials, equipment required, and procedures to ensure that the tracers are safe for human consumption. Vitamin A stable isotope tracers can be obtained commercially and are usually labeled with either deuterium [2H] or carbon-13 [13C] with >95% all-trans isomeric purity and >99% isotopic enrichment. The preparation of the vitamin A stable isotope tracers involves dissolving the tracer in edible vegetable oil through sonication and heat. Tracers need to be prepared at a concentration that is well within the volumetric limits of the delivery system. Finally, the tracer is aliquoted into sterile amber glass vials, and tracer concentrations are confirmed spectrophotometrically.

The retinol isotope dilution (RID) method estimates the total body stores (TBS) of vitamin A in humans through stable isotope dilution with the body's vitamin A pool. For this, it is critical to accurately and efficiently determine the plasma isotopic ratio of labeled to non-labeled retinol through mass spectrometry. To avoid extensive and time-consuming extraction and/or purification procedures, such as preparative HPLC and derivatization, LC-MS/MS is employed to conduct fast, sensitive, and simultaneous analysis of labeled and non-labeled retinoids. The method utilizes two levels of detection: (i) an initial mass/charge (m/z) separation of parent (precursor) ions, followed by (ii) detection of fragmented daughter (product) ions. This results in high sensitivity, with retinol detection limits as low as 6 fmol on-column. Despite the advantages of tandem mass spectrometry, a liquid chromatographic separation is required to separate retinol from retinyl esters since the terminal functional groups are lost during ionization, resulting in similar parent (m/z of 269) and daughter ion fragmentation patterns. The article describes the isolation of endogenous and labeled (13C or 2H) retinol from plasma by solvent extraction, followed by quantification by LC-MS/MS under atmospheric pressure chemical ionization (APCI) in positive ion mode.

INTRODUCTION:Micronutrient deficiencies are prevalent in West Africa, particularly among women of reproductive age (WRA) and young children. Bouillon is a promising food fortification vehicle due to its widespread consumption. This study aims to evaluate the impact of multiple micronutrient-fortified bouillon cubes, compared to control bouillon cubes (fortified with iodine only), on micronutrient status and hemoglobin concentrations among lactating and non-lactating WRA and young children in northern Ghana. METHODS:This randomized, controlled doubly-masked trial will be conducted in the Kumbungu and Tolon districts in the Northern Region of Ghana, where prior data indicate multiple micronutrient deficiencies are common. Participants will be: 1) non-pregnant non-lactating WRA (15-49 y), 2) children 2-5 y, and 3) non-pregnant lactating women 4-18 months postpartum. Eligible participants will be randomly assigned to receive household rations of one of two types of bouillon cubes: 1) a multiple micronutrient-fortified bouillon cube containing vitamin A, folic acid, vitamin B12, iron, zinc, and iodine, or 2) a control cube containing iodine only. Each participant's household will receive a ration of bouillon cubes every 2 weeks, and households will be advised to prepare meals as usual, using the study-provided cubes. The trial duration will be 9 months for non-pregnant non-lactating WRA and children, and 3 months for lactating women. The primary outcomes will be changes in biomarkers of micronutrient status and hemoglobin among WRA and children and milk micronutrient concentrations among lactating women. Secondary outcomes will include change in prevalence of micronutrient deficiency and anemia; dietary intake of bouillon and micronutrients; inflammation, malaria, and morbidity symptoms; and child growth and development. DISCUSSION:Evidence from this study will inform discussions about bouillon fortification in Ghana and West Africa. TRIAL REGISTRATION:The trial was registered on ClinicalTrials.gov (NCT05178407) and the Pan-African Clinical Trial Registry (PACTR202206868437931). This manuscript reflects protocol version 4 (August 29, 2022).

Background: Limited data are available on vitamin A kinetics and total body stores (TBS) in women. Such information can be obtained using compartmental modeling and retinol isotope dilution (RID). Objectives: Objectives were to apply population-based ("super-subject") modeling to determine retinol kinetics in nonpregnant Ghanaian women of reproductive age and to use RID to predict TBS in the group and its individuals. Methods: Women (n 1 / 4 89) ingested a dose of [2H6]retinyl acetate and blood samples (3/woman) were collected from 6 h to 91 d, with all participants sampled at 14 d, about half at either 21 or 28 d, and each at one other time. Composite data (plasma retinol fraction of dose; FDp) were analyzed using Simulation, Analysis and Modeling software to obtain kinetic parameters, TBS, and other state variables as well as model-derived values for the RID composite coefficient FaS. The latter were used in the RID equation TBS (mu mol) 1 / 4 FaS x 1/SAp (where SAp is plasma retinol specific activity) to predict TBS at various times. Results: Model-predicted TBS was 973 mu mol (n 1 / 4 87). Geometric mean RID-predicted TBS was 965, 926, and 1006 mu mol at 14, 21, and 28 d, respectively, with wide ranges [for example, 252-3848 mu mol on day 14 (n 1 / 4 86)]; TBS predictions were similar at later times. Participants had a mean 2 y of vitamin A in stores and estimated liver vitamin A concentrations in the normal range. Model-predicted vitamin A disposal rate was 1.3 mu mol/d and plasma recycling number was 37. Conclusions: Super-subject modeling provides an estimate of group mean TBS as well as group-specific values for the RID coefficient FaS; the latter can be used to confidently predict TBS by RID for individual participants in the group under study or in similar individuals at 14 d or more after isotope ingestion. Trial registration number: Trial is registered (NCT04632771) at https://clinicaltrials.gov.

A systematic review identified eight human intervention studies and thirteen animal experiments investigating associations between quinoa consumption and biomarkers of CVD risk. In humans, lipid profiles were improved following quinoa consumption compared with baseline or control. Weighted mean differences (WMD) for total- and LDL-cholesterol concentrations were -0.27 mmol/L (95% CI: -0.41, -0.12, P < .001), and -0.21 mmol/L (95% CI: -0.39, -0.03, P = .023), respectively. For triglycerides WMD were -0.08 mmol/L (95% CI: -0.13, -0.03, P = .002). Glucose concentrations were also significantly decreased by quinoa (WMD -0.14 mmol/L (95% CI: -0.39, 0.10, P = .24). Except for triglycerides, results showed no evidence of bias (Begg's and Egger's test P < .05). Results from animal experiments were more variable but overall showed similar effects for total cholesterol and triglycerides. Overall, the systematic review and meta-analysis supports potential beneficial impacts of quinoa consumption for CVD prevention but identifying mechanisms for the effects requires further investigation.

AbstractInformation on fortifiable food consumption is essential to design, monitor and evaluate fortification programmes, yet detailed methods like 24‐h recalls (24HRs) that provide such data are rarely conducted. Simplified questionnaire‐based methods exist but their validity compared with 24HRs has not been shown. We compared two simplified methods (i.e., a household food acquisition and purchase questionnaire [FAPQ] and a 7‐day semiquantitative food frequency questionnaire [SQ‐FFQ]) against 24HRs for estimating fortifiable food consumption. We assessed the consumption of fortifiable wheat flour and oil using a FAPQ and, for wheat flour only, a 7‐day SQ‐FFQ and compared the results against 24HRs. The participants included children 12−18 months (n = 123) and their mothers 18−49 years selected for a study assessing child vitamin A intake and status in Mandaluyong City, Philippines. For fortifiable wheat flour, the FAPQ estimated considerably lower mean intakes compared to 24HRs for children and mothers (2.2 vs. 14.1 g/day and 5.1 vs. 42.3 g/day, respectively), while the SQ‐FFQ estimated slightly higher mean intakes (15.7 vs. 14.1 g/day and 51.5 vs. 42.3 g/day, respectively). For fortifiable oil, the FAPQ estimated considerably higher mean intakes compared to 24HRs for children and mothers (4.6 vs. 1.8 g/day and 12.5 vs. 6.1 g/day, respectively). The SQ‐FFQ, but not the FAPQ, generated useful information on fortifiable food consumption that can inform fortification programme design and monitoring decisions in the absence of more detailed individual‐level data. Potential adaptations to improve the FAPQ, such as additional questions on foods prepared away from home and usage patterns, merit further research.

BACKGROUND:Suboptimal plasma retinol concentrations have been documented in US children with sickle cell disease (SCD) hemoglobin SS type (SCD-HbSS), but little is known about vitamin A kinetics and stores in SCD. OBJECTIVES:The objectives were to quantify vitamin A total body stores (TBS) and whole-body retinol kinetics in young people with SCD-HbSS and use retinol isotope dilution (RID) to predict TBS in SCD-HbSS and healthy peers as well as after vitamin A supplementation in SCD-HbSS subjects. METHODS:Composite plasma [13C10]retinol response data collected from 22 subjects with SCD-HbSS for 28 d after isotope ingestion were analyzed using population-based compartmental modeling ("super-subject" approach); TBS and retinol kinetics were quantified for the group. TBS was also calculated for the same individuals using RID, as well as for healthy peers (n = 20) and for the subjects with SCD-HbSS after 8 wk of daily vitamin A supplements (3.15 or 6.29 μmol retinol/d [900 or 1800 μg retinol activity equivalents/d]). RESULTS:Model-predicted group mean TBS for subjects with SCD-HbSS was 428 μmol, equivalent to ∼11 mo of stored vitamin A; vitamin A disposal rate was 1.3 μmol/d. Model-predicted TBS was similar to that predicted by RID at 3 d postdosing (mean, 389 μmol; ∼0.3 μmol/g liver); TBS predictions at 3 compared with 28 d were not significantly different. Mean TBS in healthy peers was similar (406 μmol). RID-predicted TBS for subjects with SCD-HbSS was not significantly affected by vitamin A supplementation at either dose. CONCLUSIONS:Despite differences in plasma retinol concentrations, TBS was the same in subjects with SCD-HbSS compared with healthy peers. Because 56 d of vitamin A supplementation at levels 1.2 to 2.6 times the Recommended Dietary Allowance did not increase TBS in these subjects with SCD-HbSS, further work will be needed to understand the effects of SCD on retinol metabolism. This trial was registered as NCT03632876 at clinicaltrials.gov.

Diets rich in whole grains are associated with improved health and a lower risk of non-communicable diseases, but the mechanisms through which these health benefits are conveyed are uncertain. One mechanism may be improvements in the gut environment by the delivery of fermentable substrates and associated phytochemicals to the lower gut and modification of the gut microbiome. Quinoa is included in the whole-grain category because of its structural similarities to cereals but the effects of its consumption on the gut microbiome have not been investigated to date. Our aim was to examine the impact of daily quinoa consumption on the gut microbiome in a 4-week randomised cross-over intervention separated by a 4-week wash-out period involving 28 adult males. Participants consumed either a quinoa-enriched wheat-bread roll providing 20 g quinoa flour each day, or a control wheat-only bread roll. Stool samples were collected in sterile collection tubes immediately before and at the end of each intervention period. DNA was then extracted, and the 16S rRNA V4 region of extracted DNA was amplified and sequenced. For both the control and quinoa bread periods, there were no changes at the phyla or genus level between baseline and week 4 (all p > 0.05). Diversity in the microbiome profile was not different from baseline after either intervention arms. The results show that small changes in the type of cereal consumed—substituting 20 g of refined wheat flour with whole-grain quinoa flour—was not able to significantly modulate the gut microbiome. Further studies with higher levels of quinoa or longer exposure periods are needed to ascertain if there is a dose–response effect of quinoa, and if these effects are able to translate into clinical outcomes.

Low vitamin A (VA) status is common among lactating women in low-income countries. Lactation has substantial effects on mother's metabolism and VA is required in multiple biological processes, including growth, vision, immunity, and reproduction. The objective of this pilot study was to use metabolomics profiling to conduct a broad, exploratory assessment of differences in plasma metabolites associated with low VA status versus VA adequacy in lactating women. Plasma samples from lactating women who participated in a survey in Samar, Philippines, were selected from a cross-sectional study based on plasma retinol concentrations indicating low (VA–; n = 5) or adequate (VA+; n = 5) VA status (plasma retinol <0.8 or >1.05 µmol/L). The plasma results collected from 6 metabolomics assays (oxylipins, endocannabinoids, bile acids, primary metabolomics, biogenic amines, and lipidomics) were compared by group using liquid chromatography mass spectrometry. Twenty-eight metabolites were altered in the VA– versus VA+ status groups, with 24 being lipid mediators (P < .05). These lipid mediators included lower concentrations of arachidonic acid- and eicosapentaenoic acid-derived oxylipins, as well as lysophospholipids and sphingolipids, in the VA– group (P < .05). Chemical similarity enrichment analysis identified hydroxy-eicosatetraenoic acids, hydroxy-eicosapentaenoic acids, and dihydroxy-eicosatetraenoic acids as significantly altered oxylipin clusters (P < .0001, false discovery rate [FDR] P < .0001), as well as sphingomyelins, saturated lysophosphatidylcholines, phosphatidylcholines, and phosphatidylethanolamines (P < .001, FDR P < .01). The multiassay nutritional metabolomics profiling of low VA status compared with adequacy in lactating women was characterized by reduced lipid mediator concentrations. Future studies with stronger study designs and larger sample size are needed to confirm and validate these preliminary results.

Vitamin A deficiency is a major health risk for infants and children in low- and middle-income countries. This scoping review identified, quantified, and mapped research for use in updating nutrient requirements and upper limits for vitamin A in children aged 0 to 48 months, using health-based or modelling-based approaches. Structured searches were run on Medline, EMBASE, and Cochrane Central, from inception to 19 March 2021. Titles and abstracts were assessed independently in duplicate, as were 20% of full texts. Included studies were tabulated by question, methodology and date, with the most relevant data extracted and assessed for risk of bias. We found that the most recent health-based systematic reviews and trials assessed the effects of supplementation, though some addressed the effects of staple food fortification, complementary foods, biofortified maize or cassava, and fortified drinks, on health outcomes. Recent isotopic tracer studies and modelling approaches may help quantify the effects of bio-fortification, fortification, and food-based approaches for increasing vitamin A depots. A systematic review and several trials identified adverse events associated with higher vitamin A intakes, which should be useful for setting upper limits. We have generated and provide a database of relevant research. Full systematic reviews, based on this scoping review, are needed to answer specific questions to set vitamin A requirements and upper limits.

Background: Young children exposed to high-dose vitamin A supplements (VAS) and vitamin A (VA)-fortified foods may be at risk of high VA intake and high VA total body stores (TBS). Objectives: TBS and estimated liver VA concentration were compared among children with adequate or high VA intake and different timing of exposure to VAS, and associations between estimated liver VA concentrations and biomarkers of VA toxicity were examined. Methods: Children 12-18 mo of age (n = 123) were selected for 3 groups: 1) retinol intake >600 mu g/d and VAS within the past mo, 2) retinol intake >600 mu g/d and VAS in the past 3-6 mo, and 3) VA intake 200-500 mu g retinol activity equivalents (RAE)/d and VAS in the past 3-6 mo. Dietary intake data were collected to measure VA intakes from complementary foods, breast milk, and low-dose, over-the-counter supplements. TBS were assessed by retinol isotope dilution, and VA toxicity biomarkers were measured. Main outcomes were compared by group. Results: Mean (95% CI) VA intakes excluding VAS were 1184 (942, 1426), 980 (772, 1187), and 627 (530, 724) mu g RAE/d, in groups 1-3, respectively; mean VA intake was higher in groups 1 and 2 compared with group 3 (P < 0.05). Geometric mean (GM) (95% CI) TBS were 589 (525, 661), 493 (435, 559), and 466 (411, 528) mu mol, respectively. GM TBS and GM liver VA concentrations were higher in group 1 compared with group 3 (liver VA concentration: 1.62 vs. 1.33 mu mol/g; P < 0.05). Plasma retinyl ester and 4-oxo-retinoic acid concentrations and serum markers of bone turnover and liver damage did not indicate VA toxicity. Conclusions: In this sample, most children had retinol intakes above the Tolerable Upper Intake Level (UL) and liver VA concentrations above the proposed cutoff for "hypervitaminosis A" (>1 mu mol/g liver). There was no evidence of chronic VA toxicity, suggesting that the liver VA cutoff value should be re-evaluated.

Provitamin A carotenoids (pVACs) are important contributors to vitamin A status and in reducing vitamin A deficiency in vulnerable populations. However, there are uncertainties about the effectiveness of dietary pVACs to provide sufficient amounts of vitamin A due to large variations in provitamin A bioefficacy, and if provitamin A carotenoids other than β-carotene could be utilized in biofortification programs. Although animal studies have compared the bioefficacy of β-cryptoxanthin and β-carotene in biofortified maize by measuring liver retinol concentrations after dietary exposure, it is unclear whether the higher bioavailability of β-cryptoxanthin can be offset by the lower conversion of β-cryptoxanthin to retinol, and how post-intestinal conversion of β-cryptoxanthin may contribute to the total bioefficacy of β-cryptoxanthin. Here, we present a method that, for the first time, uses stable isotope labeled β-cryptoxanthin and β-carotene to quantify bioconversion and bioefficacy of both pVACs simultaneously in human volunteers. The paper describes how positioning of the [13C] labels around the centric 15,15' double bond on either the β-carotene or β-cryptoxanthin molecule allows quantification of retinoids from both pVACs, and details the procedure for sample preparation and analysis using LC-MS/MS. Finally, we apply and discuss recent approaches to quantify bioconversion and bioefficacy of isotopically labeled β-carotene and β-cryptoxanthin in humans.