Centers for Disease Control and Prevention's 'National Report on Biochemical Indicators of Diet and Nutrition in the US Population' provides nutritional status assessment by presenting blood and urine concentrations of diet and nutrition biomarkers in a representative sample of the US population ≥1 y participating in the NHANES. Earlier reports contained information for 27 (1999-2002) and 58 (2003-2006) biomarkers. We aimed to provide an overview of the 2026 report, which covers data for ≤131 biomarkers since 1999, with 50 biomarkers from August 2021 to August 2023. Concentrations over time by cycle are stratified by dietary supplement use, reference interval concentrations focus on tail percentiles, and concentrations grouped by age, sex, and/or race and Hispanic origin demonstrate demographic differences. The report is intended for agencies and organizations involved in developing nutrition policies and monitoring their effects. It provides easily accessible, comprehensive nutritional biomarker data for the US population during the last 24 y.

Background:Tracking the prevalence of imbalances for essential nutrients over time provides insight into nutritional status changes and highlights vulnerabilities of demographic subgroups. Consolidated information for a broad range of biomarkers in the US population is missing. Objectives:We aimed to describe nutritional deficiency and excess prevalence trends for 12 biomarkers of water- and fat-soluble vitamins and iron status in the US population participating in the NHANES 1999-2023 after adjusting for demographic changes over time. Methods:We estimated the prevalence of deficiency (vitamins A, B-6, B-12, C, D, and E, as well as iron), insufficiency [vitamins B-6, B-9 (folate), B-12, C, and D], and excess (vitamins A and D as well as iron). We assessed trends as overall change in prevalence using logistic regression (unadjusted and adjusted for age, sex, and race/Hispanic origin) and as linear trend (if overall change was significant). Results:After adjusting for demographic changes, we observed small (typically <5 percentage points) temporal changes (significant linear trends) in the prevalence of deficiency and/or insufficiency for vitamin B-12 and methylmalonic acid (≥20 y), and for vitamin E and 25-hydroxyvitamin D (25OHD) (≥6 y). The prevalence of folate insufficiency (red blood cell folate (RBF) <748 nmol/L) and iron deficiency (ferritin ≤15 ng/mL) increased from 15.7% to 21.4% and from 11.6% to 17.4%, respectively, in women of reproductive age. The prevalence of excess vitamin D (25OHD >125 nmol/L) increased from <1% to 8.06% in persons ≥6 y, with the highest prevalence in adult subgroups (16.2% in supplement users, 13.2% in females, and 12.3% in non-Hispanic White persons). Conclusions:The large increases in excess 25OHD concentrations and modest increases in low ferritin and RBF concentrations merit attention and highlight the importance of providing gold standard data to continuously monitor the nutritional status of the US population and of subpopulations.

Background:The National Health and Nutrition Examination Survey (NHANES) has monitored biomarkers for 4 key nutrients important for pregnancy outcomes, neurological development, and healthy growth-folate, vitamin D, iodine, and iron-during most survey cycles from 1999 to 2000 through August 2021-August 2023 as part of the continuous NHANES. These biomarker data offer a unique opportunity to assess long-term trends in the US population. Objectives:This study aimed to describe concentration trends for 7 biomarkers/indicators after adjusting for demographic changes over time. Methods:We used the following NHANES data: serum folate (SFOL) and red blood cell folate (RBF) for persons aged ≥3 y and women of reproductive age (WRA, 12-49 y) for 1999-August 2023 (except 2017-2018), serum 25-hydroxyvitamin D (25OHD) for persons aged ≥3 y for 2001-August 2023, urine iodine (UI) for persons aged ≥3 y for 2001-2018, and serum ferritin [(FER) and FER adjusted for inflammation (FERadj)], serum soluble transferrin receptor, and body iron index (BII) for WRA and children aged 1-5 y for 2003-August 2023 (except 2011-2014). We assessed trends as overall concentration change using logistic regression (unadjusted and adjusted for age, sex, and race and Hispanic origin) and as a linear trend if the overall change was significant. Results:After adjusting for demographic changes, mean concentrations of all 7 biomarkers/indicators showed significant overall temporal changes (except for FER in children and BII in WRA) and significant linear trends (except for FERadj in children). SFOL (∼25%-30%), RBF (∼10%), and UI (∼25%-30%) concentrations decreased, whereas 25OHD (∼20%) concentrations increased. FER concentrations decreased in WRA (∼15%) and remained unchanged in children. Conclusions:Considering the long monitoring period, biomarker changes were moderate. Nonetheless, these changes highlight the need to address declining iodine, iron, and folate status in WRA and increasing vitamin D status in supplement users.

Several essential vitamins and minerals whose deficiencies are associated with metabolic and functional disorders, including increased morbidity and mortality from both communicable and noncommunicable diseases, are not being adequately addressed by large-scale, public health nutrition programs worldwide. These neglected micronutrients include thiamine, riboflavin, niacin, pyridoxine, vitamin B-12, vitamin D, vitamin K, calcium, selenium, and possibly others. In this narrative review, our objectives are to describe briefly the health implications of each of these deficiencies, summarize the limited available information on their epidemiology, and suggest possible approaches to address them. We conclude that more information, based on dietary assessments, nutritional biomarker surveys, and systematic surveillance of associated health conditions, is needed. Appropriate intervention programs, including changes in food systems to provide wider access to nutrient-rich foods, food fortification and targeted supplementation, should be implemented in settings where these deficiencies are confirmed.

BACKGROUND:The temporal variability of caffeine and caffeine metabolite concentrations in spot urine samples is unknown. Understanding this variability in both unadjusted and flow-adjusted measurements is essential to assess the suitability of spot urine sampling for measuring these compounds and estimate the number of samples required for reliable measurements. OBJECTIVES:This study aimed to examine the temporal variability of unadjusted and flow-adjusted concentrations of caffeine and its metabolites in spot urine samples, both within and between individuals, over short-to-intermediate timeframes. METHODS:We measured urine caffeine and 14 caffeine metabolite concentrations in 1749 spot samples collected from a cohort of 50 adults over a 6-wk period from the Pilot Study to Estimate Human Exposures to Pyrethroids using an Exposure Reconstruction Approach. We calculated between- and within-person variances, 95%-fold ranges, and intraclass correlation coefficients (ICCs) for unadjusted, flow rate-, creatinine-, and specific gravity-adjusted analyte concentrations over timeframes of 24 h, 1 wk, and 6 wk. Additionally, we estimated the number of spot urine samples needed for reliable measurements (ICC ≥ 0.80). RESULTS:We found that ICCs were the highest in the 24-h timeframe, with 69% of analyte × urine flow adjustment combinations yielding median ICCs ≥0.80 (range: 0.62-0.94). ICCs decreased as the timeframe increased (1-wk median ICC range: 0.49-0.84; 6-wk ICC range: 0.37-0.76). Caffeine showed no ICC improvement with urine flow adjustments, whereas dimethylxanthines showed mixed results. Later-forming metabolites consistently showed ICC improvement across all types of urine flow adjustments. CONCLUSIONS:Overall, 1-2 spot urine samples were sufficient to obtain a reliable concentration estimate (ICC ≥ 0.80) for caffeine and most caffeine metabolites during 24-h or 1-wk sampling timeframes with appropriate urine flow adjustment, with 2-3 spot urine samples being sufficient for sampling over 6 wk.

Background: Spot urine concentrations of caffeine and certain caffeine metabolites correlate well with 24-h caffeine intake and have potential use as caffeine exposure biomarkers. However, a high degree of intercorrelation exists among these urine compounds, and their correlations with intake are very similar, making the selection of a single intake biomarker challenging based on bivariate analyses alone. Objectives: To use principal components (PCs) analysis (PCA) as a dimensionality reduction tool to identify underlying correlation structures in the relationship of caffeine intake with spot urine caffeine and caffeine metabolite concentrations. Methods: We performed weighted bivariate analyses and a weighted PCA of spot urine concentrations of caffeine and 14 caffeine metabolites, and 24-h intakes of caffeine (foods, beverages, and supplements) and theobromine (foods and beverages) from 7732 participants ≥6 y in the National Health and Nutrition Examination Survey (NHANES) 2009–2014. Results: Bivariate analyses revealed intercorrelation patterns that effectively divided the urine analytes into 2 groups based on their association with either caffeine or theobromine intake. PCA yielded 2 components (PC1 and PC2) that accounted for 83.0% of the total variance. PC1 (70.9%) showed a positive correlation with all original variables and approximated a weighted sum related to the urine analyte concentrations in the original data. PC2 (12.1%) yielded 2 clusters of correlated variables resembling a weighted contrast of the caffeine and theobromine intake associations observed in the original data. Urine 1-methylxanthine showed the strongest PCA correlation with caffeine intake, followed by 5-acetylamino-6-amino-3-methyluracil, 1-methyluric acid, and 1,7-dimethyluric acid. Urine theobromine and its downstream metabolites showed PCA correlation primarily with theobromine intake, though correlation with caffeine intake was also evident. Conclusions: With PCA, we were able to identify an underlying correlation structure in the NHANES 2009–2014 data that revealed promising urine exposure biomarkers of caffeine and theobromine intake.

Aflatoxin exposure is endemic in developing countries with warm, humid climates that promote toxigenic mold growth on crops and foodstuffs. Estimating human aflatoxin exposure is key to identifying and abating contamination sources. Serum aflatoxin B1 bound to albumin lysine (AFB1-lys) is a preferred exposure biomarker, but field sample collection, processing, transportation, and storage logistics are challenging. We validated an improved LC-MS/MS method for serum AFB1-lys and applied it to three field sampling challenges: transportation/storage (elevated temperature); collection/processing (hemolysis); and sample type substitution (heparinized plasma). Our new LC-MS/MS method had a LOD of 0.03 ng/mL, accuracy (mean spike recovery) of 112%, total imprecision (replicate pool measurements) ≤5% at ≥0.2 ng/mL, and results that were 95.1% similar (mean percentage similarity) to an established method. AFB1-lys in human serum spiked with serum from aflatoxin-dosed rats was stable for 14 days at both ambient (22.5 °C) and elevated (38 °C) temperatures. Simulated hemolysis (adding 0.25–3 mg hemoglobin) did not affect AFB1-lys accuracy at ≥0.5 ng/mL but caused 10–25% signal suppression. Heparinized plasma AFB1-lys was 99.0% similar to serum but interfered with albumin measurements (bromocresol green) causing spurious low bias. Further investigation is warranted, but our findings suggest that AFB1-lys is pre-analytically robust.

ABSTRACT Aflatoxins are carcinogenic mycotoxins that contaminate a variety of crops worldwide. Acute exposure can cause liver failure, and chronic exposure can lead to stunting in children and liver cancer in adults. We estimated aflatoxin exposure across Uganda by measuring a serum biomarker of aflatoxin exposure in a subsample from the 2011 Uganda AIDS Indicator Survey, a nationally representative survey of HIV prevalence, and examined its association with geographic, demographic, and socioeconomic variables. We analysed a subsample of 985 serum specimens selected among HIV-negative participants from 10 survey-defined geographic regions for serum aflatoxin B1-lysine (AFB1-lys) by use of isotope dilution LC-MS/MS and calculated results normalised to serum albumin. We used statistical techniques for censored data to estimate geometric means (GMs), standard deviations, and percentiles. We detected serum AFB1-lys in 71.7% of specimens (LOD = 0.5 pg/mg albumin). Unadjusted GM AFB1-lys (pg/mg albumin) was 1.33 (95% CI: 1.21–1.47). Serum AFB1-lys was higher in males (GM: 1.57; 95% CI: 1.38–1.80) vs. females (GM: 1.12; 95% CI: 0.97–1.30) (P = .0019), and higher in persons residing in urban settings (GM: 2.83; 95% CI: 2.37–3.37) vs. rural (GM: 1.10; 95% CI: 0.99–1.23) (P < .0001). When we used a multivariable censored regression model to assess confounding and interactions among variables we found that survey region, gender, age, occupation, distance to marketplace, and number of meals per day were statistically significant predictors of aflatoxin exposure. While not nationally representative, our findings provide an improved understanding of the widespread burden of aflatoxin exposure throughout Uganda and identify key geographic, demographic, and socioeconomic factors that may modulate aflatoxin exposure risk.

US EPA and CDC congressional appropriations.

Phytoestrogens are estrogenic compounds that occur naturally in plants. Phytoestrogens can cross the placenta, and animal studies have found associations between in utero exposure to phytoestrogens and markers of early puberty. We investigated the association between in utero exposure to phytoestrogens and early menarche (defined as <11.5 years of age at onset) using data from a nested case-control study within the Avon Longitudinal Study of Parents and Children, a longitudinal study involving families living in the South West of England. Concentrations of six phytoestrogens were measured in maternal urine samples collected during pregnancy. Logistic regression was used to explore associations between tertiles of phytoestrogen concentrations and menarche status, with adjustment for maternal age at menarche, maternal education, pre-pregnancy body mass index (BMI), child birth order, and duration of breastfeeding. Among 367 mother-daughter dyads, maternal median (interquartile range) creatinine-corrected concentrations (in µg/g creatinine) were: daidzein 184.8 (88.8– 383.7), enterodiol 76.1 (39.1–135.8), enterolactone 911.7 (448.1–1558.0), equol 4.3 (2.8–9.0), genistein 62.1 (27.1–160.9), and O -desmethylangolensin ( O -DMA) 13.0 (4.4–34.5). In analyses comparing those in the highest tertile relative to those in the lowest tertile of in utero phytoestrogen exposure, higher enterodiol levels were inversely associated with early menarche (OR=0.45; 95% CI: 0.25–0.81), while higher O -DMA levels were associated with early menarche (odds ratio (OR) = 1.94; 95% confidence interval (CI): 1.07–3.51). These findings suggest that in utero exposure to phytoestrogens may be associated with earlier age at menarche, though the direction of association differs across phytoestrogens. A nested case-control study was conducted within the ALSPAC cohort to explore associations of prenatal maternal concentrations of various EDCs and age at menarche among the daughters. A ‘Growing and Changing’ questionnaire was developed to collect information on the offspring’s pubertal development and distributed to participants annually between the ages of 8–17 years (1999–2008), with the exception of age 12 (2003). Menarche was determined through parental- or self-report of menarche status, and, if it had occurred, month and year of occurrence so that age could be computed. From the original base population of 14,062 live births, case and control series were selected from singleton (n=11,820) female subjects (n=5,756) who had completed at least two puberty staging questionnaires between the ages of 8 and 13 (5 possible questionnaires returned; n=3,682). Girls meeting eligibility criteria were ordered according to reported age at menarche when the 13-year old data became available. A cut-off of 11.5 years was established as defining ‘early’ menarche to satisfy sample size and power needed for the case-control study. Eligible cases could complete any two questionnaires in the series, provided that one was completed after menarche, while controls had to complete the 13-year old order that menarche not by the cut-off 11.5 years. the girls

Phytoestrogens are estrogenic compounds that occur naturally in plants. Phytoestrogens can cross the placenta, and animal studies have found associations between in utero exposure to phytoestrogens and markers of early puberty. We investigated the association between in utero exposure to phytoestrogens and early menarche (defined as <11.5 years of age at onset) using data from a nested case-control study within the Avon Longitudinal Study of Parents and Children, a longitudinal study involving families living in the South West of England. Concentrations of six phytoestrogens were measured in maternal urine samples collected during pregnancy. Logistic regression was used to explore associations between tertiles of phytoestrogen concentrations and menarche status, with adjustment for maternal age at menarche, maternal education, pre-pregnancy body mass index (BMI), child birth order, and duration of breastfeeding. Among 367 mother-daughter dyads, maternal median (interquartile range) creatinine-corrected concentrations (in µg/g creatinine) were: daidzein 184.8 (88.8– 383.7), enterodiol 76.1 (39.1–135.8), enterolactone 911.7 (448.1–1558.0), equol 4.3 (2.8–9.0), genistein 62.1 (27.1–160.9), and O -desmethylangolensin ( O -DMA) 13.0 (4.4–34.5). In analyses comparing those in the highest tertile relative to those in the lowest tertile of in utero phytoestrogen exposure, higher enterodiol levels were inversely associated with early menarche (OR=0.45; 95% CI: 0.25–0.81), while higher O -DMA levels were associated with early menarche (odds ratio (OR) = 1.94; 95% confidence interval (CI): 1.07–3.51). These findings suggest that in utero exposure to phytoestrogens may be associated with earlier age at menarche, though the direction of association differs across phytoestrogens. A nested case-control study was conducted within the ALSPAC cohort to explore associations of prenatal maternal concentrations of various EDCs and age at menarche among the daughters. A ‘Growing and Changing’ questionnaire was developed to collect information on the offspring’s pubertal development and distributed to participants annually between the ages of 8–17 years (1999–2008), with the exception of age 12 (2003). Menarche was determined through parental- or self-report of menarche status, and, if it had occurred, month and year of occurrence so that age could be computed. From the original base population of 14,062 live births, case and control series were selected from singleton (n=11,820) female subjects (n=5,756) who had completed at least two puberty staging questionnaires between the ages of 8 and 13 (5 possible questionnaires returned; n=3,682). Girls meeting eligibility criteria were ordered according to reported age at menarche when the 13-year old data became available. A cut-off of 11.5 years was established as defining ‘early’ menarche to satisfy sample size and power needed for the case-control study. Eligible cases could complete any two questionnaires in the series, provided that one was completed after menarche, while controls had to complete the 13-year old order that menarche not by the cut-off 11.5 years. the girls

Background Caffeine is a widely consumed stimulant, and caffeine-containing products are increasingly available on the market. Few tools are available to capture caffeine intake, particularly among young adults. To estimate caffeine consumption in the previous 24 hours, the 24-Flour Caffeine Intake Recall (CIR-24) was modeled after the Automated Self-Administered 24-Flour Dietary Assessment Tool, using a brand-specific database of caffeine-containing foods, beverages, and supplements. Objective To evaluate the accuracy of the CIR-24 compared with caffeine concentration biomarkers in urine and a caffeinated beverage intake frequency screener (CBQ) designed to assess usual intake among a young adult population in Canada. Design/participants In all, 79 young adults, aged 18 to 29 years, provided 24-hour urine samples and completed the CIR-24 and CBQ. Main outcome measures Excretion for caffeine and eight caffeine metabolites were quantified from urine samples using high-performance liquid chromatography-polarity switching electrospray ionization-tandem quadrupole mass spectrometry with stable isotope-labeled internal standards. Statistical analyses performed Pearson correlations and weighted K coefficients were calculated for the self-report tools and caffeine biomarkers. Results The CIR-24 was significantly positively associated with all caffeine biomarkers (r(p) =0.28 to 0.52, kappa=0.39 to 0.59), and the CBQ was significantly positively associated with all but one biomarker (r(p) =0.21 to 0.40, kappa=0.32 to 0.45). The CIR-24 yielded a higher mean intake of caffeine than the CBQ There was strong linear correlation between the CIR-24 and CBQ (r(p) =0.60, P<0.001), but poor agreement in absolute caffeine consumed (t=2.83, P=0.006); quartile ranking concordance was 0.44 (P<0.001). The CIR-24 performed better than the CBQ across all biomarkers in both linear correlation and quartile ranking. Conclusions Although both the CIR-24 and CBQ performed reasonably well in capturing caffeine intake compared with urinary biomarkers of caffeine consumption, the CIR-24 had stronger agreement than the CBQ The results suggest that the CIR-24 is a promising tool for evaluating caffeine intake among this population.

Introduction Most electronic cigarettes (e-cigarettes) contain a solution of propylene glycol/glycerin and nicotine, as well as flavors. E-cigarettes and their associated e-liquids are available in numerous flavor varieties. A subset of the flavor varieties include coffee, tea, chocolate, and energy drink, which, in beverage form, are commonly recognized sources of caffeine. Recently, some manufacturers have begun marketing e-liquid products as energy enhancers that contain caffeine as an additive. Methods A Gas Chromatography-Mass Spectrometry (GC-MS) method for the quantitation of caffeine in e-liquids was developed, optimized and validated. The method was then applied to assess caffeine concentrations in 44 flavored e-liquids from cartridges, disposables, and refill solutions. Products chosen were flavors traditionally associated with caffeine (ie, coffee, tea, chocolate, and energy drink), marketed as energy boosters, or labeled as caffeine-containing by the manufacturer. Results Caffeine was detected in 42% of coffee-flavored products, 66% of tea-flavored products, and 50% of chocolate-flavored e-liquids (limit of detection [LOD] - 0.04 µg/g). Detectable caffeine concentrations ranged from 3.3 µg/g to 703 µg/g. Energy drink-flavored products did not contain detectable concentrations of caffeine. Eleven of 12 products marketed as energy enhancers contained caffeine, though in widely varying concentrations (31.7 µg/g to 9290 µg/g). Conclusions E-liquid flavors commonly associated with caffeine content like coffee, tea, chocolate, and energy drink often contained caffeine, but at concentrations significantly lower than their dietary counterparts. Estimated daily exposures from all e-cigarette products containing caffeine were much less than ingestion of traditional caffeinated beverages like coffee. Implications This study presents an optimized and validated method for the measurement of caffeine in e-liquids. The method is applicable to all e-liquid matrices and could potentially be used to ensure regulatory compliance for those geographic regions that forbid caffeine in e-cigarette products. The application of the method shows that caffeine concentrations and estimated total caffeine exposure from e-cigarette products is significantly lower than oral intake from beverages. However, because very little is known about the effects of caffeine inhalation, e-cigarette users should proceed with caution when using caffeine containing e-cigarette products. Further research is necessary to determine associated effects from inhaling caffeine.

Urine excretion of caffeine and select caffeine metabolites is common in the US population and associated with caffeine intake. In particular, caffeine metabolites resulting from metabolism via paraxanthine and/or theophylline show promise as potential caffeine intake biomarkers. The 2015–2020 Dietary Guidelines for Americans state that up to 400 mg/day of caffeine can be incorporated into a healthy eating style. In this study we performed receiver operating characteristic (ROC) analysis of unweighted data from the NHANES 2009–2010 to determine optimal cutoffs for spot urine caffeine and 14 caffeine metabolites that would maximize the correct classification of individuals with 24‐h caffeine intake >400 mg/d vs. ≤400 mg/d (from diet and supplements). We limited our study sample to subjects ≥12 y of age and excluded individuals taking prescription medications known to interfere with caffeine metabolism. For each analyte we calculated an unweighted ROC estimate for the optimal cutoff concentration, as well as estimates of sensitivity, specificity, area under the ROC curve (AUC), and an overall rate of correct classification. We calculated these estimates for the total study population, as well as stratified by age, sex and race‐ethnicity. Using our ROC estimates for optimal cutoff concentrations, we observed the highest overall rate of correct classification (72.2%) with 1,3‐dimethyluric acid (13U) (cutoff: 12.4 μmol/L; sensitivity: 69.7%; specificity: 72.2%). In addition to 13U, we also saw correct classification rates >70% for 5‐acetylamino‐6‐amino‐3‐methyluracil (AAMU) (cutoff: 98.4 μmol/L; sensitivity: 69.0%; specificity: 71.4%) and 1‐methyluric acid (1U) (cutoff: 98.1 μmol/L; sensitivity: 71.7%; specificity: 70.8%). The greatest AUC was observed for 1U (0.776; 95% CI: 0.740–0.811), with similarly high AUCs observed for AAMU (0.763; 95% CI: 0.726–0.800), 13U (0.776; 95% CI: 0.726–0.799), and 1‐methylxanthine (1X) (0.768; 95% CI: 0.733–0.804) which had correct classification rate of 68.5% (cutoff: 45.3 μmol/L; sensitivity: 0.724; specificity: 0.682). Theobromine (cutoff: 16.3 μmol/L; sensitivity: 69.7%; specificity: 49.4%) had the lowest overall rate of correct classification (50.9%), as well as the lowest AUC (0.603; 95% CI: 0.563–0.642). Depending on the analyte, we noted that the ROC estimate for optimal cutoff concentration could vary by age, sex, or race‐ethnicity. We believe that our study is the first report of using ROC analysis to determine cutoff values for spot urine caffeine and caffeine metabolite concentrations to classify individuals based on their caffeine intake, and that this approach holds promise as a means of identifying individuals whose recent caffeine intake may have exceeded a recommended upper limit of 400 mg/day.

ABSTRACT Acute aflatoxin exposure can cause death and disease (aflatoxicosis) in humans. Aflatoxicosis fatality rates have been documented to be as high as 40% in Kenya. The inclusion in the diet of calcium silicate 100 (ACCS100), a calcium montmorillonite clay, may reduce aflatoxin bioavailability, thus potentially decreasing the risk of aflatoxicosis. We investigated the efficacy, acceptability and palatability of ACCS100 in a population in Kenya with recurring aflatoxicosis outbreaks. Healthy adult participants were enrolled in this double-blinded, crossover clinical trial in 2014. Following informed consent, participants (n = 50) were randomised to receive either ACCS100 (3 g day–1) or placebo (3 g day–1) for 7 days. Treatments were switched following a 5-day washout period. Urine samples were collected daily and assessed for urinary aflatoxin M1 (AFM1). Blood samples were collected at the beginning and end of the trial and assessed for aflatoxin B1-lysine adducts from serum albumin (AFB1-lys). AFM1 concentrations in urine were significantly reduced while taking ACCS100 compared with calcium carbonate placebo (β = 0.49, 95% confidence limit = 0.32–0.75). The 20-day interval included both the placebo and ACCS100 treatments as well as a washout period. There were no statistically significant differences in reported taste, aftertaste, appearance, colour or texture by treatment. There were no statistically significant differences in self-reported adverse events by treatment. Most participants would be willing to take ACCS100 (98%) and give it to their children (98%). ACCS100 was effective, acceptable and palatable. More work is needed to test ACCS100 among vulnerable populations and to determine if it remains effective at the levels of aflatoxin exposure that induce aflatoxicosis.

Cytochrome P450 enzyme CYP2A6 is a phase I enzyme expressed primarily in the liver, accounting for approximately 4% of all P450 content. CYP2A6 plays a role in the metabolism of caffeine, nicotine, and several drugs used in the treatment of cancer and infectious diseases. CYP2A6 polymorphisms give rise to inter‐individual differences in CYP2A6 activity, and these differences can be further influenced by dietary and lifestyle factors. Lower CYP2A6 activity has been associated with a reduced risk of various cancers, and differences in CYP2A6 activity can influence smoking behavior and likelihood of cessation. Caffeine metabolite ratios (MR) have been developed to assess CYP2A6 activity, and the urine MR of 1,7‐dimethyluric acid (17U) to paraxanthine (17X) (i.e., 17U/17X) has been used to determine CYP2A6 activity. Additionally, the MR of 17U to 1‐methylxanthine (1X) (i.e., 17U/1X) has shown promise in less controlled settings where regular dietary caffeine intake and spot urine samples were used. In this study we described CYP2A6 activity in the US population ≥6 y by selected demographic, lifestyle and physiologic variables. We used spot urine molar concentrations of 17U and 1X from the cross‐sectional NHANES 2009–2010 to calculate CYP2A6 activity by use of the MR 17U/1X. We excluded the individuals with evidence of insufficient caffeine exposure to reliably calculate the CYP2A6 MR and subjects taking prescription medications known to interfere with caffeine metabolism. We performed bivariate comparisons of unadjusted geometric mean (GM) CYP2A6 MR for categorical variables (n=2197), and used Spearman correlation to examine the relation of CYP2A6 MR with continuous variables. CYP2A6 activity was higher in females vs. males (Wald F P <0.0001), highest in Hispanics, followed by non‐Hispanic whites and lowest in non‐Hispanic blacks (Wald F P <0.0001), and increased with age (Spearman ρ = 0.41, P <0.001). CYP2A6 MR was higher in individuals with evidence of inflammation (serum CRP ≥5 mg/L vs. <5 mg/L; Wald F P <0.0001), diabetes (yes vs. no based on a physician's diagnosis; Wald F P = 0.0025), and impaired kidney function (CKD 3–5 vs. CKD 1–2 and normal; Wald F P <0.0001). CYP2A6 MR was lower in smokers vs. non‐smokers (serum cotinine ≤10 ng/mL vs. >10 ng/mL; Wald F P <0.0001). CYP2A6 activity was positively associated with BMI (Spearman ρ = 0.36, P <0.0001) and was negatively associated with leisure physical activity (Spearman ρ = −0.15, P = 0.0003). The associations that we observed for CYP2A6 activity with age, sex, and race‐ethnicity, smoking status, inflammation, and physical activity were generally consistent with observations from other studies. We believe that our study is the first to report estimates of CYP2A6 activity obtained from spot urine MRs in a representative sample of the US population.

Phytoestrogens are estrogenic compounds that occur naturally in plants. Phytoestrogens can cross the placenta, and animal studies have found associations between in utero exposure to phytoestrogens and markers of early puberty. We investigated the association between in utero exposure to phytoestrogens and early menarche (defined as <11.5 years of age at onset) using data from a nested case-control study within the Avon Longitudinal Study of Parents and Children, a longitudinal study involving families living in the South West of England. Concentrations of six phytoestrogens were measured in maternal urine samples collected during pregnancy. Logistic regression was used to explore associations between tertiles of phytoestrogen concentrations and menarche status, with adjustment for maternal age at menarche, maternal education, pre-pregnancy body mass index (BMI), child birth order, duration of breastfeeding, and gestational age at sample collection. Among 367 mother-daughter dyads, maternal median (interquartile range) creatinine-corrected concentrations (in µg/g creatinine) were: genistein 62.1 (27.1–160.9), daidzein 184.8 (88.8–383.7), equol 4.3 (2.8–9.0), O-desmethylangolensin (O-DMA) 13.0 (4.4–34.5), enterodiol 76.1 (39.1–135.8), and enterolactone 911.7 (448.1–1558.0). In analyses comparing those in the highest tertile relative to those in the lowest tertile of in utero phytoestrogen exposure, higher enterodiol levels were inversely associated with early menarche (odds ratio (OR)=0.47; 95% confidence interval (CI): 0.26–0.83), while higher O-DMA levels were associated with early menarche (OR=1.89; 95% CI: 1.04–3.42). These findings suggest that in utero exposure to phytoestrogens may be associated with earlier age at menarche, though the direction of association differs across phytoestrogens.

Phenolic compounds represent a class of environmental chemicals with potentially endocrine-disrupting capabilities. We investigated longitudinal associations between childhood exposure to phenols, from both manmade and natural sources, and subsequent measures of adiposity among girls enrolled in the Breast Cancer and the Environment Research Program between 2004 and 2007. Baseline (ages 6-8 years) urinary concentrations were obtained for creatinine and phenol metabolites: enterolactone, genistein, daidzein, benzophenone-3, bisphenol A, the sum of parabens (methyl, ethyl, and propyl parabens), 2,5-dichlorophenol, and triclosan. Body mass index (weight (kg)/height (m)2), waist circumference, and percent body fat were measured at annual or semiannual examinations through 2015 (n = 1,017). Linear mixed-effects regression was used to estimate how baseline concentrations of phenols (tertile groups) were related to changes in girls' adiposity measurements from ages 7 through 15 years. Enterolactone was inversely associated with body mass index, waist circumference, and percent body fat, while 2,5-dichlorophenol was positively associated with these measurements. A nonmonotonic association was observed for triclosan and girls' adiposity; however, it was due to effect modification by baseline overweight status. Triclosan was positively associated with adiposity only among overweight girls. These results suggest that exposure to specific phenols during childhood may influence adiposity through adolescence.

To study potential environmental influences on puberty in girls, we investigated urinary biomarkers in relation to age at menarche. Phenols and phthalates were measured at baseline (6-8 years of age). Menarche was ascertained over 11 years for 1051 girls with menarche and biomarkers. Hazards ratios were estimated from Cox models adjusted for race/ethnicity and caregiver education (aHR, 95% confidence intervals [CI] for 5th vs 1st quintile urinary biomarker concentrations). 2,5-Dichlorophenol was associated with earlier menarche (aHR 1.34 [1.06-1.71]); enterolactone was associated with later menarche (aHR 0.82 [0.66-1.03]), as was mono-3-carboxypropyl phthalate (MCPP) (aHR 0.73 [0.59-0.91]); the three p-trends were <0.05. Menarche differed by 4-7 months across this range. Enterolactone and MCPP associations were stronger in girls with below-median body mass index. These analytes were also associated with age at breast development in this cohort. Findings from this prospective study suggest that some childhood exposures are associated with pubertal timing.