Development and certification of a standard reference material for vitamin D metabolites in human serum.

The National Institute of Standards and Technology (NIST), in collaboration with the National Institutes of Health's Office of Dietary Supplements (NIH-ODS), has developed a Standard Reference Material (SRM) for the determination of 25-hydroxyvitamin D [25(OH)D] in serum. SRM 972 Vitamin D in Human Serum consists of four serum pools with different levels of vitamin D metabolites and has certified and reference values for 25(OH)D(2), 25(OH)D(3), and 3-epi-25(OH)D(3). Value assignment of this SRM was accomplished using a combination of three isotope-dilution mass spectrometry approaches, with measurements performed at NIST and at the Centers for Disease Control and Prevention (CDC). Chromatographic resolution of the 3-epimer of 25(OH)D(3) proved to be essential for accurate determination of the metabolites.

Vitamin D in pregnancy and lactation in humans.

Concerns exist about the adequacy of vitamin D in pregnant and lactating women. This review assesses the evidence that maternal vitamin D status influences maternal, fetal, and breast-fed infant bone health; maternal adverse outcomes (preeclampsia, gestational diabetes, obstructed labor, and infectious disease); fetal adverse outcomes (growth, gestational age, and developmental programming); and infant adverse outcomes. The evidence for all of these outcomes is contradictory (except for maternal infectious disease) and lacking causality; thus, it is inconclusive. The 2011 Dietary Reference Intakes for vitamin D and their implications for assessing vitamin D status are discussed. An estimated 5% to 29% of American pregnant women may have inadequate vitamin D status, with the higher prevalence in African Americans. Little is known about the prevalence of inadequacy in American lactating women. Research needs are also identified, especially the need for rigorous and well-designed randomized clinical trials to determine the role of vitamin D in nonbone health outcomes in pregnancy and lactation.

Comparison of serum and red blood cell folate microbiologic assays for national population surveys.

Three laboratories participated with their laboratory-specific microbiologic growth assays (MA) in the NHANES 2007-2008 to assess whether the distributions of serum (n = 2645) and RBC folate (n = 2613) for the same one-third sample of participants were comparable among laboratories. Laboratory (L) 2 produced the highest and L1 the lowest serum and RBC folate geometric means (nmol/L) in the NHANES sample (serum: L1, 39.5; L2, 59.2; L3, 47.7; and RBC: L1, 1120; L2, 1380; L3, 1380). Each laboratory produced different reference intervals for the central 95% of the population. Pearson correlation coefficients were highest between L3 and L1 (serum, r = 0.95; RBC, r = 0.92) and lowest between L2 and L1 (serum, r = 0.81; RBC, r = 0.65). Notable procedural differences among the laboratories were the Lactobacillus rhamnosus microorganism (L1 and L3: chloramphenicol resistant, L2: wild type) and the calibrator [L1: [6S]5-methyltetrahydrofolate (5-methylTHF), L2: [6R,S] 5-formyltetrahydrofolate ([6R,S] 5-formylTHF), L3: folic acid (FA)]. Compared with 5-methylTHF as calibrator, the folate results were 22-32% higher with FA as calibrator and 8% higher with 5-formylTHF as calibrator, regardless of the matrix (n = 30 serum, n = 28 RBC). The use of different calibrators explained most of the differences in results between L3 and L1 but not between L2 and L1. The use of the wild-type L. rhamnosus by L2 appeared to be the main reason for the differences in results between L2 and the other 2 laboratories. These findings indicate how assay variations influence MA folate results and how those variations can affect population data. To ensure data comparability, better assay harmonization is needed.

Dietary supplement use in the United States, 2003-2006.

Dietary supplement use has steadily increased over time since the 1970s; however, no current data exist for the U.S. population. Therefore, the purpose of this analysis was to estimate dietary supplement use using the NHANES 2003-2006, a nationally representative, cross-sectional survey. Dietary supplement use was analyzed for the U.S. population (≥1 y of age) by the DRI age groupings. Supplement use was measured through a questionnaire and was reported by 49% of the U.S. population (44% of males, 53% of females). Multivitamin-multimineral use was the most frequently reported dietary supplement (33%). The majority of people reported taking only 1 dietary supplement and did so on a daily basis. Dietary supplement use was lowest in obese adults and highest among non-Hispanic whites, older adults, and those with more than a high-school education. Between 28 and 30% reported using dietary supplements containing vitamins B-6, B-12, C, A, and E; 18-19% reported using iron, selenium, and chromium; and 26-27% reported using zinc- and magnesium-containing supplements. Botanical supplement use was more common in older than in younger age groups and was lowest in those aged 1-13 y but was reported by ~20% of adults. About one-half of the U.S. population and 70% of adults ≥ 71 y use dietary supplements; one-third use multivitamin-multimineral dietary supplements. Given the widespread use of supplements, data should be included with nutrient intakes from foods to correctly determine total nutrient exposure.

Vitamin B-6 intake is inversely related to, and the requirement is affected by, inflammation status.

Low circulating pyridoxal 5'-phosphate (PLP) concentrations have been linked to inflammatory markers and the occurrence of inflammatory diseases. However, the implications of these findings are unclear. The measurement of PLP and C-reactive protein (CRP) in blood samples collected from participants in the 2003-2004 NHANES afforded us the opportunity to investigate this relationship in the general U.S. population. Dietary and laboratory data were available for 3864 of 5041 interviewed adults, 2686 of whom were eligible (i.e. provided reliable dietary data and were not diabetic, pregnant, lactating, or taking hormones or steroidal antiinflammatory drugs). Vitamin B-6 intake was assessed using 2 24-h diet recalls and supplement use data. After multivariate adjustment for demographics, smoking, BMI, alcohol use, antioxidant vitamin status, intakes of protein and energy, and serum concentrations of creatinine and albumin, high vitamin B-6 intake was associated with protection against serum CRP concentrations >10 mg/L compared with < or =3 mg/L. However, plasma PLP > or =20 nmol/L compared with <20 nmol/L was inversely related to serum CRP independently of vitamin B-6 intake (P < 0.001). Among participants with vitamin B-6 intakes from 2 to 3 mg/d, the multivariate-adjusted prevalence of vitamin B-6 inadequacy was <10% in participants with serum CRP < or =3 mg/L but close to 50% in those with serum CRP > 10 mg/L (P < 0.001). In conclusion, higher vitamin B-6 intakes were linked to protection against inflammation and the vitamin B-6 intake associated with maximum protection against vitamin B-6 inadequacy was increased in the presence compared to absence of inflammation.

Three-phase model harmonizes estimates of the maximal suppression of parathyroid hormone by 25-hydroxyvitamin D in persons 65 years of age and older.

The concentration or threshold of 25-hydroxyvitamin D [25(OH)D] needed to maximally suppress intact serum parathyroid hormone (iPTH) has been suggested as a measure of optimal vitamin D status. Depending upon the definition of maximal suppression of iPTH and the 2-phase regression approach used, 2 distinct clusters for a single 25(OH)D threshold have been reported: 16-20 ng/mL (40-50 nmol/L) and 30-32 ng/mL (75-80 nmol/L). To rationalize the apparently disparate published results, we compared thresholds from several regression models including a 3-phase one to estimate simultaneously 2 thresholds before and after adjusting for possible confounding for age, BMI, glomerular filtration rate, dietary calcium, and season (April-September vs. October-March) within a single data set, i.e. data from the Tufts University Sites Testing Osteoporosis Prevention/Intervention Treatment study, consisting of 181 men and 206 women (total n = 387) ages 65-87 y. Plasma 25(OH)D and serum iPTH concentrations were (mean +/- SD) 22.1 +/- 7.44 ng/mL (55.25 +/- 18.6 nmol/L) and 36.6 +/- 16.03 pg/mL (3.88 +/- 1.7 pmol/L), respectively. The 3-phase model identified 2 thresholds of 12 ng/mL (30 nmol/L) and 28 ng/mL (70 nmol/L); similar results were found from the 2-phase models evaluated, i.e. 13-20 and 27-30 ng/mL (32.5-50 and 67.5-75 nmol/L) and with previous results. Adjusting for confounding did not change the results substantially. Accordingly, the 3-phase model appears to be superior to the 2-phase approach, because it simultaneously estimates the 2 threshold clusters found from the 2-phase approaches along with estimating confidence limits. If replicated, it may be of both clinical and public health importance.

Estimation of total usual calcium and vitamin D intakes in the United States.

Our objective in this study was to estimate calcium intakes from food, water, dietary supplements, and antacids for U.S. citizens aged >or=1 y using NHANES 2003-2006 data and the Dietary Reference Intake panel age groupings. Similar estimates were calculated for vitamin D intake from food and dietary supplements using NHANES 2005-2006. Diet was assessed with 2 24-h recalls; dietary supplement and antacid use were determined by questionnaire. The National Cancer Institute method was used to estimate usual nutrient intake from dietary sources. The mean daily nutrient intake from supplemental sources was added to the adjusted dietary intake estimates to produce total usual nutrient intakes for calcium and vitamin D. A total of 53% of the U.S. population reported using any dietary supplement (2003-2006), 43% used calcium (2003-2006), and 37% used vitamin D (2005-2006). For users, dietary supplements provided the adequate intake (AI) recommendation for calcium intake for approximately 12% of those >or=71 y. Males and females aged 1-3 y had the highest prevalence of meeting the AI from dietary and total calcium intakes. For total vitamin D intake, males and females >or=71, and females 14-18 y had the lowest prevalence of meeting the AI. Dietary supplement use is associated with higher prevalence of groups meeting the AI for calcium and vitamin D. Monitoring usual total nutrient intake is necessary to adequately characterize and evaluate the population's nutritional status and adherence to recommendations for nutrient intake.

NHANES monitoring of serum 25-hydroxyvitamin D: a roundtable summary.

A roundtable to discuss monitoring of serum 25-hydroxyvitamin D [25(OH)D] in the NHANES was held in late July 2009. Topics included the following: 1) options for dealing with assay fluctuations in serum 25(OH)D in the NHANES conducted between 1988 and 2006; 2) approaches for transitioning between the RIA used in the NHANES between 1988 and 2006 to the liquid chromatography tandem MS (LC-MS/MS) measurement procedure to be used in NHANES 2007 and later; 3) approaches for integrating the recently available standard reference material for vitamin D in human serum (SRM 972) from the National Institute of Standards and Technology (NIST) into the NHANES; 4) questions regarding whether the C-3 epimer of 25-hydroxyvitamin D3 [3-epi-25(OH)D3] should be measured in NHANES 2007 and later; and 5) identification of research and educational needs. The roundtable experts agreed that the NHANES data needed to be adjusted to control for assay fluctuations and offered several options for addressing this issue. The experts suggested that the LC-MS/MS measurement procedure developed by NIST could serve as a higher order reference measurement procedure. They noted the need for a commutability study for the recently released NIST SRM 972 across a range of measurement procedures. They suggested that federal agencies and professional organizations work with manufacturers to improve the quality and comparability of measurement procedures across all laboratories. The experts noted the preliminary nature of the evidence of the 3-epi-25(OH)D3 but felt that it should be measured in 2007 NHANES and later.

Trends in blood folate and vitamin B-12 concentrations in the United States, 1988 2004.

BACKGROUND: Monitoring the folate status of US population groups over time has been a public health priority for the past 2 decades, and the focus has been enhanced since the implementation of a folic acid fortification program in the mid-1990s. OBJECTIVE: We aimed to determine how population concentrations of serum and red blood cell (RBC) folate and serum vitamin B-12 have changed over the past 2 decades. DESIGN: Measurement of blood indicators of folate and vitamin B-12 status was conducted in approximately 23,000 participants in the prefortification third National Health and Nutrition Examination Survey (NHANES III; 1988-1994) and in approximately 8000 participants in 3 postfortification NHANES periods (together covering 1999-2004). RESULTS: Serum and RBC folate concentrations increased substantially (by 119-161% and 44-64%, respectively) in each age group in the first postfortification survey period and then declined slightly (by 5-13% and 6-9%, respectively) in most age groups between the first and third postfortification survey periods. Serum vitamin B-12 concentrations did not change appreciably. Prevalence estimates of low serum and RBC folate concentrations declined in women of childbearing age from before to after fortification (from 21% to <1% and from 38% to 5%, respectively) but remained unchanged thereafter. Prevalence estimates of high serum folate concentrations increased in children and older persons from before to after fortification (from 5% to 42% and from 7% to 38%, respectively) but decreased later after fortification. CONCLUSIONS: The decrease in folate concentrations observed longer after fortification is small compared with the increase soon after the introduction of fortification. The decrease is not at the low end of concentrations and therefore does not raise concerns about inadequate status.

Dietary supplement use among infants, children, and adolescents in the United States, 1999-2002.

OBJECTIVE: To describe dietary supplement use among US children. DESIGN: Analysis of nationally representative data from the 1999-2002 National Health and Nutrition Examination Survey (NHANES). SETTING: Home interviews and a mobile examination center. PARTICIPANTS: Children from birth through 18 years who participated in NHANES (N=10,136). MAIN EXPOSURE: Frequency of use of any dietary supplement product. OUTCOME MEASURE: Prevalence of use and intake of key nutrients from supplements among children. RESULTS: In 1999-2002, 31.8% of children used dietary supplements, with the lowest use reported among infants younger than 1 year (11.9%) and teenagers 14 to 18 years old (25.7%) and highest use among 4- to 8-year-old children (48.5%). Use was highest among non-Hispanic white (38.1%) and Mexican American (22.4%) participants, lowest among non-Hispanic black participants (18.8%), and was not found to differ by sex. The type of supplement most commonly used was multivitamins and multiminerals (18.3%). Ascorbic acid (28.6%), retinol (25.8%), vitamin D (25.6%), calcium (21.1%), and iron (19.3%) were the primary supplemental nutrients consumed. Supplement use was associated with families with higher incomes; a smoke-free environment; not being certified by the US Department of Agriculture Special Supplemental Nutrition Program for Women, Infants and Children in the last 12 months; lower child body mass index; and less daily recreational screen time (television, video games, computers, etc) (P<.005). The highest prevalence of supplement use (P<.005) was in children who were underweight or at risk for underweight (P<.005). CONCLUSIONS: More than 30% of children in the United States take dietary supplements regularly, most often multivitamins and multiminerals. Given such extensive use, nutrient intakes from dietary supplements must be included to obtain accurate estimates of overall nutrient intake in children.

Folate and human reproduction.

The influence of folate nutritional status on various pregnancy outcomes has long been recognized. Studies conducted in the 1950s and 1960s led to the recognition of prenatal folic acid supplementation as a means to prevent pregnancy-induced megaloblastic anemia. In the 1990s, the utility of periconceptional folic acid supplementation and folic acid food fortification emerged when they were proven to prevent the occurrence of neural tube defects. These distinctively different uses of folic acid may well be ranked among the most significant public health measures for the prevention of pregnancy-related disorders. Folate is now viewed not only as a nutrient needed to prevent megaloblastic anemia in pregnancy but also as a vitamin essential for reproductive health. This review focuses on the relation between various outcomes of human reproduction (ie, pregnancy, lactation, and male reproduction) and folate nutrition and metabolism, homocysteine metabolism, and polymorphisms of genes that encode folate-related enzymes or proteins, and we identify issues for future research.

Vitamin D and health in the 21st century: bone and beyond. Executive summary.

Vitamin D is unique, in terms of its metabolism and physiologic features and the human reliance on both endogenous production (activation through exposure to ultraviolet light) and exogenous sources (diet, primarily fortified foods) to meet biological requirements. Recent evidence has indicated a reemergence of vitamin D-deficient rickets and an alarming prevalence of vitamin D insufficiency (ie, low circulating concentrations of 25-hydroxyvitamin D) in particular segments of the US population. Furthermore, evidence has emerged implicating vitamin D status in a range of adverse health conditions, including cancer and certain autoimmune diseases. Therefore, a conference organized by the National Institute of Child Health and Human Development and the National Institutes of Health Office of Dietary Supplements was held to explore current knowledge and to develop a research agenda to address the range of issues associated with vitamin D and health during the life cycle. These proceedings contain presentations about 1) existing data on vitamin D status in the United States and internationally, 2) the current state of knowledge regarding the biological functions of vitamin D, 3) the strength of evidence supporting reconsideration of current policies regarding vitamin D intake, 4) gaps in understanding of the factors affecting and current options for improving vitamin D status in the United States and internationally, and 5) research needs to address gaps in knowledge regarding vitamin D assessment, biological features, and requirements. This executive summary provides an overview of the conference and its conclusions.

Milk folate but not milk iron concentrations may be inadequate for some infants in a rural farming community in San Mateo, Capulhuac, Mexico.

BACKGROUND: We were interested in identifying possible nutritional reasons for growth faltering among breastfed infants in the rural farming community of San Mateo, Capulhuac, Mexico (2800 m above sea level). OBJECTIVE: We examined the prevalence of inadequate iron and folate status among lactating Otomi women and determined to what extent their iron and folate nutriture influenced the milk concentrations of these nutrients. DESIGN: Lactating women (n = 71) provided blood and milk samples and dietary information at a mean (+/- SD) of 22 +/- 13 d postpartum. Blood indexes included hemoglobin, hematocrit, serum iron, total-iron-binding capacity, ferritin, transferrin receptor, mean cell volume, plasma folate, and erythrocyte folate. RESULTS: Approximately 62% and 58% of the women had nutritional anemia defined as a hemoglobin concentration

Effect of cow milk on food folate bioavailability in young women.

BACKGROUND: The findings of animal and human studies suggest that a certain component in milk enhances food folate bioavailability. OBJECTIVE: The objective was to evaluate whether cow milk enhances the bioavailability of food folate in humans. DESIGN: Thirty-one young women were fed low-folate diets on a 4-d rotation with (n=17) or without (n=14) cow milk for 8 wk. Plasma and erythrocyte folate and plasma total homocysteine (tHcy) concentrations were monitored. RESULTS: Plasma folate concentrations steadily declined in both the milk and the nonmilk groups; however, the declines were not significantly different between groups (P=0.09). In contrast, erythrocyte folate concentrations declined only in the nonmilk group and remained unchanged in the milk group; the difference between the 2 groups was significant (P=0.01). Plasma tHcy concentrations significantly increased in the nonmilk group and remained unchanged in the milk group; the difference between the 2 groups was significant (P=0.02). CONCLUSION: The inclusion of cow milk in the diet enhanced the bioavailability of food folate as assessed by the changes in erythrocyte folate and plasma tHcy concentrations but not in plasma folate concentrations. The mechanisms of action by the components of cow milk remain to be investigated.

Portion size of common foods predicts energy intake among preschool-aged children.

This study evaluated the relationship of food intake behaviors to total energy intake among children aged 2 to 5 years old (N=5447) who participated in the Continuing Survey of Food Intakes by Individuals, 1994-1996, 1998 (CSFII 94-96, 98). Food intake behaviors examined were portion size for 10 commonly eaten foods, number of eating occasions per day, and number of foods consumed per day. Using regression models, we examined these eating behaviors as predictors of energy intakes with adjustment for body weight. Eating behaviors and body weight were positively related to energy intake, together explaining 38% to 39% of the variability. Portion size alone accounted for 17% to 19% of the variance in energy intake, whereas body weight predicted only 4%. Feeding recommendations should highlight the importance of age-appropriate portion sizes and provide guidance on the frequency of eating and number of foods consumed.

Guidelines for collection of human milk samples for monitoring and research of environmental chemicals.

This article addresses sample collection protocols for monitoring and research of environmental chemicals in human milk. The process of milk synthesis and secretion and variations in contents of constituents that may impact measurement of environmental chemicals are presented. Possible sources of variation include parity, stage of lactation, method of sampling, maternal nutritional status, and dietary intake. General principles regarding how and when to collect milk samples are provided. For any previously unstudied environmental chemical in milk, all sources of variance must be assessed before a meaningful sampling protocol can be devised.

Inorganic selenite supplementation and protection against hyperoxic injury in neonates.

This study was designed to determine if oral sodium selenite supplementation to Se-depleted rat pups furnishes protection against hyperoxic lung injury. Twelve female rats were bred and fed a Se-deficient (0.04 ppm Se) diet during pregnancy and lactation. Pups were supplemented either with 0 or 3.2 ng Se/g body weight daily from days 2 to 7. On day 4, two litters were mixed, with half of the pooled litter assigned to an air environment and the other half to an oxygen environment. Dams cross-fostered pups for 4 d. Selenite supplementation increased pup plasma and liver selenium concentration and the liver activity of glutathione peroxidase (GPx). However, lung GPx activity was more affected by oxygen exposure than selenite supplementation. While oral Se supplementation of the pups showed a tendency for decreased incidence of lung injury with oxygen exposure, this apparent effect was not statistically significant. Selenium-supplemented pups also showed a trend toward larger internal surface area and lung volume than selenium-depleted pups. These data indicate that early postnatal selenium repletion via direct oral selenite supplementation may be beneficial to rat pups against hyperoxic lung injury.

Is there a reverse J-shaped association between 25-hydroxyvitamin D and all-cause mortality? Results from the U.S. nationally representative NHANES.

CONTEXT: A reverse J-shaped association between serum 25-hydroxyvitamin D (25[OH]D) concentration and all-cause mortality was suggested in a 9-year follow-up (1991-2000) analysis of the Third National Health and Nutrition Examination Survey (NHANES III, 1988-1994). OBJECTIVE: Our objective was to repeat the analyses with 6 years additional follow-up to evaluate whether the association persists through 15 years of follow-up. PARTICIPANTS: The study included 15 099 participants aged ≥ 20 years with 3784 deaths. MAIN OUTCOME MEASURE: Relative risk (RR) of death from all causes was adjusted for age, sex, race/ethnicity, and season using 2 Poisson regression approaches: traditional categorical and cubic splines. Results were given for 9 25(OH)D levels: <20, 20 to 29, 30 to 39, 40 to 49, 50 to 59, 60 to 74, 75 to 99 (reference), 100 to 119, and ≥ 120 nmol/L. RESULTS: The reverse J-shaped association became stronger with longer follow-up and was not affected by excluding deaths within the first 3 years of follow-up. Similar results were found from both statistical approaches for levels <20 through 119 nmol/L. Adjusted RR (95% confidence interval [CI]) estimates for all levels <60 nmol/L were significantly >1 compared with the reference group. The nadir of risk was 81 nmol/L (95% CI, 73-90 nmol/L). For 25(OH)D ≥ 120 nmol/L, results (RR, 95% CI) were slightly different using traditional categorical (1.5, 1.02-2.3) and cubic splines approaches (1.2, 0.9-1.4). The association appeared in men, women, adults ages 20 to 64 years, and non-Hispanic whites but was weaker in older adults. The study was too small to evaluate the association in non-Hispanic black and Mexican-American adults. CONCLUSIONS: A reverse J-shaped association between serum 25(OH)D and all-cause mortality appears to be real. It is uncertain whether the association is causal.