Kimberly O. O’Brien

Dietary protein, calcium metabolism, and skeletal homeostasis revisited

High dietary protein intakes are known to increase urinary calcium excretion and, if maintained, will result in sustained hypercalciuria. To date, the majority of calcium balance studies in humans have not detected an effect of dietary protein on intestinal calcium absorption or serum parathyroid hormone. Therefore, it is commonly concluded that the source of the excess urinary calcium is increased bone resorption. Recent studies from our laboratory indicate that alterations in dietary protein can, in fact, profoundly affect intestinal calcium absorption. In short-term dietary trials in healthy adults, we fixed calcium intake at 20 mmol/d while dietary protein was increased from 0.7 to 2.1 g/kg. Increasing dietary protein induced hypercalciuria in 20 women [from 3.4 +/- 0.3 ( +/- SE) during the low-protein to 5.4 +/- 0.4 mmol/d during the high-protein diet]. The increased dietary protein was accompanied by a significant increase in intestinal calcium absorption from 18.4 +/- 1.3% to 26.3 +/- 1.5% (as determined by dual stable isotopic methodology). Dietary protein intakes at and below 0.8 g/kg were associated with a probable reduction in intestinal calcium absorption sufficient to cause secondary hyperparathyroidism. The long-term consequences of these low-protein diet-induced changes in mineral metabolism are not known, but the diet could be detrimental to skeletal health. Of concern are several recent epidemiologic studies that demonstrate reduced bone density and increased rates of bone loss in individuals habitually consuming low-protein diets. Studies are needed to determine whether low protein intakes directly affect rates of bone resorption, bone formation, or both.

Positive selection on a regulatory insertion-deletion polymorphism in FADS2 influences apparent endogenous synthesis of arachidonic acid

Long chain polyunsaturated fatty acids (LCPUFA) are bioactive components of membrane phospholipids and serve as substrates for signaling molecules. LCPUFA can be obtained directly from animal foods or synthesized endogenously from 18 carbon precursors via the FADS2 coded enzyme. Vegans rely almost exclusively on endogenous synthesis to generate LCPUFA and we hypothesized that an adaptive genetic polymorphism would confer advantage. The rs66698963 polymorphism, a 22-bp insertion–deletion within FADS2, is associated with basal FADS1 expression, and coordinated induction of FADS1 and FADS2 in vitro. Here, we determined rs66698963 genotype frequencies from 234 individuals of a primarily vegetarian Indian population and 311 individuals from the US. A much higher I/I genotype frequency was found in Indians (68%) than in the US (18%). Analysis using 1000 Genomes Project data confirmed our observation, revealing a global I/I genotype of 70% in South Asians, 53% in Africans, 29% in East Asians, and 17% in Europeans. Tests based on population divergence, site frequency spectrum, and long-range haplotype consistently point to positive selection encompassing rs66698963 in South Asian, African, and some East Asian populations. Basal plasma phospholipid arachidonic acid (ARA) status was 8% greater in I/I compared with D/D individuals. The biochemical pathway product–precursor difference, ARA minus linoleic acid, was 31% and 13% greater for I/I and I/D compared with D/D, respectively. This study is consistent with previous in vitro data suggesting that the insertion allele enhances n-6 LCPUFA synthesis and may confer an adaptive advantage in South Asians because of the traditional plant-based diet practice.

Impact of maternal and neonatal iron status on placental transferrin receptor expression in pregnant adolescents.

OBJECTIVE To elucidate the role of maternal and neonatal iron status on placental transferrin receptor (TfR) expression. STUDY DESIGN AND OUTCOMES: Ninety-two healthy pregnant adolescents (ages 14-18 years) were followed across pregnancy. Maternal iron status (hemoglobin, hematocrit, serum ferritin, TfR, and total body iron) was assessed in mid-gestation (21-25 wks) and at delivery in the mother and neonate. Placental TfR protein expression was assessed by western blot in placental tissue collected at delivery. RESULTS Placental TfR expression was inversely associated with maternal iron status at mid-gestation (hemoglobin p = 0.046, R(2) = 0.1 and hematocrit p = 0.005, R(2) = 0.24) and at delivery (serum ferritin p = 0.02, R(2) = 0.08 and total body iron p = 0.02, R(2) = 0.07). Mothers with depleted body iron stores had significantly greater placental expression of TfR than mothers with body iron stores greater than zero (p = 0.003). Neonatal iron stores were also inversely associated with the expression of placental TfR (p = 0.04, R(2) = 0.06). Neonates with serum ferritin values ≤ 34 μg/L had significantly greater protein expression of placental TfR compared to neonates with cord serum ferritin values >34 μg/L (p = 0.01). CONCLUSIONS Expression of placental TfR is associated with both maternal and neonatal iron demands. Increased expression of placental TfR may be an important compensatory mechanism in response to iron deficiency in otherwise healthy pregnant women.

Calcium Supplementation to Prevent Preeclampsia: Translating Guidelines into Practice in Low-Income Countries

The WHO issued a strong recommendation that pregnant women be provided calcium supplements to prevent preeclampsia. This is the first recommended nutritional intervention to prevent this condition, a leading cause of maternal mortality globally. As health systems seek to implement this new intervention, a number of issues require further clarification and guidance, including dosage regimen, supplement formulation, and alignment with other antenatal nutritional interventions. We summarize key evidence on the above points and offer our views on good practices. Most developing countries have low calcium intake, so where habitual calcium intake is unknown, calcium supplements are likely beneficial. In our view, policymakers and program planners should consider adopting doses between 1.0 and 1.5 g elemental calcium/d, depending on the local average and variation in dietary calcium intake, logistical feasibility, and acceptability in the target population. Prudent practice would entail daily administration as calcium carbonate administered in divided doses of not >500 mg elemental calcium per dose. For ease of prescribing and adherence, calcium [as with iron and folic acid (IFA)] should be administered routinely to pregnant women from the earliest contact in pregnancy until delivery. Calciums acute inhibitory effect on iron absorption translates to minimal effects in clinical studies. Therefore, to simplify the regimen and facilitate adherence, providers should not counsel that calcium and IFA pills must be taken separately. Although further research will shed more light on clinical and programmatic issues, policies can be implemented with ongoing revision as we continue to learn what works to improve maternal and newborn health.

Vitamin D status is inversely associated with anemia and serum erythropoietin during pregnancy

BACKGROUND Vitamin D and iron deficiencies frequently co-exist. It is now appreciated that mechanistic interactions between iron and vitamin D metabolism may underlie these associations. OBJECTIVE We examined interrelations between iron and vitamin D status and their regulatory hormones in pregnant adolescents, who are a group at risk of both suboptimal vitamin D and suboptimal iron status. DESIGN The trial was a prospective longitudinal study of 158 pregnant adolescents (aged ≤18 y). Maternal circulating biomarkers of vitamin D and iron were determined at midgestation (∼25 wk) and delivery (∼40 wk). Linear regression was used to assess associations between vitamin D and iron status indicators. Bivariate and multivariate logistic regressions were used to generate the OR of anemia as a function of vitamin D status. A mediation analysis was performed to examine direct and indirect relations between vitamin D status, hemoglobin, and erythropoietin in maternal serum. RESULTS Maternal 25-hydroxyvitamin D [25(OH)D] was positively associated with maternal hemoglobin at both midgestation and at delivery (P < 0.01 for both). After adjustment for age at enrollment and race, the odds of anemia at delivery was 8 times greater in adolescents with delivery 25(OH)D concentrations <50 nmol/L than in those with 25(OH)D concentrations ≥50 nmol/L (P <0.001). Maternal 25(OH)D was inversely associated with erythropoietin at both midgestation (P <0.05) and delivery (P <0.001). The significant relation observed between 25(OH)D and hemoglobin could be explained by a direct relation between 25(OH)D and hemoglobin and an indirect relation that was mediated by erythropoietin. CONCLUSIONS In this group of pregnant adolescents, suboptimal vitamin D status was associated with increased risk of iron insufficiency and vice versa. These findings emphasize the need for screening for multiple nutrient deficiencies during pregnancy and greater attention to overlapping metabolic pathways when selecting prenatal supplementation regimens.

Prepregnancy Body Mass Index and Gestational Weight Gain Have No Negative Impact on Maternal or Neonatal Iron Status

Objective: To assess the impact of maternal obesity and excessive gestational weight gain (GWG) on maternal and neonatal iron status and to explore the possible mediating role of inflammation on hepcidin. Methods: This analysis included 230 pregnant adolescents (13-18 years) enrolled in either a longitudinal or a cross-sectional study. Prepregnancy body mass index (ppBMI) and GWG were obtained from medical records. Maternal iron status (hemoglobin, serum iron, ferritin, transferrin receptor, total body iron, and hepcidin) and inflammation (interleukin-6 [IL-6] and leptin) were assessed at midgestation (26.2 ± 3.3 weeks) in the longitudinal cohort and at delivery (39.8 ± 1.3 weeks) in both study cohorts. Cord blood was collected in both studies and analyzed for iron indicators. Results: Approximately 40% of the adolescents entered pregnancy overweight or obese. Multivariate analysis identified ppBMI as a negative predictor of serum iron at midgestation (P = .009) and a positive predictor of serum hepcidin at delivery (P = .02). None of the other maternal iron status indicators were significantly associated with ppBMI or GWG. Serum IL-6 was significantly positively associated with hepcidin at delivery (P = .0001) but not at midgestation. There was a positive relationship between ppBMI and cord hemoglobin (P = .03). Conclusion: These results suggest that adiposity-related inflammation does not override the iron-mediated signals that regulate hepcidin production during pregnancy, and in this adolescent cohort, there is no strong evidence for a detrimental effect of maternal obesity and excessive weight gain on iron status in the offspring at birth.

Nutrient Inadequacy Is Prevalent in Pregnant Adolescents, and Prenatal Supplement Use May Not Fully Compensate for Dietary Deficiencies

A longitudinal study was undertaken in 156 pregnant adolescents (≤18 years old) to characterize dietary intake and to determine the degree to which prenatal supplement use compensates for dietary deficits. The adequacy of dietary intake was assessed by comparing self-reported intake from up to three 24-hour dietary recalls with the dietary reference intakes. The majority of teens did not meet the estimated average requirements (EAR) for vitamin D (93%), vitamin E (94%), Mg (90%), Fe (76%), and Ca (74%). More than half of the adolescents in each gestational window (<23 weeks; 23-30 weeks; and ≥31 weeks of gestation) self-reported daily use of prenatal supplements, but the additional supplement contributions were not sufficient to meet the EAR for Mg (90%) or Ca (54%). Pregnant adolescents are at risk for insufficient intake of several essential nutrients from diet alone in spite of adequate or excessive energy intakes. Daily use of prenatal supplements reduces the prevalence of dietary inadequacy for many ...

Vitamin D Status Affects Serum Metabolomic Profiles in Pregnant Adolescents.

Vitamin D is linked to a number of adverse pregnancy outcomes through largely unknown mechanisms. This study was conducted to examine the role of vitamin D status in metabolomic profiles in a group of 30 pregnant, African American adolescents (17.1 ± 1.1 years) at midgestation (26.8 ± 2.8 weeks), in 15 adolescents with 25-hydroxy vitamin D (25(OH)D) ≥20 ng/mL, and in 15 teens with 25(OH)D <20 ng/mL. Serum metabolomic profiles were examined using gas chromatography–mass spectrometry and liquid chromatography–tandem mass spectrometry. A novel hierarchical mixture model was used to evaluate differences in metabolite profiles between low and high groups. A total of 326 compounds were identified and included in subsequent statistical analyses. Eleven metabolites had significantly different means between the 2 vitamin D groups, after correcting for multiple hypothesis testing: pyridoxate, bilirubin, xylose, and cholate were higher, and leukotrienes, 1,2-propanediol, azelate, undecanedioate, sebacate, inflammation associated complement component 3 peptide (HWESASXX), and piperine were lower in serum from adolescents with 25(OH)D ≥20 ng/mL. Lower maternal vitamin D status at midgestation impacted serum metabolic profiles in pregnant adolescents.

Low Vitamin D is Associated With Infections and Proinflammatory Cytokines During Pregnancy

Vitamin D is known to regulate innate and adaptive immune processes at the cellular level, but the role of vitamin D status on associated inflammatory processes across pregnancy is unclear. Our primary objective was to evaluate the relationships between serum biomarkers of inflammation (interleukin [IL]-6, IL-10, tumor necrosis factor [TNF]-α), acute-phase proteins (C-reactive protein and hepcidin) and vitamin D status, 25-hydroxyvitamin D (25(OH)D) and 1,25-dihydroxyvitamin D (1,25(OH)2D), measured across pregnancy and in the neonate at birth. A second objective was to identify associations between vitamin D status and clinically diagnosed infections. In this study, 158 racially and ethnically diverse pregnant adolescents were recruited from the Rochester Adolescent Maternity Program (RAMP) in Rochester, NY. Serum 1,25(OH)2D was significantly lower in adolescents and neonates with IL-6 concentrations above the 75th percentile at delivery (P = .04) and at birth (P = .004), respectively. After adjusting for other potential covariates of inflammation, maternal serum 1,25(OH)2D was significantly positively associated with TNF-α during pregnancy (P = .02), but at delivery 1,25(OH)2D and TNF-α were inversely associated with one another (P = .02). Teens with 25(OH)D concentrations <30 ng/mL were more likely to test positive for candida (P = .002) and bacterial vaginosis (P = .02) during pregnancy. African Americans exhibited significantly lower TNF-α concentrations at both mid-gestation (P = .009) and delivery (P = .001) compared to the Caucasian adolescents. These results suggest that lower maternal vitamin D status may increase risk of infection across gestation.

Iron status of North American pregnant women: an update on longitudinal data and gaps in knowledge from the United States and Canada

Pregnant women are particularly vulnerable to iron deficiency due to the high iron demands of pregnancy. To avoid the adverse birth outcomes that are associated with maternal iron deficiency anemia, both Canada and the United States recommend universal iron supplementation for pregnant women. Although the benefits of iron supplementation in anemic women are well recognized, insufficient data are currently available on the maternal and neonatal benefits and harms of universal iron supplementation in developed countries as evidenced by the recent conclusions of the US Preventive Services Task Force on the need for further data that address existing gaps. As part of an effort to evaluate the impact of the current North American prenatal iron supplementation policy, this review highlights the lack of national data on longitudinal changes in iron status in pregnant North American women, emphasizes possible limitations with the original longitudinal hemoglobin data used to inform the current CDC reference hemoglobin values, and presents additional normative data from recent longitudinal research studies of iron status in North American pregnant women. Further longitudinal data in North American pregnant women are needed to help identify those who may benefit most from supplementation as well as to help determine whether there are adverse effects of iron supplementation in iron-replete women.