BACKGROUND: An understudied component of the diet, branched-chain fatty acids (BCFAs) are distinctive saturated fatty acids that may have an important influence on health. Human-milk fatty acid composition is known to differ worldwide, but comparative data are lacking on BCFAs. OBJECTIVE: We tested the hypotheses that concentrations of BCFAs in human milk differ between populations and are associated with maternal diet. DESIGN: We surveyed the BCFA composition of samples collected as part of a standardized, prospective study of human-milk composition. Mothers were enrolled from 3 urban populations with differing diets: Cincinnati, Ohio; Shanghai, China; and Mexico City, Mexico. Enrollment was limited to healthy mothers of term singleton infants. We undertook a cross-sectional analysis of milk from all women with samples at postpartum week 4 (n = 359; ∼120 women/site). Fatty acids were extracted from milk by using a modified Bligh-Dyer technique and analyzed by gas chromatography. Statistical analysis was performed by ANOVA and Tobit regression. For Cincinnati mothers, 24-h diet recalls were analyzed in relation to the individual BCFA concentrations measured in milk samples. RESULTS: Total BCFAs in milk differed by site, with the highest concentration in Cincinnati followed by Mexico City and Shanghai (mean ± SE: 7.90 ± 0.41, 6.10 ± 0.36, and 4.27 ± 0.25 mg/100 mL, respectively; P < 0.001). Site differences persisted after delivery mode, maternal age, and body mass index were controlled for. The individual concentrations of iso-14:0, iso-16:0, iso-18:0, anteiso-15:0, and anteiso-17:0 also differed between sites. Milk concentrations of iso-14:0 and anteiso-15:0 were associated with maternal intake of dairy; iso-16:0 was associated with maternal intakes of dairy and beef. CONCLUSIONS: BCFA concentrations in milk at 4 wk postpartum differed between mothers from Cincinnati, Shanghai, and Mexico City. Variations in human-milk BCFAs are influenced by diet. The impact of BCFAs on infant health warrants investigation.

Preterm birth (PTB) is the leading cause of infant morbidity and mortality. Genitourinary infection is implicated in the initiation of spontaneous PTB; however, examination of the urinary microbiota in relation to preterm delivery using next-generation sequencing technologies is lacking. In a case-control study nested within the Conditions Affecting Neurocognitive Development and Learning in Early Childhood (CANDLE) study, we examined associations between the urinary microbiota and PTB. A total of 49 cases (delivery < 37 weeks gestation) and 48 controls (delivery ≥ 37 weeks gestation) balanced on health insurance type were included in the present analysis. Illumina sequencing of the 16S rRNA gene V4 region was performed on urine samples collected during the second trimester. We observed no difference in taxa richness, evenness, or community composition between cases and controls or for gestational age modeled as a continuous variable. Operational taxonomic units (OTUs) classified to Prevotella, Sutterella, L. iners, Blautia, Kocuria, Lachnospiraceae, and S.marcescens were enriched among cases (FDR corrected p≤ 0.05). A urinary microbiota clustering partition dominated by S. marcescens was also associated with PTB (OR = 3.97, 95% CI: 1.19-13.24). These data suggest a limited role for the urinary microbiota in PTB when measured during the second trimester by 16S rRNA gene sequencing. The enrichment among cases in several organisms previously reported to be associated with genitourinary pathology requires confirmation in future studies to rule out the potential for false positive findings.

It is increasingly recognized that microbes that reside in and on human body sites play major roles in modifying the pathogenesis of several diseases, including cancer. However, specific microbes or microbial communities that can be mechanistically linked to cervical carcinogenesis remain largely unexplored. The purpose of the study was to examine the association between cervical microbiota and high-grade cervical intraepithelial neoplasia (CIN 2+) in women infected with high-risk (HR) human papillomaviruses (HPV) and to assess whether the cervical microbiota are associated with oxidative DNA damage as indicated by the presence of cervical cells positive for 8-hydroxy-2'-deoxyguanosine. The study included 340 women diagnosed with CIN 2+ (cases) and 90 diagnosed with CIN 1 (non-cases). Microbiota composition was determined by Illumina sequencing of the 16S rRNA gene amplified from DNA extracted from cervical mucus samples. Measures of alpha/beta-diversity were not associated with either CIN severity or oxidative DNA damage. However, a cervical mucosal community type (CT) dominated by L. iners and unclassified Lactobacillus spp was associated with CIN 2+ (OR = 3.48; 95% CI, 1.27-9.55). Sequence reads mapping to Lactobacillaceae, Lactobacillus, L. reuteri, and several sub-genus level Lactobacillus operational taxonomic units were also associated with CIN 2+ when examined independently (effect size >2.0; P < 0.05). Our 16S rRNA sequencing results need confirmation in independent studies using whole-genome shotgun sequencing and that would allow sharpening the suggested associations at finer taxonomic levels. Our results provide little evidence that DNA oxidative damage mediates the effect of the microbiome on the natural history of HPV infection and CIN severity.

Intestinal resection resulting in short bowel syndrome (SBS) carries a heavy burden of long-term morbidity, mortality, and cost of care, which can be attenuated with strategies that improve intestinal adaptation. SBS infants fed human milk, compared with formula, have more rapid intestinal adaptation. We tested the hypothesis that the major noncaloric human milk oligosaccharide 2'-fucosyllactose (2'-FL) contributes to the adaptive response after intestinal resection. Using a previously described murine model of intestinal adaptation, we demonstrated increased weight gain from 21 to 56 days (P < 0.001) and crypt depth at 56 days (P < 0.0095) with 2'-FL supplementation after ileocecal resection. Furthermore, 2'-FL increased small bowel luminal content microbial alpha diversity following resection (P < 0.005) and stimulated a bloom in organisms of the genus Parabacteroides (log2-fold = 4.1, P = 0.035). Finally, transcriptional analysis of the intestine revealed enriched ontologies and pathways related to antimicrobial peptides, metabolism, and energy processing. We conclude that 2'-FL supplementation following ileocecal resection increases weight gain, energy availability through microbial community modulation, and histological changes consistent with improved adaptation.

BACKGROUND: A dietary assessment instrument designed for use in a nationally representative pediatric population was required to examine associations between calcium intake and bone mineral accrual in a large, multicenter study. OBJECTIVE: To determine the reproducibility and intermethod reliability of a youth calcium food frequency questionnaire (FFQ) in a multiracial/ethnic sample of children and adolescents. DESIGN: Reproducibility (n=69) and intermethod reliability (n=393) studies were conducted by administering repeat FFQs and three unannounced 24-hour dietary recalls to stratified random samples of individuals participating in the Bone Mineral Density in Childhood Study. PARTICIPANTS/SETTING: Children and adolescents ages 5 to 21 years. MAIN OUTCOME MEASURES: Calcium intake estimated from the FFQ and 24-hour dietary recalls. STATISTICAL ANALYSIS: Reproducibility was assessed by the intraclass correlation coefficient (ICC). Intermethod reliability was assessed by deattenuated Pearson correlations between the FFQ and 24-hour recalls. Attenuation factors and calibration corrected effect estimates for bone density were calculated to determine the potential influence of measurement error on associations with health outcomes. RESULTS: The ICC (0.61) for repeat administrations and deattenuated Pearson correlation between the FFQ and 24-hour recalls (r=0.60) for all subjects indicated reproducibility and intermethod reliability (Pearson r=0.50 to 0.74 across sex and age groups). Attenuation factors were ≤0.50 for all sex and age groups and lower for non-Hispanic blacks (λ=0.20) and Hispanics (λ=0.26) than for non-Hispanic whites (λ=0.42). CONCLUSIONS: The Bone Mineral Density in Childhood Study calcium FFQ appears to provide a useful tool for assessing calcium intake in children and adolescents drawn from multiracial/ethnic populations and/or spanning a wide age range. However, similar to other FFQs, attenuation factors were substantially <1, indicating the potential for appreciable measurement error bias. Calibration correction should be performed and racial/ethnic differences in performance considered when analyzing and interpreting findings based on this instrument.

BACKGROUND: Little is known regarding the number of 24-hour recalls required to rank-order children and adolescents on usual intake for diet-disease studies. OBJECTIVE: To determine the within- to between-individual variance ratios and number of 24-hour recalls required to rank-order individuals on usual intake for select macro- and micronutrients in a large, multiracial/ethnic sample of children and adolescents. DESIGN: Cross-sectional survey. PARTICIPANTS/SETTING: Children and adolescents ages 6 to 17 years participating in the 2007-2008 and 2009-2010 National Health and Nutrition Examination Survey (NHANES). MAIN OUTCOME MEASURES: Variance ratios for predefined sex, age (children age 6 to 11 years, adolescents age 12 to 17 years), and racial/ethnic groups (Mexican American/Hispanic, non-Hispanic black, and non-Hispanic white). STATISTICAL ANALYSIS: Mixed-effects linear regression models were used to estimate within- and between-individual variance components for selected nutrients. The number of 24-hour recalls required to rank-order participants on usual intake (absolute values and energy-adjusted) was obtained from the nutrient variance ratios for various levels of accuracy. RESULTS: Variance ratios were more than 1 for all nutrients examined. High values (variance ratio >3) were observed for protein, saturated and unsaturated fatty acids, cholesterol, and several micronutrients. Variance ratios for absolute nutrient intakes were similar for both sexes within age groups, but higher for children than for adolescents. A total of six to nine and three to six 24-hour recalls were typically sufficient to rank-order children and adolescents, respectively, on usual intake with an accuracy of r=0.8. Additional recalls were required to achieve the same accuracy for energy-adjusted nutrients. Variance ratios were similar for adolescents across racial/ethnic groups, but highly variable in children. CONCLUSIONS: A total of six to nine 24-hour recalls may represent a reasonable trade-off between accuracy and participant burden for rank-ordering nutrient intakes in children and adolescents. Additional research is required to determine whether this may be reduced using statistical modeling-based approaches and the number of recalls children and adolescents will reliably complete.