2′FL/3FL ratios for breast milk.

<p>Relative quantitative data as raw peak areas.</p>a<p>Presence of 2″FL indicates secretor.</p>b<p>2″FL/3FL ratio of >6.5 indicates secretor.</p

HMO Urine and Plasma Level Correlations to Breast Milk Concentrations.

a<p>Pearson Correlations performed on imputed, log-transformed data, within similar data type, log (peak area) or log (mg/L).</p><p>NS, not significant; ND, not detected; NT, not tested; NC, not calculable.</p><p>CI, confidence interval; P value (two-tailed).</p

Relative quantitation of HMOs.

<p>Bar plots indicating the relative scaled intensities and Standard error of the mean (SEM) grouped according to feed. (FF vs. BF) and secretor status (BF Se+ vs. BF Se-); (for n, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0101692#pone-0101692-t001" target="_blank">Table 1</a>). ‡LNnT measurement for breast milk (HM) as LNT, LNnT Isobar; Significant differences were assessed by Mann-Whitney <i>U</i> test where *p<0.05, **p<0.01, and ***p<0.001. Quantitative comparisons between sample matrices and between oligosaccharides cannot be made; quantitative comparisons between samples within a matrix and within individual oligosaccharides are valid.</p

Schematic of Study Design.

<p>Targeted analyses generating “Absolute Quantitation” are shown in yellow. Global analyses generating “Relative Quantitation” are shown in green. Dotted arrows indicate samples and analyses from Study A used in our Follow-on Study B.</p

Absolute quantitation of HMOs.

<p>Bar plots indicating mean concentrations (mg/L) and Standard error of the mean (SEM) of 2′FL and 6′SL grouped according to feed (FF vs. BF) and secretor status (BF Se+ vs. BF Se-); (for n, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0101692#pone-0101692-t001" target="_blank">Table 1</a>). ‡6′SL not detected in any plasma samples in the Global analysis. Significant differences were assessed by Mann-Whitney U test where *p<0.05, **p<0.01, and ***p<0.001.</p

Correlation of infant plasma and urine HMO levels with breast milk concentrations.

<p>Scatter plots showing the association between individual HMO levels in breast milk and those in the corresponding infant plasma or urine.Pearson correlation statistics of log transformed data are inset. A: Absolute quantitation of 2′FL [Log (mg/L)]; B: Relative quantitation of 2′FL, 3FL, LNnT and LNFP I [Log (peak area)]. Solid lines represent best fit through the data points; (for n, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0101692#pone-0101692-t002" target="_blank">Table 2</a>). (Plasma: black circles, Urine: white diamonds, Nonsecretor subject by 2′FL/3FL ratio: asterisk). NS, not significant.</p

2′FL and 6′SL Concentrations According to Feed and Secretor Status and Relative to Breast Milk Concentration.

<p>Absolute quantitative data (mg/L).</p><p>ND: not detected (6′SL not detected in any plasma samples in the Global analysis); NA: not applicable; NS: not significant; SEM: standard error of the mean.</p><p>*Concentration relative to breast milk level; BF Se+ group only for 2′FL, BF group for 6′SL.</p

Exploratory Metabolomic Analyses Reveal Compounds Correlated with Lutein Concentration in Frontal Cortex, Hippocampus, and Occipital Cortex of Human Infant Brain

<div><p>Lutein is a dietary carotenoid well known for its role as an antioxidant in the macula, and recent reports implicate a role for lutein in cognitive function. Lutein is the dominant carotenoid in both pediatric and geriatric brain tissue. In addition, cognitive function in older adults correlated with macular and postmortem brain lutein concentrations. Furthermore, lutein was found to preferentially accumulate in the infant brain in comparison to other carotenoids that are predominant in diet. While lutein is consistently related to cognitive function, the mechanisms by which lutein may influence cognition are not clear. In an effort to identify potential mechanisms through which lutein might influence neurodevelopment, an exploratory study relating metabolite signatures and lutein was completed. Post-mortem metabolomic analyses were performed on human infant brain tissues in three regions important for learning and memory: the frontal cortex, hippocampus, and occipital cortex. Metabolomic profiles were compared to lutein concentration, and correlations were identified and reported here. A total of 1276 correlations were carried out across all brain regions. Of 427 metabolites analyzed, 257 were metabolites of known identity. Unidentified metabolite correlations (510) were excluded. In addition, moderate correlations with xenobiotic relationships (2) or those driven by single outliers (3) were excluded from further study. Lutein concentrations correlated with lipid pathway metabolites, energy pathway metabolites, brain osmolytes, amino acid neurotransmitters, and the antioxidant homocarnosine. These correlations were often brain region—specific. Revealing relationships between lutein and metabolic pathways may help identify potential candidates on which to complete further analyses and may shed light on important roles of lutein in the human brain during development.</p></div

Lutein accretion in infant brain did not correlate with age.

<p>Post-mortem infant brain tissues (age 1 to 488 days) were analyzed for lutein by HPLC. Increasing lutein concentration (pmol/g) did not correlate with age in the (A) frontal cortex (n = 29), (B) hippocampus (n = 24), or (C) occipital cortex (n = 28). FC: frontal cortex; HPC: hippocampus; OC: occipital cortex; n.s.: not-significant.</p

Fatty acid correlations with lutein in human infant brain.^a

<p>^aSubject ages ranged from 1 to 488 days old, and the median age was 100.5 days. 30 total decedents were studied and included both male (n = 21) and female (n = 9) infants. A total of 81 tissues were analyzed from three brain regions, including the frontal cortex (FC, n = 29), hippocampus (HPC, n = 24), and the occipital cortex (OC, n = 28).</p><p>^bAll values are significant (P < 0.05). Fatty acid metabolites:lutein correlations with r values ≥ |0.45| and P < 0.05 are reported.</p><p>Fatty acid correlations with lutein in human infant brain.<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0136904#t002fn001" target="_blank">^a</a></p