Interactions between Bifidobacterium and Bacteroides and human milk oligosaccharides and their associations with infant cognition.

While ample research on independent associations between infant cognition and gut microbiota composition and human milk (HM) oligosaccharides (HMOs) has been reported, studies on how the interactions between gut microbiota and HMOs may yield associations with cognitive development in infancy are lacking. We aimed to determine how HMOs and species of Bacteroides and Bifidobacterium genera interact with each other and their associations with cognitive development in typically developing infants. A total of 105 mother-infant dyads were included in this study. The enrolled infants [2.9-12 months old (8.09 ± 2.48)] were at least predominantly breastfed at 4 months old. A total of 170 HM samples from the mothers and fecal samples of the children were collected longitudinally. Using the Mullen Scales of Early Learning to assess cognition and the scores as the outcomes, linear mixed effects models including both the levels of eight HMOs and relative abundance of Bacteroides and Bifidobacterium species as main associations and their interactions were employed with adjusting covariates; infant sex, delivery mode, maternal education, site, and batch effects of HMOs. Additionally, regression models stratifying infants based on the A-tetrasaccharide (A-tetra) status of the HM they received were also employed to determine if the associations depend on the A-tetra status. With Bacteroides species, we observed significant associations with motor functions, while Bif. catenulatum showed a negative association with visual reception in the detectable A-tetra group both as main effect (value of p = 0.012) and in interaction with LNFP-I (value of p = 0.007). Additionally, 3-FL showed a positive association with gross motor (p = 0.027) and visual reception (p = 0.041). Furthermore, significant associations were observed with the interaction terms mainly in the undetectable A-tetra group. Specifically, we observed negative associations for Bifidobacterium species and LNT [breve (p = 0.011) and longum (p = 0.022)], and positive associations for expressive language with 3'-SL and Bif. bifidum (p = 0.01), 6'-SL and B. fragilis (p = 0.019), and LNFP-I and Bif. kashiwanohense (p = 0.048), respectively. Our findings suggest that gut microbiota and HMOs are both independently and interactively associated with early cognitive development. In particular, the diverse interactions between HMOs and Bacteroides and Bifidobacterium species reveal different candidate pathways through which HMOs, Bifidobacterium and Bacteroides species potentially interact to impact cognitive development in infancy.

Joint analyses of human milk fatty acids, phospholipids, and choline in association with cognition and temperament traits during the first 6 months of life.

Early dietary exposure via human milk nutrients offers a window of opportunity to support cognitive and temperament development. While several studies have focused on associations of few pre-selected human milk nutrients with cognition and temperament, it is highly plausible that human milk nutrients synergistically and jointly support cognitive and behavioral development in early life. We aimed to discern the combined associations of three major classes of human milk nutrients with cognition and temperament during the first 6 months of life when human milk is the primary source of an infant's nutrition and explore whether there were persistent effects up to 18 months old. The Mullen Scales of Early Learning and Infant Behavior Questionnaires-Revised were used to assess cognition and temperament, respectively, of 54 exclusively/predominantly breastfed infants in the first 6 months of life, whose follow-ups were conducted at 6-9, 9-12, and 12-18 months old. Human milk samples were obtained from the mothers of the participants at less than 6 months of age and analyzed for fatty acids [total monounsaturated fatty acids, polyunsaturated fatty acid, total saturated fatty acid (TSFA), arachidonic acid (ARA), docosahexaenoic acid (DHA), ARA/DHA, omega-6/omega-3 polyunsaturated fatty acids ratio (n-6/n-3)], phospholipids [phosphatidylcholine, phosphatidylethanolamine (PE), phosphatidylinositol (PI), sphingomyelin], and choline [free choline, phosphocholine (PCho), glycerophosphocholine]. Feature selection was performed to select nutrients associated with cognition and temperament. The combined effects of selected nutrients were analyzed using multiple regression. A positive association between the arachidonic acid (ARA) and surgency was observed (p = 0.024). A significant effect of DHA, n-6/n-3, PE, and TSFA concentrations on receptive language (R 2 = 0.39, p = 0.025) and the elevated ARA, PCho, and PI with increased surgency (R 2 = 0.43, p = 0.003) was identified, suggesting that DHA and ARA may have distinct roles for temperament and language functions. Furthermore, the exploratory association analyses suggest that the effects of human milk nutrients on R.L. and surgency may persist beyond the first 6 months of life, particularly surgency at 12-18 months (p = 0.002). Our study highlighted that various human milk nutrients work together to support the development of cognition and temperament traits during early infancy.

Human milk 3'-Sialyllactose is positively associated with language development during infancy.

BACKGROUND: Genetic polymorphisms leading to variations in human milk oligosaccharide (HMO) composition have been reported. Alpha-Tetrasaccharide (A-tetra), an HMO, has been shown to only be present (>limit of detection; A-tetra+) in the human milk (HM) of women with blood type A, suggesting genetic origins determining the presence or absence (A-tetra-) of A-tetra in HM. OBJECTIVES: This study aimed to determine whether associations exist between HMO concentrations and cognitive development, and whether the associations vary between A-tetra+ and A-tetra- groups in children (<25 months old). METHODS: We enrolled typically developing children (2-25 months old; mean, 10 months old) who were at least partially breastfed at the study visit. The Mullen Scales of Early Learning (MSEL) were used as the primary outcome measure to assess early cognitive development. Linear mixed effects models were employed by stratifying children based on A-tetra levels (A-tetra+ or A-tetra-) to assess associations between age-removed HMO concentrations and both MSEL composite scores and the 5 subdomain scores. RESULTS: A total of 99 mother-child dyads and 183 HM samples were included (A-tetra+: 57 samples, 33 dyads; A-tetra-: 126 samples, 66 dyads). No significant association was observed between HMOs and MSEL when all samples were analyzed together. The composite score and 3'-sialyllactose (3'-SL) levels were positively associated [P = 0.002; effect size (EF), 13.12; 95% CI, 5.36-20.80] in the A-tetra + group. This association was driven by the receptive (adjusted P = 0.015; EF, 9.95; 95% CI, 3.91-15.99) and expressive (adjusted P = 0.048; EF, 7.53; 95% CI, 2.51-13.79) language subdomain scores. Furthermore, there was an interaction between 3'-SL and age for receptive language (adjusted P = 0.03; EF, -14.93; 95% CI, -25.29 to -4.24). CONCLUSIONS: Our study reports the association of 3'-SL and cognition, particularly language functions, in typically developing children who received HM containing detectable A-tetra during infancy.

Early-Life Nutrition and Cognitive Development: Imaging Approaches.

Brain development in the first years of life is the most dynamic and perhaps the most important phase of brain maturation. While it is widely recognized that nutrition plays a key role in early brain development, particular nutrients will most likely differentially affect distinct aspects of brain development. The critical dosage windows and time frames for various nutrients at different stages of brain development are likely dissimilar. Therefore, efforts have been devoted to identifying potential associations between nutrients and early brain development. However, behavioral assessments are typically employed as the outcome measures, which are known to suffer from low sensitivity and the inability to provide neural substrates underlying brain functional maturation. In contrast, magnetic resonance imaging is capable of providing detailed anatomical and functional information - an ideal tool to characterize brain functional development and nutrition. Our team has developed strategies that enable imaging of typically developing children from birth to teens without sedation. Quantitative assessments of brain structural and functional development during the first years of life have been accomplished, which reveal important features of early brain development. These developed tools will most likely substantially enhance our ability to rigorously characterize the interplay between nutrients and early brain development.

Brain functional development separates into three distinct time periods in the first two years of life.

Recently, resting functional MRI has provided invaluable insight into the brain developmental processes of early infancy and childhood. A common feature of previous functional development studies is the use of age to separate subjects into different cohorts for group comparisons. However, functional maturation paces vary tremendously from subject to subject. Since this is particularly true for the first years of life, an alternative to physical age alone is needed for cluster analysis. Here, a data-driven approach based on individual brain functional connectivity was employed to cluster typically developing children who were longitudinally imaged using MRI without sedation for the first two years of life. Specifically, three time periods were determined based on the distinction of brain functional connectivity patterns, including 0-1 month (group 1), 2-7 months (group 2), and 8-24 (group 3) of age, respectively. From groups 1 to 2, connection density increased by almost two-fold, local efficacy (LE) is significantly improved, and there was no change in global efficiency (GE). From groups 2 to 3, connection density increased slightly, LE showed no change, and a significant increase in GE were observed. Furthermore, 27 core brain regions were identified which yielded clustering results that resemble those obtained using all brain regions. These core regions were largely associated with the motor, visual and language functional domains as well as regions associated with higher order cognitive functional domains. Both visual and language functional domains exhibited a persistent and significant increase within domain connection from groups 1 to 3, while no changes were observed for the motor domain. In contrast, while a reduction of inter-domain connection was the general developmental pattern, the motor domain exhibited an interesting "V" shape pattern in its relationship to visual and language associated areas, showing a decrease from groups 1 to 2, followed by an increase from groups 2 to 3. In summary, our results offer new insights into functional brain development and identify 27 core brain regions critically important for early brain development.

The UNC/UMN Baby Connectome Project (BCP): An overview of the study design and protocol development.

The human brain undergoes extensive and dynamic growth during the first years of life. The UNC/UMN Baby Connectome Project (BCP), one of the Lifespan Connectome Projects funded by NIH, is an ongoing study jointly conducted by investigators at the University of North Carolina at Chapel Hill and the University of Minnesota. The primary objective of the BCP is to characterize brain and behavioral development in typically developing infants across the first 5 years of life. The ultimate goals are to chart emerging patterns of structural and functional connectivity during this period, map brain-behavior associations, and establish a foundation from which to further explore trajectories of health and disease. To accomplish these goals, we are combining state of the art MRI acquisition and analysis techniques, including high-resolution structural MRI (T1-and T2-weighted images), diffusion imaging (dMRI), and resting state functional connectivity MRI (rfMRI). While the overall design of the BCP largely is built on the protocol developed by the Lifespan Human Connectome Project (HCP), given the unique age range of the BCP cohort, additional optimization of imaging parameters and consideration of an age appropriate battery of behavioral assessments were needed. Here we provide the overall study protocol, including approaches for subject recruitment, strategies for imaging typically developing children 0-5 years of age without sedation, imaging protocol and optimization, a description of the battery of behavioral assessments, and QA/QC procedures. Combining HCP inspired neuroimaging data with well-established behavioral assessments during this time period will yield an invaluable resource for the scientific community.

MR fingerprinting enables quantitative measures of brain tissue relaxation times and myelin water fraction in the first five years of life.

Quantitative assessments of normative brain development using MRI are of critical importance to gain insights into healthy neurodevelopment. However, quantitative MR imaging poses significant technical challenges and requires prohibitively long acquisition times, making it impractical for pediatric imaging. This is particularly relevant for healthy subjects, where imaging under sedation is not clinically indicated. MR Fingerprinting (MRF), a novel MR imaging framework, provides rapid, efficient, and simultaneous quantification of multiple tissue properties. In this study, a 2D MR Fingerprinting method was developed that achieves a spatial resolution of 1 × 1 × 3 mm3 with rapid and simultaneous quantification of T1, T2 and myelin water fraction (MWF). Phantom experiments demonstrated that accurate measurements of T1 and T2 relaxation times were achieved over a wide range of T1 and T2 values. MRF images were acquired cross-sectionally from 28 typically developing children, 0 to five years old, who were enrolled in the UNC/UMN Baby Connectome Project. Differences associated with age of R1 (=1/T1), R2 (=1/T2) and MWF were obtained from several predefined white matter regions. Both R1 and R2 exhibit a marked increase until ∼20 months of age, followed by a slower increase for all WM regions. In contrast, the MWF remains at a negligible level until ∼6 months of age for all predefined ROIs and gradually increases afterwards. Depending on the brain region, rapid increases are observed between 6 and 12 months to 6-18 months, followed by a slower pace of increase in MWF. Neither relaxivities nor MWF were significantly different between the left and right hemispheres. However, regional differences in age-related R1 and MWF measures were observed across different white matter regions. In conclusion, our results demonstrate that the MRF technique holds great potential for multi-parametric assessments of normative brain development in early childhood.