Growth and Morbidity of Gambian Infants are Influenced by Maternal Milk Oligosaccharides and Infant Gut Microbiota.

Human milk oligosaccharides (HMOs) play an important role in the health of an infant as substrate for beneficial gut bacteria. Little is known about the effects of HMO composition and its changes on the morbidity and growth outcomes of infants living in areas with high infection rates. Mother's HMO composition and infant gut microbiota from 33 Gambian mother/infant pairs at 4, 16, and 20 weeks postpartum were analyzed for relationships between HMOs, microbiota, and infant morbidity and growth. The data indicate that lacto-N-fucopentaose I was associated with decreased infant morbidity, and 3'-sialyllactose was found to be a good indicator of infant weight-for-age. Because HMOs, gut microbiota, and infant health are interrelated, the relationship between infant health and their microbiome were analyzed. While bifidobacteria were the dominant genus in the infant gut overall, Dialister and Prevotella were negatively correlated with morbidity, and Bacteroides was increased in infants with abnormal calprotectin. Mothers nursing in the wet season (July to October) produced significantly less oligosaccharides compared to those nursing in the dry season (November to June). These results suggest that specific types and structures of HMOs are sensitive to environmental conditions, protective of morbidity, predictive of growth, and correlated with specific microbiota

Growth and Morbidity of Gambian Infants are Influenced by Maternal Milk Oligosaccharides and Infant Gut Microbiota.

Human milk oligosaccharides (HMOs) play an important role in the health of an infant as substrate for beneficial gut bacteria. Little is known about the effects of HMO composition and its changes on the morbidity and growth outcomes of infants living in areas with high infection rates. Mother's HMO composition and infant gut microbiota from 33 Gambian mother/infant pairs at 4, 16, and 20 weeks postpartum were analyzed for relationships between HMOs, microbiota, and infant morbidity and growth. The data indicate that lacto-N-fucopentaose I was associated with decreased infant morbidity, and 3'-sialyllactose was found to be a good indicator of infant weight-for-age. Because HMOs, gut microbiota, and infant health are interrelated, the relationship between infant health and their microbiome were analyzed. While bifidobacteria were the dominant genus in the infant gut overall, Dialister and Prevotella were negatively correlated with morbidity, and Bacteroides was increased in infants with abnormal calprotectin. Mothers nursing in the wet season (July to October) produced significantly less oligosaccharides compared to those nursing in the dry season (November to June). These results suggest that specific types and structures of HMOs are sensitive to environmental conditions, protective of morbidity, predictive of growth, and correlated with specific microbiota

Additional file 1: Figure S1. of Deletion of the type-1 interferon receptor in APPSWE/PS1ΔE9 mice preserves cognitive function and alters glial phenotype

Generation and confirmation of the APPSWE/PS1ΔE9 x IFNAR1−/− mouse. A) To generate a colony of C57BL/6 APPSWE/PS1ΔE9 x IFNAR1−/− mice with appropriate IFNAR1−/− littermate controls, heterozygous APPSWE/PS1ΔE9 mice were bred to mice homozygous for the IFNAR1 gene disruption (neomycin insert; IFNAR1−/−). The subsequent litters were termed F1 progeny and were expected to yield mice heterozygous for both the APPSWE/PS1ΔE9 mutation and IFNAR1-neoE5 allele (25 % male and 25 % female according to Mendelian inheritance). Upon genetic confirmation, APPSWE/PS1ΔE9 (het) x IFNAR1+/− mice were then interbred to generate the first APPSWE/PS1ΔE9 x IFNAR1−/− mice from F2 progeny (6.25 % male and 6.25 % female according to Mendelian inheritance). APPSWE/PS1ΔE9 (het) x IFNAR1−/− mice were then interbred to generate F3 progeny consisting of APPSWE/PS1ΔE9 x IFNAR1−/− (75 %) and IFNAR1−/− (25 %) littermates. B) Genotyping results for the initial progeny containing APPSWE/PS1ΔE9 x IFNAR1−/− and littermate IFNAR1−/− mice is displayed. Genotyping PCR was performed using a combined APPSWE/PS1ΔE9 and IFNAR1−/−. For each mouse, genotyping for APPSWE, PS1ΔE9 and IFNAR1 expression was performed in separate reactions with internal control (IC) amplification (APPSWE reactions only). The expected band sizes were as follows: APPSWE: 377 bp, PS1ΔE9: 608 bp, wildtype IFNAR1: 351 bp, IFNAR1-neomycin (IFNAR1−/−): 985 bp and IC: 324 bp. APP positive bands in lanes 2, 8 and 12 are comprised of both APPSWE and IC bands as indicated in the fig. C) Immunoblotting of Tris–HCl soluble cortical brain extracts, using mAb WO-2, revealed effective APP overexpression in APPSWE/PS1ΔE9 x IFNAR1−/− mice that was indistinguishable from levels in APPSWE/PS1ΔE9 mice. (TIF 6948 kb

Additional file 2: Table S1. of Deletion of the type-1 interferon receptor in APPSWE/PS1ΔE9 mice preserves cognitive function and alters glial phenotype

Statistical table (See additional statistical table file). Statistical analysis information is displayed for all data displayed within the accompanying figures. Power values were determined post-hoc based upon population effect size, sample size and group number. An α value of 0.05 was used to set the type-1 error rate in statistical comparisons. p values are given as exact values or adjusted multiplicity variants where an ANOVA and multiple comparisons post-hoc test was applied to the data. n represents the number of individual animals or primary cell cultures derived from separate animals used in each experiment. N/A: not applicable. (XLSX 49 kb

Additional file 4: Figure S3. of Deletion of the type-1 interferon receptor in APPSWE/PS1ΔE9 mice preserves cognitive function and alters glial phenotype

Aβ1-42 does not induce a p-Stat-3 mediated response in either wildtype or IFNAR1−/− primary glial cultures. A) Representative immunoblot of primary wildtype and IFNAR1−/− glial cultures treated with 10 μM Aβ1-42 for 24–72 h using anti-p-Stat-3. B) Densitometry of p-Stat-3 levels in primary wildtype and IFNAR1−/− glial cultures treated with 10 μM Aβ1-42 for 24–72 h. For densitometry, total Stat-3 levels were normalized to the β-actin loading control and p-Stat-3 intensity was calculated relative to this value (p-Stat-3/(Stat-3/β-actin). All intensity values of Aβ1-42 treated groups are expressed as fold change relative to the genotype-specific vehicle control (average of which is represented by the dashed line). Immunodetection of β-actin was used to ascertain loading quantities. Data are displayed as mean ± SEM (n = 3 per genotype). See Additional file 2: Table S1 for further analysis. (TIF 4277 kb

Additional file 3: Figure S2. of Deletion of the type-1 interferon receptor in APPSWE/PS1ΔE9 mice preserves cognitive function and alters glial phenotype

IFNβ mRNA transcript levels are unaltered in cortical tissue 9 month old APPSWE/PS1ΔE9 mice. Q-PCR of cortical tissue isolated from 9 month old wildtype and APPSWE/PS1ΔE9 mice analyzing IFNβ mRNA levels. For Q-PCR, all samples were normalized back to the Ct value of the housekeeping gene GAPDH (ΔCt). The mRNA of the variant genotype groups were then expressed relative to their gene-specific wildtype littermate controls (fold change, ΔΔCt). Data are displayed as box plots box plots described in the statistical analysis section in Materials and Methods (n = 7 per genotype) See Additional file 2: Table S1 for further analysis. (TIF 213 kb

Matrix Selection for the Visualization of Small Molecules and Lipids in Brain Tumors Using Untargeted MALDI-TOF Mass Spectrometry Imaging

Matrix-assisted laser desorption/ionization mass spectrometry imaging allows for the study of metabolic activity in the tumor microenvironment of brain cancers. The detectable metabolites within these tumors are contingent upon the choice of matrix, deposition technique, and polarity setting. In this study, we compared the performance of three different matrices, two deposition techniques, and the use of positive and negative polarity in two different brain cancer types and across two species. Optimal combinations were confirmed by a comparative analysis of lipid and small-molecule abundance by using liquid chromatography–mass spectrometry and RNA sequencing to assess differential metabolites and enzymes between normal and tumor regions. Our findings indicate that in the tumor-bearing brain, the recrystallized α-cyano-4-hydroxycinnamic acid matrix with positive polarity offered superior performance for both detected metabolites and consistency with other techniques. Beyond these implications for brain cancer, our work establishes a workflow to identify optimal matrices for spatial metabolomics studies