Project description:Intestinal microbiota colonization is important for intestinal development and health of preterm infants, especially those with extremely low birth weight. Recent studies indicated for a dynamic crosstalk between that gut microbiota and DNA methylome of host intestinal cells. Thereby, we sought to determine the epigenomic and metagenomic consequences of suppression of microbiota colonization in the intestine of preterm neonates to gain insights into biological pathways that shape the interface between the gut microbiota and the preterm intestinal cells. We examined 14 preterm piglets by comparing the conventional preterm neonates with those ones treated with oral antibiotics for genome wide DNA methylation and 16S rDNA microbiome. Our results demonstrated an extensive genome-wide DNA methylation changes in response to the suppression of intestinal microbe colonization, especially genes involved in neonatal immune response signaling and glycol-metabolism pathways were identified. Our study highlights several key genes that might predispose preterm neonates to NEC risk due to their key roles involved in the immune-metabolic networks. Our study not only provided rich omic-data to interpret molecular program in relation with microbiota-associated methylome-proteome network changes, but also confer clinical usage of key gene markers for potential early diagnostics of NEC of preterm neonates.
Project description:Endothelial colony-forming cells were isolated and expanded from the mononuclear cell fraction (MNC) obtained from the cord blood of term (CT) and preterm (PT) neonates, and characterized as previously described (Vassallo PF et al 2014). The objective of the study was to identify differentially expressed genes between CT and PT neonates. ECFC were harvested from cultures dishes at passage 3 and total RNA was extracted using the mirVana miRNA Isolation Kit (Ambion), according to the manufacturerâs recommendations. Cy3-CTP labeled RNA was prepared according to standard Agilent protocol from 400ng total RNA. The hybridization was performed for 17 hrs at 65°C. cRNA labeling and hybridization performance were performed and all parameters checked were found within the manufacturers specifications. Arrays were scanned as described in the manufacturersâ protocol. Signal intensities on 20 bit tiff images were calculated by Feature Extraction software (FE, Version 8.5; Agilent Technologies). Data analyses were conducted with GeneSpring GX software (Vers.13.1.1; Agilent Technologies)
Project description:Delivery of the baby before 37 weeks of completed gestation is deemed to be preterm birth. Admission of these preterm infants to the conventional neonatal intensive care unit (NICU) to manage their fragile physiology also tends to cause considerable stress that impedes the baby’s normal development. A recent innovation in neonatal care is the mother-neonatal ICU (MNICU), where the mother is not a mere visitor but has her bed inside the MNICU by the baby’s side. This study enrolled 200 low birth weight preterm babies (gestational age 28-37 weeks; weight 0.800–1.8 kg) randomized to MNICU & NICU. Saliva was collected from the 200 preterm neonates at the time of admission and discharge. We measured cortisol levels in the saliva of these samples, as the hormone is an established biomarker for stress. Salivary proteomics was also performed on 12 pairs of salivary samples chosen on basis of significant growth and development of these neonates. To study the differential proteome signature in these conditions MS-based data independent acquisition identified and quantified 342 and 308 protein groups, respectively. Differential protein analysis of these proteins revealed 41 differentially expressed proteins (DEPs). Pathway enrichment of unique DEPs in MNICU vs NICU comparison revealed improvement in immunity and metabolism-associated pathways at discharge. Quantitative analysis of the standard proteins from arrival and discharge groups revealed differential expression of these proteins. In differential expression analysis, we identified 28 upregulated and 16 downregulated proteins in neonates admitted to MNICU compared to NICU. A similar analysis for neonates at discharge identified 22 upregulated and 19 downregulated proteins. Further pathway enrichment of differential proteins unique to each group; 22 DEPs were present only in the arrival group, and 19 were in the discharge group. The data show a marked, statistically significant improvement in the overall well-being of preterm children admitted to MNICU and provided KMC compared to the NICU group. Thus, our study provides evidence in favor of easily available, cost-effective care that can make huge difference in the outcome of preterm neonates, particularly in low-income settings.
Project description:Necrotizing enterocolitis (NEC) is an acute and life-threatening gastrointestinal disorder afflicting preterm infants, which is currently unpreventable. Fecal microbiota transplantation (FMT) is a promising preventative therapy, but potential bacterial infection raise concern. Removal of bacteria from donor feces may reduce this risk while maintaining the NEC-preventive effects. We aimed to assess preclinical efficacy and safety of bacteria-free fecal filtrate transfer (FFT). Using fecal material from healthy suckling piglets, we administered FMT rectally, or cognate FFT either rectally or oro-gastrically to formula-fed preterm, cesarean-delivered piglets as a model for preterm infants, We compared gut pathology and related safety parameters with saline controls, and analyzed ileal mucosal transcriptome to gauge the host e response to FMT and FFT treatments relative to control. Results showed that oro-gastric FFT prevented NEC, whereas FMT did not perform better than control. Moreover, FFT but not FMT reduced intestinal permeability, whereas FMT animals had reduced body weight increase and intestinal growth. Global gene expression of host mucosa responded to FMT but not FFT with increased and decreased bacterial and viral defense mechanisms, respectively. In conclusion, as preterm infants are extremely vulnerable to enteric bacterial infections, rational NEC-preventive strategies need incontestable safety profiles. Here we show in a clinically relevant animal model that FFT, as opposed to FMT, efficiently prevents NEC without any recognizable side effects. If translatable to preterm infants, this could lead to a change of practice and in turn a reduction in NEC burden.
Project description:The involvement of the gut microbiota on weight-gain and its relationship with childhood undernutrition and growth has been reported. Thus, the gut microbiota constitutes a potential therapeutic target for preventing growth impairment. However, our knowledge in this area is limited. In this study we aimed at evaluating the relationship among early microbiota, growth, and development in preterm infants. To this end we assessed the levels of specific microorganisms by qPCR, and those of short chain fatty acids by mean of gas-chromatography, in feces from 63 preterm newborns and determined their weight-gain during the first months. The statistical analyses performed indicate an influence of the intestinal microbiota in weight-gain, with the levels of some microorganisms showing a significant association with the weight-gain of the infant. The levels of specific microbial groups during the first days of life were found to affect weight gain by the age of 1 month. Moreover, clustering of the infants on the basis of the microbiota composition at 1 month of age rendered groups which showed differences in weight z-scores. Our results suggest an association between the gut microbiota composition and weight-gain in preterm infants at early life and point out potential microbial targets for favoring growth and maturation in these infants.
Project description:To characterize the molecular profile of the chorioamniotic membranes of preterm neonates with and without neurocognitive impairment at 18-24 months and (2) to determine if neonates who developed neurocognitive impairment can be identified at birth. Paired two-group design