GP, participated in the study design. MRO supevised the work, defined the study design and carried out Akt activator the writing of the manuscript. All authors read and approved the final manuscript.”
“Background Necrotizing enterocolitis
(NEC) is an acute inflammatory disease that affect the intestinal tract of neonates [1]. It remains one of the most common gastrointestinal emergencies in newborn neonates [2]. Onset of NEC occurs within the first three months of life and neonates who are of low birth weight and under 28 week gestation are the most susceptible [3]. The ileum and the proximal colon are the frequently affected although any segments of the gastrointestinal tract can be involved [4]. The course of NEC is multifactorial and the most important elements is prematurity, enteral feeding, bacterial colonization and an inappropriate pro-inflammatory response [5]. It is believed selleckchem that immaturities of these functions due to age predispose the premature infant to intestinal injury and inappropriate responses to injury. The bacterial role in NEC still needs to be clarified. Suggestions such as an imbalance
of the gastrointestinal microbiota, overgrowth of potential pathogenic bacteria, and LY294002 research buy ischemia causing mucosal lesions that gives the bacteria systemic access have been followed but so far no specific pathogens have been identified. Correlation of NEC with bacteria has been suggested by analysing faecal samples, however, this analysis of faecal samples is often far from the affected site and may not be representative [5–11]. The use of formalin-fixed paraffin-embedded tissue samples give an opportunity to investigate a unique stock of archival disease-specific material. The method is challenged to access the limited and fragmented bacterial DNA present in the tissue. To characterize the bacterial population in the formalin-fixed NEC tissue laser-capture-micro-dissection
(LCM) combined with fluorescence in situ hybridization (FISH), clonidine using a bacteria ribosomal RNA (rRNA)-targeting oligonucleotide probe, was used [12]. The bacterial 16S rRNA gene was PCR amplified and sequenced by pyrosequencing. The bacterial distribution was verified and visualized within the lumen and mucus of the intestinal tissues with fluorescent in situ hybridization (FISH) with group and species specific probes targeting individual microbial cells (Table 1). The aim of this study was to investigate the microbial composition and the relative number of bacteria in affected intestinal tissue samples surgically removed from neonates diagnosed with NEC and to relate this with the patient data such as antibiotic treatment.