Project description:A phylogenetic microarray targeting 66 families described in the human gut microbiota has been developped aud used to monitor the gut microbiota's structure and diversity. The microarray format provided by Agilent and used in this study is 8x15K. A study with a total of 4 chips was realized. Arrays 1 and 2: Hybridization with 100ng of labelled 16S rRNA gene amplicons from a mock community sample and 250ng of labelled 16S rRNA gene amplicons from 1 faecal sample. Each Agilent-030618 array probe (4441) was synthetized in three replicates. Arrays 3 and 4: Hybridization with 250ng of labelled 16S rRNA gene amplicons from 2 faecal samples. Each Agilent-40558 array probe (4441) was synthetized in three replicates.
Project description:Age-dependent changes of the gut-associated microbiome have been linked to increased frailty and systemic inflammation. This study found that age-associated changes of the gut microbiome of BALB/c and C57BL/6 mice could be reverted by co-housing of aged (22 months old) and adult (3 months old) mice for 30-40 days or faecal microbiota transplantation (FMT) from adult into aged mice. This was demonstrated using high-throughput sequencing of the V3-V4 hypervariable region of bacterial 16S rRNA gene isolated from faecal pellets collected from 3-4 months old adult and 22-23 months old aged mice before and after co-housing or FMT.
Project description:In this study, we performed a comparative analysis of gut microbiota composition and gut microbiome-derived bacterial extracellular vesicles (bEVs) isolated from patients with solid tumours and healthy controls. After isolating bEVs from the faeces of solid tumour patients and healthy controls, we performed spectrometry analysis of their proteomes and next-generation sequencing (NGS) of the 16S gene. We also investigated the gut microbiomes of faeces from patientsand controls using 16S rRNA sequencing. Machine learning was used to classify the samples into patients and controls based on their bEVs and faecal microbiomes.
Project description:The search for factors beyond the radiotherapy dose that could identify patients more at risk of developing radio-induced toxicity is essential to establish personalised treatment protocols for improving the quality-of-life of survivors. To investigate the role of the intestinal microbiota in the development of radiotherapy-induced gastrointestinal toxicity, the MicroLearner observational cohort study characterised the intestinal microbiota of 136 (discovery) and 79 (validation) consecutive prostate cancer patients at baseline radiotherapy. Gastrointestinal toxicity was assessed weekly during RT using CTCAE. An average grade >1.3 over time points was used to identify patients suffering from persistent acute toxicity (endpoint). The intestinal microbiota of patients was quantified from the baseline faecal samples using 16S rRNA gene sequencing technology.
Project description:IL22 induces antimicrobial peptides which influnce microbiota. We used 16s rRNA gene sequencing (16s DNA-seq) to analyze the microbiota with Fc or IL-22Fc treatment.
Project description:Chronic acid suppression by proton pump inhibitor (PPI) has been hypothesized to alter the gut microbiota via a change in intestinal pH. To evaluate the changes in gut microbiota composition by long-term PPI treatment. Twenty-four week old F344 rats were fed with (n = 5) or without (n = 6) lansoprazole (PPI) for 50 weeks. Then, profiles of luminal microbiota in the terminal ileum were analyzed. Pyrosequencing for 16S rRNA gene was performed by genome sequencer FLX (454 Life Sciences/Roche) and analyzed by metagenomic bioinformatics.
Project description:We compared the microbiota of paired mouse caecal contents and faeces by applying a multi-omic approach, including 16S rDNA sequencing, shotgun metagenomics, and shotgun metaproteomics. The aim of the study was to verify whether faecal samples are a reliable proxy for the mouse colonic luminal microbiota, as well as to identify changes in taxonomy and functional activity between caecal and faecal microbial communities, which have to be carefully considered when using stool as sample for mouse gut microbiota investigations.
Project description:<p>Objective: The gut microbiota plays a crucial role in rectal adenocarcinoma (READ). Nonetheless, little is known about corresponding changes in gut microbiota–host interactions. This study aimed to identify the structure of the gut microbiota and the characteristics of faecal metabolites in patients with READ.</p><p>Methods: Faecal samples were collected from 33 individuals with READ and 34 healthy controls (HC), with post-neoadjuvant treatment faecal samples also collected from the 5 READ patients, and 16S rRNA gene amplification sequencing and untargeted liquid chromatography-mass spectrometry metabolomics analysis were performed. Methods such as weighted correlation network analysis (WGCNA) and molecular docking to determine target gene–related information, and to explore the effects of guanidinoacetic acid (GAA) on colorectal cancer cell lines through in vitro experiments.</p><p>Results: Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetes were the most abundant phyla in patients with READ and healthy controls (HC). At the genus level, Peptostreptococcus exhibited marked enrichment in the READ cohorts, whereas Faecalibacterium was dominant in the HC group. The predicted microbial functional analysis indicated a significant increase in the metabolism of betaine, guanidinoacetic acid (GAA), L-tryptophan, arachidonic acid, and arachidic acid. Moreover, by comparing the faecal metabolites between the HC and READ groups,Pearson’s correlation analysis demonstrated significant interactions between six microbiota taxa and nine metabolites, suggesting specific human metabolic pathways. The cellular function results demonstrated that GAA promoted the proliferation, invasion, and migration of colorectal cells while inhibiting apoptosis. Further analysis revealed that GAA may might promote CRC progression of colorectal cancer by affecting molecules such as KLB, CA2, CSTG, CYP4F12, and GZBM.</p><p>Conclusion: This study provides a foundational guide for the systematic and multidimensional evaluation of the contribution of the gut microbiota and its metabolites to READ and identifies a metabolite (namely, GAA) that may be of important in the occurrence, development, and treatment of READ, offering a new perspective for further research on READ.</p>