Project description:Sexual dimorphism is one of the important topic in mammal species because appearance of males and females is obvious different in all mammal species. In addition to this, molecular mechanisms also very different each other. Furthermore, it is important to employ a variety of tissues in RNA-seq experiment because recent studies imply gene expression pattern are highly tissue specific. Although previous related studies discovered numerous sexually dimorphic mechanism in mammal species, but still, many mechanisms are undiscovered in case of non-model organisms. One of the representative mammal organism is a cattle which is less researched about sexual dimorphism. For investigate bovine sexual dimorphism, we generated two-way factorial designed 40 samples RNA-seq data composed with two factors such as gender and tissues. Two statistical approaches are employed for identifying bovine sexually dimorphic genes using such two-way factorial designed RNA-seq data. As a result, we observed that detected sexually dimorphic genes exhibited strong tissue specific pattern, but fat tissue showed relatively small tissue specificity than the others. In addition, we observed that sex-related genes are shared in two mammal species such as cattle and rat through qRT-PCR experiments. Finally, we investigated pros and cons of two statistical approaches for complex structured RNA-seq analysis.
Project description:Aging is associated with declining immunity and inflammation as well as alterations in the gut microbiome with a decrease of beneficial microbes and increase in pathogenic ones. The aim of this study was to investigate aging associated gut microbiome in relation to immunologic and metabolic profile in a non-human primate (NHP) model. 12 old (age>18 years) and 4 young (age 3-6 years) Rhesus macaques were included in this study. Immune cell subsets were characterized in PBMC by flow cytometry and plasma cytokines levels were determined by bead based multiplex cytokine analysis. Stool samples were collected by ileal loop and investigated for microbiome analysis by shotgun metagenomics. Serum, gut microbial lysate and microbe-free fecal extract were subjected to metabolomic analysis by mass-spectrometry. Our results showed that the old animals exhibited higher inflammatory biomarkers in plasma and lower CD4 T cells with altered distribution of naïve and memory T cell maturation subsets. The gut microbiome in old animals had higher abundance of Archaeal and Proteobacterial species and lower Firmicutes than the young. Significant enrichment of metabolites that contribute to inflammatory and cytotoxic pathways was observed in serum and feces of old animals compared to the young. We conclude that aging NHP undergo immunosenescence and age associated alterations in the gut microbiome that has a distinct metabolic profile.
Project description:A metaproteomics analysis was conducted on the infant fecal microbiome to characterize global protein expression in 8 samples obtained from infants with a range of early-life experiences. Samples included breast-, formula- or mixed-fed, mode of delivery, and antibiotic treatment and one set of monozygotic twins. Although label-free mass spectrometry-based proteomics is routinely used for the identification and quantification of thousands of proteins in complex samples, the metaproteomic analysis of the gut microbiome presents particular technical challenges. Among them: the extreme complexity and dynamic range of member taxa/species, the need for matched, well-annotated metagenomics databases, and the high inter-protein sequence redundancy/similarity between related members. In this study, a metaproteomic approach was developed for assessment of the biological phenotype and functioning, as a complement to 16S rRNA sequencing analysis to identify constituent taxa. A sample preparation method was developed for recovery and lysis of bacterial cells, followed by trypsin digestion, and pre-fractionation using Strong Cation Exchange chromatography. Samples were then subjected to high performance LC-MS/MS. Data was searched against the Human Microbiome Project database, and a homology-based meta-clustering strategy was used to combine peptides from multiple species into representative proteins. Bacterial taxonomies were also identified, based on species-specific protein sequences, and protein metaclusters were assigned to pathways and functional groups. The results obtained demonstrate the applicability of this approach for performing qualitative comparisons of human fecal microbiome composition, physiology and metabolism, and also provided a more detailed assessment of microbial composition in comparison to 16S rRNA.
2019-03-29 | PXD006033 | Pride
Project description:Viral metagenomics of pig fecal samples
Project description:Leaf-cutting ants of the genera Acromyrmex and Atta live in mutualistic symbiosis with a basidiomycete fungus (Leucocoprinus gongylophorus), which they cultivate as fungal gardens in underground nest chambers. The ants provide the fungus with a growth substrate consisting of freshly cut leaf fragments. After new leaf fragments are brought into the nest, the ants chew them into smaller pieces and apply droplets of fecal fluid to the leaf pulp before depositing this mixed substrate in the fungus garden and inoculating it with small tufts of mycelium from older parts of the garden. Previous work has shown that the fecal fluid contains a range of digestive enzymes including proteases, amylases, chitinases, cellulases, pectinases, hemicellulases and laccases, and that most of these enzymes are produced by the fungal symbiont in specialized structures called gongylidia that the ants eat. After ingestion, the enzymes apparently pass unharmed through the alimentary channel of the ants and end up in the fecal fluid. Most likely this complex system is an adaptation of the ant-fungus symbiosis to a herbivorous lifestyle, as the ancient ancestors of the ants and the fungus lived as hunter-gatherers and saprotrophs, respectively. The promise of fecal fluid for getting insight into the molecular adaptations that enables the ant-fungus holosymbiont to live as a herbivore, led us to investigate the fecal fluid proteome using LC-MS/MS in order to get a more comprehensive picture of the repertoire of proteins present.