Project description:Plants represent the nutritional basis of virtually all life on earth and protein-rich foods from crop plants are a global megatrend essential for sustaining an increasing human population and counteracting climate change. While the genomes of crops are increasingly elucidated, little is known about crop proteomes – the entirety of proteins that execute and control nearly every aspect of life. To address this shortcoming we optimized a protocol for mapping the proteome of different crops such as Solanum lycopersicum (tomato) fruit and included four technical replicates and three biological replicates from different tomato plants to demonstrate the robustness of the workflow.
Project description:EMP500 - http://www.earthmicrobiome.org/emp500/ The Earth Microbiome Project is a systematic attempt to characterize global microbial taxonomic and functional diversity for the benefit of the planet and humankind. The Earth Microbiome Project (EMP) is a massively collaborative effort to characterize microbial life on this planet. We use DNA sequencing and mass spectrometry of crowd-sourced samples to understand patterns in microbial ecology across the biomes and habitats of our planet. The EMP is a comprehensive example of open science, leveraging a collaborative network of 500+ investigators, supporting pre-publication data sharing, and crowdsourcing data analysis to enable universal principles to be explored. The standardized collection, curation, and analysis are enabling a robust interpretation of ecological trends.ls for metagenomic sequencing and assembly, with the goal of applying this workflow to a range of environmental samples, combined with metabolomic profiling. Our goal was to assemble a set of ~500 fresh environmental samples across a range of habitats, with the help of the EMP network of collaborators. We are doing traditional EMP amplicon sequencing, metagenomic sequencing (with assembly-free and assembly-based analysis), and metabolomics on these 500 samples. A biobank of frozen aliquots of samples is being maintained at UCSD (soil samples at PNNL) for future methods testing and analysis.
Project description:Phthalates are ubiquitous pollutants in the environment; however, the mechanisms of phthalate-associated reproductive disorders in men are not fully understood. The aim of this study is to investigate associations between urinary phthalate metabolite concentrations and sperm DNA methylation. The study was conducted on 697 men from three prospective pregnancy cohorts: Longitudinal Investigation of Fertility and the Environment (LIFE) Study, Sperm Environmental Epigenetics and Development Study (SEEDS), and Environment and Reproductive Health (EARTH) Study.
Project description:Harsh environmental conditions including microgravity and radiation during prolonged spaceflights are known to alter hepatic metabolism. Our studies have focused on the analysis of possible changes in metabolic pathways in livers of mice which experienced 30 days of spaceflight with and without an additional re-adaption period of 7 days compared to control mice on Earth. Utilizing shotgun mass spectrometry and label-free quantification, we performed proteomic profiling to investigate mice livers from the spaceflight project “Bion-M 1”. No significant alterations in protein levels were observed between control mice liver and spaceflight mice which is possibly caused by insufficient fold change detection combined with high variances within the groups. In contrast, our results show that more than a third of the quantified protein levels are altered comparing the liver proteome of mice with and without re-adaption time after their spaceflight. Proteins related to amino acid metabolism showed higher levels after re-adaption, which may indicate higher rates of gluconeogenesis. Members of the peroxisome proliferator-activated receptor pathway reconstitute their level after 7 days due to a decrease in fold change, which indicates decreased signs of non-alcoholic fatty liver disease. Moreover, bile acid secretion regenerates on Earth due to reconstitution of related transmembrane proteins and elevated levels of the drug-metabolising enzymes belonging to the CYP superfamily decrease 7 days after the spaceflight. Thus, our study demonstrates reconstitution of pharmacological response and early signs of non-alcoholic fatty liver disease recover within 7 days, whereas glucose uptake should be monitored due to alterations in gluconeogenesis.
Project description:Genome-wide transcriptional profiling showed that reducing gravity levels in the International Space Station (ISS) causes important alterations in Drosophila gene expression intimately linked to imposed spaceflight-related environmental constrains during Drosophila metamorphosis. However, simulation experiments on ground testing space-related environmental constraints, show differential responses. Curiously, although particular genes are not common in the different experiments, the same GO groups including a large multigene family related with behavior, stress response and organogenesis are over represented in them. A global and integrative analysis using the gene expression dynamics inspector (GEDI) self-organizing maps, reveals different degrees in the responses of the transcriptome when using different environmental conditions or microgravity/hypergravity simulation devices These results suggest that the transcriptome is finely tuned to normal gravity. In regular environmental conditions the alteration of this constant parameter on Earth can have mild effects on gene expression but when environmental conditions are far from optimal, the gene expression is much more intense and consistent effects.
Project description:Serial analysis of gene expression (SAGE) was used to get a global view of the gene profile in human hippocampus. A library were generated from control hippocampus, obtained by rapid autopsy. Keywords: hippocampus human inventory genes Control hippocampus (used to construct the SAGE control library) was obtained from a 48 years old man without history of seizures or other neurological diseases and no brain abnormalities at autopsy and histologically normal hippocampus. Autopsy was performed within 4 hours after death. Tissue was snap-frozen and stored at –80 0C until use. Total RNA was isolated from control hippocampus and hippocampal surgical specimens, using the Trizol method according to the manufacturer’s instructions (Invitrogen - Life Technologies, The Netherlands). Part of the anterior hippocampus of control (including sectors CA1- CA4 and dentate gyrus, DG) was used. Poly(A)+ RNA isolation, cDNA synthesis and all subsequent steps of the SAGE procedure were essentially performed as described (Velculescu et al., 1995) with minor modifications given previously (Michiels et al., 1999).