EGAS00001001597-sc-20160414 - samples
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ABSTRACT:
PROVIDER: EGAD00001002014 | EGA |
REPOSITORIES: EGA
Similar Datasets
Project description:Isolated populations have unique population genetics characteristics that can help boost power in genetic association studies for complex traits. Leveraging these advantageous characteristics requires an in-depth understanding of parameters that have shaped sequence variation in isolates. This study performs a comprehensive investigation of these parameters using low-depth whole genome sequencing (WGS) across multiple isolates.
| EGAS00001001597 | EGA
Project description:The field of phosphoproteomics has been revolutionized in the last years, with enhanced efficiency and sensitivity, now enabling deep phosphoproteomic analysis single-shot from minimal peptide inputs. In this study, we rigorously evaluate the effects of various experimental parameters in automated Zirconium-IMAC (Zr-IMAC) based phosphoproteomic sample preparation with focus on low peptide input samples. Assessing metrics such as number of identified phosphopeptides, selectivity, phosphosite localization scores and relative purification of multiply phosphorylated phosphopeptides, we were able to pinpoint key influencing variables. Determination of the optimal glycolic acid concentration in the loading buffer, percentage of ammonium hydroxide in the elution buffer, peptide to beads ratio, binding time, sample volume and loading buffer volume, allowed us to confidently localize up to ~15,800 phosphopeptides using 30 µg of peptides as starting material. Furthermore, we evaluated how sequential enrichment can boost the depth of the analysis, and most importantly, whether pooling fractions into a single-LCMS analysis also increases the phosphoproteome depth. As a result, we present a novel strategy based on incremental addition of beads and subsequent fraction pooling, which can boost by 17% the phosphoproteome coverage when compared to standard enrichment.
2024-03-26 | PXD045601 | Pride
Project description:We compared multiple strains of lab trophozoites to recent clinical isolates. Clinical isolates were grown in xenic media, and maintained many characteristics of the cyst stage of devlopment Keywords: Stage conversion
2007-03-01 | GSE6648 | GEO
Project description:A major goal of systems biology is the development of models that accurately predict responses of a cell or organism to perturbation. Constructing such models requires collection of dense measurements of system states, yet transformation of the data into predictive constructs remains a challenge. As a first step towards modeling human immunity, we have analyzed immune parameters in depth both at baseline and in response to perturbation with influenza vaccination. Peripheral blood cell transcriptomes, serum titers, frequencies of 126 cell subpopulations, and B cell responses were assessed before and after vaccination in 63 individuals and used to develop a systematic, computational framework to dissect inter- and intra-individual variation and build predictive models of post-vaccination antibody responses. Strikingly, independent of age and pre-existing antibody titers, accurate models could be constructed using pre-perturbation parameters alone, which were validated using data from independent baseline time-points. Most of the parameters contributing to prediction delineated temporally-stable baseline differences across individuals, raising the prospect of immune responsiveness prediction before intervention. According to CHI protocol 09-H1-0239, PBMC samples from 63 healthy voluntiers were collected 7 days prior to vaccination, immediately before vaccination (day0), and at 3 time points (day1, day7 and day70) post vaccination. The CHI Consortium
2014-04-10 | E-GEOD-47353 | biostudies-arrayexpress
Project description:Dynamic tyrosine phosphorylation is fundamental to a myriad of cellular processes. However, the inherently low abundance of tyrosine phosphorylation in the proteome and the inefficient enrichment of phosphotyrosine(pTyr)-containing peptides has led to poor pTyr peptide identification and quantitation, critically hindering researchers’ ability to elucidate signaling pathways regulated by tyrosine phosphorylation in systems where cellular material is limited. The most popular approaches to wide-scale characterization of the tyrosine phosphoproteome use pTyr enrichment with pan-specific, anti-pTyr antibodies from a large amount of starting material. Methods that decrease the amount of starting material and increase the characterization depth of the tyrosine phosphoproteome while maintaining quantitative accuracy and precision would enable the discovery of tyrosine phosphorylation networks in rarer cell populations. To achieve these goals, a novel method leveraging the multiplexing capability of tandem mass tags (TMT) and the use of trigger channels – cells treated with the promiscuous tyrosine phosphatase inhibitor pervanadate (PV) – selectively increased the relative abundance of pTyr-containing peptides. After PV facilitated selective fragmentation of pTyr-containing peptides, TMT reporter ions delivered sensitive quantitation of each peptide for the experimental samples while the quantitation from PV trigger channels was ignored. This method yielded up to 6.3-fold boost in quantification depth of statistically significant data derived from contrived ratios, compared to TMT without trigger channels or intensity-based label-free (LF) quantitation while maintaining quantitative accuracy and precision, allowing quantitation of over 2300 unique pTyr peptides from only 1 mg of T cell receptor-stimulated Jurkat T cells. The trigger channel strategy can potentially be applied in analyses of other post-translational modifications where treatments that broadly boost the levels of those modifications across the proteome are available.
2020-03-24 | PXD016192 | Pride
Project description:A major goal of systems biology is the development of models that accurately predict responses to perturbation. Constructing such models requires the collection of dense measurements of system states, yet transformation of data into predictive constructs remains a challenge. To begin to model human immunity, we analyzed immune parameters in depth both at baseline and in response to influenza vaccination. Peripheral blood mononuclear cell transcriptomes, serum titers, cell subpopulation frequencies, and B cell responses were assessed in 63 individuals before and after vaccination and were used to develop a systematic framework to dissect inter- and intra-individual variation and build predictive models of postvaccination antibody responses. Strikingly, independent of age and pre-existing antibody titers, accurate models could be constructed using pre-perturbation cell populations alone, which were validated using independent baseline time points. Most of the parameters contributing to prediction delineated temporally stable baseline differences across individuals, raising the prospect of immune monitoring before intervention.
2014-04-10 | GSE47353 | GEO
Project description:Natural S. cerevisiae isolates were evaluated for tolerance to a complex mixture of lignocellulosic inhibitors. Two isolates, with intermediate and superior tolerance characteristics, were investigated to determine the proteomic changes required for tolerance to lignocellulosic inhibitors.
2018-11-02 | PXD010868 | Pride
Project description:Global transcript profiles of different Arabidopsis thaliana genotypes were examined at 0h, 12h, 24h or 48h post infection with one of three different isolates (races) of the oomycete pathogen Hyaloperonospora parasitica (Hp). The three different isolates of Hp were used for this study, were HpEmoy2, HpHiks1 and HpEmco5 (see Eulgem et al., 2004, Plant Physiol. 135: 1129-1144 for more information). Experimental parameters varied in this study were Arabidopsis genotype, Hp isolate and time after infection.
2007-01-29 | E-TABM-96 | biostudies-arrayexpress
Project description:We compared multiple strains of lab trophozoites to recent clinical isolates. Clinical isolates were grown in xenic media, and maintained many characteristics of the cyst stage of devlopment Keywords: Stage conversion E. histolytica trophozoites of three different strains (HM1:IMSS, Rahman, and 200:NIH) and grown under multiple conditions (Long term axenic growth in TYI, Growth in low glucose media, Mouse adapted parasites and in vivo growth in a mouse model) were compared to recent clinical isolates that had grown in xenic culture for 1-8 weeks.
2007-03-01 | E-GEOD-6648 | biostudies-arrayexpress
Project description:Purpose : The goal of this study was to use RNA Seq to validate transcriptional data of two clinical isolates focussing on a subset of 74 transcript that were selected specifically for Nanostring analysis. Methods : mRNA profiles were generated for the clinical isolates FRD1 and CI224_M, in duplicate, by deep sequencing. Strains were grown for 8 hours in LB medium at 37C prior to RNA harvest. Ribosomal RNA was removed using the Ribi-Zero rRNA Removal Kit (Epicentre). mRNA reads were trimmed and mapped to the PAO1 NC_002516 reference genome from NCBI using the ClC Genomics Workbench platform and defaut parameters.
2016-08-09 | GSE83773 | GEO