Project description:We used a cDNA microarray previously defined for the marine sentinel organism Mytilus galloprovincialis (MytArray1.0) to evaluate the effects of nanomolar doses of combined metal salts (50, 100 and 200 nM mixtures of Cd, Cu and Hg) after 48 hours of mussel exposure. Pointing to the mussel gills, first target of toxic water contaminants and actively proliferating tissue, we found significant dose-related increases of cells with micronuclei and other nuclear abnormalities in the treated mussels, with differences in the bioconcentration of the three metals determined in the mussel pulp by atomic absorption spectrometry. Following gill RNA purification and DNA microarray analysis, individual gene expression profiles revealed some transcriptional changes at the 50 nM dose, and substantial increases of differentially expressed genes at the 100 and 200 nM doses with roughly similar amounts of up- and down-regulated signals. The functional annotation of transcripts with consistent expression trends and significantly altered at least in one dose point disclosed the complexity of the induced cell response. Three-condition experiment (50, 100 and 200 nM doses), individual treated mussel vs. pooled control mussels (N=5), 3 biological replicates with dye-swap competitive hybridization (array A and B, 2 technical replicates/array).
Project description:Sentinel lymph node, the first node draining the primary tumor, is a key component of tumor microenvironment promoting immune tolerance. In melanoma, sentinel node is one of the most important prognostic factors and the most frequent site of regional metastasis. To unravel the immunomodulatory pathways that are triggered by melanoma cells in the draining node that allow tumor spreading, transcriptional profiles associated to disease progression were analyzed in archival sentinel node biopsies (SNB). Gene expression profiles of melanoma-positive and negative SNB selected to maximize the differences in terms of disease stage and course, revealed subgroups correlating with regional node involvement and disease progression within positive biopsies. Transcriptional profiles revealed that genes showing differential expression between tumor-positive SNB with/without disease progression were mainly related to inflammatory response and that were mostly down regulated in patients with poor prognosis. TNFRSF8 encoding CD30 showing up regulation in SNB from patients with progressing disease displayed higher expression by immunohistochemical staining compared to SNB from non progressing patients. Subpopulations of CD30 positive CD4/CD8 double negative and CD4 Foxp3/PD-1 CD147 positive T cells were identified by flow cytometry analysis in metastatic nodes, suggesting a potential role of regulatory and tolerogenic T cells in melanoma progression.
Project description:We evaluated and correlated gene expression profiles of pre-treatment endoscopic biopsies in a pre-defined patient population of resectable oesophageal adenocarcinoma who had received neoadjuvant platinum and fluoropyrimidine based chemotherapy with clinical outcome, the primary end-point being overall survival. At the time of pre-treatment staging EUS, tumour biopsies were collected (approximately 1.5 mm3). and immediately snap frozen and stored in liquid nitrogen. Total RNA was extracted from the biopsies and amplified using a T7 bacteriophage RNA polymerase based linear amplification protocol, then expression profiling was carried out. The expression levels of 47 genes selected by biological function, fold change and degree of significance were further validated using quantitative reverse transcription-PCR Keywords: Tumour vs normal comparison
Project description:Liver cancer is one of the most lethal cancers worldwide. Liquid biopsy provides a noninvasive approach in detecting and monitoring cancer biomarkers to overcome current limitations associated with tissue biopsies, comprising the analysis of circulating tumor-derived material. In this study, we profiled plasma cell-free RNA-seq to identify recurrently dysregulated RNA biomarkers for the liquid biopsy of cancer.
Project description:Identification of early diagnostic and prognostic biomarkers measurable in liquid biopsies remains a major challenge in oncology today. OMICs technologies cannot be applied on clinically precious human material as this is required in its integrity for routine pathology investigation. Hence all OMICs discovery today is done on residual and not clinically relevant material. In the study, we present a novel, non-destructive, procedure named EXPEL, which uses rapid, pressure-assisted, interstitial fluid extrusion, allowing for full clinical pathology and OMICs analysis (proteins, metabolites, miRNA and DNA) on the same specimen. To demonstrate this we engage in a simultaneous biomarker discovery and routine diagnostic analysis of human colorectal cancer and liver metastases. Biomarkers discovered with EXPEL methodology can be readily detected in human sera, allowing for unprecedented translation to the clinic. EXPEL method enables for the first time both clinicians and scientist to explore identical, fresh, clinical material regardless of origin and size.
Project description:Identification of early diagnostic and prognostic biomarkers measurable in liquid biopsies remains a major challenge in oncology today. OMICs technologies cannot be applied on clinically precious human material as this is required in its integrity for routine pathology investigation. Hence all OMICs discovery today is done on residual and not clinically relevant material. In the study, we present a novel, non-destructive, procedure named EXPEL, which uses rapid, pressure-assisted, interstitial fluid extrusion, allowing for full clinical pathology and OMICs analysis (proteins, metabolites, miRNA and DNA) on the same specimen. To demonstrate this we engage in a simultaneous biomarker discovery and routine diagnostic analysis of human colorectal cancer and liver metastases. Biomarkers discovered with EXPEL methodology can be readily detected in human sera, allowing for unprecedented translation to the clinic. EXPEL method enables for the first time both clinicians and scientist to explore identical, fresh, clinical material regardless of origin and size.
Project description:Endogenously determined inter-individual differences in growth rate of bivalve molluscs have been widely analyzed at different organizational levels. Most studies have focused on the characterization of the physiological differences between fast (F) and slow (S) growing individuals. Although several genes have been described to be up regulated on fast growing individuals, the molecular basis underlying the mechanisms at the origin of growth variation is still poorly understood. In the present study we reared mussel spat of the species Mytilus galloprovincialis under diets below the pseudofaeces threshold (BP) and above the pseudofaeces threshold (AP). After 3 months, F and S mussels on each condition were selected, so that 4 experimental groups were obtained: FBP, SBP, FAP and SAP. We hypothesized that nurturing conditions during their growing period would modify the molecular basis of growth rate differences. However, results of feeding experiments showed that F mussels displayed higher clearance and ingestion rates and higher efficiencies of food selection prior to ingestion, as well as higher gill surface areas, irrespective of the rearing nutritional environment. To decipher molecular mechanism at the origin of growth variation, gills of the 4 mussel groups were dissected, and used for transcriptome analysis with a custom Agilent single channel microarray. Gene expression analysis revealed i) a low number (12) of genes differentially expressed associated to maintenance condition differences and ii) 117 genes differentially expressed when comparing fast and slow growing mussels (FBP + FAP vs. SBP + SAP). We further investigated this comparison: GO terms and KEGG pathway association of the differentially expressed genes allowed us to analyze the functions involved on the differentially expressed encoding. Transcriptomic differences between F and S mussels were mainly based on the up-regulation of response to stimulus, growth and cell activity Biological Process GO terms. Regarding the KEGG terms, carbohydrate metabolism and Krebs cycle were found to be up-regulated in F mussels whereas biosynthetic processes were up-regulated in S mussels. Among the differentially expressed genes that are annotated, the following ones were found to be up regulated in F mussels: i) Mucin, related to mucus secretion, known to be crucial in food acquisition and pre-ingestive selection processes in bivalves, ii) genes related to growth such as Myostatin or Insulin-like growth factor, iii) genes involved in feeding activity, such as Fibrocystin or Dynein and iv) genes involved in the energetic metabolism; Citrate synthase. S mussels mainly over-expressed genes related to immune system and defence (Leucine-rich repeat-containing protein, Metalloendopeptidase, Small heat shock protein 24, Multidrug resistance,…).The present results suggest that differences in feeding activity and in the allocation of metabolic energy between growth groups could account for the differences in growth rate in spat of Mytilus galloprovincialis. In accordance with their higher feeding rates and growth, fast growing mussels were found to mainly over-express genes involved in the development and maintenance of such activities, however, slow growing mussels needed to expend energy in immune and defence processes to ensure survival at the expense of growth rate.
2018-10-10 | GSE120975 | GEO
Project description:Bias in liquid-based air sampling