Transcription profiling of human healthy peripheral blood cells isolated from male participants at rest
ABSTRACT: These gene expression profiles were measured to create a broad and balanced control for any project that examines gene expression changes in men exposed to a defined stimulus (see series 5105). Experiment Overall Design: 9 healthy men (aged 21 to 44) gave written agreement to participate to this study. At the time of blood withdrawal all participants were non-smokers, with normal BMI and did not obtain any medicamentation. We collected 12ml of whole blood and isolated T-lymphocytes out of 4ml and monocytes out of 8ml whole blood.
Project description:Exercise has an measureable impact on gene expression profiles of white blood cells. While subpopulations of white blood cells undergo specific changes in answer to physical load we focused on pan T-lymphocytes and Monocytes. These cells are in direct contact to organs influenced by exercise and additionally should be affected by alterations of nutrient and gas composition in blood, which occur as a consequence of exercise. We screened the gene expression profiles of top athletes at the beginning and the end of the Tour de France 2005 and at at the annual rest period in order to find functional processes that are differentially expressed in answer to the longtime and high load training and competition. Therefor we filtered out genes with significant changes in gene expression more than 1.5fold between two of the three conditions and build overlap lists of this list and GO (gene ontology) lists and KEGG (kyoto encyclopedia of genes and genomes) lists. We observed a strong decline in immune functions and an equal decline in metabolic processes. Experiment Overall Design: 12 professional cyclists from two german top teams gave written agreement to participate to this study. We collected 12ml of whole blood at each time point and isolated T-lymphocytes out of 4ml and monocytes out of 8ml whole blood. The experimental groups are divided into rest (6 replicates in Tcells/ 7 replicates in Monocytes), start (10 replicates in each cell type) and final (8 replicates in Tcells/ 9 replicates in Monocytes). This means that we collected samples at the beginning and the end of the Tour de France and during the annual rest period of the athletes.
Project description:These gene expression profiles were measured to create a broad and balanced control for any project that examines gene expression changes in men exposed to a defined stimulus (see series 5105). Experiment Overall Design: 9 healthy men (aged 21 to 44) gave written agreement to participate to this study. At the time of blood withdrawal all participants were non-smokers, with normal BMI and did not obtain any medicamentation. We collected 12ml of whole blood and isolated T-lymphocytes out of 4ml and monocytes out of 8ml whole blood.
Project description:Lung cancers are a heterogeneous group of diseases with respect to biology and clinical behavior. So far, diagnosis and classification are based on histological morphology and immunohistological methods for discrimination between two main histologic groups: small cell lung cancer (SCLC) and non-small cell lung cancer which account for 20% and 80% of lung carcinomas, respectively. While SCLCs express properties of neuroendocrine cells, NSCLCs, which are divided into the three major subtypes adenocarcinoma, squamous cell carcinoma and dedifferentiated large cell carcinoma, show different characteristics such as the expression of certain keratins or production of mucin and lack neuroedocrine differentiation. The molecular pathogenesis of lung cancer involves the accumulation of genetic und epigenetic alterations including the activation of proto-oncogenes and inactivation of tumor suppressor genes which are different for lung cancer subgroups. The development of microarray technologies opened up the possibility to quantify the expression of a large number of genes simultaneously in a given sample. There are several recent reports on expression profiling on lung cancers but the analysis interpretation of the results might be difficult because of the heterogeneity of cellular components. A contamination of the tumor sample with normal epithelia, blood vessels, stromal cells, leucocytes and tumor necrosis may confound the true expression profile of the tumor. The use of laser capture microdissection (LCM) greatly improves the sample preparation for microarray expression analysis. Consequently, we used advanced technology including LCM and microarray analysis. In detail, we examined gene expression profiles of tumor cells from 29 previously untreated patients with lung cancer (10 adenocarcinomas (AC), 10 squamous cell carcinomas (SCC), 9 small cell lung cancer (SCLC)) in comparison to normal lung tissue (LT) of 5 control patients without tumor. Bronchoscopical biopsies from the primary lung tumor were taken before treatment. Biopsies were cut into 8µm sections and from each section cancer cells were isolated using laser capture microdissection in order to obtain pure samples of tumor cells. Total RNA was extracted, reversely transcribed, in-vitro transcribed, labelled and hybridized to the array. For expression analysis, microarrays covering 8793 defined genes (Human HG Focus Array, Affymetrix) were used. Following quality control, array data were normalized and analysed for significant differences using variance stabilizing transformation (VSN) and significance analysis of microarrays (SAM), respectively. Based on differentially expressed genes cancer samples could be clearly separated from non cancer samples using hierarchical clustering. Comparing AC, SCC and SCLC with normal lung tissue, we found 205, 335 and 404 genes, respectively, that were at least 2-fold differentially expressed with an estimated false discovery rate < 2.6%. Each histological subtype showed a distinct expression profile. Further, using a genetic programming approach we constructed a classificator to discriminate AC, SCC, NT and SCLC. To this end, the 50 genes with the greatest signal-to-noise ratio were selected to train the classificator. By leave-one-out cross validation all 34 samples were correctly classified in this training set. In order to validate the 50-gene-classificator on a test set, further 13 microdissected lung cancer samples were used and correctly classified in concordance to pathologic finding. In conclusion, the different lung cancer subtypes have distinct molecular phenotypes which reflect biological characteristics of the tumor cells and which might be the basis for development of targeted therapy. Moreover, gene expression profiling and genetic programming is a suitable tool for classification and discrimination of different histological subtypes in lung cancer in comparison to normal lung tissue. Experiment Overall Design: Comparison of gene expression profiles of normal lung tissues, adenocarcinomas, squamous cell carcinomas and small cell lung cancers.
Project description:To identify genes associated with lung cancer progression, we examined gene expression profiles of tumor cells from 20 patients with primary, untreated non-small cell lung cancer (10 adenocarcinomas (AC) and 10 squamous cell carcinomas (SCC)) in comparison to lung tissue of 23 patients with stage IIIB or stage IV non-small cell lung cancer (15 AC and 8 SCC). Bronchoscopical biopsies from patient with recurrent lung tumor were taken after initial treatment. Cancer cells were isolated using laser capture microdissection in order to obtain pure samples of tumor cells. For expression analysis, microarrays covering 8793 defined genes (Human HG Focus Array, Affymetrix) were used. Array data were normalized and analysed for significant differences using variance stabilizing transformation (VSN) and significance analysis of microarrays (SAM), respectively. Genes were considered to be up- or down-regulated when the ratio between primary and recurrent tumor samples were at least 1.5-fold differentially expressed with an estimated false discovery rate: < 5%. Based on differentially expressed genes, primary cancer samples could be separated from recurrent tumor samples. We identified 115 and 124 significantly regulates genes in AC and SCC, respectively. For example, in recurrent AC we found increased expression of genes related to the wingless (FZD6, RYK, MYC) and calcium (CALM1, ATB2B1, S100A2) signalling pathways which might play a role in metastasis of tumor cells. Other differentially expressed genes were related to cell cycle (CCND1, CDK2), transcription factors (TTF1, TAF2, YY1), nuclear mRNA splicing and mRNA processing (SFRS1, HNRPL), protein-nucleus import (NUTF2, KPNB1, NUP50) and chromatin modification (HIST1H4C, SMARCC1). In SCC, we found an increased expression of CTNNB1, an important mediator in wingless signalling pathway. Among the down-regulated genes in SCC, the utmost fraction belonged to genes coding for ubiquitin mediated proteolysis (UCHL1, PSMA3, COPS6) and ribosomal proteins (RPS26, RPL7A, RPS15). Other down regulated genes were related to transcription factors (TCEA2, TAF10), nuclear mRNA splicing and mRNA processing (SNRPD2, HNRPM). In conclusion, a distinct pattern of gene expression is found during the progression from primary carcinoma to recurrent NSCLC. Our microarray-based expression profiling revealed interesting novel candidate genes and pathways that may contribute to lung cancer progression. Experiment Overall Design: - 20 patients with primary, untreated non-small cell lung cancer (10adenocarcinomas (AC) and 10 squamous cell carcinomas (SCC)) in comparison to lung tissue of 23 patients with stage IIIB or stage IV non-small cell lung cancer (15 AC and 8 SCC) Experiment Overall Design: - Human HG Focus Array, Affymetrix) were used Experiment Overall Design: - Array data were normalized and analysed for significant differences using variance stabilizing transformation (VSN) and significance analysis of microarrays (SAM) Experiment Overall Design: - Genes were considered to be up- or down-regulated when the ratio between primary and recurrent tumor samples were at least 1.5-fold differentially expressed with an estimated false discovery rate: < 5%
Project description:In this study we used the Affymetrix Barley 1 GeneChip to investigate transcriptome responses of barley cv. Morex to low temperature, including triplicated measurements of cold, freeze/thaw cycles and de-acclimation over 33 days. Experiment Overall Design: Plants were grown at 20ºC for seven days and subject to a symmetrical cycle of acclimation, cold, freeze-thaw, and deacclimation. Chilling began by decreasing the temperature overnight from 20ºC to 4ºC at a rate of 1.3ºC•h-1 and maintaining temperatures of 4 ºC in the day and 2ºC at night for 5 days. Freeze-thaw cycling lasted 12 days with day temperatures of 4ºC and night temperatures gradually decreasing from -2ºC the first night to -4ºC for three nights and -10ºC for four nights, then recovering to -4ºC for three nights and -2ºC for one night. This treatment was designed to allow daily freeze-thaw cycling and protein synthesis. Chilling conditions (4ºC day, 2ºC night) were resumed for five days, followed by deacclimation with increasing temperature to 20ºC overnight and maintaining for three days. Sampling was done at four different times, each at the 11th hour of light to avoid circadian effects: 1) before chilling treatment, 2) five days after initiation of chilling treatment, 3) eight days into freeze-thaw treatment and 4) three days into de-acclimation.
Project description:The effect of ahg1 and ahg3 on the gene expression profiles is similar but some genes are differentially affected. Experiment Overall Design: To investigate the effect of ahg1 and ahg3 mutations, the genome wide gene expression profiles of germinating seedlings were examined. These mutations occurred in close but distinct PP2C genes. Stratified seeds were transferred to plates containing 0.5 uM ABA and allowed germinate at 23 °C under 16hr light 8hr dark conditions. After two days samples were collected for RNA extraction.
Project description:BACKGROUND AND OBJECTIVES: Low-grade B-cell lymphomas include several subtypes of tumors with different degrees of histological, biological or clinical features. Differential diagnosis is frequently compromised by the lack of specific cytogenetic or molecular features. As a consequence, therapies remain in many lymphoma types largely based in common protocols with largely variable success. Our objectives were to describe and to compare the genomic profile of a series of samples from the most prevalent low-grade lymphoma subtypes; all of them systematically analyzed with the same approach. DESIGN-AND-METHODS: We carried out a high-resolution genomic DNA analysis (44K probes) in 87 low grade B cell lymphoma tumor samples that unambiguously presented the clinical picture, analytical features, and peripheral blood morphology and phenotype described for each entity. RESULTS: The genomic integrity of the samples was heavily compromised (80% of the tumors presented at least one aberration). This phenomenon also involved lymphoplasmocytic and marginal zone lymphomas that have not been previously studied by this genomic approach. New aberrations have been described for almost every subtype. We have also generated reports of the extension of the genomic instability, detecting distinct patterns of genomic instability within subtypes. INTERPRETATION-AND-CONCLUSIONS: The genomic profile of each subgroup showed substantive differences. The bioinformatic analysis of the data detected a set of new aberrations which were present in the majority of the subgroups and that pointed out to specific pathways, such as NF-kB (gains that involved REL, BCL11A and COMMD1) or DNA repair checkpoint pathways (deletion of 16q24 involving CDT1). Experiment Overall Design: Comparative experiment: pheripheral blood pool of ten healthy female donors: CONTROL vs. 87 Low grade Lymphoma tumour samples
Project description:Expression profiling of Drosophila mir-8 homozygous mutant pupae at a single developmental stage (72 hours APF). Homozygous mutant and wild type pupae were collected at 72 hours APF (After Puparium Formation). Three independent collections were performed. Total RNA was extracted and amplified. Mutant and respective wildtype samples were hybridized together to custom cDNA arrays (DGC1 and 2 EST collections).
Project description:Ewing’s Sarcoma (ES) is characterized by specific chromosomal translocations, the most common being t(11;22)(q24;q12). Additionally, other types of genetic abnormalities may occur and be relevant to explain the variable tumoural biology and clinical outcome. We have carried out a high-resolution array CGH and expression profiling on 25 ES tumour samples to characterize the DNA copy number aberrations (CNA) occurring in these tumours and to determine their association with gene expression profiles (GEO Series accession number: GSE8303) and their clinical outcome. CNA were observed in 84% of the cases. We observed a median number of 3 aberrations per case. Besides numerical chromosomal changes, smaller aberrations were found and defined at chromosomes 5p, 7q and 9p. All CNA were compiled to define the smallest overlapping regions of imbalance (SORI). Thirty five SORI were delimited. Bioinformatic analyses were conducted to identify subgroups according to the pattern of genomic instability obtained. Unsupervised and supervised clustering analyses (using SORI as variables) segregated the tumours into two distinct groups: one genomically stable (≤ 3 CNA) and another genomically unstable one (> 3 CNA). The genomic unstable group showed a statistically significant shorter overall survival and was more refractory to chemotherapy. This report elucidates data about genomic instability in ES, based on CNA and expression profiling for the first time, and shows that a genomically unstable group of Ewing tumours is correlated with a significant poor prognosis. Experiment Overall Design: Comparative experiment: pheripheral blood pool of ten healthy female donors: CONTROL vs. 26 Ewing Sarcoma Tumour Samples. Experiment Overall Design: Note that although 26 samples were profiled, only 25 were analysed (sample #11 did not pass quality control).
Project description:Internal sugar and light specific dependent regulation of leaf gene expression was addressed by changing [CO2] to lower than compensation point [CO2] in combination with light or prolonged darkness. Plants were grown on soil in a 12/12 h light/dark rhythm at 20°C day and night and under normal [CO2]. 5 weeks after germination, the above-ground rosettes of the non-flowering plants were harvested, 12 plants per sample. Plants were harvested 4hrs after the end of night (i) under low (< 50 ppm) [CO2] and 150 µE fluorescent light , (ii) under normal [CO2] and light, and, (iii) under low [CO2] and prolonged darkness. The low [CO2] treatment started 30 min before the end of night and stopped with harvesting. Experiment Overall Design: 2 biological replicates of plants under 3 treatments (4hrs low CO2+light, 4hrs low CO2+prolonged darkness, and 4hrs normal CO2+light).