Project description:Prostate basal epithelial cultures generated from patient normal and cancer tissue were treated with LTP for 3 minutes and RNA harvested at 2hrs post-LTP. Microarrays were used to analyze whole transcriptome changes between untreated and LTP-treated prostate cells.
Project description:The histopathological and molecular heterogeneity of prostate cancer and the limited availability of human tumor tissue make unraveling the mechanisms of prostate carcinogenesis a challenging task. Our goal was to develop an ex vivo model that could be reliably utilized to define a prognostic signature based on gene expression profiling of cell cultures that maintained the tumor phenotype. To this end, we derived epithelial cultures from tissue explanted from 59 patients undergoing radical prostatectomy or cistoprostatectomy because of Prostate Benign Hyperplasia/Prostate Cancer or Bladder Carcinoma. Patient selection criteria were absence of hormonal neo-adjuvant treatment before surgery and diagnosis of clinically localized disease. Using this unique experimental material we analyzed expression of 22.500 transcripts on the Affymetrix Human U133A Gene Chips platform. Cultures from normal/hyperplastic tissues with a prevalent luminal phenotype, and from normal prostate epithelial tissue with basal phenotype (PrEC) served as controls. We have established a large number of prostate primary cultures highly enriched in the secretory phenotype. From them we derived an epithelial-restricted transcriptional signature that: 1) differentiated normal from tumor cells; and 2) clearly separated cancer derived lines into two distinct groups which correlated with indolent and aggressive clinical behavior of the disease. Our findings provide:1) a method to expand human primary prostate carcinoma cells with a luminal phenotype; 2) a powerful experimental model to study primary prostate cancer biology; and 3) a novel means to characterize these tumors from a molecular genetic standpoint for prognostic and/or predictive purposes. Keywords: prostate cancer; gene profile; primary cultures; prognosis; molecular pathology.
Project description:xtracellular matrix from prostate tissue samples and from cultures of primary fibroblasts of the same samples from individuals was analysed by mass spectrometry.
Project description:Aims: To investigate the inactivation kinetics of Bacillus cereus vegetative cells upon exposure to low-temperature nitrogen gas plasma and to reveal the mode of inactivation by transcriptome profiling. Methods and results: Exponentially growing B. cereus cells were filtered and put on agar plates. The plates, carrying the filters with the vegetative cells, were placed into nitrogen gas plasma at atmospheric pressure and cold temperature (~37°C). After different exposure times the cells were harvested for RNA extraction and enumeration. The RNA was used to perform whole transcriptome profiling using DNA microarrays. The transcriptome profile showed a large overlap with profiles obtained from conditions generating reactive oxygen species inside B. cereus. However, excess radicals such as peroxynitrite, hydroxyl and or superoxide were not found. Conclusions: Antibacterial activity of nitrogen gas plasmas is not based on UV radiation but on the formation of reactive oxygen or nitrogen species in the plasma jet rather than inside the targeted cells. Significance and impact of the study: This study represents the first investigation of differential gene expression on a genome-wide scale in B. cereus following nitrogen gas plasma exposure. This study may help to design cheap, safe, and effective plasma decontamination devices.
Project description:The histopathological and molecular heterogeneity of prostate cancer and the limited availability of human tumor tissue make unraveling the mechanisms of prostate carcinogenesis a challenging task. Our goal was to develop an ex vivo model that could be reliably utilized to define a prognostic signature based on gene expression profiling of cell cultures that maintained the tumor phenotype. To this end, we derived epithelial cultures from tissue explanted from 59 patients undergoing radical prostatectomy or cistoprostatectomy because of Prostate Benign Hyperplasia/Prostate Cancer or Bladder Carcinoma. Patient selection criteria were absence of hormonal neo-adjuvant treatment before surgery and diagnosis of clinically localized disease. Using this unique experimental material we analyzed expression of 22.500 transcripts on the Affymetrix Human U133A Gene Chips platform. Cultures from normal/hyperplastic tissues with a prevalent luminal phenotype, and from normal prostate epithelial tissue with basal phenotype (PrEC) served as controls.,We have established a large number of prostate primary cultures highly enriched in the secretory phenotype. From them we derived an epithelial-restricted transcriptional signature that: 1) differentiated normal from tumor cells; and 2) clearly separated cancer derived lines into two distinct groups which correlated with indolent and aggressive clinical behavior of the disease. ,Our findings provide:1) a method to expand human primary prostate carcinoma cells with a luminal phenotype; 2) a powerful experimental model to study primary prostate cancer biology; and 3) a novel means to characterize these tumors from a molecular genetic standpoint for prognostic and/or predictive purposes.
Project description:Aims: To investigate the inactivation kinetics of Bacillus cereus vegetative cells upon exposure to low-temperature nitrogen gas plasma and to reveal the mode of inactivation by transcriptome profiling. Methods and results: Exponentially growing B. cereus cells were filtered and put on agar plates. The plates, carrying the filters with the vegetative cells, were placed into nitrogen gas plasma at atmospheric pressure and cold temperature (~37°C). After different exposure times the cells were harvested for RNA extraction and enumeration. The RNA was used to perform whole transcriptome profiling using DNA microarrays. The transcriptome profile showed a large overlap with profiles obtained from conditions generating reactive oxygen species inside B. cereus. However, excess radicals such as peroxynitrite, hydroxyl and or superoxide were not found. Conclusions: Antibacterial activity of nitrogen gas plasmas is not based on UV radiation but on the formation of reactive oxygen or nitrogen species in the plasma jet rather than inside the targeted cells. Significance and impact of the study: This study represents the first investigation of differential gene expression on a genome-wide scale in B. cereus following nitrogen gas plasma exposure. This study may help to design cheap, safe, and effective plasma decontamination devices. Plasma treated sampes compared with nitrogen gas flow treated samples and plasma treated samples compared to untreated control samples. Nitrogen flow samples in duplicate, plasma treated samples 4 replicates. Replicates hybridized with dyes swapped.
Project description:Diurnal temperature cycling is an intrinsic characteristic of many exposed microbial ecosystems. However, its influence on yeast physiology and transcriptome has not been studied in detail. In this study, 24-h sinoidal temperature cycles, oscillating between 12 and 30°C, were imposed on anaerobic, glucose-limited chemostat cultures of Saccharomyces cerevisiae. After three diurnal temperature cycles (DTC), concentrations of glucose, and extracellular metabolites, as well as CO2-production rates showed regular, reproducible circadian rhytms. DTC also led to waves of transcriptional activation and repression, which involved one sixth of the yeast genome. A substantial fraction of these DTC-responsive genes appeared to primarily respond to changes in glucose concentration. Elimination of known glucose-responsive genes revealed overrepresentation of previously identified temperature-responsive genes as well as genes involved in cell cycle and de novo purine biosynthesis. Analyses of budding index and flow cytomery demonstrated that DTC led to a partial synchronization of the cell cycle of the yeast populations in the chemostat cultures, which was lost upon release from DTC. Comparison of DTC results with data from steady-state cultures showed that DTC was sufficiently slow to allow S. cerevisiae chemostat cultures to almost completely acclimatize their transcriptome and physiology at the DTC temperature maximum, and to approach acclimation at the DTC temperature minimum.
Project description:Mass spectrometry-based proteomics has been successfully used to characterize biofluids, most notably blood, for biomarker discovery or to evaluate patients’ underlying diseases. Despite immense progress, challenges such as blood’s large dynamic range or low throughput remain for those types of studies. In this work, we used cutting-edge proteomics technologies to construct labelled and label-free l workflows, capable of quantifying approximately 2,000 proteins in biofluids. With just 70µL of blood and a single depletion strategy, we conducted an analysis of a cohort comprising 64 individuals, comparing prostate cancer patients to healthy donors. The results revealed dozens of differentially expressed proteins in both plasma and serum. Notably, we identified the upregulation of Prostate Specific Antigen (PSA), a well-known biomarker for prostate cancer, in the cancer cohort, along with the discovery of potential novel markers. Most of the differentially regulated proteins were consistently quantified using both the lower-throughput and higher-throughput workflows. Our bioinformatics analysis of the prostate cancer cohort data suggests that some of the newly discovered proteins may be secreted differentially into the bloodstream, making them potential candidates for disease markers.