Project description:Chronic constant hypoxia (CCH), such as in pulmonary diseases or high altitude, and chronic intermittent hypoxia (CIH), such as in sleep apnea, can lead to major changes in the heart. The molecular mechanisms underlying these cardiac alterations are not well understood. We hypothesized that analysis on the changes in gene expression could help to delineate such mechanisms. In addition, the differences that can be anticipated between CCH and CIH could be potentially dissected. Current study used CCH and CIH mouse models combined with cDNA microarrays to determine the changes of gene expression in CCH or CIH mice heart. Keywords = heart Keywords = hypoxia Keywords = mouse Keywords = microarray Keywords: time-course
Project description:Rationale: Despite the deleterious effects associated with elevated carbon dioxide (CO2), or hypercapnia, it has been hypothesized that CO2 can protect the lung from injury. However, the effects of chronic hypercapnia on the neonatal lung are unknown. Objectives: To determine whether chronic hypercapnia alters alveolar development and to identify genes that could potentially contribute to hypercapnia-mediated lung protection. Methods: Newborn mouse litters were exposed to 8% CO2, 12% CO2 or room air for 2 weeks. Lungs were excised and analyzed for morphometric alterations. Gene expression changes were assessed by microarray techniques and RT-PCR, and gene products by western blotting. Results: The alveolar walls of CO2-exposed mice appeared thinner than those of controls. In addition, genes from a variety of functional categories were differentially expressed with hypercapnia, including those involved with cell growth/maintenance, signal transduction, protein metabolism, ion transport, stress response and inflammation. In particular and of major interest, gene expression was increased for surfactant proteins (SP) A and D, epithelial Na+ channel, GATA binding protein 6 and fibroblast growth factor receptor 2. In addition, SP-A and SP-D protein expression was increased with hypercapnia. Conclusions: Our results lead us to conclude that: 1) There are potentially a number of gene families which may contribute to hypercapnia-mediated lung protection, and 2) up-regulation of SP-A and SP-D may play a role as anti-inflammatory or antioxidant agents. Based on our genomic results, the effects of CO2 depend on the level to which the lung is exposed. Total 5 arrays including 2 dye-swaps were performed for the 8% CO2 treated animals. Five individual animals were analyzed. Total 4 arrays including 2 dye-swaps were performed for the 12% CO2 treated animals. Four individual animals were analyzed.
Project description:The complete genome sequence of the P. vivax Sal-1 strain allowed the design of a first version array representing 1 oligo/2 kb of coding sequences (http://zblab.sbs.ntu.edu.sg/vivax/index.html). Here, proof-of-principle experiments using total RNA of parasites obtained from the Sal-1 strain, from P. falciparum and from parasites obtained directly from two human patients are presented. To determine the extent of cross-hybridization of P. falciparum with P. vivax, and to determine overlaps in expression profiles of the P. vivax Sal1 monkey-adapted strain vs wild isolates, single dual hybridization analyses were performed.
Project description:A set of 22 expts. aimed at identifying splicing events dependent upon on the Spt4-5 transcription elongation factors in yeast. Four spt mutants and an mRNA capping mutant were analyzed four times each, including biological and technical (dye-swap) replicates. Two wt vs wt expts. were also performed. Keywords = transcription/spt5/spt4/splicing/DEDS
Project description:The goal of this study is to identify P. vivax genes whose expression is dependent on the intact spleen in experimental infections in Aotus monkeys. These studies were carried-out at the facilities of the “Fundación Centro de Primates de la Universidad del Valle”, Cali, Colombia and in the “Barcelona Centre for International Health Resarch” - CRESIB, Barcelona, Spain. This protocol was approved from the Ethical Committees of both Centres. A total of 4 Aotus lemurinus griseimembra naive animals were used in these experiments. Three animals were splenectomized whereas another had an intact spleen. A donor monkey was infected with P. vivax Sal-I strain and after peak parasitemias appeared a time-series infections into Sp-1, Sp-2, Sp-3, and Sp+2 animals were performed. Parasites from each infection were obtained from peripheral blood, monkey leukocytes were removed by MidMacs and only purified schizont stages were used for RNA extractions. Dual-hybridizations Cy3/Cy5 comparing the global expression of parasites obtained from different infections (Cy5) with a reference pool PvSp-1 obtained from splenectomized monkeys from CDC (PvSp-1) were perfomed using an Agilent's 60-mer platform representing the complete coding genome of P. vivax (1 oligonucleotide/2 kb coding sequences) GPL6667
Project description:Human HaCaT keratinocytes were cultured to confluence (day10). Differentiated confluent cells were irradiated at 0.5 Gy or 2 Gy. RNA from irradiated cells was compared to RNA from non-irradiated reference cells in a dye-swap hybridization procedure 24 h after irradiation. Keywords = human keratinocytes, gamma irrdiation
Project description:Human keratinocytes were cultured to confluence (day 10). Differentiated confluent cells were irradiated at 0.5 and 2 Gy. RNA from irradiated cells was compared to RNA from non-irradiated reference cells in a dye-swap procedure 24 h after irradiation. Keywords = Human keratinocytes, gamma irradiation
Project description:We used cDNA microarray technology to compare the genome-wide expression profiles of a wild type strain (BY4700) (E02, E04, E06) or the isogenic strain deleted of GLN3 and GAT1 genes (E01, E03, E05) grown in YNB medium with glutamine as nitrogen source (M.Gln) against the wild type strain grown in M.Gln after addition of rapamacyn (20 min) (E01, E02), or M.proline (M.Pro) (E03, E04) or after a two hours shift from M.Gln to M.Pro (E05, E06), all growth conditions known to modify the expression of genes involved in nitrogen utilization. These microarrays allowed to identify the set of genes that were up or down-regulated in response to the quality of the nitrogen source. To evaluate whether the majority of genes responding to the nitrogen source were dependent on Gln3 and Gat1, we compared the expression profiles of the wild type strain and of the isogenic strain deleted of GLN3 and GAT1 genes (03167b: ura3, gln3â, gat1â), when both strains were grown on M.Gln + rapamycin (E07), or M.Pro (E08) or after a shift from M.Gln to M.Pro (E09). We also used an independent means of identifying Gln3-Gat1 regulated genes by comparing the expression profiles in wild type and ure2â (4709âURE2) strains on M.Gln medium (E10). The hybridization signal was measured using a GSM418 laser scanner. Image analysis for each array was processed using the GenePix Pro 4.0 (Axon Instruments, Inc.) software package, which measures fluorescence intensity pairs for each gene. Following image acquisition, a visual inspection of the individual spots on each microarray (size, signal-to-noise ratio, background level, and spot uniformity) completed the flagging (present/not present, good/bad) of the data. To maximize sensitivity two scans were made, one at high laser power and high PMT (Photo Multiplier Tube) gain to detect the faintest spots, and a second one using low laser power and a PMT gain avoiding saturation. The values for spots presenting ⥠5% saturation in the first scan were calculated based on an extrapolation after linear regression analysis of the intensities from both scans. These data were then imported in the GeneSpring 7.1 (Silicon Genetics) software package, applying a per spot per chip intensity-dependent (Lowess) normalization for further analysis.