MicroRNA profiling by array of mouse lung from individuals sensitised with chicken egg OVA and then challenged with aerosolised OVA
ABSTRACT: specific pathogen-free female BALB/c mice aged 7-8 weeks (Animal Resources Centre, Perth, Western Australia) were systemically sensitised by intraperitoneal injection of 50 µg of alum-precipitated chicken egg OVA (Grade V, ?98% pure, Sigma Australia) 21 and 7 days before inhalational challenge, then exposed to aerosolised OVA in a whole body inhalation exposure chamber (Unifab Corporation, Kalamazoo, MI). Chronic low-level challenge involved exposure to ?3 mg/m3 aerosolised OVA for 30 minutes/day on 3 days/week for up to 6 weeks. Particle concentration within the chamber was continuously monitored using a DustTrak 8520 instrument (TSI, St Paul, MN). All experimental procedures complied with the requirements of the Animal Care and Ethics Committee of the University of New South Wales (reference numbers: 06/119B and 08/09B). Mice were sacrificed after 1,2,4 and 6 weeks of OVA exposure. Control groups included naïve mice and mice that were not sensitised but were challenged for 6 weeks with aerosolised OVA.
BACKGROUND: The role of microRNAs (miRNAs) in regulating gene expression is currently an area of intense interest. Relatively little is known, however, about the role of miRNAs in inflammatory and immunologically-driven disorders. In a mouse model, we have previously shown that miRNAs are potentially important therapeutic targets in allergic asthma, because inhibition of miR-126, one of a small subset of miRNAs upregulated in the airway wall, effectively suppressed Th2-driven airway inflammation ...[more]
Project description:In this study we explored the antiviral gene expression induced in the CNS of MHV-infected mice, by performing whole-genome expression profiling. Three different mouse strains (BALB/c, 129SvEv and 129SvEv IFNAR-/- mice), differing in their susceptibility to infection with MHV, were used.
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Project description:Relative levels of RNA transcripts were compared between anterior and posterior wing bud thirds from stage HH24 normal and talpid3 mutant chicken embryos using chicken Affymetrix chips. Data collected with Affymetrix scanner was normalized using the Plier algorithm within the expression console package from Affymetrix and log2 transformed. 5 replicates of anterior third normal wing buds, 4 replicates of posterior third of normal wing buds and 4 replicates each of anterior and posterior thirds of talpid3 wing buds at stage HH24 were examined.
Project description:Bovine rotavirus (BRV) and bovine coronavirus (BCV) infect intestinal villous epithelium in young cattle. A surgical model was adapted for neonatal calves in which a region of the jejunum was isolated from the digestive tract but lymph drainage, enervation and blood flow were maintained. Replicate sections of intestine (loops) were infected with either BRV or BCV and adjacent segments were injected with phosphate-buffered saline. Tissues were collected 18 hours post-infection. Four animals were used for BRV infection studies, and three animals were used for BCV infection studies. Microarray analyses provided a global evaluation of host gene expression patterns following BRV and BCV infection and changes in gene expression were validated by qRT-PCR analyses.
Project description:MCF7 cells at 80% confluency were treated with 5 micromolar azacytidine with gene expression profiling determined at 0, 1, 2, 3, 4, 6 and 12 hours of exposure to compound. Keywords: time course Approximately 5,000,000 million cells were plated into 10 cm plates from a common pool. Cells were grown in DMEM supplemented with 10% foetal calf serum until 80% confluency was attained. Cells were then exposed to 5 micromolar azacytidine prepared as a 10 millimolar stock in DMSO. Each time point was represented by three biological replicates. Total RNA was extracted from the cells using the trizol method and RNA samples were validated for integrity throughout the isolation and extraction procedure. All gene expression profiles showed similar distributions with similar raw median intensities.