MicroRNA profiling of human peripheral blood leukocytes before and after stimulation with E. coli lipopolysaccharide
ABSTRACT: To gain insight into miRNAs involved in the regulation of the human innate immune response, we screened for differentially expressed miRNAs in circulating leukocytes in an in vivo model of acute inflammation triggered by E. coli lipopolysaccharide (LPS) infusion. Leukocyte RNA was isolated from venous blood samples obtained from healthy male volunteers before and 4 hours after LPS-infusion. After fluorescence labeling, RNA samples were hybridized to microarrays containing capture probes for measuring the abundance of more than 600 human miRNAs (Exiqon).
Project description:Two experiments were preformed, one for evaluating the effect of E2 exposure in males and a second experiment for comparing gene expression in the liver of non-vitellogenic females with that of vitellogenic females. The experiments designs were as follows:<br>1) Four months old zebrafish were divided into three groups consisting of 8 fish in each group: i) males (N=8) were exposed to E2 (Sigma-Aldrich, Israel) by immersion for 48 h (group E). The concentration used was 5 ?g/L (18 nM), as 3-4 ng/ml was determined to be the E2 natural concentration in the plasma of adult vitellogenic female ZF (Heiden et al 2006); ii) untreated males(N=8); iii) untreated vitellogenic females(N=8). Four replicate samples were prepared for each group and each replicate consisted of a pooled sample from livers of two fish. <br>2) In order to reveal the differences between vitellogenic and non-vitellogenic females, a second experiment was designed. One month old zebrafish were divided into two groups consisting of 32 fish in each group. Due to the small size of the liver, pooled samples from eight fish were prepared for each of the four replicates in the expression studies. The groups consisted of: i) fish that were kept under a light/dark cycle of 14/10 h for 5 weeks (N=32). with ovaries in vitellogenic stage; ii) fish that were kept under a light/dark cycle of 6/18 h for the same time period (N=32) with non-vitellogenic ovaries.
Project description:U87 cell spheroids were harvested from collagen on day 0 (no migration), or day 3 (extensive migration). The RNA extracted from these cells was used to identify microRNA alterations that occur during glioma cell line migration.
Project description:We identify a panel of microRNAs that are differentially expressed during both spontaneous and LPS-induced DC maturation and show the M-CSF receptor (M-CSFR) as a key target for microRNA-mediated regulation. MicroRNA-22, -34a and -155 are up-regulated in mature GM-CSF-generated DC and mediate M-CSFR mRNA and protein down-regulation.
Project description:H69 cells were cultured in H69 medium with 1 ng/ml lipopolysaccharide(LPS, for smaples 04, 05 and 06) or without LPS(for samples 01, 02 and 03) for 8 hours and then collected for array analysis. <br>
Project description:The purpose of the present study was to elucidate in more details the molecular mechanisms of neutrophil-mediated inflammation. We therefore investigated the time-dependent stimulatory potential of LPS from Escherichia coli on cytokine response in neutrophil-like HL-60 cells. <br><br>To get a general overview on the total mRNA changes in DMSO-differentiated HL-60 cells, we performed whole-transcript analysis of LPS-stimulated dHL-60 cells after 2h and 6h of LPS treatment. The samples were collected from three independent experiments from three different passages in cell culture. Non-stimulated dHL-60 cells served as control. We identified the differentially expressed cytokine genes implicated in the human inflammatory response and prominent for their role in neutrophil-mediated inflammatory processes.
Project description:Male C57B1/CBA mice were exposed to mainstream tobacco smoke (MTS) from two cigarettes daily, 5 days/week for 6 or 12 weeks. Mice were sacrificed immediately, or six weeks following, the last cigarette. High density DNA microarrays were used to characterize global gene expression changes in whole heart.<br><br>Western blot and ELISA assays were used to determine levels of Cyp1A1 protein in heart microsomes and total and active PAI-1 protein from tissue extracts.
Project description:Mice were acclimatized for 2 weeks and randomly assigned to treatment and control groups (five per group). Animals were given a single dose of 250, 50, 5 and 0.5 mg/kg isoproterenol by oral gavage alongside 0.9% saline (vehicle control group)and sacrificed after eight hours. Total mouse liver RNA was isolated and used for the microarry analysis on custom oligonucleotide DNA microarray, the HC ToxArray V1.2.<br><br>