Project description:LPS (10 μg/50 μl PBS; E. coli Serotype 055:B5, Sigma-Aldrich) was administered onto the nares. Baseline levels of genes in mice treated with vehicle (Hanks Balanced Salt solution (HBSS) and PBS), Intranasal S100s (10 μg/50 μl HBSS) were given 2 h before LPS; control mice received equal volumes of PBS and HBSS. Mice were sacrificed 4 h post LPS inhalation.
Project description:To evaluate the change of gene expression at cerebral infarction by the treatment of IV-human umblical cord derived mesenchymal stem cell. RNA was isolated from the ipsilateral hemisphere to MCAo in rats. At 72 h post-MCAo, the ipsilateral hemisphere subjected to MCAo was used for mRNA microarray. RNA was isolated from the ipsilateral hemisphere to MCAo in rats without MCAo (control group, n=5), rats treated with 1x106 IV-hUMSC (hUMSC group, n=6) and saline (saline group, n=5) at 24h post-MCAo.
Project description:Identify differentially expressed genes related to the neurodegenerative process in a new animal model of hepatic encephalopathy (HE). The animal model consists on the simulation of several bouts of HE in PCA rats, being the precipitant factors of the episodes ammonia (NH3) and/or lipopolisaccharide (LPS) or saline.
Project description:Identify differentially expressed genes related to the neurodegenerative process in a new animal model of hepatic encephalopathy (HE). The animal model consists on the simulation of several bouts of HE in PCA rats, being the precipitant factors of the episodes ammonia (NH3) and/or lipopolisaccharide (LPS) or saline. Regular administration (one every two weeks up to 10 infussion) of NH3 (20 μl/min along 3h) and/or LPS (5mg/kg) or in PCA rats. Sham rats were used as a surgery control.
Project description:To identify possible target molecules for acute alcohol intoxication therapy, we used microarray analysis to compare cerebella gene expression profiles of control and acute alcohol-intoxicated rats. We first established a model of acute alcohol intoxication in SD rats, and then used rat cDNA microarray to profile mRNA expression in the cerebella of alcohol-intoxicated rats (experimental group) and saline-treated rats (control group).
Project description:m-ICcl2 cells (Bens et al., Am. J. Physiol., 1996) were grown for 6 days on collagen coated cell culture flasks in medium as recommended (see ference above). Medium was changed every 48 hours. The cells were mycoplasma tested (PCR) and found negative. The medium and PBS was endotoxin tested (ELISA) and found to be negative (< 50pg/mL). Unstimulated cells (control) were left cultured in medium only. Lipopolysaccharide (100 ng/mL in water, D31m4 E. coli LPS from List Biochemicals) was sonicated (10 min_ prior to use. Cells were stimulated 6 hours with 100 ng/mL LPS and harvested or stimulated with LPS (100 ng/mL) and washed with warm phosphate buffered saline (PBS) after 6 hours, followed by medium change every 48 hours. Cells were harvested after 96 hours (6 hours incubated in the presence of LPS, 90 hours after stimulation). RNA was isolated after repeated (3x) washing with PBS using Trizol reagent (1 mL/75 cm2) using the standard protocol and resuspended in Millipore water. Quantification was done by spectroscopy and the RNA was stored at -80C.
Project description:T helper type 2 (Th2) responses are crucial for defense against infections by helminths and are responsible for the development of allergic reactions that can lead to severe clinical disorders, such as asthma or anaphylaxis, and ultimately to death. The induction of Th2 responses requires a specific activation process, triggered by specialized dendritic cells (DCs), by which naive CD4+ Th0 cells acquire the capacity to produce Th2 cytokines. However, the mechanistic basis of the functional specialization enabling DCs for the initiation of Th2 responses has remained elusive. Here we show that interleukin-4 (IL-4), a cytokine produced by basophils, mast cells and Th2-polarized CD4+ T helper cells, exerting a crucial function during anti-helminths and allergic Th2 responses, has a key role in the licensing/conditioning of DCs for the induction of Th2 responses, by bloking their potential to produce Th1-driving cytokines, such as IL-12, IL-18 and IL-23. Microarray analyses (duplicates) were for two types of comparisons: 1. moDCs stimulated with LPS from Escherichia coli versus C-moDCs non stimulated (control). 2. moDCs stimulated with LPS from Escherichia coli in presence of IL4 versus C-moDCs non stimulated (control).
Project description:LPS (10 μg/50 μl PBS; E. coli Serotype 055:B5, Sigma-Aldrich) was administered onto the nares. Baseline levels of genes in mice treated with vehicle (Hanks Balanced Salt solution (HBSS) and PBS), Intranasal S100s (10 μg/50 μl HBSS) were given 2 h before LPS; control mice received equal volumes of PBS and HBSS. Mice were sacrificed 4 h post LPS inhalation. Expression of inflammatory genes was evaluated with the RT-qPCR array. Relative quantities of mRNA in duplicate samples were obtained using the LightCycler® 480 Software 1.5 and the Efficiency-Method.