ABSTRACT
BACKGROUND AND OBJECTIVE: Lipid metabolism dysregulation has been implicated in the pathogenesis of IPF; however, the roles of most lipid metabolites in lung fibrosis remain unexplored. Therefore, we aimed to identify changes in lipid metabolites in the lung tissues of IPF patients and determine their roles in pulmonary fibrosis. METHODS: Free fatty acids in the lung tissues of IPF patients and controls were quantified using a metabolomic approach. The roles of free fatty acids in fibroblasts or epithelial cells treated with TGF-ß1 were evaluated using fibrotic markers. The antifibrotic role of stearic acid was also assessed in a bleomycin-induced lung fibrosis mouse model. Protein levels in cell lysates or tissues were measured by western blotting. RESULTS: The levels of stearic acid were lower in IPF lung tissues than in control lung tissues. Stearic acid significantly reduced TGF-ß1-induced α-SMA and collagen type 1 expression in MRC-5 cells. Furthermore, stearic acid decreased the levels of p-Smad2/3 and ROS in MRC-5 cells treated with TGF-ß1 and disrupted TGF-ß1-induced EMT in Beas-2B cells. Stearic acid reduced the levels of bleomycin-induced hydroxyproline in a mouse model. CONCLUSION: Changes in the free fatty acid profile, including low levels of stearic acid, were observed in IPF patients. Stearic acid may exert antifibrotic activity by regulating profibrotic signalling.
Subject(s)
Bleomycin/pharmacology , Fibroblasts/metabolism , Idiopathic Pulmonary Fibrosis , Lung/physiology , Stearic Acids/chemistry , Transforming Growth Factor beta1/chemistry , Animals , Bleomycin/chemistry , Humans , Idiopathic Pulmonary Fibrosis/drug therapy , Idiopathic Pulmonary Fibrosis/genetics , Mice , Signal Transduction/physiology , Transforming Growth Factor beta1/metabolismABSTRACT
AcrR, the toxic-compounds-response regulator, regulates motility in microorganisms, presumably to escape from toxic environments. In this study, the genome-wide target genes of AcrR were investigated in a ΔacrR mutant strain by microarray analysis. In the absence of AcrR, the transcription of most flagella/motility genes was highly increased. In addition, flagella formation was increased in this mutant strain. Motility assays revealed that AcrR modulates swimming motility, but not swarming.
Subject(s)
Cell Movement/genetics , Escherichia coli Proteins/genetics , Escherichia coli/genetics , Escherichia coli/physiology , Mutation/genetics , Repressor Proteins/genetics , Flagella/genetics , Flagella/physiology , Mutation/physiologyABSTRACT
Catalytic CO oxidation was activated at low temperature by injecting O3 as an additive. It was empirically confirmed that CO removal rate was dramatically enhanced by supplying a small amount of O3, and the reaction temperature was almost half that required for CO oxidation when using a catalyst only. By optimizing the concentration of O3, catalytic CO oxidation could be achieved within 1 min at low operational temperature. The removal rate of CO was sensitive to the concentration of O3, and a deduced reaction mechanism is discussed to explain how catalytic CO oxidation is activated but subsequently deactivated at higher O3 concentration. Moreover, the presence of C3H8 and C3H6 were considered to evaluate the effects of each gas on the enhancement of CO removal rate by O3. Finally, the rate of CO removal was evaluated with increasing O3 concentration for practical applications such as the cold-start problem in automobile engines.
Subject(s)
Carbon Monoxide/isolation & purification , Ozone/chemistry , Catalysis , Equipment Design , Hydrocarbons/chemistry , Oxidation-Reduction , Platinum , Temperature , Vehicle EmissionsABSTRACT
Acriflavine resistance regulator (AcrR), a local transcription factor, regulates the expression of the acrRAB genes associated with the AcrAB-TolC multidrug efflux pump. Screening of organic solvent tolerance (OST) with the overexpression of 13 genes in Escherichia coli revealed that the overexpression of acrR improved OST. Overexpression of AcrR in a background strain of wild-type E. coli and in the OST strain LMB015 (ΔfadR ΔmarR; acrR (+) and ΔfadR ΔmarR acrR (+) strain, respectively) significantly increased cell growth in the presence of n-hexane/cyclohexane, which attenuated the membrane reduction capacity of the wild-type strain below 50 % of the control level. This was recovered to control levels in the acrR (+) strain. Quantitative real-time PCR analysis of RNA from the wild-type, ΔacrR, and acrR (+) strains showed that AcrR represses the transcription of marRAB and soxRS, and its own gene cluster, acrRAB. Electrophoretic mobility shift assay demonstrated that AcrR binds directly to the promoter region of acrRAB, marAB, and soxRS, indicating that AcrR acts on global regulators to affect mar-sox-rob regulon. In the acrR (+) strain, soxS expression was significantly upregulated compared with the wild-type. The OST of the acrR (+) strain was completely lost in the ΔsoxS acrR (+) strain, indicating that SoxS mediated OST improvement in the acrR (+) strain. The observation that all genes associated with marRAB and soxRS are upregulated in the ΔacrR strain, and that there is only moderate induction of soxS (and marB) in the acrR (+) strain, provides insight into how acrR overexpression confers bacterial OST and the mar-sox-rob regulon control network.
Subject(s)
Drug Tolerance , Escherichia coli Proteins/biosynthesis , Escherichia coli/drug effects , Gene Expression Regulation, Bacterial , Organic Chemicals/toxicity , Repressor Proteins/biosynthesis , Solvents/toxicity , Trans-Activators/biosynthesis , DNA, Bacterial/metabolism , Electrophoretic Mobility Shift Assay , Escherichia coli/genetics , Escherichia coli/growth & development , Escherichia coli/physiology , Escherichia coli Proteins/genetics , Gene Deletion , Gene Expression , Gene Expression Profiling , Protein Binding , Real-Time Polymerase Chain Reaction , Regulon , Repressor Proteins/genetics , Trans-Activators/geneticsABSTRACT
To activate a catalyst efficiently at low temperature by plasma for environmental control, we developed a hybrid reactor that combines plasma with a honeycomb-structured catalyst in a practical manner. The reactor developed generated stable cold plasma at atmospheric pressure because of the dielectric and conductive nature of the honeycomb catalyst by consuming low amounts of power. In this reactor, the applied voltage and temperature determined the balance between the oxidation and adsorption by the plasma and catalyst. The synergistic reaction of the plasma and catalyst was more effective at low temperatures, resulting in a reduction in a lowered light-off temperature.
Subject(s)
Air Pollution/prevention & control , Vehicle Emissions , Aluminum Oxide/chemistry , Atmospheric Pressure , Catalysis , Plasma Gases , Platinum/chemistry , TemperatureABSTRACT
The improvement of bacterial tolerance to organic solvents is a main prerequisite for the microbial production of biofuels which are toxic to cells. For targeted genetic engineering of Escherichia coli to increase organic solvent tolerances (OSTs), we selected and investigated a total of 12 genes that participate in relevant mechanisms to tolerance. In a spot assay of 12 knockout mutants with n-hexane and cyclohexane, the genes fadR and marR were finally selected as the two key genes for engineering. Fatty acid degradation regulon (FadR) regulates the biosynthesis and degradation of fatty acids coordinately, and the multiple antibiotic resistance repressor (MarR) is the repressor of the global regulator MarA for multidrug resistance. In the competitive growth assay, the ΔmarR mutant became dominant when the pooled culture of 11 knockout mutants was cultivated successively in the presence of organic solvent. The increased OSTs in the ΔmarR and ΔfadR mutants were confirmed by a growth experiment and a viability test. The even more highly enhanced OSTs in the ΔfadR ΔmarR double mutant were shown compared with the two single mutants. Cellular fatty acid analysis showed that the high ratio of saturated fatty acids to unsaturated fatty acids plays a crucial role in OSTs. Furthermore, the intracellular accumulation of OST strains was significantly decreased compared with the wild-type strain.
Subject(s)
Bacterial Proteins/genetics , Escherichia coli Proteins/genetics , Escherichia coli/drug effects , Escherichia coli/genetics , Gene Deletion , Organic Chemicals/pharmacology , Repressor Proteins/genetics , Bacterial Proteins/metabolism , Drug Resistance, Bacterial , Escherichia coli/metabolism , Escherichia coli Proteins/metabolism , Gene Expression Regulation, Bacterial/drug effects , Genes, Regulator , Repressor Proteins/metabolism , Up-RegulationABSTRACT
An acid pellet, obtained as a side fraction from a conventional gonadotropin purification pathway, has been found to contain the bulk of the pituitary lactogenic hormones (growth hormone or GH and prolactin or PRL). This discarded side fraction has been utilized to obtain buffalo lactogenic hormones (buGH and buPRL), simultaneously, and in bulk. The immunoreactivities of the purified semi-pure buffalo GH and PRL (APECS and APP-I, respectively) preparations were compared by direct binding ELISA with semi-pure standard buGH and PRL (ECS and EP-I, respectively) and were found to be as pure as standard semi-pure buGH and buPRL. When checked by direct binding ELISA using buGH and buPRL antisera, it was observed that APECS and APP-I were not cross-immunoreactive. SDS-PAGE and western blot analysis of APECS and APP-I showed major bands located at the same positions as in the case of standard semi-pure preparations (20 kDa for APECS and 23 kDa for APP-I). The semi-purified buGH and buPRL (APECS and APP-I) were converted to a highly purified preparation by chromatographing them via Sephacryl S-200 gel-filtration chromatography.
