PURPOSE: The oil spill from the Heibei Spirit in December 2007 contaminated the Yellow Coast of South Korea. We evaluated the respiratory effects of that spill on children who lived along the Yellow Coast. METHODS: Of 662 children living in the area exposed to the oil spill, 436 (65.9%) were enrolled as subjects. All subjects completed a modified International Study of Asthma and Allergies in Childhood questionnaire. A health examination, including a skin prick test, pulmonary function test, and methacholine bronchial provocation test (MBPT), was administered. The children were assigned to two groups: those who lived close to the oil spill area and those who lived far from the oil spill area. RESULTS: The children who lived close to the oil spill area showed a significantly lower forced expiratory volume in one second (FEV1), an increased prevalence of 'asthma ever' (based on a questionnaire), and 'airway hyperresponsiveness' (based on the MBPT) than those who lived far from the oil spill area (FEV1; P=0.011, prevalence of 'asthma ever' based on a questionnaire; P=0.005, prevalence of 'airway hyperresponsiveness' based on the MBPT; P=0.001). The onset of wheezing after the oil spill was significantly higher in children who lived close to the oil spill area than in those who lived far from the oil spill area among the 'wheeze ever' group (P=0.002). In a multiple logistic regression analysis, male sex, family history of asthma, and residence near the oil spill area were significant risk factors for asthma (sex [male/female]: odds ratio [OR], 2.54; 95% confidence interval [CI], 1.31-4.91; family history of asthma [No/Yes]: OR, 3.77; 95% CI, 1.83-7.75; exposure group [low/high]; OR, 2.43; 95% CI, 1.27-4.65). CONCLUSIONS: This study suggests that exposure to an oil spill is a risk factor for asthma in children.
Asthma ; Bronchial Provocation Tests ; Child ; Forced Expiratory Volume ; Humans ; Hypersensitivity ; Industrial Oils ; Korea ; Logistic Models ; Male ; Methacholine Chloride ; Odds Ratio ; Petroleum Pollution ; Prevalence ; Republic of Korea ; Respiratory Function Tests ; Respiratory Sounds ; Risk Factors ; Skin ; Surveys and Questionnaires

PURPOSE: The oil spill from the Heibei Spirit in December 2007 contaminated the Yellow Coast of South Korea. We evaluated the respiratory effects of that spill on children who lived along the Yellow Coast. METHODS: Of 662 children living in the area exposed to the oil spill, 436 (65.9%) were enrolled as subjects. All subjects completed a modified International Study of Asthma and Allergies in Childhood questionnaire. A health examination, including a skin prick test, pulmonary function test, and methacholine bronchial provocation test (MBPT), was administered. The children were assigned to two groups: those who lived close to the oil spill area and those who lived far from the oil spill area. RESULTS: The children who lived close to the oil spill area showed a significantly lower forced expiratory volume in one second (FEV1), an increased prevalence of 'asthma ever' (based on a questionnaire), and 'airway hyperresponsiveness' (based on the MBPT) than those who lived far from the oil spill area (FEV1; P=0.011, prevalence of 'asthma ever' based on a questionnaire; P=0.005, prevalence of 'airway hyperresponsiveness' based on the MBPT; P=0.001). The onset of wheezing after the oil spill was significantly higher in children who lived close to the oil spill area than in those who lived far from the oil spill area among the 'wheeze ever' group (P=0.002). In a multiple logistic regression analysis, male sex, family history of asthma, and residence near the oil spill area were significant risk factors for asthma (sex [male/female]: odds ratio [OR], 2.54; 95% confidence interval [CI], 1.31-4.91; family history of asthma [No/Yes]: OR, 3.77; 95% CI, 1.83-7.75; exposure group [low/high]; OR, 2.43; 95% CI, 1.27-4.65). CONCLUSIONS: This study suggests that exposure to an oil spill is a risk factor for asthma in children.
Asthma ; Bronchial Provocation Tests ; Child ; Forced Expiratory Volume ; Humans ; Hypersensitivity ; Industrial Oils ; Korea ; Logistic Models ; Male ; Methacholine Chloride ; Odds Ratio ; Petroleum Pollution ; Prevalence ; Republic of Korea ; Respiratory Function Tests ; Respiratory Sounds ; Risk Factors ; Skin ; Surveys and Questionnaires

Increased expression of a number of proinflammatory genes, including IL-8, is associated with inflammatory conditions such as asthma. Glucocorticoid receptor (GR)beta, one of the GR isoforms, has been suggested to be upregulated in asthma associated with glucocorticoid insensitivity and to work as a dominant negative inhibitor of wild type GRalpha. However, recent data suggest that GRbeta is not a dominant negative inhibitor of GRalpha in the transrepressive process and has its own functional role. We investigated the functional role of GRbeta expression in the suppressive effect of glucocorticoids on tumor necrosis factor (TNF)-alpha-induced IL-8 release in an airway epithelial cell line. GRbeta expression was induced by treatment of epithelial cells with either dexamethasone or TNF-alpha. GRbeta was able to inhibit glucocorticoid-induced transcriptional activation mediated by binding to glucocorticoid response elements (GREs). The suppressive effect of dexamethasone on TNF-alpha-induced IL-8 transcription was not affected by GRbeta overexpression, rather GRbeta had its own weak suppressive activity on TNF-alpha-induced IL-8 expression. Overall histone deacetylase activity and histone acetyltransferase activity were not changed by GRbeta overexpression, but TNF-alpha-induced histone H4 acetylation at the IL-8 promoter was decreased with GRbeta overexpression. This study suggests that GRbeta overexpression does not affect glucocorticoid-induced suppression of IL-8 expression in airway epithelial cells and GRbeta induces its own histone deacetylase activity around IL-8 promoter site.
Acetylation ; Cell Line, Tumor ; Dexamethasone/pharmacology ; Epithelial Cells/metabolism ; *Gene Expression Regulation ; Histones/*metabolism ; Humans ; Interleukin-8/*genetics/metabolism ; Receptors, Glucocorticoid/genetics/*metabolism ; Transcriptional Activation ; Transfection ; Tumor Necrosis Factor-alpha/*antagonists & inhibitors/pharmacology

Increased expression of a number of proinflammatory genes, including IL-8, is associated with inflammatory conditions such as asthma. Glucocorticoid receptor (GR)beta, one of the GR isoforms, has been suggested to be upregulated in asthma associated with glucocorticoid insensitivity and to work as a dominant negative inhibitor of wild type GRalpha. However, recent data suggest that GRbeta is not a dominant negative inhibitor of GRalpha in the transrepressive process and has its own functional role. We investigated the functional role of GRbeta expression in the suppressive effect of glucocorticoids on tumor necrosis factor (TNF)-alpha-induced IL-8 release in an airway epithelial cell line. GRbeta expression was induced by treatment of epithelial cells with either dexamethasone or TNF-alpha. GRbeta was able to inhibit glucocorticoid-induced transcriptional activation mediated by binding to glucocorticoid response elements (GREs). The suppressive effect of dexamethasone on TNF-alpha-induced IL-8 transcription was not affected by GRbeta overexpression, rather GRbeta had its own weak suppressive activity on TNF-alpha-induced IL-8 expression. Overall histone deacetylase activity and histone acetyltransferase activity were not changed by GRbeta overexpression, but TNF-alpha-induced histone H4 acetylation at the IL-8 promoter was decreased with GRbeta overexpression. This study suggests that GRbeta overexpression does not affect glucocorticoid-induced suppression of IL-8 expression in airway epithelial cells and GRbeta induces its own histone deacetylase activity around IL-8 promoter site.
Acetylation ; Cell Line, Tumor ; Dexamethasone/pharmacology ; Epithelial Cells/metabolism ; *Gene Expression Regulation ; Histones/*metabolism ; Humans ; Interleukin-8/*genetics/metabolism ; Receptors, Glucocorticoid/genetics/*metabolism ; Transcriptional Activation ; Transfection ; Tumor Necrosis Factor-alpha/*antagonists & inhibitors/pharmacology

Angiotensin II (Ang II) stimulates migration of vascular smooth muscle cell (VSMC) in addition to its contribution to contraction and hypertrophy. It is well established that Rho GTPases regulate cellular contractility and migration by reorganizing the actin cytoskeleton. Ang II activates Rac1 GTPase, but its upstream guanine nucleotide exchange factor (GEF) remains elusive. Here, we show that Ang II-induced VSMC migration occurs in a betaPIX GEF-dependent manner. betaPIX-specific siRNA treatment significantly inhibited Ang II-induced VSMC migration. Ang II activated the catalytic activity of betaPIX towards Rac1 in dose- and time-dependent manners. Activity reached a peak at 10 min and declined close to a basal level by 30 min following stimulation. Pharmacological inhibition with specific kinase inhibitors revealed the participation of protein kinase C, Src family kinase, and phosphatidylinositol 3-kinase (PI3-K) upstream of betaPIX. Both p21-activated kinase and reactive oxygen species played key roles in cytoskeletal reorganization downstream of betaPIX-Rac1. Taken together, our results suggest that betaPIX is involved in Ang II-induced VSMC migration.
1-Phosphatidylinositol 3-Kinase/metabolism ; Angiotensin II/*metabolism ; Animals ; *Cell Movement ; Cells, Cultured ; Guanine Nucleotide Exchange Factors/genetics/*metabolism ; Muscle, Smooth, Vascular/cytology ; Myocytes, Smooth Muscle/*cytology ; NADPH Oxidase/metabolism ; Protein Kinase C/metabolism ; RNA, Small Interfering/genetics ; Rats ; Rats, Sprague-Dawley ; p21-Activated Kinases/metabolism ; rac1 GTP-Binding Protein/metabolism ; src-Family Kinases/metabolism

Cyclooxygenase-2 (COX-2) has been reported to be associated with tumor development and progression as well as to protect cells from apoptosis induced by various cellular stresses. Through a tetracycline-regulated COX-2 overexpression system, we found that COX-2 inhibits detachment-induced apoptosis (anoikis) in a human bladder cancer cell line, EJ. We also found that the inhibition of anoikis by COX-2 results from activation of the PI-3K/Akt pathway as evidenced by suppression of the COX-2 effect on anoikis by a PI-3K inhibitor, LY294002. Furthermore, COX-2 enhanced Mcl-1 expression in the anoikis process, implying that Mcl-1 also may be involved in mediating the survival function of COX-2. Together, these results suggest that COX-2 inhibits anoikis by activation of the PI-3K/Akt pathway and probably by enhancement of Mcl-1 expression in human bladder cancer cells. This anti- anoikis effect of COX-2 may be a part of mechanisms to promote tumor development and progression.
1-Phosphatidylinositol 3-Kinase/*metabolism ; Anoikis/*physiology ; Bladder Neoplasms/*metabolism/pathology ; Enzyme Activation ; Humans ; Neoplasm Proteins/*metabolism ; Prostaglandin-Endoperoxide Synthase/*metabolism ; Protein-Serine-Threonine Kinases/*metabolism ; Proto-Oncogene Proteins/*metabolism ; Proto-Oncogene Proteins c-bcl-2/*metabolism ; Research Support, Non-U.S. Gov't ; Signal Transduction ; Transfection ; Tumor Cells, Cultured

TNF-alpha, a trimeric cytokine, was known to inhibit differention of preadipocytes to adipocytes. In the present study, we investigated signal mediators working downstream of TNF-alpha using murine 3T3-L1 cells. TNF-alpha induced activation of both c-jun NH2-terminal kinase (JNK) and nuclear transcription factor-kappaB (NF-kappaB) in 3T3-L1 cells. Blockage of these two mediators activities by specific inhibitors, SP600125 and Ad-IkappaBalpha-SR restored adipogenesis differentiation suggesting their involvement in the inhibited differentiation of 3T3-L1 cells by TNF-alpha. Consistent with previous studies, peroxisome proliferator-activated receptor gamma (PPARgamma) a key transcriptional regulator was remarkably reduced by TNF-alpha treatment. Compared with adipogenesis, however, SP600125, a chemical JNK inhibitor hardly relieved TNF-alpha effect on PPARgamma expression whereas S32A/S36A mutant of IkappaBalpha considerably recovered PPARgamma expression, indicating that two signal mediators exploit separable main routes to achieve reduced adipogenesis. These results suggest that inhibition of 3T3-L1 cells differentiation by TNF-alpha is partly implemented through NF-kappaB and one of its downstream effectors be PPARgamma.
3T3-L1 Cells ; Adipocytes/cytology/drug effects ; Animals ; Cell Differentiation/*drug effects ; Gene Expression Regulation/*drug effects ; Mice ; NF-kappa B/*metabolism ; Promoter Regions (Genetics)/genetics ; RNA, Messenger/genetics/metabolism ; Receptors, Cytoplasmic and Nuclear/*genetics ; Support, Non-U.S. Gov't ; Transcription Factors/*genetics ; Tumor Necrosis Factor/*pharmacology

BACKGROUND: NF-κB is the most important transcriptional factor in Il-8 gene expression. Triptolide is a new compound that recently has been shown to inhibit NF-κB activation. The purpose of this study is to investigate how triptolide inhibits NF-κB-dependent IL-8 gene transcription in lung epithelial cells and to pilot the potential for the clinical application of triptolide in inflammatory lung diseases. METHODS: A549 cells were used and triptolide was provided from Pharmagenesis Company (Palo Alto, CA). In order to examine NF-κB-dependent IL-8 transcriptional activity, we established stable A549 IL-8-NF-κB-luc. cells and performed luciferase assays. IL-8 gene expression was measured by RT-PCR and ELISA. A Western blot was done for the study of IκBα degradation and as electromobility shift assay was done to analyze NF-κB DNA binding. p65 specific transactivation was analyzed by a cotransfection study using a Gal4-p65 fusion protein expression system. To investigate the involvement of transcriptional coactivators, we perfomed a transfection study with CBP and SRC-1 expression vectors. RESULTS: We observed that triptolide significantly suppresses NF-κB-dependent IL-8 transcriptional activity induced by IL-1β and PMA. RT-PCR showed that triptolide represses both IL-1β- and pMA-induced IL-8 mRNA expression and ELISA confirmed this triptolide-mediated IL-8 suppression at the protein level. However, triptolide did not affect IκBα degradation and NF-κB DNA binding. In a p65-specific transactivation study, triptolide significantly suppressed Gal4-p65TA1 and Gal4-p65TA2 activity suggesting that triptolide inhibits NF-κB activation by inhibiting p65 transactivation. However, this triptolide-mediated inhibition of p65 transactivation was not rescued by the overexpression of CBP or SRC-1, thereby excluding the role of transcriptional coactivators. CONCLUSIONS: Triptolide is a new compound that inhibits NF-κB-dependent IL-8 transcriptional activation by inhibiting p65 transactivation, but not by an IκBα-dependent mechanism. This suggests that triptolide may have a therapeutic potential for inflammatory lung diseases.
Blotting, Western ; DNA ; Enzyme-Linked Immunosorbent Assay ; Epithelial Cells* ; Gene Expression ; Interleukin-8* ; Luciferases ; Lung Diseases ; Lung* ; NF-kappa B* ; RNA, Messenger ; Transcriptional Activation ; Transfection

Glucocorticoid receptor (GR) functions as a suppressor of inflammation by inhibiting the expression of many cytokine gene activated by NF-κB. The goal of this study is to investigate the mechanism by which GR repress NF-κB activation in lung epithelial cells. We used A549 and BEAS-2B lung epithelial cell lines. Using IgGκ-NF-κB luciferase reporter gene construct, we found that dexamethasone significantly suppressed TNF-α-induced NF-κB activation and the overexpression of GR showed dose-dependent reduction of TNF-α-induced NF-κB activity in both cell lines. However, DNA binding of NF-κB induced by TNF-α in electromobility shift assay was not inhibited by dexamethasone. Super shift assay with anti-p65 antibody demonstrated the existence of p65 in NF-κB complex induced by TNF-α Western blot showed that IκBα degradation induced by TNF-α was not affected by dexamethasone and IκBκ was not induced by dexamethasone, neither. To evaluate p65 specific transactivation, we adopted co-transfection study of Ga14-p65TA1 or TA2 fusion protein expression system together with 5xGa14-luciferase vector. Co-transfection of GR with Ga14-p65TA1 or TA2 repressed luciferase activity profoundly to the level of 10-20% of p65TA1- or TA2-induced transcriptional activity. And this transrepressional effect was abolished by co-transfection of CBP or SRC-1 expression vectors. These results suggest that Gr-mediated transrepression of NF-κB in lung epithelial cells is through competing for binding to limiting amount of transcriptional coactivators, CBP or SRC-1.
Blotting, Western ; Cell Line ; Dexamethasone* ; DNA ; Epithelial Cells* ; Genes, Reporter ; Inflammation ; Luciferases ; Lung ; NF-kappa B* ; Receptors, Glucocorticoid ; Transcriptional Activation*

No abstract available.
Cell Cycle*