Quercetin prevents necrotic cell death induced by co-exposure to benzo(a)pyrene and UVA radiation.

The phototoxicity of low-energy ultraviolet radiation, such as UVA, can be enhanced by the presence of photosensitizing agents. Hence, co-exposure of cells to benzo[a]pyrene (BaP), a widespread environmental carcinogen and photosensitizing agent, and UVA may synergistically induce DNA damage. In this study, exposure of cells to various concentrations of BaP for 1h followed by UVA irradiation (2J/cm(2)) increased DNA damage and decreased cell viability. Expression of apoptosis-related proteins (caspase-9, caspase-3, PARP, and Bax) and hypodiploid DNA content (sub-G(1)) were not changed. LDH release into the culture medium increased in a dose-dependent manner with BaP under UVA irradiation, suggesting that cell death due to BaP/UVA co-treatment occurred via necrosis. Intracellular reactive oxygen species (ROS) levels were increased significantly in the co-exposed cells, and treatment with the polyphenol quercetin, but not with sodium azide or N-acetylcysteine, decreased ROS levels and increased cell viability in BaP/UVA-treated cells. In conclusion, UVA irradiation combined with BaP synergistically promoted necrosis of A549 cells by increasing intracellular ROS levels, and quercetin prevented BaP-enhanced phototoxicity due to UVA irradiation.

Association of the NQO1, MPO, and XRCC1 polymorphisms and chromosome damage among workers at a petroleum refinery.

Benzene is a leukemogen, and exposure to benzene is an occupational hazard in the petroleum refining industries. The effects of genetic polymorphisms in the NQO1 (rs1800566), MPO (rs2333227), and XRCC1 (rs25487) genes on benzene-induced chromosome abnormalities were assessed in 108 benzene-exposed workers and 33 office workers. The mean benzene exposure for exposed workers was 0.51 ppm for full-shift workers, and the time-weighted average ranged from 0.004 to 4.25 ppm. The frequencies of micronuclei (MN) and chromosome aberrations (CA) were significantly higher in workers exposed to benzene than unexposed controls. Exposed workers with the T/T genotype for NQO1 showed significant 1.9-fold (95% CI = 1.5-2.3) and 2.6-fold (95% CI = 1.7-3.9) increases in MN and CA frequencies, respectively, compared to controls with C/C and C/T genotypes, after adjusting for age, smoking status, and alcohol intake. Among exposed workers, subjects with the combination of MPO G/G and XRCC1 Arg/Gln or Gln/Gln showed a significantly higher CA frequency compared to those with the combination of MPO G/A or A/A and XRCC1 Arg/Arg genotypes. These results indicated that the genotoxicity induced by a chronic benzene exposure is modulated by genes involved in both DNA repair and benzene metabolic pathways.

Reoxygenation following hypoxia activates DNA-damage checkpoint signaling pathways that suppress cell-cycle progression in cultured human lymphocytes.

Cellular responses to DNA damage after hypoxia and reoxygenation (H/R) were examined in human lymphocytes. Cultured lymphocytes exposed to H/R showed a lower cytokinesis block proliferation index and a higher frequency of micronuclei in comparison to control cells. Western blots showed that H/R exposure induced p53 expression; however, p21 and Bax expression did not increase, indicating that H/R did not affect p53 transactivational activity. Phosphorylation of p53 (Ser15), Chk1 (Ser345), and Chk2 (Thr68) was also observed, suggesting that H/R activates p53 through checkpoint signals. In addition, H/R exposure caused the phosphorylation and negative regulation of Cdc2 and Cdc25C, proteins that are involved in cell-cycle arrest at the G2/M checkpoint. The S-phase checkpoint, regulated by the ATM-p95/NBS1-SMC1 pathway, was also triggered in H/R-exposed lymphocytes. These results demonstrate that H/R exposure triggers checkpoint signaling and induces cell-cycle arrest in cultured human lymphocytes.

Desferrioxamine (DFX) has genotoxic effects on cultured human lymphocytes and induces the p53-mediated damage response.

Desferrioxamine (DFX), which is an iron chelator, mimics hypoxia by enhancing HIF1-alpha accumulation and upregulating inflammatory mediators. DFX is usually beneficial, with preventive effects related primarily to its ability to scavenge reactive oxygen species. However, toxic effects on skeletal and ocular organs have been reported. The cytokinesis block micronucleus test and alkaline single-cell gel (Comet) assay were used to evaluate the genotoxic effects of DFX on human blood lymphocytes. Cultured human lymphocytes treated with 130microM DFX for various periods of time showed significant differences in the incidence of micronucleated binucleate cells, as well as in the length and moment of the comet tail. Western blot analysis using antibodies to proteins involved in the p53-mediated response to DNA damage revealed that p53 was accumulated and DNA damage checkpoint kinases were activated in lymphocytes treated with DFX. On the other hand, the p53 downstream target proteins p21 and bax were not affected, which indicates that DFX does not promote the transactivational activity of p53. Apoptosis assays demonstrated DFX-induced apoptosis of lymphocytes via the caspase cascade. The observed increase in the sub-G1 fraction and enhanced caspase-3 activity indicate that DFX can promote apoptosis in human lymphocytes, and these results were confirmed by protein immunoblot analysis. As apoptotic cell death is preceded by the collapse of the mitochondrial membrane potential, we also measured the mitochondrial membrane potential (Deltapsi(m)) using DiOC6, which is a fluorescent membrane potential probe. The fluorescence intensity of DiOC6 in lymphocytes was significantly reduced in a time-dependent manner after DFX treatment. Taken together, these results indicate that DFX activates p53-mediated checkpoint signals and induces apoptosis via mitochondrial damage in human peripheral blood lymphocytes.

Hypoxia/reoxygenation-induced cytotoxicity in cultured human lymphocytes.

Reactive oxygen species (ROS) generated after exposure to hypoxia and reoxygenation (H/R) play a pivotal role in the stimulation of cell death. In this study, we explored H/R-induced cytotoxicity in human lymphocytes. Compared to cells under normoxic conditions, H/R-treated cells exhibited significantly decreased viability and increased DNA breakage. Western blotting analysis demonstrated that H/R-induced the accumulation of p53 and p63 proteins. H/R also led to the activation of caspase-3 and -9, accompanied by the cleavage of PARP (poly(ADP-ribose)polymerase). Because apoptosis is usually accompanied by ROS generation and collapse of the mitochondrial membrane potential (MMP, Deltapsi(m)), we examined ROS and MMP levels in H/R-treated lymphocytes. Cells subjected to H/R exhibited significantly increased ROS and decreased MMP, compared with normoxic cells. Taken together, these results indicate that H/R treatment of human lymphocytes induces rapid ROS generation and MMP collapse, which triggers apoptosis.

[Does living nearby a garbage dumping site degrade the quality of life? A case study based on Shin-dong Myeon residents, Chun-cheon Si].

OBJECTIVES: This study aims to examine if a garbage dumping site has real and negative influence on the quality of life (QOL) for the nearby residents. The net effects of the residential distance from the garbage dumping site and from the garbage truck route were investigated for five domains of the QOL. METHODS: Two hundred fifty seven Shin-dong Myeon residents, Chun-cheon Si, participated in a self-administrated survey. The Shin-dong Myeon garbage dumping site began operating in 1996. ANCOVA with generalized linear models and multiple regression analysis were performed. RESULTS: Descriptive analyses show that a residence nearby a garbage dumping site is negatively associated with the physical and environmental domains of the QOL. The residential distance from the garbage truck route does not exert any significant effect on various domains of QOL, except for the environmental domain. On the multivariate analysis, the residents living near the garbage dumping site tended to have a significantly negative QOL in the physical and environmental domains. However, the distance from the garbage truck route did not show a significant nor substantial effect on the QOL. The demographic and socioeconomic control variables are associated with a number of the QOL domains, and their patterns are consistent with the general expectations. CONCLUSIONS: The results indicated that a garbage dumping site is considered to be an environmental hazard among the nearby residents according to the lower scores on the physical and environmental domains of the QOL. The findings from this study provide comprehensive\ understanding on the residents' QOL, and they may help politicians and policy makers make decisions for appropriate interventions.