Metal enriched quasi-ultrafine particles from stainless steel gas metal arc welding induced genetic and epigenetic alterations in BEAS-2B cells.

Recent evidence has supported welding fume (WF)-derived ultrafine particles (UFP) could be the driving force of their adverse health effects. However, UFP have not yet been extensively studied and are currently not included in present air quality standards/guidelines. Here, attention was focused on the underlying genetic and epigenetic mechanisms by which the quasi-UFP (Q-UFP, i.e., ≤ 0.25 µm) of the WF emitted by gas metal arc welding-stainless steel (GMAW-SS) exert their toxicity in human bronchial epithelial BEAS-2B cells. The Q-UFP under study showed a monomodal size distribution in number centered on 104.4 ± 52.3 nm and a zeta potential of -13.8 ± 0.3 mV. They were enriched in Fe > Cr > Mn > Si, and displayed a relatively high intrinsic oxidative potential. Dose-dependent activation of nuclear factor erythroid 2-related factor 2 and nuclear factor-kappa B signaling pathway, glutathione alteration, and DNA, protein and lipid oxidative damage were reported in BEAS-2B cells acutely (1.5 and 9 µg/cm2, 24 h) or repeatedly (0.25 and 1.5 µg/cm2, 3 × 24 h) exposed to Q-UFP (p < 0.05). Alterations of the Histone H3 acetylation were reported for any exposure (p < 0.05). Differentially regulated miRNA and mRNA indicated the activation of some critical cell signaling pathways related to oxidative stress, inflammation, and cell cycle deregulation towards apoptosis. Taken together, these results highlighted the urgent need to better evaluate the respective toxicity of the different metals and to include the Q-UFP fraction of WF in current air quality standards/guidelines relevant to the occupational settings.

Mitochondrial alterations triggered by repeated exposure to fine (PM2.5-0.18) and quasi-ultrafine (PM0.18) fractions of ambient particulate matter.

Nowadays ambient particulate matter (PM) levels still regularly exceed the guideline values established by World Health Organization in most urban areas. Numerous experimental studies have already demonstrated the airway toxicity of the fine fraction of PM (FP), mainly triggered by oxidative stress-induced airway inflammation. However, only few studies have actually paid close attention to the ultrafine fraction of PM (UFP), which is likely to be more easily internalized in cells and more biologically reactive. Mitochondria are major endogenous sources of reactive oxygen species (ROS) through oxidative metabolism, and coordinate many critical cellular signaling processes. Mitochondria have been often studied in the context of PM toxicity and generally associated with apoptosis activation. However, little is known about the underlying adaptation mechanisms that could occur following exposure at sub-apoptotic doses of ambient PM. Here, normal human bronchial epithelial BEAS-2B cells were acutely or repeatedly exposed to relatively low doses (5 µg.cm-2) of FP (PM2.5-0.18) or quasi-UFP (Q-UFP; PM0.18) to better access the critical changes in mitochondrial morphology, functions, and dynamics. No significant cytotoxicity nor increase of apoptotic events were reported for any exposure. Mitochondrial membrane potential (ΔΨm) and intracellular ATP content were also not significantly impaired. After cell exposure to sub-apoptotic doses of FP and notably Q-UFP, oxidative phosphorylation was increased as well as mitochondrial mass, resulting in increased production of mitochondrial superoxide anion. Given this oxidative boost, the NRF2-ARE signaling pathway was significantly activated. However, mitochondrial dynamic alterations in favor of accentuated fission process were observed, in particular after Q-UFP vs FP, and repeated vs acute exposure. Taken together, these results supported mitochondrial quality control and metabolism dysfunction as an early lung underlying mechanism of toxicity, thereby leading to accumulation of defective mitochondria and enhanced endogenous ROS generation. Therefore, these features might play a key role in maintaining PM-induced oxidative stress and inflammation within lung cells, which could dramatically contribute to the exacerbation of inflammatory chronic lung diseases. The prospective findings of this work could also offer new insights into the physiopathology of lung toxicity, arguably initiate and/or exacerbate by acutely and rather repeated exposure to ambient FP and mostly Q-UFP.

Study of in vitro and in vivo genotoxic effects of air pollution fine (PM2.5-0.18) and quasi-ultrafine (PM0.18) particles on lung models.

Air pollution and particulate matter (PM) are classified as carcinogenic to humans. Pollutants evidence for public health concern include coarse (PM10) and fine (PM2.5) particles. However, ultrafine particles (PM0.1) are assumed to be more toxic than larger particles, but data are still needed to better understand their mechanism of action. In this context, the aim of our work was to investigate the in vitro and in vivo genotoxic potential of fine (PM2.5-018) and quasi ultra-fine (PM0.18) particles from an urban-industrial area (Dunkirk, France) by using comet, micronucleus and/or gene mutation assays. In vitro assessment was performed with 2 lung immortalized cell lines (BEAS-2B and NCI-H292) and primary normal human bronchial epithelial cells (NHBE) grown at the air-liquid interface or in submerged conditions (5 µg PM/cm2). For in vivo assessment, tests were performed after acute (24 h, 100 µg PM/animal), subacute (1 month, 10 µg PM/animal) and subchronic (3 months, 10 µg PM/animal) intranasal exposure of BALB/c mice. In vitro, our results show that PM2.5-018 and PM0.18 induced primary DNA damage but no chromosomal aberrations in immortalized cells. Negative results were noted in primary cells for both endpoints. In vivo assays revealed that PM2.5-018 and PM0.18 induced no significant increases in DNA primary damage, chromosomal aberrations or gene mutations, whatever the duration of exposure. This investigation provides initial answers regarding the in vitro and in vivo genotoxic mode of action of PM2.5-018 and PM0.18 at moderate doses and highlights the need to develop standardized specific methodologies for assessing the genotoxicity of PM. Moreover, other mechanisms possibly implicated in pulmonary carcinogenesis, e.g. epigenetics, should be investigated.

Toxicological effects of ambient fine (PM2.5-0.18) and ultrafine (PM0.18) particles in healthy and diseased 3D organo-typic mucocilary-phenotype models.

The knowledge of the underlying mechanisms by which particulate matter (PM) exerts its health effects is still incomplete since it may trigger various symptoms as some persons may be more susceptible than others. Detailed studies realized in more relevant in vitro models are highly needed. Healthy normal human bronchial epithelial (NHBE), asthma-diseased human bronchial epithelial (DHBE), and COPD-DHBE cells, differentiated at the air-liquid interface, were acutely or repeatedly exposed to fine (i.e., PM2.5-0.18, also called FP) and quasi-ultrafine (i.e., PM0.18, also called UFP) particles. Immunofluorescence labelling of pan-cytokeratin, MUC5AC, and ZO-1 confirmed their specific cell-types. Baselines of the inflammatory mediators secreted by all the cells were quite similar. Slight changes of TNFα, IL-1ß, IL-6, IL-8, GM-CSF, MCP-1, and/or TGFα, and of H3K9 histone acetylation supported a higher inflammatory response of asthma- and especially COPD-DHBE cells, after exposure to FP and especially UFP. At baseline, 35 differentially expressed genes (DEG) in asthma-DHBE, and 23 DEG in COPD-DHBE, compared to NHBE cells, were reported. They were involved in biological processes implicated in the development of asthma and COPD diseases, such as cellular process (e.g., PLA2G4C, NLRP1, S100A5, MUC1), biological regulation (e.g., CCNE1), developmental process (e.g., WNT10B), and cell component organization and synthesis (e.g., KRT34, COL6A1, COL6A2). In all the FP or UFP-exposed cell models, DEG were also functionally annotated to the chemical metabolic process (e.g., CYP1A1, CYP1B1, CYP1A2) and inflammatory response (e.g., EREG). Another DEG, FGF-1, was only down-regulated in asthma and specially COPD-DHBE cells repeatedly exposed. While RAB37 could help to counteract the down-regulation of FGF-1 in asthma-DHBE cells, the deregulation of FGR, WNT7B, VIPR1, and PPARGC1A could dramatically contribute to make it worse in COPD-DHBE cells. Taken together, these data contributed to support the highest effects of UFP versus FP and highest sensitivity of asthma- and notably COPD-DHBE versus NHBE cells.

Individual exposure level following indoor and outdoor air pollution exposure in Dakar (Senegal).

The consequences of indoor and outdoor air pollution on human health are of great concern nowadays. In this study, we firstly evaluated indoor and outdoor air pollution levels (CO, CO2, NO, NO2, PM10) at an urban site in Dakar city center and at a rural site. Then, the individual exposure levels to selected pollutants and the variations in the levels of biomarkers of exposure were investigated in different groups of persons (bus drivers, traders working along the main roads and housemaids). Benzene exposure levels were higher for housemaids than for bus drivers and traders. High indoor exposure to benzene is probably due to cooking habits (cooking with charcoal), local practices (burning of incense), the use of cleaning products or solvent products which are important emitters of this compound. These results are confirmed by the values of S-PMA, which were higher in housemaids group compared to the others. Urinary 1-HOP levels were significantly higher for urban site housemaids compared to semirural district ones. Moreover, urinary levels of DNA oxidative stress damage (8-OHdG) and inflammatory (interleukin-6 and -8) biomarkers were higher in urban subjects in comparison to rural ones. The air quality measurement campaign showed that the bus interior was more polluted with PM10, CO, CO2 and NO than the market and urban or rural households. However, the interior of households showed higher concentration of VOCs than outdoor sites confirming previous observations of higher indoor individual exposure level to specific classes of pollutants.

Air pollution-derived PM2.5 impairs mitochondrial function in healthy and chronic obstructive pulmonary diseased human bronchial epithelial cells.

In order to clarify whether the mitochondrial dysfunction is closely related to the cell homeostasis maintenance after particulate matter (PM2.5) exposure, oxidative, inflammatory, apoptotic and mitochondrial endpoints were carefully studied in human bronchial epithelial BEAS-2B, normal human bronchial epithelial (NHBE) and chronic obstructive pulmonary disease (COPD)-diseased human bronchial epithelial (DHBE) cells acutely or repeatedly exposed to air pollution-derived PM2.5. Some modifications of the mitochondrial morphology were observed within all these cell models repeatedly exposed to the highest dose of PM2.5. Dose- and exposure-dependent oxidative damages were reported in BEAS-2B, NHBE and particularly COPD-DHBE cells acutely or repeatedly exposed to PM2.5. Nuclear factor erythroid 2-p45 related factor 2 (NRF2) gene expression and binding activity, together with the mRNA levels of some NRF2 target genes, were directly related to the number of exposures for the lowest PM2.5 dose (i.e., 2 µg/cm2), but, surprisingly, inversely related to the number of exposures for the highest dose (i.e., 10 µg/cm2). There were dose- and exposure-dependent increases of both nuclear factor kappa-B (NF-κB) binding activity and NF-κB target cytokine secretion in BEAS-2B, NHBE and particularly COPD-DHBE cells exposed to PM2.5. Mitochondrial ROS production, membrane potential depolarization, oxidative phosphorylation, and ATP production were significantly altered in all the cell models repeatedly exposed to the highest dose of PM2.5. Collectively, our results indicate a cytosolic ROS overproduction, inducing oxidative damage and activating oxygen sensitive NRF2 and NF-kB signaling pathways for all the cell models acutely or repeatedly exposed to PM2.5. However, one of the important highlight of our findings is that the prolonged and repeated exposure in BEAS-2B, NHBE and in particular sensible COPD-DHBE cells further caused an oxidative boost able to partially inactivate the NRF2 signaling pathway and to critically impair mitochondrial redox homeostasis, thereby producing a persistent mitochondrial dysfunction and a lowering cell energy supply.

Genetic and epigenetic alterations in normal and sensitive COPD-diseased human bronchial epithelial cells repeatedly exposed to air pollution-derived PM2.5.

Even though clinical, epidemiological and toxicological studies have progressively provided a better knowledge of the underlying mechanisms by which air pollution-derived particulate matter (PM) exerts its harmful health effects, further in vitro studies on relevant cell systems are still needed. Hence, aiming of getting closer to the human in vivo conditions, primary human bronchial epithelial cells derived from normal subjects (NHBE) or sensitive chronic obstructive pulmonary disease (COPD)-diseased patients (DHBE) were differentiated at the air-liquid interface. Thereafter, they were repeatedly exposed to air pollution-derived PM2.5 to study the occurrence of some relevant genetic and/or epigenetic endpoints. Concentration-, exposure- and season-dependent increases of OH-B[a]P metabolites in NHBE, and to a lesser extent, COPD-DHBE cells were reported; however, there were more tetra-OH-B[a]P and 8-OHdG DNA adducts in COPD-DHBE cells. No increase in primary DNA strand break nor chromosomal aberration was observed in repeatedly exposed cells. Telomere length and telomerase activity were modified in a concentration- and exposure-dependent manner in NHBE and particularly COPD-DHBE cells. There were a global DNA hypomethylation, a P16 gene promoter hypermethylation, and a decreasing DNA methyltransferase activity in NHBE and notably COPD-DHBE cells repeatedly exposed. Changes in site-specific methylation, acetylation, and phosphorylation of histone H3 (i.e., H3K4me3, H3K9ac, H3K27ac, and H3S10ph) and related enzyme activities occurred in a concentration- and exposure-dependent manner in all the repeatedly exposed cells. Collectively, these results highlighted the key role played by genetic and even epigenetic events in NHBE and particularly sensitive COPD-DHBE cells repeatedly exposed to air pollution-derived PM2.5 and their different responsiveness. While these specific epigenetic changes have been already described in COPD and even lung cancer phenotypes, our findings supported that, together with genetic events, these epigenetic events could dramatically contribute to the shift from healthy to diseased phenotypes following repeated exposure to relatively low doses of air pollution-derived PM2.5.

Continuous cerebroventricular administration of dopamine: A new treatment for severe dyskinesia in Parkinson's disease?

In Parkinson's disease (PD) depletion of dopamine in the nigro-striatal pathway is a main pathological hallmark that requires continuous and focal restoration. Current predominant treatment with intermittent oral administration of its precursor, Levodopa (l-dopa), remains the gold standard but pharmacological drawbacks trigger motor fluctuations and dyskinesia. Continuous intracerebroventricular (i.c.v.) administration of dopamine previously failed as a therapy because of an inability to resolve the accelerated dopamine oxidation and tachyphylaxia. We aim to overcome prior challenges by demonstrating treatment feasibility and efficacy of continuous i.c.v. of dopamine close to the striatum. Dopamine prepared either anaerobically (A-dopamine) or aerobically (O-dopamine) in the presence or absence of a conservator (sodium metabisulfite, SMBS) was assessed upon acute MPTP and chronic 6-OHDA lesioning and compared to peripheral l-dopa treatment. A-dopamine restored motor function and induced a dose dependent increase of nigro-striatal tyrosine hydroxylase positive neurons in mice after 7days of MPTP insult that was not evident with either O-dopamine or l-dopa. In the 6-OHDA rat model, continuous circadian i.c.v. injection of A-dopamine over 30days also improved motor activity without occurrence of tachyphylaxia. This safety profile was highly favorable as A-dopamine did not induce dyskinesia or behavioral sensitization as observed with peripheral l-dopa treatment. Indicative of a new therapeutic strategy for patients suffering from l-dopa related complications with dyskinesia, continuous i.c.v. of A-dopamine has greater efficacy in mediating motor impairment over a large therapeutic index without inducing dyskinesia and tachyphylaxia.

Differential responses of healthy and chronic obstructive pulmonary diseased human bronchial epithelial cells repeatedly exposed to air pollution-derived PM4.

While the knowledge of the underlying mechanisms by which air pollution-derived particulate matter (PM) exerts its harmful health effects is still incomplete, detailed in vitro studies are highly needed. With the aim of getting closer to the human in vivo conditions and better integrating a number of factors related to pre-existing chronic pulmonary inflammatory, we sought to develop primary cultures of normal human bronchial epithelial (NHBE) cells and chronic obstructive pulmonary disease (COPD)-diseased human bronchial epithelial (DHBE) cells, grown at the air-liquid interface. Pan-cytokeratin and MUC5AC immunostaining confirmed the specific cell-types of both these healthy and diseased cell models and showed they are closed to human bronchial epithelia. Thereafter, healthy and diseased cells were repeatedly exposed to air pollution-derived PM4 at the non-cytotoxic concentration of 5 µg/cm2. The differences between the oxidative and inflammatory states in non-exposed NHBE and COPD-DHBE cells indicated that diseased cells conserved their specific physiopathological characteristics. Increases in both oxidative damage and cytokine secretion were reported in repeatedly exposed NHBE cells and particularly in COPD-DHBE cells. Diseased cells repeatedly exposed had lower capacities to metabolize the organic chemicals-coated onto the air-pollution-derived PM4, such as benzo[a]pyrene (B[a]P), but showed higher sensibility to the formation of OH-B[a]P DNA adducts, because their diseased state possibly affected their defenses. Differential profiles of epigenetic hallmarks (i.e., global DNA hypomethylation, P16 promoter hypermethylation, telomere length shortening, telomerase activation, and histone H3 modifications) occurred in repeatedly exposed NHBE and particularly in COPD-DHBE cells. Taken together, these results closely supported the highest responsiveness of COPD-DHBE cells to a repeated exposure to air pollution-derived PM4. The use of these innovative in vitro exposure systems such as NHBE and COPD-DHBE cells could therefore be consider as a very useful and powerful promising tool in the field of the respiratory toxicology, taking into account sensitive individuals.

Trace elements in e-liquids - Development and validation of an ICP-MS method for the analysis of electronic cigarette refills.

Electronic cigarette use has rapidly increased in recent years. In assessing their safety, and in view of coming regulations, trace elements (TE) are among the potentially toxic compounds required to be evaluated in electronic cigarette refill fluids ("e-liquids"). An analytical method using inductively coupled plasma with mass spectrometric detection (ICP-MS) was developed and rigorously validated in order to determine concentrations of 15 TE in 54 e-liquids from a French brand. Despite a significant matrix effect from the main e-liquid constituents, and difficulties related to the current lack of reference materials, our method demonstrated satisfactory linearity, precision and robustness, and permitted the quantification of low concentrations of these 15 elements: lower limits of quantification (LLQ) obtained were ≤4 ppb for all elements except for Ni, Cu and Zn (16 ppb, 20 ppb and 200 ppb, respectively). All TE concentrations in all tested samples were <510 ppb, mostly near or below the LLQs. This method is transposable and is timely for laboratories seeking to meet a prospective demand in light of current or future regulations.

Effects of engineered iron nanoparticles on the bryophyte, Physcomitrella patens (Hedw.) Bruch & Schimp, after foliar exposure.

The effects of iron nanoparticles on bryophytes (Physcomitrella patens) were studied following foliar exposure. We used iron nanoparticles (Fe-NP) representative of industrial emissions from the metallurgical industries. After a characterization of iron nanoparticles and the validation of nanoparticle internalization in cells, the effects (cytotoxicity, oxidative stress, lipid peroxidation of membrane) of iron nanoparticles were determined through the axenic culturing of Physcomitrella patens exposed at five different concentrations (5 ng, 50 ng, 500 ng, 5 µg and 50 µg per plant). Following exposure, the plant health, measured as ATP concentrations, was not impacted. Moreover, we studied oxidative stress in three ways: through the measure of reactive oxygen species (ROS) production, through malondialdehyde (MDA) production and also through glutathione regulation. At concentrations tested over a short period, the level of ROS, MDA and glutathione were not significantly disturbed.

Low-level environmental exposure to lead and renal adverse effects: a cross-sectional study in the population of children bordering the Mbeubeuss landfill near Dakar, Senegal.

This study deals with the health effects within a child population, neighbouring a landfill. After detecting metals in soil and air samples collected in the surroundings of the landfill and in a control site, we have studied: (i) levels of lead (Pb) and exposure biomarkers in blood and urine, (ii) oxidative stress biomarkers and (iii) renal injury by applying a set of early effect biomarkers. Levels of Pb were higher in the exposed site (i.e. 1129 mg/kg and 640 ng/m(3) in soil and air samples, respectively) versus those in the control site (i.e. 14.3 mg/kg and 9.3 ng/m(3) in soil and air samples, respectively). Pb impregnation and levels of delta-aminolevulinic acid in urine were influenced by the living site that shows the prevailingly alarming situation in the Mbeubeuss landfill. Malondialdehyde changes indicated Pb-induced excessive production of reactive oxygen species. Lactate dehydrogenase activities and proteinuria were found to be higher in the children living in the exposed site. These evidences may reveal the usefulness of these two effect biomarkers to monitor the kidney injury entailed by relatively low-environmental exposure to Pb. Overall, these results show that the Mbeubeuss landfill constitutes a real source of environmental and health risk, be it living or working on site, of the surrounding population, predominantly for children.

Mutagenicity and genotoxicity of PM2.5 issued from an urbano-industrialized area of Dunkerque (France).

Epidemiological studies have demonstrated the link between chronic exposure to particulate matter (PM), especially particles with an aerodynamic diameter lesser than 2.5 µm (PM(2.5) ), and lung cancer. Mechanistic investigations focus on the contribution of the various genotoxicants adsorbed onto the particles, and more particularly on polycyclic aromatic hydrocarbons or nitroaromatics. Most of the previous studies dealing with genotoxic and/or mutagenic measurements were performed on organic extracts obtained from PM(2.5) collected in polluted areas. In contrast, we have evaluated genotoxic and mutagenic properties of urbano-industrial PM(2.5) (PM) collected in Dunkerque (France). Thermally desorbed PM(2.5) (dPM) was also comparatively studied. Suspensions of PM and dPM (5-50 µg per plate) were tested in Salmonella tester strains TA98, TA102 and YG1041 ± S9mix. Significant mutagenicity was observed for PM in YG1041 ± S9 mix. In strain TA102 - S9mix, a slight, but not significant dose-response increase was observed, for both PM and dPM. Genotoxic properties of PM and dPM were evaluated by the measurement of (1) 8-OHdG in A549 cells and (2) bulky DNA adducts on A549 cells and on human alveolar macrophages (AMs) in primary culture. A dose-dependant formation of 8-OHdG adducts was observed on A549 cells for PM and dPM, probably mainly attributed to the core of the particles. Bulky DNA adducts were observed only in AMs after exposure to PM and dPM. In conclusion, using relevant exposure models, suspension of PM(2.5) induces a combination of DNA-interaction mechanisms, which could contribute to the induction of lung cancer in exposed populations.

Behavior of Trifolium repens and Lolium perenne growing in a heavy metal contaminated field: Plant metal concentration and phytotoxicity.

The use of a vegetation cover for the management of heavy metal contaminated soils needs prior investigations on the plant species the best sustainable. In this work, behaviors of Trifolium repens and Lolium perenne, growing in a metal-polluted field located near a closed lead smelter, were investigated through Cd, Pb and Zn-plant metal concentrations and their phytotoxicity. In these plant species, metals were preferentially accumulated in roots than in shoots, as follow: Cd>Zn>Pb. Plant exposure to such metals induced oxidative stress in the considered organs as revealed by the variations in malondialdehyde levels and superoxide dismutase activities. These oxidative changes were closely related to metal levels, plant species and organs. Accordingly, L. perenne seemed to be more affected by metal-induced oxidative stress than T. repens. Taken together, these findings allow us to conclude that both the plant species could be suitable for the phytomanagement of metal-polluted soils.

Environmental lead exposure and its relationship to traffic density among Senegalese children: a cross-sectional study.

Leaded-gasoline is probably the primary source of lead (Pb) exposure in Dakar (Senegal). The present cross-sectional study was undertaken to investigate the levels of Pb in Senegalese children and to present helpful data on the relationship between Pb levels and changes in biological markers of heme biosynthesis and oxidative stress. A total of 330 children, living since birth either in rural or urban areas (ie, Khombole (n = 162) and Dakar (n = 168), respectively) were included. During this cross-sectional study, the mean blood (B)-Pb level in all children was 7.32 +/- 5.33 microg/dL, and was influenced by the area of residence and gender. In rural children, 27 subjects (16.7%), 18 boys (19.6%) and nine girls (12.9%), had a B-Pb level > 10 microg Pb/dL, whereas 99 urban children (58.9%), respectively, 66 boys (71.8%) and 33 girls (43.4%), had alarmingly high B-Pb levels. Accordingly, urine delta-aminolevulinic acid levels were higher in children living in the urban area than in the rural areas (P < 0.001), and closely correlated with the B-Pb levels (P < 0.01). Moreover, glutathione peroxidase (GPx) activity, selenium (Se) level, glutathione reductase (GR) activity, and glutathione status were significantly influenced by area of residence and/or by gender. GPx activity and Se level were not only negatively correlated with B-Pb levels, but also positively correlated together (P < 0.01). Taken together, the present results allow us to conclude that urban children have higher B-Pb levels than rural children, and that of these children, boys have higher B-Pb levels than girls, leading thereby to alterations of heme biosynthesis and pro-oxidant/antioxidant balance. We also suggest that exposure to Pb and the Pb-induced adverse effects merits attention and that the development of preventive actions are of increasing importance in Senegal.

Environmental lead exposure and its relationship to traffic density among Senegalese children: a pilot study.

In Senegal, as in many developing countries, traffic density is increasing in urban areas; in Dakar more than 50% of vehicles use gasoline. Yet the extent and real magnitude of the problem has neither been recognized nor assessed in these countries. Systemic data assessment of lead pollution and people's exposure are not well known in Senegal. This study was also designed to determine the impregnation levels of the lead released by the exhaust of cars and the changes of some early biological markers in Senegalese children. Blood lead (BPb) levels showed that all the children enrolled were exposed. However, lead exposure levels (from 34.7 to 145.8 microg/L) were less important for children living in rural areas (60.9+/-18.3 microg/L) than for those living in urban areas (106.7+/-16.9 microg/L). These changes could be correlated to the difference in the automobile traffic between both these regions (P < 0.001). BPb mean levels found in boys were higher than those in girls (P < 0.05). Despite elevated BPb levels, all values for blood zinc protoporphyrin and urine delta-aminolevulinic acid were within physiological ranges. In addition, variations in some biological markers of oxidative stress and renal disorders were seen; however, they must be confirmed by a future epidemiological study.

Involvement of oxidative sress in the toxicity of 4-monochlorophenol in Hep G2 cells in culture.

Chlorophenols, mainly used as biocides, are compounds with a wide spectrum of toxic effects, including teratogenic and carcinogenic actions. The aim of this study was to examine possible 4-monochlorophenol (4-MCP) toxicity related to metabolic pathways, which may implicate semiquinones and reactive oxygen species (ROS), in human Hep G2 cells. The effects of 4-MCP were performed through cytotoxicity assays (viability, ATP level), metabolic activities (4-MCP intracellular concentration, NADPH cytochrome P-450 reductase (Cyt P-450 red.) and glutathione-S-transferase activities, CYP 3A7 mRNA expression) and oxidative stress (superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase activities, glutathione status, malondialdehyde concentration, CYP 2E1 mRNA expression). According to the literature, in this work Hep G2 cells were incubated in the continuous presence of 4-MCP at 350 microM over 24 or 48 h. Results showed statistically significant decreases in ATP levels (24 or 48 h, P < 0.05) versus controls. The 4-MCP intracellular concentrations increased as early as 8-24 h and then decreased (P < 0.01). Decreases in Cyt. P-450 red. (24 h, P < 0.05), catalase (24 h, P < 0.05; 48 h, P < 0.01), glutathione peroxidase activities (48 h, P < 0.05) and reduced glutathione concentrations (48 h, P < 0.05) were observed. In addition, exposure to 4-MCP increased mRNA expressions of CYP 3A7 (24 h, P < 0.05; 48 h, P < 0.01) and CYP 2E1 (24 h, P < 0.01) versus controls. Taken together, these results suggest that 4-MCP metabolites could induce oxidative stress conditions in Hep G2 cells.

Antioxidant defense disruption by polycyclic aromatic hydrocarbons-coated onto Fe(2)O(3) particles in human lung cells (A549).

We addressed the hypothesis that in vitro short-term exposure to hematite (Fe(2)O(3)) and polycyclic aromatic hydrocarbons (PAHs) is more deleterious by virtue of their combinations being able to cause higher oxidative stress conditions in human lung cells (A549), than either chemical alone. Lipid peroxidation (malondialdehyde; MDA), antioxidant enzyme activities (superoxide dismutase; SOD, glutathione peroxidase; GPx, glutathione reductase; GR), glutathione status (reduced glutathione; GSH, oxidized glutathione; GSSG) and alpha-tocopherol (alpha-Toc) consumption were studied in cells exposed to Fe(2)O(3), benzo(a)pyrene (B(a)P) or pyrene, alone or in association. We found that increases in GSSG/GSH (P<0.01) and in alpha-Toc consumption (P<0.01) counteracted Fe(2)O(3)-induced lipid peroxidation. Exposure to B(a)P did not induce oxidative injury because of the involvement of non-enzymatic antioxidants in cell homeostasis. Pyrene did not induce free radicals (FR)-induced injury. Exposure to PAHs-coated onto Fe(2)O(3) particles damaged both the enzymatic (i.e. increases in SOD and GR activities; P<0.01) and the non-enzymatic (i.e. increases in GSSG/GSH; P<0.001, alpha-Toc consumption; P<0.01) antioxidant defenses, thereby allowing lipid peroxidation (i.e. MDA production; P<0.05). Exposure to PAHs-coated onto Fe(2)O(3) particles induced not only higher lipid peroxidation (i.e. MDA production; P<0.05) but also higher antioxidant alterations (i.e. SOD and GR activities; P<0.05, GSSH/GSH; P<0.01 or P<0.05) than either chemical alone. Several mechanisms could account for this result, enhanced uptake of Fe(2)O(3) and/or greater availability of PAHs. Hence, our results indicate that exposure to PAHs-coated onto Fe(2)O(3) particles is more deleterious in lungs than either chemical alone.

Pulmonary induction of proinflammatory mediators following the rat exposure to benzo(a)pyrene-coated onto Fe2O3 particles.

Epidemiological evidence firmly implicated an interactive effect between Fe2O3 and benzo(a)pyrene (B(a)P) in causing lung cancer. However, despite intensive investigation, the mechanism involved is not precisely established. Since the accumulation of reactive oxygen intermediates (ROI)-mediated damage and/or immune-induced injury might be a possible cause of lung cancer, we studied the oxidative and the inflammatory effects of Fe2O3 (3 mg), B(a)P (3 mg) or B(a)P (3 mg)-coated onto Fe2O3 (3 mg) particles on this relevant organ target in Sprague-Dawley rats. We investigated lipid peroxidation (malondialdehyde; MDA) and secretion of some inflammatory mediators (tumor necrosis factor-alpha, TNF-alpha; interleukin-1 beta, IL-1beta; nitric oxide, NO) in lungs. In addition, mRNA expressions of TNF-alpha, IL-1beta and inducible nitric oxide synthase (iNOS) were evaluated. Our results show that exposure to Fe2O3 and B(a)P, alone or in association, induced 2-fold increases in MDA production suggesting thereby oxidative stress conditions (P<0.01). Exposure to Fe2O3, B(a)P or B(a)P-coated onto Fe2O3 particles significantly increased both mRNA expression and/or synthesis of inflammatory mediators. The main findings of this work were that the association of Fe2O3 and B(a)P induces more pronounced induction of inflammatory mediators (IL-1beta secretion, P<0.01; IL-1beta mRNA expression, P<0.01; iNOS mRNA expression, P<0.05) than B(a)P by itself. Hence, our results may explain why concurrent exposure to Fe2O3 and B(a)P is more deleterious in lungs than exposure to B(a)P alone.

Benzo(a)pyrene-coated onto Fe(2)O(3) particles-induced lung tissue injury: role of free radicals.

Lipid peroxidation (as malondialdehyde; MDA), activities of some antioxidant enzymes (as superoxide dismutase; SOD, glutathione peroxidase; GPx, glutathione reductase; GR), glutathione status, and oxidative DNA damage (as 8-hydroxy-2'-deoxyguanosine; 8-OHdG) were investigated in the lungs of rats exposed to hematite (Fe(2)O(3); 3 mg), benzo(a)pyrene (B(a)P; 3 mg), or B(a)P (3 mg)-coated onto Fe(2)O(3) particles (3 mg). Approximately 2-fold increases in MDA production were seen in animals exposed to Fe(2)O(3), B(a)P, or B(a)P-coated onto Fe(2)O(3) particles (P<0.01). Decreases in SOD activities were observed in rats treated with Fe(2)O(3) (1.66-fold, P<0.01), B(a)P (1.66-fold, P<0.001) or B(a)P-coated onto Fe(2)O(3) particles (1.43-fold, P<0.01). GPx and GR activities could not be detected. No alteration of the glutathione status was observed. Significant increases in the 8-OHdG formation occurred in response to exposure to B(a)P (2.0-fold, P<0.01) or B(a)P-coated onto Fe(2)O(3) particles (23.7-fold, P<0.001). Our results demonstrate also that Fe(2)O(3) generates free radical (FR)-induced lung injury and is not an inert carrier. We established that exposure to B(a)P or B(a)P-coated onto Fe(2)O(3) particles resulted in lipid peroxidation and SOD inactivation, thereby leading to oxidative damages in DNA. The main findings of this work was that B(a)P-coated onto Fe(2)O(3) particles caused higher lung concentrations of 8-OHdG than B(a)P by itself. Hence, our data may explain why exposure to B(a)P-coated onto Fe(2)O(3) particles resulted in a decreased latency and an increased incidence of lung tumors in rodents compared to exposure to B(a)P.