Cellular and Acellular Assays for Measuring Oxidative Stress Induced by Ambient and Laboratory-Generated Aerosols.
Ontology highlight
ABSTRACT:
SUBMITTER: Ng NL
PROVIDER: S-EPMC7266377 | biostudies-literature | 2019 Mar
REPOSITORIES: biostudies-literature
Publications
Cellular and Acellular Assays for Measuring Oxidative Stress Induced by Ambient and Laboratory-Generated Aerosols.
Ng N L NL Tuet W Y WY Chen Y Y Fok S S Gao D D Tagle Rodriguez M S MS Klein M M Grosberg A A Weber R J RJ Champion J A JA
Research report (Health Effects Institute) 20190301 197
<h4>Introduction</h4>Many studies have established associations between exposure to air pollution, or atmospheric particulate matter (PM), and adverse health effects. An increasing array of studies have suggested oxidative stress as a possible mechanism by which PM-induced health effects arise, and as a result, many chemical and cellular assays have been developed to study PM-induced oxidant production. Although significant progress has been made in recent years, there are still many gaps in thi ...[more]
Similar Datasets
Project description:BackgroundModern microbiology laboratories are designed to protect workers and the environment from microbial aerosols produced during microbiological procedures and accidents. However, there is only limited data available on the aerosols generated from common microbiology procedures.MethodsA series of common microbiological procedures were undertaken with high concentration spore suspensions while air samplers were operated to sample the aerosols generated. Surface contamination from droplets was visualized using sodium fluorescein within the suspension. A total of 36 procedures were studied using different sample volumes (0.1-10 mL) and two spore suspension titers (107 and 109 colony forming units [cfu]/mL).ResultsThe aerosol concentrations generated varied from 0 to 13,000 cfu/m3. There was evidence to suggest that titer, volume, and poor use of equipment were significant factors in increased aerosol generation from some of the procedures. A risk assessment undertaken using the data showed that any aerosol generated from these processes would be contained within a correctly operating biological safety cabinet. Therefore, with these procedures, the operator and the environment would not require any additional protective measures such as respiratory protective equipment or a negative pressure laboratory to prevent aerosol exposure or release.ConclusionsAerosol generation from common laboratory processes can be minimized by reducing sample volumes and concentrations if possible. Training laboratory staff in good microbiological techniques would further mitigate aerosols generated from common laboratory processes.
| S-EPMC9150131 | biostudies-literature
Project description:Biomass burning aerosol (BBA) contributes significantly to the global aerosol burden, yet its chemical nature and ice nucleation activities (INAs) are unconstrained due to the heterogeneity in biomass sources and complex evolution of atmospheric aging processes. This study comprehensively investigates the chemical composition and INA of BBA generated through laboratory-controlled burns with different biomasses and burning conditions. Both freshly emitted and photochemically aged BBA produced from different processes exhibit distinct and reproducible chemical compositions. However, the INA of BBA shows substantial variability at mixed-phase cloud temperatures and cannot be predicted by the chemical variability of the enriched carbonaceous materials. This indicates the negligible role of carbonaceous materials in determining the INA of BBA. Using laboratory data, we further evaluate the impact of BBA on atmospheric ice nucleation using particulate matter mass concentration and particle equivalent spherical radius. The estimated ice nucleating particle (INP) concentrations contributed by laboratory-produced BBA are lower than those observed during BBA pollution in field studies. This discrepancy is likely attributed to co-lofted mineral particles during real-world biomass burning, such as ash or soil particles, rather than carbonaceous-rich particles from combustion. We encourage further research to quantify the contribution of mineral particles to the INP concentrations of BBA.
| S-EPMC11823448 | biostudies-literature
Project description:Electronic cigarettes (e-cigs) have fast increased in popularity but the physico-chemical properties and toxicity of the generated emission remain unclear. Reactive oxygen species (ROS) are likely present in e-cig emission and can play an important role in e-cig toxicity. However, e-cig ROS generation is poorly documented. Here, we generated e-cig exposures using a recently developed versatile exposure platform and performed systematic ROS characterization on e-cig emissions using complementary acellular and cellular techniques: 1) a novel acellular Trolox-based mass spectrometry method for total ROS and hydrogen peroxide (H2O2) detection, 2) electron spin resonance (ESR) for hydroxyl radical detection in an acellular and cellular systems and 3) in vitro ROS detection in small airway epithelial cells (SAEC) using the dihydroethidium (DHE) assay. Findings confirm ROS generation in cellular and acellular systems and is highly dependent on the e-cig brand, flavor, puffing pattern and voltage. Trolox method detected a total of 1.2-8.9nmol H2O2eq./puff; H2O2 accounted for 12-68% of total ROS. SAEC cells exposed to e-cig emissions generated up to eight times more ROS compared to control. The dependency of e-cig emission profile on e-cig features and operational parameters should be taken into consideration in toxicological studies.
| S-EPMC5848214 | biostudies-literature
Project description:BackgroundElectronic cigarette (EC) aerosols contain unique compounds in addition to toxicants and carcinogens traditionally found in tobacco smoke. Studies are warranted to understand the public health risks of ECs.ObjectiveThe aim of this study was to determine the genotoxicity and the mechanisms induced by EC aerosol extracts on human oral and lung epithelial cells.MethodsCells were exposed to EC aerosol or mainstream smoke extracts and DNA damage was measured using the primer anchored DNA damage detection assay (q-PADDA) and 8-oxo-dG ELISA assay. Cell viability, reactive oxygen species (ROS) and total antioxidant capacity (TAC) were measured using standard methods. mRNA and protein expression were evaluated by RT-PCR and western blot, respectively.ResultsEC aerosol extracts induced DNA damage in a dose-dependent manner, but independently of nicotine concentration. Overall, EC aerosol extracts induced significantly less DNA damage than mainstream smoke extracts, as measured by q-PADDA. However, the levels of oxidative DNA damage, as indicated by the presence of 8-oxo-dG, a highly mutagenic DNA lesion, were similar or slightly higher after exposure to EC aerosol compared to mainstream smoke extracts. Mechanistically, while exposure to EC extracts significantly increased ROS, it decreased TAC as well as the expression of 8-oxoguanine DNA glycosylase (OGG1), an enzyme essential for the removal of oxidative DNA damage.ConclusionsExposure to EC aerosol extracts suppressed the cellular antioxidant defenses and led to significant DNA damage. These findings emphasize the urgent need to investigate the potential long-term cancer risk of exposure to EC aerosol for vapers and the general public.
| S-EPMC5436899 | biostudies-literature
Project description:Evaluation of the semi-continuous Monitor for AeRosols and GAses in ambient air (MARGA, Metrohm Ap-plikon B.V.) was conducted with an emphasis on examination of accuracy and precision associated with processing of chromatograms. Using laboratory standards and atmospheric measurements, analytical accuracy, precision and method detection limits derived using the commercial MARGA software were compared to an alternative chromatography procedure consisting of a custom Java script to reformat raw MARGA conductivity data and Chromeleon (Thermo Scientific Dionex) software for peak integration. Our analysis revealed issues with accuracy and precision resulting from misidentification and misintegration of chromatograph peaks by the MARGA automated software as well as a systematic bias at low concentrations for anions. Reprocessing and calibration of raw MARGA data using the alternative chromatography method lowered method detection limits and re-duced variability (precision) between parallel sampler boxes. Instrument performance was further evaluated during a 1-month intensive field campaign in the fall of 2014, including analysis of diurnal patterns of gaseous and particulate water-soluble species (NH3, SO2, HNO3, N H 4 + , S O 4 2 - and N O 3 - , gas-to-particle partitioning and particle neutralization state. At ambient concentrations below ~ 1 µg m-3, concentrations determined using the MARGA software are biased +30 and +10 % for N O 3 - and S O 4 2 - , respectively, compared to concentrations determined using the alternative chromatography procedure. Differences between the two methods increase at lower concentrations. We demonstrate that positively biased N O 3 - and S O 4 2 - measurements result in overestimation of aerosol acidity and introduce nontrivial errors to ion balances of inorganic aerosol. Though the source of the bias is uncertain, it is not corrected by the MARGA online single-point internal LiBr standard. Our results show that calibra-tion and verification of instrument accuracy by multilevel external standards is required to adequately control analytical accuracy. During the field intensive, the MARGA was able to capture rapid compositional changes in PM2.5 due to changes in meteorology and air mass history relative to known source regions of PM precursors, including a fine N O 3 - aerosol event associated with intrusion of Arctic air into the southeastern US.
| S-EPMC6192433 | biostudies-literature
Project description:BackgroundEpidemiological studies have suggested that elevated concentrations of particulate matter (PM) are strongly associated with the incidence of atherosclerosis, however, the underlying cellular and molecular mechanisms of atherosclerosis by PM exposure and the components that are mainly responsible for this adverse effect remain to be established. In this investigation, we evaluated the effects of ambient PM on vascular smooth muscle cell (VSMC) behavior. Furthermore, the effects of polycyclic aromatic hydrocarbons (PAHs), major components of PM, on VSMC migration and the underlying mechanisms were examined.ResultsVSMC migration was significantly increased by treatment with organic matters extracted from ambient PM. The total amount of PAHs contained in WPM was higher than that in SPM, leading to higher ROS generation and VSMC migration. The increased migration was successfully inhibited by treatment with the anti-oxidant, N-acetyl-cysteine (NAC). The levels of matrix metalloproteinase (MMP) 2 and 9 were significantly increased in ambient PM-treated VSMCs, with MMP9 levels being significantly higher in WPM-treated VSMCs than in those treated with SPM. As expected, migration was significantly increased in all tested PAHs (anthracene, ANT; benz(a)anthracene, BaA) and their oxygenated derivatives (9,10-Anthraquinone, AQ; 7,12-benz(a)anthraquinone, BAQ, respectively). The phosphorylated levels of focal adhesion kinase (FAK) and formation of the focal adhesion complex were significantly increased in ambient PM or PAH-treated VSMCs, and these effects were blocked by administration of NAC or α-NF, an inhibitor of AhR, the receptor that allows PAH uptake. Subsequently, the levels of phosphorylated Src and NRF, the downstream targets of FAK, were altered with a pattern similar to that of p-FAK.ConclusionsPAHs, including oxy-PAHs, in ambient PM may have dual effects that lead to an increase in VSMC migration. One is the generation of oxidative stress followed by MMP upregulation, and the other is actin reorganization that results from the activation of the focal adhesion complex.
| S-EPMC9026692 | biostudies-literature
Project description:Today air pollution caused by particulate matter (PM) is a global issue, especially in densely populated and high-traffic cities. The formation of reactive oxygen species (ROS) by various toxicological studies is considered as one of the important effects caused by airborne particles that can lead to adverse effects on human health. In this study, to answer the question of whether particle size affects oxidative potential (OP), we searched the main databases, including PubMed, Scopus, Embase, and Web of Science, and defined search strategy based on the MESH terms for the above-mentioned search engines. All articles published until 2021 were searched. An ANOVA was run using R software to show the correlation between the size distributions of particulate matter and oxidative potential (base on mass and volumetric units) in ambient air. As expected, the regression results showed that the relationship between particle size and OP values for the studies based on mass-logarithm has a significant difference in the different distribution size categories, which was related to the difference between the <2.5 and < 1 categories. However, ANOVA analysis did not show a significant difference in the volumetric OP logarithm in the different distribution size categories. In this study, it was found that sizes higher than 2.5 μm did not have much effect on human health, and it is recommended that future research focus on PM2.5.Supplementary informationThe online version contains supplementary material available at 10.1007/s40201-021-00768-w.
| S-EPMC9163285 | biostudies-literature
Project description:IntroductionChemical elements and their toxicity were evaluated in electronic cigarette (EC) solvents, fluids, and aerosols.Aims and methodsElement identification and quantification in propylene glycol (PG), glycerin (G), refill fluids before and after use, and aerosols was done using inductively coupled plasma optical emission spectrometry. Cytotoxicity and oxidative stress were evaluated using in vitro assays.ResultsSeven elements were present in PG, G, and popular refill fluids, and they transferred to aerosols made with ECs. Selenium was in all products (0.125-0.292 mg/L), while arsenic, aluminum, and tin were frequently in solvent and refill fluid samples at lower concentrations. Iron, chromium, copper, nickel, zinc, and lead were only detected in fluid after EC use, indicating they came from heated atomizers. Elements transferred most efficiently to aerosols made with second-/third-generation ECs. Of the elements in fluid, selenium and arsenic were the most cytotoxic to human bronchial epithelial cells (BEAS-2B) and pulmonary fibroblasts in the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay. Selenium increased superoxide production in mitochondria and nucleoli and elevated selenoprotein H in nucleoli of BEAS-2B cells at concentrations found in EC aerosols (10 nM or 0.002 mg/L).ConclusionsElements in EC aerosols came from both e-fluids and atomizing units. Within second-/third-generation products, transfer became more efficient as power increased. In vitro responses occurred at concentrations of selenium found in some EC aerosols. Human exposure to chemical elements in ECs could be reduced by regulating (decreasing) allowable EC power and by improving the purity of PG and G.ImplicationsPG, G, refill fluids, and e-fluids contained potentially toxic chemical elements that transferred to aerosols. Transfer was more efficient in second- and third-generation EC products and increased as power increased. Selenium and arsenic were the most cytotoxic of the elements tested in the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay. Selenium tetrachloride-induced oxidative stress in BEAS-2B cells, but not in human pulmonary fibroblasts. All fluids contained selenium above the concentration that induced oxidative stress in human bronchial epithelial cells. Selenium increased superoxide in mitochondria and nucleoli and increased selenoprotein H, a redox responsive DNA-binding protein that is upregulated by superoxide and an indicator of nucleolar stress. EC users are exposed to elements in aerosols, which may with chronic exposure contribute to diseases associated with oxidative stress.
| S-EPMC7737483 | biostudies-literature
Project description:The distribution of contractile forces generated in cytoskeletal stress fibers (SFs) contributes to cellular dynamic functions such as migration and mechanotransduction. Here we describe a novel (to our knowledge) method for measuring local tensions in SFs based on the following procedure: 1), known forces of different magnitudes are applied to an SF in the direction perpendicular to its longitudinal axis; 2), force balance equations are used to calculate the resulting tensions in the SF from changes in the SF angle; and 3), the relationship between tension and applied force thus established is extrapolated to an applied force of zero to determine the preexisting tension in the SF. In this study, we measured tensions in SFs by attaching magnetic particles to them and applying known forces with an electromagnetic needle. Fluorescence microscopy was used to capture images of SFs fluorescently labeled with myosin II antibodies, and analysis of these images allowed the tension in the SFs to be measured. The average tension measured in this study was comparable to previous reports, which indicates that this method may become a powerful tool for elucidating the mechanisms by which cytoskeletal tensions affect cellular functions.
| S-EPMC3127176 | biostudies-literature
Project description:Flavored tobacco products are increasing in popularity but remain unregulated, with the exception of the ban on flavored conventional cigarettes. Lack of regulation of cigarillos and little cigars allows vendors to have their own version of popular flavors, each with different chemical components. A new classification system was created for flavored cigars in order to easily communicate our results with the scientific community. To understand the physicochemical characteristics of flavored little cigars and cigarillo smoke, size distribution and concentration of particulate matter in smoke were determined. Acellular reactive oxygen species generation was measured as an indirect measurement of the potential to cause oxidative stress in cells. In addition, cigarillo smoke extract treatment on bronchial epithelial (Beas-2b) cells were assessed to determine the flavor-induced cellular toxicity. Flavored cigars/cigarillos showed significant variability in the tested parameters between flavors as well as brands of the same flavor, but most of the cigars showed higher potential of generating oxidative stress, than research grade cigarettes. Flavored cigars produced maximum particle concentrations at 1.0μm and 4.0 μm compared with 3R4F reference research cigarettes. A differential cytotoxic response was observed with cigarillo smoke extract treatments, with "fruits/candy" and "drinks" being the most toxic, but were not more cytotoxic than smoke from cigarettes. These cigarillos with flavors must be well characterized for toxicity in order to prevent adverse effects caused by exposure to flavor chemicals. Our study provides insight into understanding the potential health effects of flavor-infused cigars/cigarillos and the need for the regulation of those flavoring chemicals in these products. Future research is directed to determine the flavoring toxicity of little cigars and cigarillos in vivo studies.
| S-EPMC6905550 | biostudies-literature