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Tar content per cigarette of various brands, the dry weight of tar trapped on the Cambridge filter after combustion of one cigarette and the EC50 values of cCSEs for inhibition of MTS reduction activity.
Source publication
Cigarette smoke consists of tar and gas phase: the latter is toxicologically important because it can pass through lung alveolar epithelium to enter the circulation. Here we attempt to establish a standard method for preparation of gas phase extract of cigarette smoke (CSE). CSE was prepared by continuously sucking cigarette smoke through a Cambrid...
Citations
... SPME is a simple, low-cost, and solvent-free sample preparation technique (in this case, cigarette smoke compound) [25]. Tar is a part of a particular phase that can be assessed for its levels by measuring the total particulate matter (TPM) obtained from the weight difference of Cambridge filter paper (CFP) before and after smoking and then subtracting it with nicotine and water levels [26]. ...
Cigarette smoke contains carcinogenic residues such as volatile organic compounds and polycyclic aromatic hydrocarbons that stay on surfaces and can be released into the air causing stains and unpleasant smells. The use of chemical cleaners and deodorizers may cause other health and environmental consequences. Eco-enzyme from organic waste such as fruit peels and vegetables left over are one of the organic innovations that are commonly used to remove odor. In this study, we introduce the use of eco-enzyme in the form of spray and evaluate the effectiveness of eco-enzyme spray to eliminate cigarette smoke odor. This research was an experimental organoleptic test involving 20 human subjects, female, older than 18 years old, and have normal olfactory function to rate the cigarette smoke odor intensity after being sprayed with eco-enzyme. Data were analyzed with the Kruskal-Wallis test and continued with Post-Hoc test. We found that the difference in the concentration of eco-enzymes made a significant difference in the intensity of the smell of cigarette smoke (p=0,000, p<0,05), with the highest average score 3.95 at a concentration of 1:3. In conclusion, the eco-enzyme spray is capable to eliminate cigarette smoke odor using 1:3 concentration.
... Reference cigarettes (Code 3R4F) were procured from the Center for Tobacco Reference Products, Kentucky Tobacco Research & Development Center, Lexington, Kentucky, USA. For preparation of the cigarette smoke extract (CSE), a modified method based on a previous report was used (Higashi et al., 2014). Briefly, smoke emissions from 10 cigarettes suctioned in using slow vacuum through an in-house Cigarette Smoke Extractor set-up ( Supplementary Fig. 1) were allowed to pass through sterile PBS (10 ml) for 10 min. ...
Carbon nanotubes (CNTs) are emerging environmental and occupational toxicants known to induce lung immunotoxicity. While the underlying mechanisms are evolving, it is yet unknown whether inhaled CNTs would cause abnormalities in gut microbiota (dysbiosis), and if such microbiota alteration plays a role in the modulation of CNT-induced lung immunotoxicity. It is also unknown whether co-exposure to tobacco smoke will modulate CNT effects. We compared the effects of lung exposure to multi-wall CNT, cigarette smoke extract (CSE), and their combination (CNT + CSE) in a 4-week chronic toxicity mouse model. The exposures induced differential perturbations in gut microbiome as evidenced by altered microbial α- and β- diversity, indicating a lung-to-gut communication. The gut dysbiosis due to CNTs, unlike CSE, was characterized by an increase in Firmicutes/Bacteroidetes ratio typically associated with proinflammatory condition. Notably, while all three exposures reduced Proteobacteria, the CNT exposure and co-exposure induced appearance of Tenericutes and Cyanobacteria, respectively, implicating them as potential biomarkers of exposure. CNTs differentially induced certain lung proinflammatory mediators (TNF-α, IL-1β, CCL2, CXCL5) whereas CNTs and CSE commonly induced other mediators (CXCL1 and TGF-β). The co-exposure showed either a component-dominant effect or a summative effect for both dysbiosis and lung inflammation. Depletion of gut microbiota attenuated both the differentially-induced and commonly-induced (TGF-β) lung inflammatory mediators as well as granulomas implying gut-to-lung communication and a modulatory role of gut dysbiosis. Taken together, the results demonstrated gut dysbiosis as a systemic effect of inhaled CNTs and provided the first evidence of a bidirectional gut-lung crosstalk modulating CNT lung immunotoxicity.
... [6][7][8] Long-term exposure to CS leads to the accumulation of reactive oxygen species (ROS) in airway epithelial cells, followed by plasma membrane damage and the release of damage-associated molecular patterns (DAMPs), which in turn trigger inflammation and active cell death pathways. 9,10 Although many types of cells and mediators are involved in the pathophysiological process of COPD, 11 data from the literature have indicated the key role of Nrf2, which is strongly upregulated by oxidants. 2 Nrf2 is a critical factor in maintaining the oxidation/ antioxidant balance. ...
Background
Nuclear factor E2-related factor 2 (Nrf2) is involved in oxidative stress and lung inflammation and regulates the etiology of chronic obstructive pulmonary disease (COPD). Ferroptosis is characterized by the accumulation of lipid reactive oxygen species (ROS) via ferrous ion-dependent Fenton reactions and is involved in COPD. However, the role of Nrf2 in ferroptosis and its epigenetic regulation in the pathogenesis of COPD remain unclear.
Methods
Ferroptosis was detected by 4-HNE, MDA, C11BODIPY, DCFH-DA, Peals’ staining and CCK-8 assays. qPCR and Western blotting were performed to examine the Nrf2 levels in peripheral lung tissues, primary epithelial cells collected from patients with COPD and subjects with normal pulmonary function (never-smoker [control-NS]; smoker [control-S]), and cigarette smoke extract (CSE)-treated human bronchial epithelial (HBE) cells. ELISA was used to quantify IL-8 and IL-1β levels. Methylation of the Nrf2 promoter was analyzed by bisulfite sequencing and pyrosequencing.
Results
Ferroptosis was involved in COPD and glutathione peroxidase 4 (GPX4) expression was downregulated in the COPD group. Reactive oxygen species (ROS), lipid peroxides and MDA were increased, but GPX4 and SOD were exhausted in CSE-treated HBE cells. The production of IL-1β and IL-8 was promoted in HBE cells in response to CSE but could be reversed by the ferroptosis inhibitor fer-1. The Nrf2 level was significantly decreased in the COPD group compared with the control-S and control-NS groups. Increased Nrf2 expression enhanced GPX4 and SOD levels and inhibited ferroptosis and proinflammatory cytokines in the supernatant. Inhibition of GPX4 reversed the effect of Nrf2 overexpression and promoted ferroptosis. Two specific CpG sites within the Nrf2 promoter were hypermethylated in the COPD group. Similarly, CSE-treated HBE cells exhibited hypermethylation of the Nrf2 gene.
Conclusion
Nrf2 expression was downregulated in the lungs of COPD patients due to hypermethylation of the Nrf2 promoter, inhibiting Nrf2/GPX4 and ferroptosis, which is related to the initiation and progression of COPD. Targeting Nrf2/GPX4 may inhibit ferroptosis, which could provide strategies to delay or treat COPD.
... Higher responses for carbonyl sensors were observed for houses 3 and 8, which both had a negative score on PC2. These homes also reported smoking in the kitchen, an activity that is known to generate numerous volatile carbonyl compounds [29]. House 9 separated from other houses on PC2 whereby the use of cleaning and personal care products influenced this separation. ...
Indoor air quality monitoring as it relates to the domestic setting is an integral part of human exposure monitoring and health risk assessment. Hence there is a great need for easy to use, fast and economical indoor air quality sensors to monitor the volatile organic compound composition of the air which is known to be significantly perturbed by the various source emissions from activities in the home. To meet this need, paper-based colorimetric sensor arrays were deployed as volatile organic compound detectors in a field study aiming to understand which activities elicit responses from these sensor arrays in household settings. The sensor array itself is composed of pH indicators and aniline dyes that enable molecular recognition of carboxylic acids, amines and carbonyl-containing compounds. The sensor arrays were initially deployed in different rooms in a single household having different occupant activity types and levels. Sensor responses were shown to differ for different room settings on the basis of occupancy levels and the nature of the room emission sources. Sensor responses relating to specific activities such as cooking, cleaning, office work, etc were noted in the temporal response. Subsequently, the colorimetric sensor arrays were deployed in a broader study across 9 different households and, using multivariate analysis, the sensor responses were shown to correlate strongly with household occupant activity and year of house build. Overall, this study demonstrates the significant potential for this type of simple approach to indoor air pollution monitoring in residential environments.
... The insufficient efficacy of administering pure NFB led us to develop alternative approaches. Among the various kinds of tobacco smoke mimetics, cigarette smoke extract (CSE) is another agent widely accepted for use in research [22]. We speculated that CSE may be more effective in modulating the aortic phenotype than nicotine alone, due to its complex ingredient composition. ...
... Cigarette smoking extract (CSE) was made by bubbling smoke of nine cigarettes (3R4F, Kentucky Tobacco Research & Development Center, Lexington, KY) through 3 ml PBS. CSE was injected i.p. daily, at a dose of 0.1 ml per mouse [22,52]. The day of the first tamoxifen injection was considered to be day 0. Aortic samples were surgically harvested for morphological analysis. ...
Tobacco smoke is an established risk factor for thoracic aortic aneurysms and dissections (TAAD). However, little is known about its underlying mechanisms due to the lack of validated animal models. The present study developed a mouse model that may be utilized to investigate exacerbation of TAAD formation by mimetics of tobacco smoke. TAADs were created via inducible deletion of smooth muscle cell-specific Tgfbr2 receptors. Using this model, the first set of experiments evaluated the efficacy of nicotine salt (34.0 mg/kg/day), nicotine free base (NFB, 5.0 mg 90-day pellets), and cigarette smoke extract (0.1 ml/mouse/day). Compared with their respective control groups, only NFB pellets promoted TAAD dilation (23 ± 3% vs. 12 ± 2%, P = 0.014), and this efficacy was achieved at a cost of >50% acute mortality. Infusion of NFB with osmotic minipumps at extremely high, but nonlethal, doses (15.0 or 45.0 mg/kg/day) failed to accelerate TAAD dilation. Interestingly, costimulation with β-aminopropionitrile (BAPN) promoted TAAD dilation and aortic rupture at dosages of 3.0 and 45.0 mg/kg/day, respectively, indicating that BAPN sensitizes the response of TAADs to NFB. In subsequent analyses, the detrimental effects of NFB were associated with clustering of macrophages, neutrophils, and T-cells in areas with structural destruction, enhanced matrix metalloproteinase- (MMP-) 2 production, and pathological angiogenesis with attenuated fibrosis in the adventitia. In conclusion, modeling nicotine exacerbation of TAAD formation requires optimization of chemical form, route of delivery, and dosage of the drug as well as the pathologic complexity of TAADs. Under the optimized conditions of the present study, chronic inflammation and adventitial mal-remodeling serve as critical pathways through which NFB exacerbates TAAD formation.
... Cigarette smoke extract preparation followed a previously reported method with some modifications (Higashi et al., 2014(Higashi et al., , 2016. One piece of cigarette (Septwolves, Longyan Cigarette Factory, Fujian, China) was burned in a 500 ml bottle with a modified 50-ml syringe-driven apparatus. ...
Background: Cigarette smoking is a major risk factor for bronchoalveolar epithelial cell (BAEC) injury. Understanding the relevant pathogenesis is important for the treatment of cigarette smoke–related chronic airway diseases such as chronic obstructive pulmonary disease.
Methods: In this study, BAECs were cultured in 5% cigarette smoke extract (CSE) or regular culture medium for 24 h. Differentially expressed genes (DEGs) were detected by next-generation RNA sequencing (RNA-seq) and validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Bioinformatic analysis was performed on DEGs. Co-treated BAECs with 5% CSE and the ferroptosis inhibitor, ferrostatin-1 was applied to observe the role of ferroptosis.
Results: In the CSE group, 210 upregulated genes and 159 downregulated genes were identified compared with the control group. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that the DEGs were related to oxidative stress and ferroptosis. Ferroptosis-related genes were further verified by qRT-PCR. The mRNA level of GPX4 decreased; the mRNA levels of ACSL4, FTH1 and SLC7A11 increased (p < 0.05). Pretreatment with the ferroptosis inhibitor ferrostatin-1 mitigated CSE-induced ROS accumulation and inflammatory mediator expression in BAECs (p < 0.05).
Conclusion: CSE treatment altered ferroptosis-related gene expression patterns in cultured BAECs. Inhibition of ferroptosis reduced the inflammatory response of CSE-treated BAECs. These data provide a better understanding of the underlying molecular mechanisms of CSE-related lung injury.
... It can induce several airway pathologies such as asthma and COPD (1,2,7,8). CSE is a widely-used model for studying the effects of tobacco smoke in vitro and in vivo (31)(32)(33)(34)(35). ...
Cigarette smoking is a major risk factor for pulmonary diseases, including chronic obstructive pulmonary disease (COPD) and cancer. Cigarette smoke is reported to contain over 4,000 chemical compounds. Therefore, it needs to study the effects of cigarette smoke extract (CSE) administration on intracellular calcium concentration. In this study, we investigated how CSE influences intracellular calcium concentration in human lung adenocarcinoma A549 cells. The CSE concentrations used (0.4, 2, 3%) did not influence cell viability. However, at these CSE concentrations, calcium influx transient receptor potential vanilloid 4 (TRPV4) and transient receptor potential vanilloid 6 (TRPV6) proteins significantly increased, whereas calcium efflux sodium-calcium exchanger (NCX1) and plasma membrane Ca2+ ATPase (PMCA1) proteins significantly decreased from those of the control cells. The 3% CSE treatment produced an intracellular calcium concentration higher than that of the control treatment through methods of co-transfection of pGP-CMV-GCaMP6f/CMV-R-GECO1.2 and Rhod-4 Assay. CSE induced concentration-dependent increments in hypoxia-inducible factor (HIF)-1α and HIF-2α protein levels. Moreover, phosphorylation of ERK and Akt was induced by CSE treatment. Also, mitochondrial marker B-cell lymphoma 2 (Bcl-2) protein level decreased and Bcl-2-associated X (Bax) protein level increased following CSE treatment. Also, endoplasmic reticulum (ER) stress markers BiP, CHOP, p-SAPK, and p-eIF2α levels were increased by CSE treatment. These results suggest that CSE may increase the concentration of intracellular calcium, thus increasing mitochondrial and ER stress.
... In order to identify the active component in CSE, we examined the action of MVK on the invasiveness of Colon-26 cells, and we confirmed that MVK significantly reduced the invasiveness of cells. 9) In the present study, we searched for novel α,β-unsaturated carbonyl compounds in CSE to examine the anticancer action by utilizing the reactivity with GSH by the Michael addition. First, CSE was mixed with GSH and the reaction solution was analyzed using a high-resolution mass spectrometer to determine the elemental composition of newly formed peaks, and then we selected candidate compounds. ...
... 12) Most recently, Hatai et al. reported that MVK inhibited the invasiveness of mouse rectal carcinoma Colon-26 cells. 9) In this study, we searched for effective and unidentified α,β-unsaturated aldehydes other than ACR, CA, and MVK. It is well-known that α,β-unsaturated carbonyl compounds readily form adducts with GSH by Michael addition. ...
α,β-Unsaturated carbonyl compounds readily form adducts with SH or NH2 residues, which are nucleophilic agents, by Michael addition. Glutathione (GSH) is a tripeptide that contains an SH residue and functions as an antioxidant. We demonstrated previously that acrolein (ACR), crotonaldehyde (CA), and methyl vinyl ketone (MVK) are present in nicotine- and tar-removed cigarette smoke extract (CSE) and reacted with GSH in B16-BL6 mouse melanoma cells to form GSH-ACR, GSH-CA, and GSH-MVK adducts, suggesting a possible mechanism for CSE-induced cytotoxicity. In this study, we searched for novel α,β-unsaturated carbonyl compounds other than ACR, CA, and MVK. We selected candidate compounds in CSE based on accurate mass values generated using LC/MS analysis of products formed between CSE and GSH, and identified these using GC/MS analysis and library screening. As a result, we isolated trans-2-methyl-2-butenal, 2-methyl-2-cyclopenten-1-one, 3-methyl-2-cyclopenten-1-one, and furfural, which were poorly reactive with GSH and only very weakly inhibited growth of Colon-26 mouse carcinoma cells and BALB/3T3 clone A31 mouse normal cells. We also isolated 2-cyclopenten-1-one, trans-2-pentenal, 3-methyl-2-butenal and ethyl vinyl ketone, which were highly reactive with GSH and significantly inhibited the growth of both cell lines. Our data suggest that the reactivity of compounds in CSE with GSH may be positively correlated with the effect on inhibiting cell growth. Notably, trans-2-pentenal showed marked inhibition of carcinoma cells growth, whereas this compound exhibited little inhibitory effect on normal cells. trans-2-Pentenal may be a potent candidate or seed for antitumor agents.
Graphical Abstract Fullsize Image
... The absence of any standardized method to prepare as well as compare the extracts from different laboratories [31] has further compounded the issue of variabilities in the prepared extracts. We propose using a UV spectrophotometry based method for standardizing the prepared extract. ...
Tobacco smoking and exposure to biomass smoke are the major sources of exposure to harmful chemicals and particulate matter
which may lead to cardiovascular and lung diseases. Our understanding of the pathophysiology of these diseases and the role of
smoke constituents in the development of these diseases has evolved over time, but is still limited due to the difficulty in mimicking
human smoke exposure in vitro. In this study, we developed an adaptable smoke extraction system to generate smoke extracts from
cigarettes, bidis (Indian cigarettes), and biomass (cow dung). These extracts were then standardized using UV spectroscopy. A strong
positive correlation was observed between the number of cigarettes, bidis or amount of biomass burnt and the absorbance at 400nm.
Reproducibility and internal consistency was confirmed between multiple preparations of the smoke extracts using Cronbach’s Alpha
(α) and intraclass correlation analysis (ICC) of the complete absorption spectra. Variation in the absorption spectra were mostly
observed between 350 and 550 nm and thus a further analysis of the variation in the absorbance within this range was performed.
Internal consistency was again found to be excellent as indicated by α value of > 0.9 for all the three extracts. ICC measurements also
indicated high reproducibility for the absorbance of all three sources of extracts within this range. A dose-dependent decrease in cell
viability was observed in RAW 264.7 and A549 cells after treatment with these extracts for 4h as measured by the MTT cell viability
assay. Thus, we show the development of a simple smoke extraction system, method for extract standardization and its effect on cell
viability with extracts from different sources with these data
... Total nicotine delivery was quantified by HPLC-UV based on previous methods. 19 −21 Previously, α fb determination by NMR was done using a concentric NMR tube insert containing pure e-cigarette eliquid, 11 condensed aerosol, or cigarette smoke, 18 which was inturn surrounded by lock solvent, DMSO-d 6 . Duell et al. calculated α fb by comparing relative chemical shift differences between methyl and aromatic protons of Nic and NicH + standards in glycerol/propylene glycol with commercial eliquids. ...
Heat-not-burn products, e.g. IQOS, are becoming popular as alternative tobacco products. The nicotine protonation state in aerosols has addiction implications due to differences in absorption kinetics and harshness. The nicotine free-base fraction (αfb) ranges from 0-1. Herein we report αfb for IQOS aerosols using exchange-average ¹H NMR chemical shift of the nicotine methyl protons in the bulk aerosol. This was verified by HS-SPME-GCMS, with total nicotine delivery quantified by HPLC-UV. αfb ≈ 0 for all products tested, lower than JUUL; likely a result of proton transfer from acetic acid and/or other additives in the largely aqueous aerosol. Others have reported higher αfb values (0.057) for these products, however, their methods were subject to error due to solvent perturbation effects. The αfb values presented herein were generated using an ¹H NMR method that avoids this issue.
