Pleural effusion (PE) is a common clinical manifestation associated with advanced stages of both malignant and non-malignant diseases. PE frequently occurs in advanced non-small cell lung cancer (NSCLC) and contributes to tumor progression. NSCLC accounts for more than 85% of the lung cancers and remains a problem worldwide due to its late diagnosis and low rate of response to treatment. Extracellular vesicles (EVs) present in PE are emerging as key mediators of intercellular communication, capable of transferring oncogenic signals through their molecular cargo. Among these molecules, microRNAs (miRNAs) are increasingly recognized as important drivers of cancer progression. miR-21 is a representative onco-miRNA, involved in lung cancer progression; moreover EV-miR-21 upregulation at the pre-dissemination stage promotes cancer cell survival in the pleural cavity. This study compares, for the first time, the functional role of EVs isolated from malignant PE in NSCLC patients (NSCLC-PE-EVs) with those isolated from PE in patients with congestive heart failure (CHF-PE-EVs), focusing on their ability to modulate lung cancer cell behavior. The effects of these EVs were evaluated on COLO699 lung adenocarcinoma cells with proliferation, migration, and gene expression assays. NSCLC-PE was found to contain approximately twice the amount of EVs compared to CHF-PE. NSCLC-PE-EVs were enriched in the oncogenic miR-21-5p, while CHF-PE-EVs had higher levels of the tumor-suppressive miR-126-3p. Only NSCLC-PE-EVs induced dose-dependent increases in COLO699 cell proliferation and migration, consistent with elevated miR-21-5p expression. Functional studies confirmed that miR-21-5p mediates these effects by downregulating PTEN and PDCD4, and by upregulating MMP9 expression. Our findings show that NSCLC-PE-EVs promote malignant phenotypes in lung cancer cells via the transfer of miR-21-5p.

OBJECTIVE:Monoclonal gammopathy of undetermined significance (MGUS) is a preneoplastic disease that often precedes multiple myeloma. The multistep evolutionary pattern of multiple myeloma is driven by genetic instability, a pro-inflammatory and immunosuppressive microenvironment, and tumor growth. Inflammation has long been recognized as a factor in both the onset and progression of cancer. PATIENTS AND METHODS:In this study, interleukin-18 plasma levels were compared in patients with multiple myeloma and monoclonal gammopathy of undetermined significance, as well as in a group of healthy controls. RESULTS:Our study shows that monoclonal gammopathy of undetermined significance patients have lower levels of interleukin-18 than healthy controls (521.657 ± 168.493 pg/ml vs. 1,266.481 ± 658.091 pg/ml for controls, p < 0.001). Thus, we discovered a significant difference in interleukin-18 levels between multiple myeloma patients and controls (418.177 ± 197.837 pg/ml; p = 0.001). CONCLUSIONS:In our work, we identified a reduction of interleukin-18 in monoclonal gammopathies. Furthermore, in this paper, we aimed to evaluate the existing literature on the potential mechanisms of action of this pro-inflammatory cytokine in the development of these diseases.

High concentrations of H2O2, indicative of increased oxidative stress in the lung, are observed in the exhaled breath of individuals affected by different respiratory diseases. Therefore, measuring H2O2 in breath exhale represents a promising and non-invasive approach for monitoring the onset and progression of these diseases. Herein, we have developed an innovative, inexpensive, and easy-to-use device for the measurement of H2O2 in exhaled breath. The device is based on a silver layer covered with an electrodeposited thin film of chitosan, that ensures the wettability of the sensor in a humid atmosphere. Sensors were calibrated in the aerosol phase using both phosphate buffer solution and cell culture medium. In the buffer, a sensitivity of 0.110 ± 0.0042 μA μM−1 cm−2 (RSD: 4%) and a limit of detection of 30 μM was calculated, while in the cell culture medium, a sensitivity of 0.098 ± 0.0022 μA μM−1 cm−2 (RSD 2%) and a limit of detection of 40 μM were obtained. High selectivity to different interfering species was also verified. Sensors were further tested versus an aerosol phase obtained by nebulizing the culture medium derived from human bronchial epithelial cells that had been exposed to pro-oxidant and antioxidant treatments. The results were comparable with those obtained using the conventional cytofluorimetric method. Finally, sensors were tested in real breath exhalate samples and even after undergoing physical deformations. Data presented herein support that in future applications, these sensors can be integrated into face masks allowing for easy breath monitoring.

Paper Received: 18 January 2023; Revised: 26 March 2023; Accepted: 7 June 2023 Please cite this article as: Bruno M.G., Patella B., Aiello G., Torino C., Vilasi A., Cipollina C., Di Vincenzo S., Pace E., O'Riordan A., Inguanta R., 2023, Wearable Sensor for Real-time Monitoring of Hydrogen Peroxide in Simulated Exhaled Air, Chemical Engineering Transactions, 100, 655660 DOI:10.3303/CET23100110 CHEMICAL ENGINEERING TRANSACTIONS

Macrophage (M) polarization with a prevalence in M2 has been shown to have a relevant impact on the pathogenesis of upper and lower airway allergic diseases. IL-6, IL-1b and IL-8 are among the cytokines produced by M1 macrophages while TGF-b is included among the cytokines produced by M2 macrophages. Tyndallization is a thermal process able to inactivate microorganisms. Formulations containing inactivated microorganisms and/or their components, defined as postbiotics, can exert beneficial effects on human chronic respiratory diseases via modulation of immune responses. The present study explores the effects on macrophage polarization by a blend of tyndallized bacteria (TB) containing Lactobacillus Casei, Lactobacillus Acidophilus, Lactobacillus Plantarum, Streptococcus Thermophilus. Human macrophage-like THP1 cells were exposed to different concentrations of TB (106, 5x106, 107, 5x107, 108 CFU/ml) and then cell viability (MTS), TB phagocytosis and gene expression and release of IL-1b, IL-6, IL-8 and TGF b were assessed. The obtained results demonstrated that TB were well tolerated at all the tested concentrations and were phagocyted after 30 minutes in a dose-dependent manner. In addition, treatment with TB was able to: a) increase the release of IL-1b; b) increase the gene expression of IL-6 and IL-8; c) decrease the gene expression of TGFb. In conclusion, this study demonstrates that TB are capable to orientate M polarization toward M1, a functional phenotype that protects from allergic diseases and that can control viral infections thus preventing disease exacerbations.

Background: Lung cancer, the leading cause of morbidity and mortality worldwide, frequently affects patients with history of Chronic Obstructive Pulmonary Disease (COPD). Cigarette smoke exposure is considered the main risk factor for COPD and lung cancer. Cigarette smoke, increasing oxidative stress and modulating epithelial–mesenchymal transition (EMT) processes in cancer cells, favours cancer progression. Formoterol (FO), a long-acting β2-agonist widely used for COPD treatment, exerts anti-oxidant activities. Aim: this study explored in lung adenocarcinoma cell line (A549) whether FO was able to counteract the effects of cigarette smoke extract (CSE) exposure on oxidative stress and EMT processes. Methods: A549 were stimulated with CSE and FO alone or combined, ROS and mitochondrial superoxide were evaluated by flow-cytometry and EMT markers (E-cadherin, SNAIL1) were evaluated by flow-cytometry and Real-Time PCR. Results: CSE increased production of ROS and mitochondrial superoxide, decreased E-cadherin and increased SNAIL1. FO reverted all these phenomena in CSE stimulated A549 cells. Conclusions: the present study provides intriguing evidence that FO might exert anti-cancer effects reverting oxidative stress and some EMT processes due to cigarette smoke exposure in lung adenocarcinoma cells. These evidences must be validated in future clinical studies to support a repositioning of FO as add on treatment for lung cancer management.

A strong association exists among inflammation and lung cancer. Oncogenic miR-21 is a target of the pro-inflammatory transcription factor NF-κB, while NF-κB-miR-21 interplay has a relevant role in the amplification of inflammatory responses that promote cancer progression. A new fish oil integrally extracted from anchovy fillet leftovers (“AnchoisOil”) and microencapsulated in mesoporous silica particles (“Omeg@Silica”) is significantly more effective than fish oil alone in anti-proliferative and pro-apoptotic effects in non-small cell lung cancer (NSCLC) cell lines. The biomoecular mechanisms that explain this efficacy are not yet understood. The present study was aimed to investigate the biomolecular mechanisms by which Omeg@Silica microparticles exert anti-cancer effects in NSCLC by exploring miR-21, NF-kB and IL-8 expression and release in these cells. We treated NSCLC cell (adenocarcinoma A549, and muco-epidermoid NCI-H292) with AnchoisOil dispersed in ethanol (10 and 15 mg/ml), or encapsulated in silica (Omeg@Silica: FOS in graphs and data plots), evaluating the expression/release of miRNA-21, IL-8 and NF-kB. On both NSCLC cell lines the Omeg@Silica microparticles were more effective than AnchoisOil fish oil alone in reducing IL-8 gene expression. In A549, the microencapsulated fish oil was more effective in decreasing miRNA-21, IL-8 protein release and the expression of NF-kB. These new results suggest that Omeg@Silica microparticles could exert their anti-cancer activity, counteracting tumor-related inflammation, by the inhibition of NF-kB/miR-21/IL-8 interplay in lung adenocarcinoma cells.

The recurrence of pleural effusions is a common event in lung cancer patients. Pleural inflammation generates an ideal microenvironment to attract cancer cells and to promote their growth. Pleural fluids (PF), as other biological fluids, contain extracellular vesicles (EVs). EVs carry proteins, metabolites, lipids, and nucleic acids, including microRNAs (miRNAs) that can induce phenotypic and functional reprogramming of target cells. Our study is designed to assess the roles of EVs contained in PF of patients with lung cancer (cancer PF-EVs) comparing the effects of these vesicles to EVs isolated by pleural fluid of patients with congestive heart failure (CHF PF-EVs) on target cells, focusing on the roles of mir-126-3p and mir-21 5p. Pleural EVs (PF-EVs) are isolated by commercial kit, from 5 ml of PF. The miRNAs contained in PF-EVs are analyzed by qRT-PCR. The effects of cancer PF-EVs and CHF PF-EVs on proliferation of lung cancer cells are evaluated by MTS assay. In cancer PF the amount of EVs is about 2-fold higher than CHF. We also observed that the levels of miR-21-5p and miR-126-3p contained in PF-EVs are different in cancer respect to CHF. MiR-126 amount, normalized per number of EVs, in CHF is lower than in cancer. Although miR-21 levels in cancer PF-EV and CHF PF-EV are comparable, globally the levels of miR-21 in cancer PF are higher that CHF-PF. Interestingly cancer PF-EVs have a proliferative effect on lung cancer cells in a dose-dependent manner. Our results indicated that cancer PF-EVs might contribute to cancer expansion and pleural EV-miRNAs can be useful as potential biomarker for lung cancer prognosis.

Increased oxidative burden contributes to the pathogenesis of most inflammatory diseases and is associated with aging and chronic inflammation. Macrophages contribute to the generation of reactive oxygen species (ROS) within inflamed tissues. Currently, ROS generation is measured using fluorescent probes and colorimetric/fluorimetric biochemical assays. Hydrogen peroxide (H2O2) diffuses through the cell membrane and can be monitored in the extracellular space. Herein, we present a sensor for H2O2 detection released by cells in culture supernatants. H2O2 sensing performance was evaluated using chronoamperometric detection. A sensitivity of 0.0641 mu A mu M-1 cm(-2) with a limit of detection of 6.55 mu M and excellent selectivity against many interferents was found. H2O2 release was also measured in conditioned medium from human THP-1 macrophages exposed to pro-oxidant and anti-oxidant treatments. The results were compared with those obtained by flow cytometry using the same cells stained with carboxy-H(2)DCFDA and MitoSOX Red, which detect intracellular ROS and mitochondrial superoxide, respectively. The addition of pro-oxidants lipopolysaccharide (LPS) and nigericin resulted in a significant increase in the cathodic current due to the H2O2 reduction, indicating an increased release of H2O2. The addition of 17-oxo-DHA, which inhibits LPS- and nigericin-dependent responses, decreased the LPS- and nigericin-induced release of H2O2. All the results obtained with the sensor were consistent with those obtained using flow cytometry. The operation of the sensor directly in the cell culture growth medium had no impact on cell viability. The sensor is highly sensitive, fast, and cost effective, and it can potentially be used for real time monitoring of oxidative stress.

A dysregulated epithelial–mesenchymal transition (EMT) contributes to tumor progression, fibrosis and lung tissue remodeling in Chronic Obstructive Pulmonary Disease (COPD). Cigarette smoke, a risk factor for all these processes, induces oxidative stress, mitochondrial damage and drives cell senescence by reducing FoxO3, an anti-aging factor. Mitochondrial dysfunction is involved in aging and increases lactate generation, which is correlated with EMT induction. Here, we investigated the cigarette smoke extract (CSE) effects on mitochondrial status and cell metabolism and on the expression of EMT markers in lung epithelial cells, focusing on the role of FoxO3 in this process. A549 cells were exposed to CSE and TGF-β1. Reactive oxygen species (ROS), mitochondrial superoxide, intracellular ATP, E-cadherin and FoxO3 expression were assessed. Extracellular lactate was measured as readout of the glycolytic flux. FoxO3 expression was silenced using siRNA and E-cadherin gene expression was evaluated. Some key results were confirmed in primary bronchial epithelial cells (PBEC). Our results showed that CSE and TGF-β1 reduced E-cadherin and FoxO3 expression in A549 and this was confirmed in PBEC. FoxO3 silencing decreased E-cadherin in A549. EMT induced by CSE in A549 was associated with ROS and mitochondrial superoxide increase, lactate release and intracellular ATP decrease. These data suggest that cigarette smoke-induced EMT is mediated in part by a decrease of FoxO3, in lung epithelial cells. This is accompanied metabolic reprogramming where mitochondrial damage is associated with an increase of glycolysis, most likely as a compensatory mechanism.

Notch-1 intervenes in the reparative processes of mucosa by controlling cell proliferation, differentiation and stem cell maintenance. Cigarette smoke alters airway epithelial homeostasis. The present study explored whether: Smokers showed altered Notch-1 expression; and whether in bronchial epithelial cells (16HBE): a) cigarette smoke extracts (CSE) altered the expression of Notch-1, of its ligand Jagged-1 (Jag-1) and the nuclear translocation of Notch-1; b) Notch-1 signaling activation as well as CSE modified Ki67, PCNA, p21, IL-33 expression, cell proliferation and repair processes. Notch-1 expression was assessed in the epithelium from large airway surgical samples from non-smoker and smoker subjects by immunohistochemistry.16HBE were cultured with/without CSE and Jag-1. A Notch-1 inhibitor (DAPT) was used as control. The expression of Notch-1, Jag-1, Ki67, PCNA, p21, IL-33 and cell proliferation (by CFSE) were all assessed by flow cytometry. Notch-1 nuclear expression was evaluated by immunofluorescence and western blot analysis. Repair processes were assessed by wound assay. Smokers had cytoplasmic but not nuclear Notch-1 expression. Although CSE increased Notch-1 expression, it counteracted Notch-1 signaling activation since it reduced Jag-1 expression and Notch-1 nuclear translocation. Notch-1 signaling activation by Jag-1 increased Ki67, PCNA and repair processes but reduced intracellular IL-33 and p21 expression without affecting cell proliferation. DAPT counteracted the effects of Notch-1 activation on PCNA and IL-33. CSE increased Ki67, PCNA, p21 and IL-33 expression but reduced cell proliferation and repair processes. In conclusion, cigarette smoke exposure, limiting Notch-1 signaling activation and hindering repair processes, amplifies injury processes in bronchial epithelial cells.

Inflammation and cellular senescence (also called inflammaging) are involved in the pathogenesis of premature lung aging, a key driver of chronic obstructive pulmonary disease (COPD). Downregulation of histone deacetylases and FoxO3 expression, activation of the ERK 1/2 pathway and IL-8 increase are hallmarks of lung inflammaging. The effects of Budesonide (BUD), Aclidinium (ACL) and Formoterol (FO) on lung inflammaging are unknown. This study was aimed to assess the effects of BUD, ACL and FO in bronchial epithelial cells exposed to cigarette smoke extract (CSE) by evaluating: a) Expression of TLR4 and survivin and LPS binding by flow cytometry; b) expression of HDAC2, HDAC3, SIRT1 and FoxO3 and activation of the ERK 1/2 pathway by western blot; c) IL-8 mRNA levels and release by Real Time-PCR and ELISA, respectively. Reported results show that CSE increased TLR4 and survivin, LPS binding, ERK 1/2 activation, IL-8 release and mRNA levels but decreased SIRT1, HDAC2, HDAC3 and FoxO3 nuclear expression. Combined therapy with BUD, ACL and FO counteracted the effects of CSE on LPS binding, FoxO3 nuclear expression, ERK 1/2 activation, survivin and IL-8 release and mRNA levels. These findings suggest a new role of combination therapy with BUD, ACL and FO in counteracting inflammaging processes induced by cigarette smoke exposure.

BackgroundAlterations in the nasal epithelial barrier homeostasis and increased interleukin 33 (IL-33) expression contribute to the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP). AimsAs Notch-1 signaling is crucial in repair processes of mucosa, the current study assessed Notch-1/Jagged-1 signaling and IL-33 in the epithelium of nasal polyps biopsies from allergic (A-CRSwNP; n=9) and not allergic (NA-CRSwNP; n=9) subjects by immunohistochemistry. We also assessed, in a model of nasal epithelial cells, the effects of stimulation of Notch-1 with Jagged-1 on the expression of IL-33 (by flow cytometry, immunofluorescence, and immunocytochemistry), Jagged-1 (by flow cytometry), andp-CREB transcription factor (by western blot analysis). ResultsEx vivo(a) in normal epithelium, the expression of Notch-1 and IL-33 were higher in NA-CRSwNP than in A-CRSwNP; (b) in metaplastic epithelium, the expression of Notch-1, Jagged-1, and IL-33 were higher in NA-CRSwNP than in A-CRSwNP; (c) in hyperplastic epithelium, the expression of Notch-1, Jagged-1, and IL-33 were higher in A-CRSwNP than in NA-CRSwNP; and (d) in basal epithelial cells, no differences were observed in the expression of Jagged-1, IL-33, and Notch-1. The expression of Notch-1 significantly correlated with the expression of IL-33. In vitro, stimulation of Notch-1 with Jagged-1 induced the expression of (a) Jagged-1; (b) IL-33; and (c) p-CREB transcription factor. The inhibitor of Notch-1, DAPT, reduced all the effects of Jagged-1 on nasal epithelial cells. ConclusionsThe data herein provided support, for the first time, a putative role of Notch-1/Jagged-1 signaling in the overexpression of IL-33 in the epithelium of nasal polyps from patients with CRSwNP.

Background: Cigarette smoke is considered a risk factor for lung and colorectal cancer. A convincing link between epithelial-to-mesenchymal transition (EMT) with colorectal cancer progression and therapeutic resistance has emerged. Deregulated expression of E-Cadherin and Claudin-1 and increased miR-21 expression and invasiveness represent hallmarks of EMT. The effects of cigarette smoke exposure on EMT in colorectal adenocarcinoma cells are largely unknown. Aim: The aim of the study is to evaluate the effect of cigarette smoke extract (CSE) on miR-21, Claudin-1 and E-Cadherin, molecules associated to EMT in colorectal cancer cells. Methods: A human colorectal adenocarcinoma cell line (Caco-2) was treated with CSE at different concentration (5% and 10%) and for different time points (3 h and 24 h). Metabolic activity (by MTS assay), cell necrosis/cell apoptosis (evaluating Propidium Iodide/Annexin V expression by flow cytometry), miR-21, Claudin-1 and E-Cadherin gene expression were evaluated by Real time PCR. Cell permeability, actin polymerization and cancer cell migration was assessed by Trans-Epitelial Electrical Resistance (TEER), Phalloidin expression and matrigel system, respectively. Results: CSE at all the tested concentrations and at all time points reduced cell necrosis. CSE at 10% increased miR-21 and reduced the metabolic activity, cell necrosis, Claudin-1 and E-cadherin mRNA at 3 h. Cell permeability, actin polymerization and cancer cell migration were all increased upon CSE exposure. Conclusion: These results showed that CSE increasing miR-21, Claudin-1 and E-Cadherin and enhancing the aggressiveness of cancer cells, may concur to colorectal cancer progression.

Background: The addition of long-acting beta2-agonists (LABAs) to corticosteroids improves asthma control. Cigarette smoke exposure, increasing oxidative stress, may negatively affect corticosteroid responses. The anti-inflammatory effects of formoterol (FO) and fluticasone propionate (FP) in human bronchial epithelial cells exposed to cigarette smoke extracts (CSE) are unknown.Aims: This study explored whether FP, alone and in combination with FO, in human bronchial epithelial cell line (16-HBE) and primary bronchial epithelial cells (NHBE), counteracted some CSE-mediated effects and in particular some of the molecular mechanisms of corticosteroid resistance.Methods: 16-HBE and NHBE were stimulated with CSE, FP and FO alone or combined. HDAC3 and HDAC2 activity, nuclear translocation of GR and NF-kappa B, pERK1/2/tERK1/2 ratio, IL-8, TNF-alpha, IL-1 beta mRNA expression, and mitochondrial ROS were evaluated. Actin reorganization in neutrophils was assessed by fluorescence microscopy using the phalloidin method.Results: In 16-HBE, CSE decreased expression/activity of HDAC3, activity of HDAC2, nuclear translocation of GR and increased nuclear NF-kappa B expression, pERK 1/2/tERK1/2 ratio, and mRNA expression of inflammatory cytokines. In NHBE, CSE increased mRNA expression of inflammatory cytokines and supernatants from CSE exposed NHBE increased actin reorganization in neutrophils. FP combined with FO reverted all these phenomena in CSE stimulated 16-HBE cells as well as in NHBE cells.Conclusions: The present study provides compelling evidences that FP combined with FO may contribute to revert some processes related to steroid resistance induced by oxidative stress due to cigarette smoke exposure increasing the anti-inflammatory effects of FP.

Background: Cigarette smoke, the principal risk factor for chronic obstructive pulmonary disease (COPD), negatively influences the effectiveness of the immune system's response to a pathogen. The antibiotic ceftaroline exerts immune-modulatory effects in bronchial epithelial cells exposed to cigarette smoke.Aims and methods: The present study aims to assess the effects of ceftaroline on TLR2 and TLR4 expression, LPS binding and TNF-alpha and human beta defensin (HBD2) release in an undifferentiated and PMA-differentiated human monocyte cell line (THP-1) exposed or not to cigarette smoke extracts (CSE). TLR2, TLR4, and LPS binding were assessed by flow cytometry, TNF-alpha and HBD2 release were evaluated by ELISA.Results: The constitutive expression of TLR2 and TLR4 and LPS binding were higher in differentiated compared to undifferentiated THP-1 cells. In undifferentiated THP-1 cells, CSE increased TLR2 and TLR4 protein levels, LPS binding and TNF-a release and reduced HBD2 release and ceftaroline counteracted all these effects. In differentiated THP-1, CSE did not significantly affect TLR2 and TLR4 expression and LPS binding but reduced HBD2 release and increased TNF-a release. Ceftaroline counteracted the effects of CSE on HBD2 release in differentiated THP-1.Conclusion: Ceftaroline counteracts the effect of CSE in immune cells by increasing the effectiveness of the innate immune system. This effect may also assist in reducing pathogen activity and recurrent exacerbations in COPD patients.

Background: Cigarette smoke may accelerate cellular senescence by increasing oxidative stress. Altered proliferation and altered expression of anti-aging factors, including SIRT1 and FoxO3, characterise cellular senescence. The effects of carbocysteine on the SIRT1/FoxO3 axis and on downstream molecular mechanisms in human bronchial epithelial cells exposed to cigarette smoke are largely unknown.Aims: Aim of this study was to explore whether carbocysteine modulated SIRT1/FoxO3 axis, and downstream molecular mechanisms associated to cellular senescence, in a bronchial epithelial cell line (16-HBE) exposed to cigarette smoke.Methods: 16HBE cells were stimulated with/without cigarette smoke extracts (CSE) and carbocysteine. Flow cytometry and clonogenic assay were used to assess cell proliferation; western blot analysis was used for assessing nuclear expression of SIRT1 and FoxO3. The nuclear co-localization of SIRT1 and FoxO3 was assessed by fluorescence microscopy. Beta galactosidase (a senescence marker) and SIRT1 activity were assessed by specific staining and colorimetric assays, respectively. ChiP Assay and flow cytometry were used for assessing survivin gene regulation and protein expression, respectively.Results: CSE decreased cell proliferation, the nuclear expression of SIRT1 and FoxO3 and increased beta galactosidase staining. CSE, reduced SIRT1 activity and FoxO3 localization on survivin promoter thus increasing survivin expression. In CSE stimulated bronchial epithelial cells carbocysteine reverted these phenomena by increasing cell proliferation, and SIRT1 and FoxO3 nuclear expression, and by reducing beta galactosidase staining and survivin expression.Conclusions: The study shows for the first time that carbocysteine may revert some senescence processes induced by oxidative stress due to cigarette smoke exposure. (C) 2016 Elsevier Inc. All rights reserved.