15 research outputs found
Changing DNA damage in peripheral blood mononuclear cells by Comet Assay IV parameters from never-smoking control individuals through current smokers or ex-smokers with normal lung function and patients with stabile chronic obstructive pulmonary disease (COPD) of the Global Initiative for Chronic Obstructive Lung Disease (GOLD) stages I-IV to patients with a COPD exacerbation, evaluated with ordinal logistic regression analysis.
*<p>Adjusted for age, gender, body mass index, smoking history (pack-years), status of a current smoker and forced expiratory volume in one second % predicted. <sup>#</sup>Novel derived parameters.</p
Characteristics of the individuals involved in the measurement of DNA damage by Comet Assay IV and poly(ADP-ribose) polymerase (PARP) activity in peripheral blood mononuclear cells.
<p>Data are presented as mean ± SEM or n (%). <sup>*</sup>To test the equality of the data across the study groups, Kruskal-Wallis test and Pearson’s chi-square test was applied for numeric and nominal variables, respectively; in case of smoking-related variables, non-smoking controls were omitted from the analysis.</p
Characteristics of the individuals involved in the measurement of poly(ADP-ribose) polymerase (PARP)-1 mRNA expression in peripheral blood mononuclear cells.
<p>Data are presented as mean ± SEM or n (%). <sup>*</sup>To test the equality of the data across the study groups, Kruskal-Wallis test and Pearson’s chi-square test was applied for numeric and nominal variables, respectively; in case of smoking-related variables, non-smoking controls were omitted from the analysis.</p
The level of damage to DNA in peripheral blood mononuclear cells.
<p>a) Evaluation by comet assay. b) Tail migration/cell length ratio measured with Comet Assay IV. c) Tail length/cell length ratio measured with Comet Assay IV. The study groups are designated as follows: (1) non-smoking, non-obstructive control individuals (n = 7); (2) non-obstructive smokers (n = 4); (3) patients with mild chronic obstructive pulmonary disease (COPD) (n = 3); (4) patients with moderate COPD (n = 7); (5) patients with severe COPD (n = 5); (6) patients with very severe COPD (n = 4); (7) patients with COPD exacerbation requiring hospitalisation (n = 6). Presented as mean ± SEM.</p
Changing poly(ADP-ribose) polymerase (PARP) activity and PARP-1 mRNA expression in peripheral blood mononuclear cells from never-smoking control individuals through current smokers or ex-smokers with normal lung function and patients with stabile chronic obstructive pulmonary disease (COPD) of the Global Initiative for Chronic Obstructive Lung Disease (GOLD) stages I-IV to patients with a COPD exacerbation, evaluated with ordinal logistic regression analysis.
*<p>Adjusted for age, gender, body mass index, smoking history (pack-years), status of a current smoker and forced expiratory volume in one second % predicted.</p
Principal component analysis of the metabolic state of primary normal human bronchial epithelial cells cultivated in air-liquid interface after exposure to (A) e-cigarette liquid (ECL) (100 μM by nicotine) and (B) 10 μg/mL cigarette smoke condensate (CSC) (black solid lines).
<p>The time points at the arrow turns are 1, 2, 5, 7, and 13 h. Long dashes: Addition of 10 μM O-methyl-l-tyrosinyl-γ-l-glutamyl-l-cysteinylglycine to the cells having been exposed to ECL for 1 h (the arrow starts at 2 h time point), short dashes: Addition of 2 mM N-acetylcysteine to the cells having been exposed to ECL for 1 h (the arrow starts at 2 h time point). For each treatment and for any time point, n = 3 (gray circles). Error bars indicate standard errors of means. PComp1 and PComp2 are the two principal components describing the highest fraction of the effect of cells’ metabolome.</p
E-Cigarette Affects the Metabolome of Primary Normal Human Bronchial Epithelial Cells
<div><p>E-cigarettes are widely believed to be safer than conventional cigarettes and have been even suggested as aids for smoking cessation. However, while reasonable with some regards, this judgment is not yet supported by adequate biomedical research data. Since bronchial epithelial cells are the immediate target of inhaled toxicants, we hypothesized that exposure to e-cigarettes may affect the metabolome of human bronchial epithelial cells (HBEC) and that the changes are, at least in part, induced by oxidant-driven mechanisms. Therefore, we evaluated the effect of e-cigarette liquid (ECL) on the metabolome of HBEC and examined the potency of antioxidants to protect the cells. We assessed the changes of the intracellular metabolome upon treatment with ECL in comparison of the effect of cigarette smoke condensate (CSC) with mass spectrometry and principal component analysis on air-liquid interface model of normal HBEC. Thereafter, we evaluated the capability of the novel antioxidant tetrapeptide O-methyl-l-tyrosinyl-γ-l-glutamyl-l-cysteinylglycine (UPF1) to attenuate the effect of ECL. ECL caused a significant shift in the metabolome that gradually gained its maximum by the 5<sup>th</sup> hour and receded by the 7<sup>th</sup> hour. A second alteration followed at the 13<sup>th</sup> hour. Treatment with CSC caused a significant initial shift already by the 1<sup>st</sup> hour. ECL, but not CSC, significantly increased the concentrations of arginine, histidine, and xanthine. ECL, in parallel with CSC, increased the content of adenosine diphosphate and decreased that of three lipid species from the phosphatidylcholine family. UPF1 partially counteracted the ECL-induced deviations, UPF1’s maximum effect occurred at the 5<sup>th</sup> hour. The data support our hypothesis that ECL profoundly alters the metabolome of HBEC in a manner, which is comparable and partially overlapping with the effect of CSC. Hence, our results do not support the concept of harmlessness of e-cigarettes.</p></div
Mass spectrum of primary normal human bronchial epithelial cells cultivated in air-liquid interface after exposure to e-cigarette liquid (ECL) (100 μM by nicotine) and 10 μg/mL cigarette smoke condensate (CSC) consisting of 1,822 distinct mass-to-charge signals.
<p>The figure represents the proportions of the spectrum that were significantly (p<i><</i>0.05) affected by addition of ECL (392 signals) or CSC (569 signals) during the first 7 h. There were 138 signals that were significantly affected by both stimuli.</p
The numbers of mass spectrometry signals (mass-to-charge ratios) from the lysate of primary normal human bronchial epithelial cells cultured in air-liquid interface significantly (p<i><</i>0.05) altered by e-cigarette liquid (ECL) (100 μM by nicotine) or 10 μg/mL cigarette smoke condensate (CSC) at the respective time points.
<p>The numbers of signals in case of which 10 μM O-methyl-l-tyrosinyl-γ-l-glutamyl-l-cysteinylglycine (UPF1)/2 mM N-acetylcysteine (NAC) significantly reduced the effect of the respective stimuli are shown in parentheses.</p><p>The numbers of mass spectrometry signals (mass-to-charge ratios) from the lysate of primary normal human bronchial epithelial cells cultured in air-liquid interface significantly (p<i><</i>0.05) altered by e-cigarette liquid (ECL) (100 μM by nicotine) or 10 μg/mL cigarette smoke condensate (CSC) at the respective time points.</p
Time dynamics of the expression of the metabolites of primary normal human bronchial epithelial cells (HBEC) cultured in air-liquid interface being affected by e-cigarette liquid (ECL) (100 μM by nicotine) (black lines) and 10 μg/mL cigarette smoke condensate (CSC) (grey lines) for 13 h.
<p>Solid lines: HBEC treated with ECL (solid black lines) or with CSC (solid grey lines). Long black dashes: HBEC treated with ECL and 10 μM O-methyl-l-tyrosinyl-γ-l-glutamyl-l-cysteinylglycine (UPF1) (added at 1h). Long grey dashes: HBEC treated with CSC and 10 μM UPF1 (added at 1h). Short black dashes: HBEC treated with ECL and 2 mM N-acetylcysteine (NAC) (added at 1h). Short grey dashes: HBEC treated with CSC and 2 mM NAC (added at 1h). For each treatment and for any time point, n = 3. Error bars indicate standard errors of means. (A) arginine ([M+H]<sup>+</sup> = 175); (B) adenosine diphosphate ([M-H]<sup>-</sup> = 426); (C) phosphatidylcholine (18:0/20:4) ([M+H]<sup>+</sup> = 811); (D) α-ketoglutarate ([M-H]<sup>-</sup> = 145). *p < 0.05, ECL-exposed cells versus untreated cells; #p < 0.05, ECL- and UPF1-exposed cells versus untreated cells; ^p < 0.05, ECL- and NAC-exposed cells versus untreated cells; ¤p < 0.05, CSC-exposed cells versus untreated cells; ~p < 0.05, CSC- and UPF1-exposed cells versus untreated cells; “p < 0.05, CSC- and NAC-exposed cells versus untreated cells.</p