Smoking Subjects Research Articles

Beneficial effects of omega-3 fatty acids in the proteome of high-density lipoprotein proteome

BackgroundOmega-3 poly-unsaturated fatty acids (ω-3 PUFAs) have demonstrated to be beneficial in the prevention of cardiovascular disease, however, the mechanisms by which they perform their cardiovascular protection have not been clarified. Intriguingly, some of these protective effects have also been linked to HDL. The hypothesis of this study was that ω-3 PUFAs could modify the protein cargo of HDL particle in a triglyceride non-dependent mode. The objective of the study was to compare the proteome of HDL before and after ω-3 PUFAs supplemented diet.MethodsA comparative proteomic analysis in 6 smoker subjects HDL before and after a 5 weeks ω-3 PUFAs enriched diet has been performed.ResultsAmong the altered proteins, clusterin, paraoxonase, and apoAI were found to increase, while fibronectin, α-1-antitrypsin, complement C1r subcomponent and complement factor H decreased after diet supplementation with ω-3 PUFAs. Immunodetection assays confirmed these results. The up-regulated proteins are related to anti-oxidant, anti-inflammatory and anti-atherosclerotic properties of HDL, while the down-regulated proteins are related to regulation of complement activation and acute phase response.ConclusionsDespite the low number of subjects included in the study, our findings demonstrate that ω-3 PUFAs supplementation modifies lipoprotein containing apoAI (LpAI) proteome and suggest that these protein changes improve the functionality of the particle.

Association of SNP Rs6903956 on Chromosome 6p24.1 with Angiographical Characteristics of Coronary Atherosclerosis in a Chinese Population

ObjectiveTo explore the association between rs6903956 and severity of coronary artery disease (CAD) in a Chinese population.MethodsA cohort of 1075 consecutive patients who underwent coronary arteriography for suspected or known coronary atherosclerosis was enrolled in our study. Coronary atherosclerosis severity was defined by Gensini's Score System and counts of diseased vessels.ResultsGensini score frequencies and counts of diseased vessels differed among GG, AG, AA genotype groups at the rs6903956 locus (p = 0.025 for Gensini score frequencies vs. p = 0.024 for counts of diseased vessels, respectively). A univariate logistic regression analysis revealed that the genotype distribution of this SNP was associated significantly with angiographical characteristics of coronary atherosclerosis risk (p = 0.030, odds ratio (OR) = 1.444, 95% confidence interval (CI) = 1.036∼2.013 for AG vs. GG; p = 0.021, OR = 5.896, 95% CI = 1.299∼26.750 for AA vs. GG and p = 0.007, OR = 1.564, 95% CI = 1.132∼2.162 for combined (AG+AA) vs. GG). A multivariate logistic regression analysis indicated that the genotype distribution of the rs6903956 polymorphism be associated significantly with the angiographical characteristics of coronary atherosclerosis risk (p = 0.004, OR = 1.578, 95% CI = 1.155∼2.154 for GG vs. AG vs. AA; p = 0.013, OR = 1.541, 95% CI = 1.097∼2.163 for GG vs. GA+ AA). A stratification analysis revealed that male subjects and smoking subjects had a higher frequency of the rs6903956 heterozygous mutant among higher Gensini score subjects than among lower Gensini score subjects (p = 0.023, OR = 1.579, 95% CI = 1.064∼2.344 for male subgroup; p = 0.005, OR = 2.075, 95% CI = 1.249∼3.448 for smoking subgroup).ConclusionsAllele A is a risk factor for CAD and the G-to-A allele substitution may underlie the association between rs6903956 and CAD.

Structural alterations of foreskin caused by chronic smoking may explain high levels of urethral reconstruction failure using foreskin flaps

The aim of the present study was to perform a stereological and biochemical analysis of the foreskin of smoker subjects. Foreskin samples were obtained from 20 young adults (mean = 27.2 years old) submitted to circumcision. Of the patients analyzed, one group (n = 10) had previous history of chronic smoking (a half pack to 3 packs per day for 3 to 13 years (mean = 5.8 ± 3.2). The control group included 10 nonsmoking patients. Masson 's trichrome stain was used to quantify the foreskin vascular density. Weigert's resorcin-fucsin stain was used to assess the elastic system fibers and Picrosirius red stain was applied to study the collagen. Stereological analysis was performed using the Image J software to determine the volumetric densities. For biochemical analysis, the total collagen was determined as µg of hydroxyproline per mg of dry tissue. Means were compared using the unpaired t-test (p < 0.05). Elastic system fibers of smokers was 42.5 % higher than in the control group (p = 0.002). In contrast, smooth muscle fibers (p = 0.42) and vascular density (p = 0.16) did not show any significant variation. Qualitative analysis using Picrosirius red stain with polarized light evidenced the presence of type I and III collagen in the foreskin tissue, without significant difference between the groups. Total collagen concentration also did not differ significantly between smokers and non-smokers (73.1 µg/mg ± 8.0 vs. 69.2 µg/mg ± 5.9, respectively, p = 0.23). The foreskin tissue of smoking patients had a significant increase of elastic system fibers. Elastic fibers play an important role in this tissue's turnover and this high concentration in smokers possibly causes high extensibility of the foreskin. The structural alterations in smokers' foreskins could possibly explain the poor results in smoking patients submitted to foreskin fasciocutaneous flaps in urethral reconstruction surgery.

Analysis of aberrant methylation on promoter sequences of tumor suppressor genes and total DNA in sputum samples: a promising tool for early detection of COPD and lung cancer in smokers

BackgroundChronic obstructive pulmonary disease (COPD) is a disorder associated to cigarette smoke and lung cancer (LC). Since epigenetic changes in oncogenes and tumor suppressor genes (TSGs) are clearly important in the development of LC. In this study, we hypothesize that tobacco smokers are susceptible for methylation in the promoter region of TSGs in airway epithelial cells when compared with non-smoker subjects. The purpose of this study was to investigate the usefulness of detection of genes promoter methylation in sputum specimens, as a complementary tool to identify LC biomarkers among smokers with early COPD.MethodsWe determined the amount of DNA in induced sputum from patients with COPD (n = 23), LC (n = 26), as well as in healthy subjects (CTR) (n = 33), using a commercial kit for DNA purification, followed by absorbance measurement at 260 nm. The frequency of CDKN2A, CDH1 and MGMT promoter methylation in the same groups was determined by methylation-specific polymerase chain reaction (MSP). The Fisher’s exact test was employed to compare frequency of results between different groups.ResultsDNA concentration was 7.4 and 5.8 times higher in LC and COPD compared to the (CTR) (p < 0.0001), respectively. Methylation status of CDKN2A and MGMT was significantly higher in COPD and LC patients compared with CTR group (p < 0.0001). Frequency of CDH1 methylation only showed a statistically significant difference between LC patients and CTR group (p < 0.05).ConclusionsWe provide evidence that aberrant methylation of TSGs in samples of induced sputum is a useful tool for early diagnostic of lung diseases (LC and COPD) in smoker subjects.Virtual slidesThe abstract MUST finish with the following text: Virtual Slides The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1127865005664160

Prediction Equations for Single-Breath Diffusing Capacity in Subjects Aged 65 to 85 Years

In senior subjects, diffusing capacity of the lung for carbon monoxide (Dlco) is interpreted using prediction equations derived from primarily younger adult populations. Our objectives were to provide reference equations for single-breath Dlco for a cohort of healthy, never-smoking, white, European adults between 65 and 85 years of age and to compare the predicted values of this sample with those from other studies involving middle-aged adults. Reference equations were derived from a randomly selected sample from the general population of 431 healthy never smoker subjects aged 65 to 85 years (262 women and 169 men). Spirometry, lung volume determinations by plethysmography, and single-breath Dlco (corrected for hemoglobin) were performed following the American Thoracic Society/European Respiratory Society guidelines. Reference values and lower and upper limits of normal were derived using a piecewise polynomial model. In addition to age, our reference equations confirmed the height and body size dependence of Dlco and diffusing capacity for alveolar volume (Dlco/Va) in older subjects. Practically all of the reference values obtained by extrapolating reference equations of middle-aged adults underestimated the true diffusing capacity of the healthy elderly volunteers. Middle-aged reference equations underestimated Dlco by 2.1% to 22.3% in women and 2.8% to 37.8% in men. In addition, Dlco/Va was overestimated up to 18% and 39.8% in women and men, respectively, whereas other equations underestimated Dlco/Va up to 22.2% and 11.9% in women and men, respectively. These results underscore the importance of using prediction equations appropriate to the origin and age characteristics of the subjects being studied.

Extended NO analysis in health and disease

Extended NO analysis is a promising tool in different diseases where NO metabolism is altered. One single exhalation cannot give insight to the NO production in the respiratory system; rather the use of multiple exhalation flows can give the alveolar levels (CANO), airway wall concentration (CawNO) and the diffusion rate of NO (DawNO). Increased values of CANO are shown in COPD, systemic sclerosis, hepatopulmonary syndrome and in severe asthma. In asthma the CawNO and DawNO are increased leading to an increase in bronchial NO flux (J′awNO). Low levels of J′awNO are seen in cystic fibrosis, primary ciliary dyskinesia and in smoking subjects. More studies are needed to evaluate the clinical usefulness of the extended NO analysis, similar to what has been done in systemic sclerosis where a cut-off value has been identified predicting pulmonary function deterioration.

Extracellular matrix composition in COPD

Extracellular matrix (ECM) composition has an important role in determining airway structure. We postulated that ECM lung composition of chronic obstructive pulmonary disease (COPD) patients differs from that observed in smoking and nonsmoking subjects without airflow obstruction. We determined the fractional areas of elastic fibres, type-I, -III and -IV collagen, versican, decorin, biglycan, lumican, fibronectin and tenascin in different compartments of the large and small airways and lung parenchyma in 26 COPD patients, 26 smokers without COPD and 16 nonsmoking control subjects. The fractional area of elastic fibres was higher in non-obstructed smokers than in COPD and nonsmoking controls, in all lung compartments. Type-I collagen fractional area was lower in the large and small airways of COPD patients and in the small airways of non-obstructed smokers than in nonsmokers. Compared with nonsmokers, COPD patients had lower versican fractional area in the parenchyma, higher fibronectin fractional area in small airways and higher tenascin fractional area in large and small airways compartments. In COPD patients, significant correlations were found between elastic fibres and fibronectin and lung function parameters. Alterations of the major ECM components are widespread in all lung compartments of patients with COPD and may contribute to persistent airflow obstruction.

Antielastin B-cell and T-cell immunity in patients with chronic obstructive pulmonary disease

RationaleAntielastin autoimmunity has been hypothesised to drive disease progression in chronic obstructive pulmonary disease (COPD). The proposed mechanism is currently disputed by conflicting data. The authors aimed to explore antibody...

Association of Mitochondrial DNA Variations with Lung Cancer Risk in a Han Chinese Population from Southwestern China

Mitochondrial DNA (mtDNA) is particularly susceptible to oxidative damage and mutation due to the high rate of reactive oxygen species (ROS) production and limited DNA-repair capacity in mitochondrial. Previous studies demonstrated that the increased mtDNA copy number for compensation for damage, which was associated with cigarette smoking, has been found to be associated with lung cancer risk among heavy smokers. Given that the common and “non-pathological” mtDNA variations determine differences in oxidative phosphorylation performance and ROS production, an important determinant of lung cancer risk, we hypothesize that the mtDNA variations may play roles in lung cancer risk. To test this hypothesis, we conducted a case-control study to compare the frequencies of mtDNA haplogroups and an 822 bp mtDNA deletion between 422 lung cancer patients and 504 controls. Multivariate logistic regression analysis revealed that haplogroups D and F were related to individual lung cancer resistance (OR = 0.465, 95%CI = 0.329–0.656, p<0.001; and OR = 0.622, 95%CI = 0.425–0.909, p = 0.014, respectively), while haplogroups G and M7 might be risk factors for lung cancer (OR = 3.924, 95%CI = 1.757–6.689, p<0.001; and OR = 2.037, 95%CI = 1.253–3.312, p = 0.004, respectively). Additionally, multivariate logistic regression analysis revealed that cigarette smoking was a risk factor for the 822 bp mtDNA deletion. Furthermore, the increased frequencies of the mtDNA deletion in male cigarette smoking subjects of combined cases and controls with haplogroup D indicated that the haplogroup D might be susceptible to DNA damage from external ROS caused by heavy cigarette smoking.

Interactions between CYP1A1 polymorphisms and cigarette smoking are associated with the risk of hepatocellular carcinoma: evidence from epidemiological studies

The Cytochrome P-450 1A1 (CYP1A1) gene has been implicated in the etiology of hepatocellular carcinoma (HCC). However, the results have been inconsistent. In this study, a meta-analysis was performed to clarify the associations of polymorphisms in CYP1A1 gene with HCC risk. Published literature from PubMed, Embase, CNKI and Wanfang Data were retrieved. Pooled odds ratio (OR) with 95% confidence interval (CI) was calculated using fixed- or random-effects model. Eight studies (1,752 cases and 2,279 controls) for Ile-Val polymorphism and eight studies (933 cases and 1,449 controls) for MspI polymorphism were identified. The results showed that there was no statistically significant association between the Ile-Val polymorphism and HCC risk under all genetic models (co-dominant model: Val/Val vs. Ile/Ile: OR = 1.62, 95% CI 0.96-2.72 and Ile/Val vs. Ile/Ile: OR = 1.15, 95% CI 0.87-1.52; dominant model: OR = 1.25, 95% CI 0.92-1.70; recessive model: OR = 1.48, 95% CI 0.99-2.21). The MspI polymorphism was also not associated with HCC risk (co-dominant model: m2m2 vs. m1m1: OR = 1.09, 95% CI 0.83-1.42 and m1m2 vs. m1m1: OR = 1.30, 95% CI 1.05-1.61; dominant model: OR = 1.20, 95% CI 0.99–1.45; recessive model: OR = 0.94, 95% CI 0.74-1.18). However, the significant associations were found between both the Ile–Val and MspI polymorphisms and HCC risk among the cigarette smoking subjects (Ile-Val: OR = 1.40, 95% CI 1.06-1.85; MspI: OR = 2.65, 95% CI 1.47-4.77). The present meta-analysis indicated that the MspI and Ile-Val polymorphisms of CYP1A1 may play important roles in increasing susceptibility to smoking-related HCC.

Impact of Low-and High-Density Lipoprotein Cholesterol Levels on Carotid Intima-Media Thickness Differs by Smoking Status in Middle-Aged Men

Cigarette smoking is a strong risk factor for atherosclerotic disease; however, it remains unclear whether the impact of other risk factors differs by smoking status. The aim of this study was to investigate this issue, especially with regard to low-density and high-density lipoprotein (LDL/HDL) levels. In total, 448 healthy, middle-aged men (aged 37 to 61) participated in this study. Smoking habits were recorded, carotid intima-media thickness (IMT) was measured by B-mode ultrasound, and serum lipids and other biochemical parameters were determined from fasted blood samples. Among the overall subjects, multivariate regression analyses showed that IMT was significantly associated with age (p < 0.0001 for mean IMT, p= 0.002 for max IMT), body mass index (BMI, mean IMT, p= 0.028), LDL-C levels (mean/max IMT, p= 0.001), HDL-C levels (max IMT, p= 0.022) and current smoking habit (mean IMT, p=0.012). Subgroup analyses according to smoking status revealed that LDL-C levels were significantly associated with mean/max IMT in current smokers (p=0.001) but not in ex- or nonsmokers (never smoked subjects). After adjusting for age, BMI, systolic blood pressure, hemoglobin A1c and serum lipids, mean IMT respectively increased and decreased progressively across LDL-C and HDL-C quartiles (p= 0.004 and 0.045) in the overall subjects. These associations were observed in current smokers (p= 0.01) but not in ex- or nonsmokers for LDL-C, and were observed in ex- and nonsmokers (p= 0.025, 0.017, respectively) but not in current smokers for HDL-C. The impact of LDL-C/HDL-C levels on carotid IMT differs by smoking status. These observations imply that distinct mechanisms are involved in the (anti) atherogenesis of LDL/HDL according to smoking status.

Non-Invasive Biomarkers of Lung Inflammation in Smoking Subjects

Cigarette smoking is the most important risk factor for the development of chronic obstructive pulmonary disease (COPD) and lung cancer, but only a part of smoking subjects develop these respiratory pathologies. Therefore, it is necessary to find sensible parameters to detect early lung alterations due to chronic tobacco smoke exposure. Long-term cigarette smoking is associated with a persistent inflammatory response in the lung that leads to tissue injury and dysfunction. Bronchoscopy and bronchial biopsies are the gold standard techniques for assessing pulmonary inflammation, but are invasive and not routinely used. Cellular analysis of induced sputum and measurement of fraction of exhaled nitric oxide (F(E)NO) are validated non-invasive techniques for assessing respiratory inflammation. Measurement of biomolecules in sputum supernatants and exhaled breath condensate (EBC) are used as a research tool, but require standardization of procedures and, generally, analytical validation. Electronic nose differentiates healthy smokers from healthy nonsmokers based on breath volatile organic compounds (VOC) patterns. These techniques are potentially useful for identifying biomarkers of pulmonary inflammation and oxidative stress. Induced sputum, F(E)NO, EBC and electronic nose are suitable for longitudinal sampling, thereby facilitating monitoring of lung damage process. This approach could enable an early identification of subgroups of healthy smokers at higher risk for tobacco-induced lung damage and prompt planning of secondary prevention strategies.

High serum peroxynitrite level is an early effect of chronic cigarette smoking

Background: Nicotine dependence is a chronic and relapsing disease. Oxidant components of cigarette smoke reduce nitric oxide (NO) production and/or inhibit its function. Objectives: The aim of this study was to assess serum NO, peroxynitrite ONOO and C-reactive protein in smoker individuals using the Fagerström test for nicotine dependence. Materials and Methods: A total number of 50 male smoker subjects and 20 non-smoker subjects serving as controls were recruited in this study. The Fagerström test of nicotine dependence was used to assess the level of nicotine dependence. Venous blood was obtained from each subject and serum was separated for the following tests: C-reactive protein, NO and ONOO. Results: The mean values of C-reactive proteins in the smokers were less than 6 mg/L and there was no significant difference in the C-reactive protein level between levels of nicotine dependence according to the Fagerström test scoring. A non-significant low serum NO level was observed in smokers, which did not correlate with C-reactive protein. A significant high serum ONOO level was observed in smokers compared with non-smoker subjects, which did not correlate with C-reactive protein. Conclusions: The study concludes that serum ONOO level may serve as an initial marker to point the impact of smoking on human beings. Its elevation preceded the decline in serum NO level and the evidence of inflammation.

S52 Association of microtubule instability with defective phagocytosis in COPD

Acute exacerbations of COPD are the commonest cause of acute medical admissions in the UK and ∼50% are associated with bacterial infection. Alveolar macrophages (AM) normally clear inhaled bacteria but...

Epidemiology, radiology, and genetics of nicotine dependence in COPD

BackgroundCigarette smoking is the principal environmental risk factor for developing COPD, and nicotine dependence strongly influences smoking behavior. This study was performed to elucidate the relationship between nicotine dependence, genetic susceptibility to nicotine dependence, and volumetric CT findings in smokers.MethodsCurrent smokers with COPD (GOLD stage ≥ 2) or normal spirometry were analyzed from the COPDGene Study, a prospective observational study. Nicotine dependence was determined by the Fagerstrom test for nicotine dependence (FTND). Volumetric CT acquisitions measuring the percent of emphysema on inspiratory CT (% of lung <-950 HU) and gas trapping on expiratory CT (% of lung <-856 HU) were obtained. Genotypes for two SNPs in the CHRNA3/5 region (rs8034191, rs1051730) previously associated with nicotine dependence and COPD were analyzed for association to COPD and nicotine dependence phenotypes.ResultsAmong 842 currently smoking subjects (335 COPD cases and 507 controls), 329 subjects (39.1%) showed high nicotine dependence. Subjects with high nicotine dependence had greater cumulative and current amounts of smoking. However, emphysema severity was negatively correlated with the FTND score in controls (ρ = -0.19, p < .0001) as well as in COPD cases (ρ = -0.18, p = 0.0008). Lower FTND score, male gender, lower body mass index, and lower FEV1 were independent risk factors for emphysema severity in COPD cases. Both CHRNA3/5 SNPs were associated with FTND in current smokers. An association of genetic variants in CHRNA3/5 with severity of emphysema was only found in former smokers, but not in current smokers.ConclusionsNicotine dependence was a negative predictor for emphysema on CT in COPD and control smokers. Increased inflammation in more highly addicted current smokers could influence the CT lung density distribution, which may influence genetic association studies of emphysema phenotypes.Trial registrationClinicalTrials (NCT): NCT00608764

The clinical features of the overlap between COPD and asthma

BackgroundThe coexistence of COPD and asthma is widely recognized but has not been well described. This study characterizes clinical features, spirometry, and chest CT scans of smoking subjects with both COPD and asthma.MethodsWe performed a cross-sectional study comparing subjects with COPD and asthma to subjects with COPD alone in the COPDGene Study.Results119 (13%) of 915 subjects with COPD reported a history of physician-diagnosed asthma. These subjects were younger (61.3 vs 64.7 years old, p = 0.0001) with lower lifetime smoking intensity (43.7 vs 55.1 pack years, p = 0.0001). More African-Americans reported a history of asthma (33.6% vs 15.6%, p < 0.0001). Subjects with COPD and asthma demonstrated worse disease-related quality of life, were more likely to have had a severe COPD exacerbation in the past year, and were more likely to experience frequent exacerbations (OR 3.55 [2.19, 5.75], p < 0.0001). Subjects with COPD and asthma demonstrated greater gas-trapping on chest CT. There were no differences in spirometry or CT measurements of emphysema or airway wall thickness.ConclusionSubjects with COPD and asthma represent a relevant clinical population, with worse health-related quality of life. They experience more frequent and severe respiratory exacerbations despite younger age and reduced lifetime smoking history.Trial registrationClinicalTrials.gov: NCT00608764

Comparison of Mouth Level Exposure to ‘Tar’ and Nicotine in Smokers of Normal and Superslim King Size Cigarettes in Romania

Abstract Filter analysis methodology can be used to estimate smokers’ mouth level exposure (MLE) to ‘tar’ and nicotine from spent cigarette filters. Variation in smoking behaviour and cigarette design are known to have an impact on the range of ‘tar’ and nicotine MLE. Most ‘King Size’ (KS) cigarettes have a circumference of about 25 mm and are 83-84 mm long, with filters 20-27 mm in length. Recently, a slimmer format, described as a ‘King Size Superslim’ (KSSS), with a circumference of 17 mm, has become popular in several countries. Although several studies have estimated the MLE to ‘tar’ and nicotine of smokers of KS cigarettes, there have been no studies of KSSS smokers reported to date. KSSS cigarettes from Romania were chosen for this study with a single product to represent each 1, 4 and 7 mg machine-smoked ISO ‘tar’ yields. Since these products have filters containing activated carbon, comparisons were made with a single product at each ISO ‘tar’ level of KS cigarettes with both activated carbon filters (King Size Carbon - KSC), to assess the effect of format on MLE, and cellulose acetate filters (King Size Non-carbon - KSNC) to assess the effect of filter carbon on MLE. A target population of 50 to 70 male and female smokers (subjects) aged 21 to 50 years of each product were recruited in Romania, with self-reported consumption of 15-25 cigarettes per day. The subjects gave written informed consent before commencing the study. Mean MLE to ‘tar’ and nicotine obtained by the smokers of all products were greater than ISO pack ‘tar’ and nicotine as reported in numerous studies. Regarding MLE to ‘tar’, there were no significant differences between the KSC, KSNC and KSSS smokers within each of the 1, 4 and 7 mg machine-smoked ISO ‘tar’ yield groups, but there were significant differences between the ISO pack ‘tar’ groups: 1 mg &lt; 4 mg &lt; 7 mg. Regarding MLE to nicotine, KSC and KSSS smokers obtained similar yields within an ISO pack ‘tar’ group. KSNC smokers obtained lower MLE nicotine yields than the KSC and KSSS smokers at both 4 mg and 7 mg ISO pack ‘tar’, corresponding to lower blend nicotine levels. No gender differences were observed.

Genotoxic effects of chlorpyrifos, cypermethrin, endosulfan and 2,4‐D on human peripheral lymphocytes cultured from smokers and nonsmokers

Pesticides often cause environmental pollution and adverse effects on human health. We have chosen four structurally different pesticides (endosulfan, an organochlorine pesticide; chlorpyrifos, an organophosphate insecticide; cypermethrin, type II pyrethroid insecticide, and 2,4-dichlorophenoxyacetic acid, a chlorinated aromatic hydrocarbon acid pesticide) to examine and compare their effects on DNA damage in acutely cultured human lymphocytes by the comet assay. In addition, possible differences in response between smoking and nonsmoking subjects were also investigated. Venous blood samples were obtained from healthy male nonsmoker (n = 7) and smoker (n = 8) donors. Primary cultures of lymphocytes were prepared and test groups were treated with three different concentrations (1, 5, and 10 μM) of endosulfan, chlorpyrifos, cypermehrin, and 2,4-D. DNA damage was assessed by alkaline comet assay. We determined an increase in the ratio of DNA migration in human lymphocyte cell cultures as a result of treatment with cypermethrin, 2,4-D and chlorpyrifos at high concentration. Endosulfan had no significant genotoxic effect even at 10 μM concentration. We suggest that chlorpyrifos and cypermethrin are more potentially genotoxic than endosulfan and 2,4-D. Our findings also indicate that the only significant DNA damage between smokers and nonsmokers was observed in the 2,4-D-treated group.

Contrasting behavioral effects of acute nicotine and chronic smoking in detoxified alcoholics

Background Current literature suggests that acute nicotine administration provides a compensatory mechanism by which alcoholics might alleviate attentional deficits. In contrast, chronic smoking is increasingly recognized as negatively affecting neurobehavioral integrity. These opposing effects have not been simultaneously examined. Thus, we sought to a) extend previous work by exploring the effects of acute nicotine effects on vigilance components of attention and replicate previous findings suggesting that treatment-seeking alcoholics experience benefit to a greater extent than do other groups; and b) to examine the impact of chronic smoking on these tasks and across subgroups. Methods Substance abusing participants (N = 86) were recruited and subgrouped on the basis of dependency criteria as either alcoholics, alcoholics with co-morbid stimulant dependence, or stimulant dependent individuals. Groups of cigarette-smoking (N = 17) and non-smoking (N = 22) community controls were recruited as comparison groups. Smoking subjects were assigned a placebo, low, or high dose nicotine patch in a double-blind placebo controlled fashion. Non-smoking controls were administered either a placebo or low dose. Testing occurred after dose stabilization. Results General linear models indicated greater sensitivity to acute nicotine administration among alcoholics than other groups when controlling for the effect of intensity of smoking history, as reflected by pack-years. Pack-years correlated negatively with performance measures in alcoholics but not stimulant abusing subgroups or smoking controls. Finally, regression analyses demonstrated that pack-years predicted poorer performance only for the alcoholic subgroup. Conclusions These results support previous work finding a compensatory effect of acute nicotine administration on attentional performance in alcoholics and reinforce the consideration of recent nicotine use as a confound in neurocognitive studies of alcoholics. Of particular interest is the finding that smoking history as reflected in pack-years predicted poorer performance, but only among alcoholics. Further systematic study of these opposing effects among alcoholics and other groups using a broader array of tasks is needed.

RETRACTED: In vitro anticholinergic drugs affect CD8+ peripheral blood T-cells apoptosis in COPD

Novel pharmacological strategies are aimed at the resolution of systemic inflammation in COPD potentiating peripheral blood T-cell (PBT-cell) apoptosis. Although muscarinic acetylcholine receptors (mAChRs) M3 and choline-acetyltransferase (ChAT) participate in the airway inflammation of COPD, their role in PBT-cell apoptosis remains unexplained. We evaluated in PBT-cells from COPD patients, smoker (S) and control (C) subjects: (1) apoptosis (by annexin V binding), (2) mAChR M3 and ChAT expression, acetylcholine (ACh)-binding; (3) choline levels in serum and PBT-cells extracts. We tested the effects of Tiotropium (Spiriva®) and hemicholinium-3 (HCh-3) on apoptosis, NFκB pathway, caspases 3 and 8 activity and choline levels, in PBT-cells from COPD patients. We showed that: (1) apoptosis, mAChR M3 and ChAT expression and the CD3+ and CD8+ ACh-binding are increased in PBT-cells from COPD patients when compared to C subjects, while CD4+/CD8+ ratio of ACh-binding to PBT cells was reduced in COPD; (2) choline levels are higher in serum and PBT-cells extracts from COPD patients than in S and C; (3) Tiotropium and HCh-3 reduced CD4+ and increased CD8+ apoptosis via caspases 3 and 8 activities and via IκB mediated mechanisms in COPD patients. This study suggests the involvement of non-neuronal components of cholinergic system in the regulation of PBT-cell apoptosis in COPD and demonstrates that Tiotropium regulates CD4+ and CD8+ PBT-cell apoptosis. It provides novel putative pharmacological targets for the resolution of systemic inflammation in COPD.