Smokers Research Articles

Cigarette Smoking Impairs Cardiorespiratory and Metabolic Response at Peak Incremental Exercise and during Recovery in Young, Physically Active Adults

ABSTRACT Purpose: Cigarette smoking (CS) induces systemic changes that impair cardiorespiratory and muscular function both at rest and during exercise. Although these abnormalities are reported in sedentary, middle-aged smokers (SM) with pulmonary disease, few and controversial studies focused on young, physically active SM at the early stage of smoking history. This study aimed at assessing the impact CS on cardiorespiratory and metabolic response during an incremental test and the subsequent recovery in young, physically active SM without known lung or cardiovascular disease. Methods: After pulmonary function evaluation, 12 SM (age: 22±2 yr; body mass: 75±8 kg; stature: 1.78±0.06 m; 12±4 cigarette/day for 6±2 years; mean±SD) and 12 non-smokers (CTRL; age: 23±1 yr; body mass: 76±8 kg; stature: 1.79±0.08 m) matched for age and exercise habits, underwent an exhaustive incremental step test (25 W/2 min) on a cycle ergometer. Pulmonary O2 uptake (V’O2), expiratory ventilation (V’E), heart rate (f H) responses and lactate concentration were assessed during the test and subsequent recovery. Results: Despite similar static lung volumes, SM reported lower peak expiratory flow (-23%; P = 0.003) and maximal voluntary ventilation (-10%; P = 0.003). At submaximal exercise, no differences in the cardiorespiratory and metabolic were noted between the two groups. However, SM exhibited ventilatory (P < 0.01) and lactate thresholds at lower work rates (P = 0.01). At peak exercise, SM exhibited lower V’O2 (-8%; P = 0.02), mechanical power (-11%; P = 0.02) and V’E (-9%; P = 0.01). During recovery, SM showed longer time constants (τ) in V’O2 (+52%; P = 0.002), V’E (+19%; P = 0.027) and f H (+21%; P = 0.022) and smaller f H at 30 s of recovery (HRR30; -31%; P = 0.032). Conclusions: These results are compatible with an early CS-related impairment of the cardiorespiratory and metabolic function even in young individuals with relatively short smoking history.

A Pilot Cross-Sectional Study of Immunological and Microbiome Profiling Reveals Distinct Inflammatory Profiles for Smokers, Electronic Cigarette Users, and Never-Smokers.

Smokers (SM) have increased lung immune cell counts and inflammatory gene expression compared to electronic cigarette (EC) users and never-smokers (NS). The objective of this study is to further assess associations for SM and EC lung microbiomes with immune cell subtypes and inflammatory gene expression in samples obtained by bronchoscopy and bronchoalveolar lavage (n = 28). RNASeq with the CIBERSORT computational algorithm were used to determine immune cell subtypes, along with inflammatory gene expression and microbiome metatranscriptomics. Macrophage subtypes revealed a two-fold increase in M0 (undifferentiated) macrophages for SM and EC users relative to NS, with a concordant decrease in M2 (anti-inflammatory) macrophages. There were 68, 19, and 1 significantly differentially expressed inflammatory genes (DEG) between SM/NS, SM/EC users, and EC users/NS, respectively. CSF-1 and GATA3 expression correlated positively and inversely with M0 and M2 macrophages, respectively. Correlation profiling for DEG showed distinct lung profiles for each participant group. There were three bacteria genera-DEG correlations and three bacteria genera-macrophage subtype correlations. In this pilot study, SM and EC use were associated with an increase in undifferentiated M0 macrophages, but SM differed from EC users and NS for inflammatory gene expression. The data support the hypothesis that SM and EC have toxic lung effects influencing inflammatory responses, but this may not be via changes in the microbiome.

Lung mitochondrial DNA copy number, inflammatory biomarkers, gene transcription and gene methylation in vapers and smokers.

SummaryBackgroundMitochondrial DNA copy number (mtCN) maintains cellular function and homeostasis, and is linked to nuclear DNA methylation and gene expression. Increased mtCN in the blood is associated with smoking and respiratory disease, but has received little attention for target organ effects for smoking or electronic cigarette (EC) use.MethodsBronchoscopy biospecimens from healthy EC users, smokers (SM), and never-smokers (NS) were assessed for associations of mtCN with mtDNA point mutations, immune responses, nuclear DNA methylation and gene expression using linear regression. Ingenuity pathway analysis was used for enriched pathways. GEO and TCGA respiratory disease datasets were used to explore the involvement of mtCN-associated signatures.FindingsmtCN was higher in SM than NS, but EC was not statistically different from either. Overall there was a negative association of mtCN with a point mutation in the D-loop but no difference within groups. Positive associations of mtCN with IL-2 and IL-4 were found in EC only. mtCN was significantly associated with 71,487 CpGs and 321 transcripts. 263 CpGs were correlated with nearby transcripts for genes enriched in the immune system. EC-specific mtCN-associated-CpGs and genes were differentially expressed in respiratory diseases compared to controls, including genes involved in cellular movement, inflammation, metabolism, and airway hyperresponsiveness.InterpretationSmoking may elicit a lung toxic effect through mtCN. While the impact of EC is less clear, EC-specific associations of mtCN with nuclear biomarkers suggest exposure may not be harmless. Further research is needed to understand the role of smoking and EC-related mtCN on lung disease risks.FundingThe National Cancer Institute, the National Heart, Lung, and Blood Institute, the Food and Drug Administration Center for Tobacco Products, the National Center For Advancing Translational Sciences, and Pelotonia Intramural Research Funds.

Abstract 751: Lung mitochondrial DNA copy number variations: E-cig users, smokers, and never-smokers

Abstract Background: Electronic cigarettes (e-cigs) are one of the most popular tobacco products in the US. Little is known regarding their pulmonary effects. E-cigs induce similar oxidant reactivity as cigarette smoke and promote oxidative damage/inflammation in airway cells. Given that mtDNA is more prone to oxidative stress than nuclear DNA because of a less effective proofreading system, mtDNA alterations may be important indicators of e-cigs' toxic effects. Clinically, mitochondrial DNA alteration is an emerging biomarker of respiratory diseases. Methods: We compared mtDNA copy number (mtDNA-CN) from lung brushings in a cross-sectional bronchoscopy study of healthy young adults, e-cig users (EC)(n=15), non-smokers, non-EC users (NS)(n=43), and smokers (SM)(n=26). We examined associations of mtDNA-CN with immune response (differential cell counts and cytokines in bronchoalveolar lavage), DNA methylation and gene expression brushings. Associations for: 1) EC vs NS vs SM, and 2) tobacco product users (EC+SM) vs NS for MtDNA-CN with immune response, methylation, and expression were made using linear regression. Further, significant features by group interactions were followed up by within-group tests. False Discovery Rate (FDR) at 0.1 was considered significant. Ingenuity pathway analysis was used to identify the most significantly enriched pathways/molecular functions/diseases. Results: MtDNA-CN was not significantly different among the three groups (P=0.06). MtDNA-CN was higher in SM than NS (P=0.02), and in tobacco product users than NS (P=0.02); EC mtDNA-CN tended to be intermediate between the 2 other groups. There were significantly positive associations of IL-2 and IL-4 with mtDNA-CN in EC, but not in SM or NS (Interaction FDR=0.06 for both). We found 147 transcripts (60 genes) and 1,153 CpGs (713 genes) to be significantly associated with mtDNA-CN in all three groups. The most common canonical pathway of the signatures for both expression and methylation were immune responses. The top molecular and cellular functions for both included cell death and survival. Ten transcripts (LINC01184, SNU13, RPL35A, COLCA1, HLA-DRB1, LOC105379655, TRIM9, TCIRG1, CLPB, MIR2114) and 3,929 CpGs (top: ULK4, STARD13, HLCS, FLT1, TMEM91, CYP2J2) were associated with mtDNA-CN in E-cig users only. Some of these genes are known to play a role in lung diseases, including cancer. For the signatures associated in all groups, we found many more significant signatures (236 vs 147 transcripts and 40,830 vs 1,153 CpGs) in the two group vs. three group comparisons, respectively. Conclusion: While the sample size was small, this study is the first to suggest that mtDNA-CN is a site of pulmonary toxic effects. We found associations of mtDNA-CN with inflammatory markers among EC users, and with a number of biological signatures, particularly genes related to immune response, in the lungs of EC, SM, and NS, but differently by groups for some. Citation Format: Kellie M. Mori, Joseph P. McElroy, Daniel Y. Weng, Sangwoon Chung, Sarah A. Reisinger, Kevin L. Ying, Quentin A. Nickerson, Theodore M. Brasky, Mark D. Wewers, Jo L. Freudenheim, Peter G. Shields, Min-Ae Song. Lung mitochondrial DNA copy number variations: E-cig users, smokers, and never-smokers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 751.

Differences in biomarkers of inflammation and immune responses in chronic smokers and moist snuff users

BackgroundCigarette smoking is a major risk factor for cancer and other diseases. While smoking induces chronic inflammation and aberrant immune responses, the effects of smokeless tobacco products (STPs) on immune responses is less clear. Here we evaluated markers related to immune regulation in smokers (SMK), moist snuff consumers (MSC) and non-tobacco consumers (NTC) to better understand the effects of chronic tobacco use. Materials and methodsSeveral markers associated with immune regulation were measured in peripheral blood mononuclear cells (PBMCs) from SMK (n = 40), MSC (n = 40), and NTC (n = 40) by flow cytometry. ResultsRelative to NTC, seven markers were significantly suppressed in SMK, whereas in MSC, only one marker was significantly suppressed. In a logistic regression model, markers including granzyme B+ lymphocytes, perforin+ lymphocytes, granzyme B+ CD8+T cells, and KLRB1+ CD8+ T cells remained as statistically significant predictors for classifying the three cohorts. Further, cell-surface receptor signaling pathways and cell–cell signaling processes were downregulated in SMK relative to MSC; chemotaxis and LPS-mediated signaling pathways, were upregulated in SMK compared to MSC. A network of the tested markers was constructed to visualize the immunosuppression in SMK relative to MSC. ConclusionMoist snuff consumption is associated with significantly fewer perturbations in inflammation and immune function biomarkers relative to smoking. ImpactThis work identifies several key immunological biomarkers that differentiate the effects of chronic smoking from the use of moist snuff. Additionally, a molecular basis for aberrant immune responses that could render smokers more susceptible for infections and cancer is provided.

Maternal Weight Variation in Different Intrauterine Environments: An Important Role of Hypertension.

Different intrauterine environments may influence the maternal prepregnancy body weight (BW) variation up to 6 months postpartum. The objective of the present study was to verify the association of sociodemographic, obstetric, nutritional, and behavioral factors with weight variation in women divided into four groups: hypertensive (HM), diabetic (DM), smokers (SM), and control mothers (CM). It was a convenience sample of 124 postpartum women recruited from 3 public hospitals in the city of Porto Alegre, state of Rio Grande do Sul, Brazil, between 2011 and 2016. Multiple linear regressions and generalized estimating equations (GEE) were conducted to identify the factors associated with maternal weight variation. For all GEE, the maternal weight measurements were adjusted for maternal height, parity, educational level, and the type of delivery, and 3 weight measurements (prepregnancy, preceding delivery, and 15 days postpartum) were fixed. A hierarchical model closely associated the maternal diagnosis of hypertension and a prepregnancy body mass index (BMI) classified as overweight with maternal weight gain measured up to the 6th month postpartum (the difference between the maternal weight at 6 months postpartum and the prepregnancy weight). These results showed that the BW of the HM group and of overweight women increased ∼ 5.2 kg 6 months postpartum, compared with the other groups. Additionally, women classified as overweight had a greater BW variation of 3.150 kg. This evidence supports the need for specific nutritional guidelines for gestational hypertensive disorders, as well as great public attention for overweight women in the fertile age.

Frequency of Porphyromonas gingivalis fimA in smokers and nonsmokers after periodontal therapy.

Porphyromonas gingivalis is one of the most important Gram-negative anaerobe bacteria involved in the pathogenesis of periodontitis. P. gingivalis has an arsenal of specialized virulence factors that contribute to its pathogenicity. Among them, fimbriae play a role in the initial attachment and organization of biofilms. Different genotypes of fimA have been related to length of fimbriae and pathogenicity of the bacterium. Objectives The aim of this study was to identify 5 types of fimA genotype strains in smokers and nonsmokers with periodontitis, before and after periodontal therapy.Material and Methods Thirty-one patients with periodontitis harboring P. gingivalis were selected: 16 nonsmokers (NS) and 15 smokers (SM). Clinical and microbiological parameters were evaluated at baseline and 3 months after periodontal treatment, namely: plaque index, bleeding on probe, probing depth, gingival recession and clinical attachment level. The frequency of P. gingivalis and fimA genotype strains were determined by polymerase chain reaction.Results Type I fimA was detected in the majority of SM and NS at baseline, and the frequency did not diminish after 3 months of treatment. The frequency of type II genotype was higher in SM than NS at baseline. After 3 months, statistical reduction was observed only for types II and V fimA genotypes in SM. The highest association was found between types I and II at baseline for NS (37.5%) and SM (53.3%).Conclusion The most prevalent P. gingivalis fimA genotypes detected in periodontal and smoker patients were genotypes I and II. However, the presence of fimA genotype II was higher in SM. Periodontal treatment was effective in controlling periodontal disease and reducing type II and V P. gingivalis fimA.

Pathway Analysis of Global Metabolomic Profiles Identified Enrichment of Caffeine, Energy, and Arginine Metabolism in Smokers but Not Moist Snuff Consumers.

Existing US epidemiological data demonstrate that consumption of smokeless tobacco, particularly moist snuff, is less harmful than cigarette smoking. However, the molecular and biochemical changes due to moist snuff consumption relative to smoking remain incompletely understood. We previously reported that smokers (SMK) exhibit elevated oxidative stress and inflammation relative to moist snuff consumers (MSC) and non-tobacco consumers (NTC), based on metabolomic profiling data of saliva, plasma, and urine from MSC, SMK, and NTC. In this study, we investigated the effects of tobacco consumption on additional metabolic pathways using pathway-based analysis tools. To this end, metabolic pathway enrichment analysis and topology analysis were performed through pair-wise comparisons of global metabolomic profiles of SMK, MSC, and NTC. The analyses identified >8 significantly perturbed metabolic pathways in SMK compared with NTC and MSC in all 3 matrices. Among these differentially enriched pathways, perturbations of caffeine metabolism, energy metabolism, and arginine metabolism were mostly observed. In comparison, fewer enriched metabolic pathways were identified in MSC compared with NTC (5 in plasma, none in urine and saliva). This is consistent with our transcriptomics profiling results that show no significant differences in peripheral blood mononuclear cell gene expression between MSC and NTC. These findings, taken together with our previous biochemical, metabolomic, and transcriptomic analysis results, provide a better understanding of the relative changes in healthy tobacco consumers, and demonstrate that chronic cigarette smoking, relative to the use of smokeless tobacco, results in more pronounced biological changes, which could culminate in smoking-related diseases.

Changes in CRP and TNF‐a after Runs at Different Intensities in Smokers Following Two Hour Cessation

The purpose of this study was to examine changes in C‐reactive protein (CRP) and TNF‐α from blood samples before and after exercise at different intensities in smokers (SM) and non‐smokers (NS) after a 2 hour smoking cessation. Our laboratory previously reported large differences in CRP and TNF‐α at Pre between SM and NS. Furthermore, there were more profound disturbances in SM following three runs at different intensities approximately 30 minutes after smoking cessation before each run (EB 2015, Boston). The previous findings indicated that SM responded more sensitively to relatively equal intensity of exercise, which suggested that high intensity aerobic exercise should be avoided in SM in the beginning phase of exercise training.MethodsA total of eighteen physically inactive subjects (eight SM and ten NS) aged 20 to 27 were recruited. Smokers were accepted if their level of exhaled carbon monoxide (eCO) was greater than 16 ppm. Each subject completed three treadmill runs at different intensities in a random order (55%, 65%, and 75% of VO2max). Running distance for all three runs was equivalent to 35‐min run at 65% VO2max. Smokers had a 2 hour smoking cessation before each run. Blood samples were collected before (Pre), immediately post (IP), and an hour following each run (1h Post).ResultsPre‐exercise CRP levels were higher in SM as compared to the NS. (2.291±.27 mg/L Vs. 2.082±.25, mean±SE, P = 0.042). As compared to Pre, no significant changes were found in CRP at IP and 1h Post following runs at three different intensities in both SM and NS. TNF‐α showed higher pre‐exercise levels in SM compared to NS (32.565±4.28 mg/L vs. 27.732±3.34, P=0.027). TNF‐α was significantly increased at IP following 75% of VO2max run in both SM and NS (SM: 32.657±4.36 → 38.797±6.33, P=0.013; NS: 27.694±3.26 → 33.466±4.35, P=0.019), but not significantly changed at IP following 55% and 65% of VO2max runs.ConclusionFindings of the current study revealed a dose‐response relationship for CRP and TNF‐a in SM and NS. The patterns of changes corresponded with previous presentation (EB, 2015) with a 30 minute smoking cessation; however, the differences between SM and NS were much smaller in this study suggesting that smokers may need to avoid cigarette smoking before exercise to minimize immune disturbance caused by combined stress from exercise and chronic smoking.Support or Funding InformationThis study was supported by Texas A&M International University Research Grant.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Cigarette cues capture attention of smokers and never-smokers, but for different reasons

BackgroundWhile the notion that smokers reliably show higher reactivity to cigarette-related versus neutral cues is both theoretically and empirically supported, it is unclear why never-smokers also show enhanced brain responses to cigarette-related cues. MethodsUsing a repetitive picture viewing paradigm, in which responses evoked by affective cues are more resistant to habituation, we assessed the effects of stimulus repetition on event-related potentials (ERPs) evoked by pleasant, unpleasant, cigarette-related, and neutral images in 34 smokers (SMO) and 34 never-smokers (NEV). We examined the early posterior negativity (EPN) and the late positive potential (LPP), two ERP components which are sensitive to a picture's motivational qualities. ResultsBefore stimulus repetition, pleasant, unpleasant, and cigarette-related cues produced greater EPN and LPP amplitudes than neutral cues in all subjects. During stimulus repetition, both components were similarly modulated by emotional arousal, such that pleasant, unpleasant, and cigarette-related cues evoked greater EPN and LPP amplitude, relative to neutral. Smoking status did not modulate these effects. While there were no group differences in self-reported stimulus ratings of valence for pleasant, unpleasant, or neutral stimuli, NEV rated cigarette-related cues as unpleasant. We observed a moderate, negative correlation between LPP amplitude and self-reported valence ratings of cigarette-related cues among NEV. ConclusionsThese data suggest that cigarette-related cues capture attentional resources of both SMO and NEV, but for different reasons. For SMO, cigarette-related cues have acquired motivational significance through repeated associations with nicotine delivery, whereas for NEV, cigarette-related cues are perceived as unpleasant.

The Impact of Combinations of Alcohol, Nicotine, and Cannabis on Dynamic Brain Connectivity.

Alcohol, nicotine, and cannabis are among the most commonly used drugs. A prolonged and combined use of these substances can alter normal brain wiring in different ways depending on the consumed cocktail mixture. Brain connectivity alterations and their change with time can be assessed using functional magnetic resonance imaging (fMRI) because of its spatial and temporal content. Here, we estimated dynamic functional network connectivity (dFNC) as derived from fMRI data to investigate the effects of single or combined use of alcohol, nicotine, and cannabis. Data from 534 samples were grouped according to their substance use combination as controls (CTR), smokers (SMK), drinkers (DRN), smoking-and-drinking subjects (SAD), marijuana users (MAR), smoking-and-marijuana users (SAM), marijuana-and-drinking users (MAD), and users of all three substances (ALL). The DRN group tends to exhibit decreased connectivity mainly in areas of sensorial and motor control, a result supported by the dFNC outcome and the alcohol use disorder identification test. This trend dominated the SAD group and in a weaker manner MAD and ALL. Nicotine consumers were characterized by an increment of connectivity between dorsal striatum and sensorimotor areas. Where possible, common and separate effects were identified and characterized by the analysis of dFNC data. Results also suggest that a combination of cannabis and nicotine have more contrasting effects on the brain than a single use of any of these substances. On the other hand, marijuana and alcohol might follow an additive effect trend. We concluded that all of the substances have an impact on brain connectivity, but the effect differs depending on the dFNC state analyzed.

Comparative Network Analysis of Patients with Non-Small Cell Lung Cancer and Smokers for Representing Potential Therapeutic Targets

Cigarette smoking is the leading cause of lung cancer worldwide. In this study, we evaluated the serum autoantibody (AAb) repertoires of non-small cell lung cancer (NSCLC) patients and smokers (SM), leading to the identification of overactivated pathways and hubs involved in the pathogenesis of NSCLC. Surface- and solution-phase biopanning were performed on immunoglobulin G purified from the sera of NSCLC and SM groups. In total, 20 NSCLC- and 12 SM-specific peptides were detected, which were used to generate NSCLC and SM protein datasets. NSCLC- and SM-related proteins were visualized using STRING and Gephi, and their modules were analyzed using Enrichr. By integrating the overrepresented pathways such as pathways in cancer, epithelial growth factor receptor, c-Met, interleukin-4 (IL-4) and IL-6 signaling pathways, along with a set of proteins (e.g. phospholipase D (PLD), IL-4 receptor, IL-17 receptor, laminins, collagens, and mucins) into the PLD pathway and inflammatory cytokines network as the most critical events in both groups, two super networks were made to elucidate new aspects of NSCLC pathogenesis and to determine the influence of cigarette smoking on tumour formation. Taken together, assessment of the AAb repertoires using a systems biology approach can delineate the hidden events involved in various disorders.

Association of body mass index (BMI) and stage in patients with non-small cell lung cancer harboring KRAS mutations.

e13071 Background: Body mass index (BMI) is inversely associated with lung cancer risk, particularly in former or current smokers. KRAS mutant non-small cell lung cancer (NSCLC) has been shown to be highly associated with smoking status. Therefore, we sought to investigate the association of BMI and stage in a cohort of patients with KRAS mutant NSCLC. Methods: Patients with KRAS mutant lung cancer who underwent tumor genotyping using next generation sequencing testing were identified. BMI( kg/m2 ) was categorized as: Underweight ( < 18.5), normal weight (18.5 – 24.9), overweight (25-29.9) and obese (≥30). Chi-square and t-tests were performed to test association between stage (I vs. II+) and BMI (4 categories), and stage and BMI as a continuous variable, respectively. Results: From 5/2013 to 9/2016, 201 patients with KRAS mutant NSLC were identified. Of these, 178 (88.6%) were current or former/ever smokers (SM) and 23 (11.4%) were never smokers (NS). In the SM group, 114 were female and 64 male with a median age at diagnosis of 66.5 years (range 45-93). In the NS group, 15 were female and 8 male with a median age at diagnosis of 71.5 years (range 51-92) (age for NS vs. SM p = 0.0439). Fifty-two (25.9%) patients presented with stage I disease, 149 (74.1%) presented with advanced (stage II-IV) disease. There was no significance in the BMI at diagnosis of SM vs. NS or by sex. There was a difference (p = 0.0430) in the BMI between patients with stage I disease (mean 28.6) and patients with stage II or greater disease (mean 26.6). This was also seen in SM with stage I vs. II+ disease (p = 0.0417); however, this was no longer significant in SM when broken out by sex or in NS alone. Conclusions: We identified a significant difference in BMI in patients with KRAS mutant NSCLC with stage I vs. advance stage disease, with a higher BMI associated with early stage disease at diagnosis. The lower BMI in patients with more advanced stage may be a confounding variable explained by comorbidities, and the lung cancer itself. It is unlikely that treatment effects were a factor as the KRAS testing was done prior to initiating therapy. Further studies looking at BMI and survival analysis in patients with mutant KRAS NSCLC are warranted.

Alterations of resting state functional network connectivity in the brain of nicotine and alcohol users

Alcohol and nicotine intake result in neurological alterations at the circuit level. Resting state functional connectivity has shown great potential in identifying these alterations. However, current studies focus on specific seeds and leave out many brain regions where effects might exist. The present study uses a data driven technique for brain segmentation covering the whole brain. Functional magnetic-resonance-imaging (fMRI) data were collected from 188 subjects:51 non-substance consumption controls (CTR), 36 smoking-and-drinking subjects (SAD), 28 drinkers (DRN), and 73 smokers (SMK). Data were processed using group independent component analysis to derive resting state networks (RSN). The resting state functional network connectivity (rsFNC) was then calculated through correlation between time courses. One-way ANOVA tests were used to detect rsFNC differences among the four groups. A total of 50 ANOVA tests were significant after multi-comparison correction. Results delineate a general pattern of hypo-connectivity in the substance consumers. Precuneus, postcentral gyrus, insula and visual cortex were the main brain areas with rsFNC reduction suggesting reduced interoceptive awareness in drinkers. In addition, connectivity reduction between postcentral and one RSN covering right fusiform and lingual gyri showed significant association with severity of hazardous drinking. In smokers, connectivity changes agreed with the idea of a shift towards endogenous information processing, represented by the DMN. Hypo-connectivity between thalamus and putamen was observed in smokers. In contrast, the angular gyrus showed hyper-connectivity with the precuneus linked to smoking and significantly correlated with nicotine dependence severity. In spite of the presence of common effects, our results suggest that particular effects of alcohol and nicotine can be separated and identified. Results also suggest that concurrent use of both substances affects brain connectivity in a complex manner, requiring careful consideration of interaction effects.

Elevated Nerve Growth Factor/p75NTR Co-localization In Smokers After Eccentric Exercise

PURPOSE: Smoking increases musculoskeletal injury risk and protracts healing. While the underlying mechanisms are poorly understood, altered inflammatory responses may contribute. We previously found at 48h post-eccentric contractions (ECC) that smokers (SM) experienced: 1) 15% greater delayed onset muscle soreness (DOMS) than non-smokers (NS); and 2) altered muscle activity of key pro-inflammatory regulator canonical nuclear factor kappa-beta (p65 NFκB; NS: -7.1±1.3 fold change from baseline; SM: -1.1±1.0 fold). Coinciding with a shift from pro- to anti-inflammatory signaling and peak DOMS, these data suggest delayed inflammatory resolution in SM. Nerve growth factor (NGF) plays a role in pain development and is stimulated by p65 NFκB. We hypothesized that NGF expression and co-localization with both high- (tyrosine receptor kinase A, TrkA) and low- (p75NTR) affinity receptors would be greater in SM than NS following ECC, and may explain greater DOMS in SM. METHODS: Young adult male SM (N=6) and NS (N=5) performed 100 maximal ECC using only the non-dominant knee extensors. Bilateral vastus lateralis biopsies were obtained at 48h post-exercise for NGF mRNA quantification via qPCR. Immunohistochemistry was performed on a subcohort (n=3/group) to quantify and localize NGF, TrkA, and p75NTR protein. RESULTS: ECC did not alter NGF mRNA (NS: 1.9±0.3 fold; SM: 2.8±0.7 fold; p=0.09) or protein levels, which were similarly induced in SM compared to NS (1.1 fold). NGF co-localization with TrkA was unrelated to exercise or smoking status (~23.2±2%). However, SM expressed 2.1 fold higher p75NTR levels than NS (p<0.05). Further, there was a significant interaction (p<0.05) whereby NGF/p75NTR co-localization after ECC decreased in NS (control: 39.6±8.1%; exercise: 31.3±11.4%) but increased in SM (control: 59.2±8.4%; exercise: 72.4±13.7%). CONCLUSIONS: Greater p75NTR expression and p75NTR/NGF co-localization may enhance nerve sensitivity in SM and further augment DOMS. Along with our previous DOMS and inflammatory findings, p75NTR/NGF in SM could delay inflammatory resolution, enhancing risk for chronic pain/disability. This work provides insights into mechanisms that may place SM at greater risk for musculoskeletal injury and impaired healing. Supported, in part, by a grant from the Department of Defense.

Attention and Recall of Point-of-sale Tobacco Marketing: A Mobile Eye-Tracking Pilot Study.

IntroductionAs tobacco advertising restrictions have increased, the retail ‘power wall’ behind the counter is increasingly invaluable for marketing tobacco products.ObjectiveThe primary objectives of this pilot study were 3-fold: (1) evaluate the attention paid/fixations on the area behind the cash register where tobacco advertising is concentrated and tobacco products are displayed in a real-world setting, (2) evaluate the duration (dwell-time) of these fixations, and (3) evaluate the recall of advertising displayed on the tobacco power wall.MethodsData from 13 Smokers (S) and 12 Susceptible or non-daily Smokers (SS) aged 180–30 from a mobile eye-tracking study. Mobile-eye tracking technology records the orientation (fixation) and duration (dwell-time) of visual attention. Participants were randomized to one of three purchase tasks at a convenience store: Candy bar Only (CO; N = 10), Candy bar + Specified cigarette Brand (CSB; N = 6), and Candy bar + cigarette Brand of their Choice (CBC; N = 9). A post-session survey evaluated recall of tobacco marketing. Key outcomes were fixations and dwell-time on the cigarette displays at the point-of-sale.ResultsParticipants spent a median time of 44 seconds during the standardized time evaluated and nearly three-quarters (72%) fixated on the power wall during their purchase, regardless of smoking status (S: 77%, SS: 67%) or purchase task (CO: 44%, CSB: 71%, CBC: 100%). In the post session survey, nearly all participants (96%) indicated they noticed a cigarette brand and 64% were able to describe a specific part of the tobacco wall or recall a promotional offer.ConclusionsConsumers are exposed to point-of-sale tobacco marketing, regardless of smoking status. FDA should consider regulations that limit exposure to point-of-sale tobacco marketing among consumers.

A cross-sectional study of biomarkers of exposure and effect in smokers and moist snuff consumers.

Cigarette smoking is a major risk factor for several chronic diseases. Epidemiological data indicate the use of smokeless tobacco (ST) is associated with significantly lower risk for smoking-related diseases compared to cigarettes. Several biomarkers of exposure (BioExp) and effect (BioEff) associated with smoking and use of moist snuff (ST) were evaluated. A single site, cross-sectional clinical study enrolled three groups of generally healthy male smokers (SMK), moist snuff consumers (MSC), and non-tobacco consumers (NTC), and several BioExp and BioEff were evaluated. Blood and urinary BioExp, including total nicotine equivalents and tobacco-specific nitrosamines, were higher in MSC compared to SMK. Biomarkers of combustion-related toxicants and cadmium were elevated in SMK. Elevated levels of some BioEff associated with oxidative stress (urinary isoprostanes and leukotriene E4), inflammation (white blood cell count), platelet activation (thromboxane metabolites), and lipid metabolism (apolipoprotein B100 and oxidized low-density lipoprotein) were observed in SMK relative to NTC and MSC (all p<0.05). The non-smoking groups (MSC and NTC) showed similar levels of combustion-related BioExp and BioEff. Higher levels of exposure to nicotine and some N'-nitrosamines may be observed in MSC, and SMK are exposed to higher levels of combustion-related toxicants. Changes in BioEff consistent with some aspects of inflammation, oxidative stress, and altered lipid metabolism were detected in SMK compared to the non-smoking groups. The biomarker data further improve our understanding of pathophysiological changes and the risk continuum associated with various tobacco products, and could be useful components of future assessments of tobacco products.

Effects of Exercise Intensity on Cortisol, Antioxidant, and DNA damage in Smokers and Non‐Smokers

The purpose of the study was to examine blood level changes of cortisol, total antioxidant status, and 8‐oxo deoxyguanisine (8‐OXO) before and after exercise at different intensities in smokers and non‐smokers.MethodsFourteen physically inactive smokers and non‐smokers were recruited. Each subject completed three treadmill runs at different intensities in a random order (60%, 70%, and 80% of VO2max). Running distance for all three runs was equivalent to a 30‐min run at 70% VO2max. Blood samples were collected at rest (PRE), immediately after (POST), and 1 hour following (1H) each run.ResultsSmokers (SM) showed higher levels of Cortisol (P=.043) and 8‐OXO (p=.024) and lower level of antioxidant (P=.001) as compared to non‐smokers (NS) (group effect). Cortisol levels were elevated in both SM and NS following the 80% VO2max run [SM (PRE:611.9±13.6nmol/L→POST:761.6±16.2, Mean±SE, P=.0001); NS(498.2±13.9→624.2±16.2, P=.0001)]. Total antioxidant increased only in SM following runs (60%:1.58±.04mM→1.76±.06, P=.0001; 70%:1.57±.04→1.72±.08, P=.001; 80%:1.59±.04→1.71±.09, P=.012). 8‐OXO elevated at POST following 70% (SM:3188±242pg/mL→4667±199, P=.001; NS:2432±181→3586± 199, P=.011 ) and 80% (SM: 3268±234→5832±488, P=.0001; NS:2473±200→3813±255, P=.003) VO2max runs.ConclusionHigher levels of cortisol and lower antioxidant status induced greater DNA damage in smokers, especially following runs at 70% and 80% VO2max, indicating that DNA damage in blood immune cells following moderate‐high and high intensity aerobic exercises is greater in Smokers than Non‐Smokers. Therefore, smokers may need to avoid high intensity aerobic exercise since combined stress from exercise and chronic smoking may aggravate immune function.

Study of cardiovascular disease biomarkers among tobacco consumers. Part 3: evaluation and comparison with the US National Health and Nutrition Examination Survey

Cardiovascular disease (CVD) biomarkers of biological effect (BoBE), including hematologic biomarkers, serum lipid-related biomarkers, other serum BoBE, and one physiological biomarker, were evaluated in adult cigarette smokers (SMK), smokeless tobacco consumers (STC), and non-consumers of tobacco (NTC). Data from adult males and females in the US National Health and Nutrition Examination Survey and a single site, cross-sectional study of healthy US males were analyzed and compared. Within normal clinical reference ranges, statistically significant differences were observed consistently for fibrinogen, C-reactive protein (CRP), hematocrit, mean cell volume, mean cell hemoglobin, hemoglobin, white blood cells, monocytes, lymphocytes, and neutrophils in comparisons between SMK and NTC; for CRP, white blood cells, monocytes, and lymphocytes in comparisons between SMK and STC; and for folate in comparisons with STC and NTC. Results provide evidence for differences in CVD BoBE associated with the use of different tobacco products, and provide evidence of a risk continuum among tobacco products and support for the concept of tobacco harm reduction.

Study of cardiovascular disease biomarkers among tobacco consumers, part 1: biomarkers of exposure

A study was conducted to evaluate biomarkers of biological effect and physiological assessments related to cardiovascular disease (CVD) among adult male cigarette smokers (SMK), moist snuff consumers (MSC) and non-consumers of tobacco (NTC). Additionally, biomarkers of tobacco and tobacco smoke exposure (BoE) were measured in spot urines and are reported here. Except for the BoE to nicotine and NNK, BoE were generally greater in SMK compared with MSC, and BoE were generally not different in comparisons of MSC and NTC. Results demonstrated that MSC had lower systemic exposures to many harmful and potentially harmful constituents than SMK, which is consistent with epidemiological data that indicate a differential in CVD risk between these groups.