Human alveolar macrophages spontaneous reduction of BSPT salt

Summary Alveolar macrophages of non smoking and smoking human adults reduce BSPT salt spontaneously. The staining obtained is located on three cell membrane systems: the endoplasmic reticulum, the Golgi apparatus and the nuclear envelope. Methylene blue MB inhibits BSPT reduction. The smokers alveolar macrophages have less positivity than those of the non smokers. The endogenous cell substrate revealed in this work is the initial common pathway of two different oxidative chains bounded to the microsome. One acts with cytochrome P 450 for chemical detoxification by hydroxylation, the other one acts with cytochrome B₅ for lipid oxidation or peroxidation and both may be connected with the cell bactericidal system.     

Changes in the levels of ACTH and cortisol after passive exposure to cigarette smoke in smokers and non-smokers

Plasma ACTH and cortisol levels were studied in smokers and non smokers, (exposed or not to smoke of the environment), after passive exposure to cigarette smoking. Non smokers, usually not exposed to smoke, show a rise in both hormones, whereas smokers and non smokers commonly exposed to smoke don't show any change in ACTH and cortisol levels. These data suggest that nicotine acts as an acute stimulus on the hypophysis-adrenal axis even passively inhaled.     

Increased sister chromatid exchange in the peripheral blood lymphocytes of young women who smoke cigarettes

Rates of sister chromatid exchange in dividing human peripheral blood lymphocytes were determined and compared between smoking and non smoking young women between the ages of 16 and 25. Chromosomes block-stained with Giemsa were also examined for chromosome aberrations. A striking difference in the frequency of sister chromatid exchange was found between young women who smoked and those who did not. Smokers scored a significantly higher, F(1) = 15.99, p = 0.0015, rate of sister chromatid exchange than non smokers. Smokers scored a higher mean of SCEs per cell (12.771, SD 3.53) than non smokers (9.712, SD 2.53). Smokers also scored a higher range of SCEs (4 to 28) as opposed to non smokers (4 to 17). No statistical difference was found between smokers and non smokers for the frequency of chromosome aberrations. The significantly higher frequency of exchange in young smoking women may indicate that initial damage to the DNA in many of these women has probably already occurred, thus causing an increased risk of developing cancer later in life.     

The implication of cigarette smoking and cessation on macrophage cholesterol efflux in coronary artery disease patients

We investigated ATP-binding cassette transporters A1/G1 expression and function in mediating cholesterol efflux by examining the macrophages of cigarette-smoking patients with coronary artery disease (CAD) before and after smoking abstinence. Peripheral blood monocyte cells were collected from nonsmokers (n = 17), non-CAD (NCAD) smokers (n = 35), and CAD smokers (n = 32) before and after 3 months of smoking cessation. We found that the ABCA1 expression level was lower in macrophages from NCAD and CAD smokers than from nonsmokers at baseline. The ABCA1 function of mediating cholesterol efflux was reduced in NCAD and CAD smokers as compared with nonsmokers. After 3 months of smoking cessation, ABCA1 expression and function were improved in CAD smokers. However, ABCG1 expression and function did not change after smoking cessation. Furthermore, ABCA1 expression was inhibited by tar in human acute monocytic leukemia cell line THP-1-derived macrophages through the inhibition of liver X receptors. Nicotine and carbon monoxide did not inhibit ABCA1 expression. Our results indicate that chronic cigarette smoking impaired ABCA1-mediated cholesterol efflux in macrophages and that tobacco abstinence reversed the function and expression of ABCA1, especially in CAD patients. It was tobacco tar, rather than nicotine or carbon monoxide, that played a major role in the tobacco-induced disturbance of cellular cholesterol homeostasis.     

Association of current smoking with airway inflammation in chronic obstructive pulmonary disease and asymptomatic smokers

Background

Inflammation in the airways and lung parenchyma underlies fixed airway obstruction in chronic obstructive pulmonary disease. The exact role of smoking as promoting factor of inflammation in chronic obstructive pulmonary disease is not clear, partly because studies often do not distinguish between current and ex-smokers.

Methods

We investigated airway inflammation in sputum and bronchial biopsies of 34 smokers with chronic obstructive pulmonary disease (9 Global initiative for Chronic Obstructive Lung Disease stage 0, 9 stage I, 10 stage II and 6 stage III) and 26 asymptomatic smokers, and its relationship with past and present smoking habits and airway obstruction.

Results

Neutrophil percentage, interleukin-8 and eosinophilic-cationic-protein levels in sputum were higher in chronic obstructive pulmonary disease (stage I-III) than asymptomatic smokers. Inflammatory cell numbers in bronchial biopsies were similar in both groups. Current smoking correlated positively with macrophages: in bronchial biopsies in both groups, and in sputum in chronic obstructive pulmonary disease. Pack-years smoking correlated positively with biopsy macrophages only in chronic obstructive pulmonary disease.

Conclusion

Inflammatory effects of current smoking may mask the underlying ongoing inflammatory process pertinent to chronic obstructive pulmonary disease. This may have implications for future studies, which should avoid including mixed populations of smokers and ex-smokers.     

Semiquantitative cytochemistry in evaluation of apoptotic capacity in bronchoalveolar lavage of smokers and patients with non-small-cell lung cancer

Apoptotic capacity of pulmonary tissue to produce or remove apoptotic cells by alveolar macrophages (ALMs) was investigated in three groups: healthy volunteers, smokers and patients with non-small-cell lung cancer (NSCLC). Differential cell counting of bronchoalveolar lavage (BAL) specimens revealed significantly higher percentages of neutrophils and eosinophils and decreased percentage of macrophages in BAL of patients with NSCLC in comparison with nonsmokers and smokers. Proportion of lymphocytes was significantly higher in patients with NSCLC than in smokers. These changes in the BAL cell profile may reflect immunology of the lung in pulmonary malignancies. BAL eosinophils were significantly lower and AMs increased in smokers in comparison with nonsmokers. This result might be understood as a consequence of changed tissue architecture of pulmonary tissue in situ, influenced by smoking. Apoptotic detection in cytocentrifuge preparations of BAL cell suspensions was evaluated by TUNEL method. Subsequent steps, adsorption, internalization and digestion of apoptotic cells by alveolar macrophages (AMs) were estimated by semiquantitative cytochemical scoring and indexing method and correlated with percent of free apoptotic cells. Significant increase of apoptotic capacity of pulmonary tissue in control smokers (289.55+/-50.77) in comparison with that of non-smokers (218.29+/-56.24) could be a consequence of stimulated digestion inside the AMs; decreased apoptotic capacity of pulmonary tissue in NSCLC (150.30+/-40.61; p<0.05), in comparison with non-smokers and smokers is in relation to a reduced phagocytosis of the apoptotic remnants, which might be either the cause or the consequence of the oncogenic process.     

ACTH and cortisol after cigarette smoke exposure during the dexamethasone suppression test in smokers and non-smokers

In this research the effect of nicotine, (smoke of cigarette), was studied in smokers and non smokers during dexamethasone inhibitory test (1 mg h 23.00). ACTH and cortisol plasma levels, physiologically suppressed at 08.00, increased, after 30 min from smoking, only in group of non smokers. These data suggest that nicotine, in non smokers, could induce a maximum stimulus on diencephalic structures, so to overcome the inhibition of dexamethasone.     

A Stochastic Model of Smoking Behavior Under a Cessation Program

A stochastic model describing the movements, resulting from a smoking cessation program, of a cohort of individuals from the smoking to the non‐smoking compartment is proposed. The distribution and first two moments of the number of smokers are derived.     

Association of 1 hydroxypyrene glucuronide in human urine with cigarette smoking and broiled or roasted meat consumption

Humans are exposed to polycyclic aromatic hydrocarbons PAHs from various occupational, dietary, environmental and medicinal sources. We measured 1 hydroxypyrene glucuronide 1 OHP gluc concentration in urines from male non smokers n = 50 , smokers of blond tobacco n = 31 , smokers of black tobacco n = 16 , and pipe smokers n = 3 . Immunoaffinity chromatography was used as a preparative step and synchronous fluorescence spectroscopy as the quantitation method. The concentration of 1 OHP gluc in urine from smokers mean SE: 1.04 0.13 pmol ml-1 urine was significantly higher than in urine from non smokers 0.55 0.05 pmol ml-1 urine by the Wilcoxon rank sum test non smokers versus all smokers, p = 0.001; vs black tobacco smokers, p = 0.001; vs blond tobacco smokers, p = 0.007 . Urinary 1 OHP gluc concentration among subjects who had consumed roasted, grilled or broiled meat within the past 24 h was elevated compared with those who had not p = 0.025 . Multiple linear regression showed significant associations of urinary 1 OHP gluc with number of cigarettes smoked p = 0.002 and consumption of roasted, grilled or broiled meat p = 0.028 . Systemic CYP1A2 activity estimated by caffeine metabolism was significantly correlated with urinary 1 OHP gluc concentration. However, this association was probably due to cigarette smoking, since adjusting for cigarette smoking by multiple linear regression made the association between urinary 1 OHP gluc and CYP1A2 phenotype non significant. These results further support the use of urinary 1 OHP gluc as a biomarker of recent pyrene exposure through inhalation or diet.     

Association of 1 hydroxypyrene glucuronide in human urine with cigarette smoking and broiled or roasted meat consumption

Humans are exposed to polycyclic aromatic hydrocarbons PAHs from various occupational, dietary, environmental and medicinal sources. We measured 1 hydroxypyrene glucuronide 1 OHP gluc concentration in urines from male non smokers n = 50, smokers of blond tobacco n = 31, smokers of black tobacco n = 16, and pipe smokers n = 3 . Immunoaffinity chromatography was used as a preparative step and synchronous fluorescence spectroscopy as the quantitation method. The concentration of 1 OHP gluc in urine from smokers mean SE: 1.04 0.13 pmol ml-1 urine was significantly higher than in urine from non smokers 0.55 0.05 pmol ml-1 urine by the Wilcoxon rank sum test non smokers versus all smokers, p = 0.001; vs black tobacco smokers, p = 0.001; vs blond tobacco smokers, p = 0.007 . Urinary 1 OHP gluc concentration among subjects who had consumed roasted, grilled or broiled meat within the past 24 h was elevated compared with those who had not p = 0.025 . Multiple linear regression showed significant associations of urinary 1 OHP gluc with number of cigarettes smoked p = 0.002 and consumption of roasted, grilled or broiled meat p = 0.028 . Systemic CYP1A2 activity estimated by caffeine metabolism was significantly correlated with urinary 1 OHP gluc concentration. However, this association was probably due to cigarette smoking, since adjusting for cigarette smoking by multiple linear regression made the association between urinary 1 OHP gluc and CYP1A2 phenotype non significant. These results further support the use of urinary 1 OHP gluc as a biomarker of recent pyrene exposure through inhalation or diet.     

Comparison of antioxidant enzymes in saliva of elderly smokers and non-smokers

Objectives: This study was designed to compare the levels of copper/zinc superoxide dismutase (Cu/Zn SOD), peroxidase (POx) and glutathione peroxidase (GSH‐Px) in saliva of smokers and those in saliva of non‐smokers. Methods: Unstimulated saliva samples were collected from 88 elderly males (65 years old or over) who visited a private dental clinic. Forty‐four subjects were current smokers (more than 20 cigarettes daily for at least 30 years) and 44 were non‐smokers. The levels of salivary thiocyanate, Cu/Zn SOD, GSH‐Px, and POx activity were measured using standard procedures. Results: The mean levels of salivary thiocyanate (SCN^?) and SOD were significantly higher (p < 0.01) in the smoking group than in the non‐smoking group, whereas the specific activity levels of POx and GSH‐Px were significantly higher (p < 0.05) in the non‐smoking group than in the smoking group. Significant correlation coefficients were found between the levels of SCN^? and SOD (r = 0.37, p < 0.001). In the non‐smoking group, a significant positive association was found between specific activity of POx and age (r = 0.33, p < 0.05). Conclusion: Measurement of SCN^? and Cu/Zn SOD in human saliva might be useful for estimating the level of oxidative stress caused by cigarette smoke. Despite increased H₂O₂ level as a defense system induced by SOD, detoxification of H₂O₂ might be deteriorated in the oral cavity of elderly smokers.     

Milk consumption, smoking and lead concentration in human hair

Trace elements in hair samples were examined in 79 medical and sports students in Vienna and 77 biology students in Rome (ages 19-31 years). Lead concentration was determined by inductively coupled plasma mass spectrometry. The effect of smoking on lead levels is confirmed. Those who consume milk and dairy products daily show a lower lead concentration than those who consume them occasionally, both overall and in smokers and non-smokers. The effects of milk consumption and smoking appear to be independent. It seems that smokers may not enjoy the protective effect of milk consumption to the same extent as non smokers.     

Clinical considerations in study designs that use cotinine as a biomarker.

Subjects enrolled in studies are not always screened for routine habits such as smoking. Personal history is not always reliable and therefore an objective biomarker is necessary to screen for smokers. The objectives of this article were to review the metabolism of nicotine and other metabolic considerations associated with smoking; to review some of the routine methods used to assess exposure to nicotine-containing products; to revisit cotinine breakpoints utilized to distinguish smokers from non-smokers during screening for clinical trials; to assess the utility of screening questions regarding smoking practices; and to recommend standards for clinical pharmacology studies. The results indicated that cotinine levels serve as a useful biomarker of tobacco exposure; racial issues may be clinically relevant in determining smoking status; cessation of smoking should occur at least 14 days prior to the start of the study; adverse effects from nicotine withdrawal such as craving, hunger and weight gain may persist for more than 6 months; potential metabolic interactions via cytochrome P2A6 and P1A2 need to be considered when designing a study; and the use of a single calibrator as a breakpoint is acceptable if a categorical outcome such as 'smoker' versus 'non-smoker' is desired. Nicotine from food products is not expected to impact assay sensitivity or to be clinically relevant; a serum cotinine concentration of 10 ng ml(-1) be employed as a breakpoint for non-smokers versus smokers; other non-invasive alternatives are collection of urine, saliva, or hair (with suggested breakpoints of 200 ng ml(-1), 5 ng ml(-1) and 0.3 ng mg(-1), respectively; screening questions be accompanied by testing for cotinine; and the inclusion of smokers in studies should be considered once the impact of smoking on the targeted population is understood.     

Clinical considerations in study designs that use cotinine as a biomarker

Subjects enrolled in studies are not always screened for routine habits such as smoking. Personal history is not always reliable and therefore an objective biomarker is necessary to screen for smokers. The objectives of this article were to review the metabolism of nicotine and other metabolic considerations associated with smoking; to review some of the routine methods used to assess exposure to nicotine-containing products; to revisit cotinine breakpoints utilized to distinguish smokers from non-smokers during screening for clinical trials; to assess the utility of screening questions regarding smoking practices; and to recommend standards for clinical pharmacology studies. The results indicated that cotinine levels serve as a useful biomarker of tobacco exposure; racial issues may be clinically relevant in determining smoking status; cessation of smoking should occur at least 14 days prior to the start of the study; adverse effects from nicotine withdrawal such as craving, hunger and weight gain may persist for more than 6 months; potential metabolic interactions via cytochrome P2A6 and P1A2 need to be considered when designing a study; and the use of a single calibrator as a breakpoint is acceptable if a categorical outcome such as 'smoker' versus 'non-smoker' is desired. Nicotine from food products is not expected to impact assay sensitivity or to be clinically relevant; a serum cotinine concentration of 10 ng ml^-1 be employed as a breakpoint for non-smokers versus smokers; other non-invasive alternatives are collection of urine, saliva, or hair (with suggested breakpoints of 200 ng ml^-1, 5 ng ml^-1 and 0.3 ng mg^-1, respectively; screening questions be accompanied by testing for cotinine; and the inclusion of smokers in studies should be considered once the impact of smoking on the targeted population is understood.     

Concomitant elevations of MMP‐9, NGAL,proMMP‐9/NGAL and neutrophil elastase in serum of smokers with chronic obstructive pulmonary disease

A growing body of evidence points towards smoking‐related phenotypic differences in chronic obstructive pulmonary disease (COPD). As COPD is associated with systemic inflammation, we determined whether smoking status is related to serum levels of matrix metalloproteinase‐9 (pro‐ and active MMP‐9), neutrophil gelatinase‐associated lipocalin (NGAL) and the proMMP‐9/NGAL complex in patients with COPD. Serum samples were collected in 100 stable‐phase COPD patients (82 smokers, 18 never‐smokers) and 28 healthy adults (21 smokers, 7 never‐smokers). Serum levels of studied factors were measured in ELISA. Our data provide the first evidence of simultaneously elevated serum levels of MMP‐9, NGAL and proMMP‐9/NGAL in COPD smokers. While the triad discriminated between smokers and non‐smokers in the COPD group, MMP‐9 and proMMP‐9/NGAL (but not NGAL) discriminated between smokers with and without COPD. Adjustment for age and smoking pack‐years did not alter the findings. Serum MMP‐9, NGAL and proMMP‐9/NGAL levels were not correlated with the GOLD stage or FEV1 decline. Furthermore, serum levels of neutrophil elastase (NE) and MMP‐3 (but not of IL‐6 and MMP‐12) were also higher in COPD smokers than in healthy smokers before and after adjustment for age and pack‐years. Among COPD smokers, levels of MMP‐9, NGAL and proMMP‐9/NGAL were positively correlated with NE (P < 0.0001) but not with the remaining factors. Gelatin zymography detected proMMP‐9 in serum samples of healthy and COPD smoking groups. Our results suggest that associated serum levels of proMMP‐9, NGAL, proMMP‐9/NGAL and NE may reflect the state of systemic inflammation in COPD related to cigarette smoking.     

A controlled cycle of tetrahydrocannabinol smoking: T and B cell rosette formation.

T and B cell rosettes were serially determined before, during, and after a four week cycle of tetrahydrocannabinol (THC) smoking in 10 hospitalized subjects. The early T cell rosettes were significantly fewer during the period of THC smoking and tended to return toward baseline after smoking, whereas no changes were seen in the seven simultaneously studied controls. Both the THC smokers and controls showed no change in their normal values for WBC, % lymphocytes, B cell rosettes (complement receptor) and late T cell rosettes. The data suggest that T cells, present in nearly normal numbers in the smokers, had reduced capacities to form early rosettes during and directly related to THC smoking. While the immunological significance of reduced early rosette formation is unclear, the data suggest that further immunological study of marijuana smokers would be indicated.     

Smoking-reIated DNA adducts and genetic polymorphism for metabolic enzymes in human lymphocytes

Smoking-related aromatic DNA adducts in lymphocytes were measured from smokers (n = 76), ex-smokers (n = 25) and non-smokers (n = 56) by the ³²P-postlabelling method, to clarify whether a genetic polymorphism for metabolic enzymes could explain the inter-individual variation of DNA adduct levels. Adduct levels were compared with respect to smoking status and polymorphic genotypes of cytochrome P4501A1 (CYP1A1) and glutathione S-transferase M1 (GTSM1). The mean adduct level (1.24 per 10⁸ nucleotides) in smokers was significantly higher than that (0.85 per 10⁸) in non-smokers. Although we expected higher adduct levels in the CYP1A1 variant or GSTM1 null subjects, the adduct level in 'GSN1 nulls' was significantly lower than that in 'GSTM1 presents' among smokers. DNA adduct levels had significant positive correlations with smoking indices such as number of cigarettes or smoking years in all subjects. In smokers only, however, no correlation was found, because there were negative correlations between adduct levels and smoking dose in GSTM1 null genotypes. CYP1A1 genotypes had no effects on adduct levels.     

Effect of Mild-to-Moderate Smoking on Viral Load,Cytokines, Oxidative Stress,and Cytochrome P450 Enzymes in HIV-Infected Individuals

Mild-to-moderate tobacco smoking is highly prevalent in HIV-infected individuals, and is known to exacerbate HIV pathogenesis. The objective of this study was to determine the specific effects of mild-to-moderate smoking on viral load, cytokine production, and oxidative stress and cytochrome P450 (CYP) pathways in HIV-infected individuals who have not yet received antiretroviral therapy (ART). Thirty-two human subjects were recruited and assigned to four different cohorts as follows: a) HIV negative non-smokers, b) HIV positive non-smokers, c) HIV negative mild-to-moderate smokers, and d) HIV positive mild-to-moderate smokers. Patients were recruited in Cameroon, Africa using strict selection criteria to exclude patients not yet eligible for ART and not receiving conventional or traditional medications. Those with active tuberculosis, hepatitis B or with a history of substance abuse were also excluded. Our results showed an increase in the viral load in the plasma of HIV positive patients who were mild-to-moderate smokers compared to individuals who did not smoke. Furthermore, although we did not observe significant changes in the levels of most pro-inflammatory cytokines, the cytokine IL-8 and MCP-1 showed a significant decrease in the plasma of HIV-infected patients and smokers compared with HIV negative non-smokers. Importantly, HIV-infected individuals and smokers showed a significant increase in oxidative stress compared with HIV negative non-smoker subjects in both plasma and monocytes. To examine the possible pathways involved in increased oxidative stress and viral load, we determined the mRNA levels of several antioxidant and cytochrome P450 enzymes in monocytes. The results showed that the levels of most antioxidants are unaltered, suggesting their inability to counter oxidative stress. While CYP2A6 was induced in smokers, CYP3A4 was induced in HIV and HIV positive smokers compared with HIV negative non-smokers. Overall, the findings suggest a possible association of oxidative stress and perhaps CYP pathway with smoking-mediated increased viral load in HIV positive individuals.