Smoke treatment triggers the release of a novel DNA damaging factor by lymphocytes

Abstact Organic fuel smoke is a hazardous agent, which pushes the cells towards“prooxidant state', leading to 4,46,400 strand breaks/cell/day as against 47,000 strand breaks/cell/day produced by constitutive oxygen radicals. This prooxidants scenario switches on a plethora of intercellular events. Here we report a novel DNA damaging factor released by lymphocytes, upon treatment with smoke condensate. Human lymphocytes, when exposed to cow dung cake smoke condensate, were found to release a low molecular weight factor into the media at 20 min of exposure. The conditioned media, displayed a propensity of inducing DNA damage in fresh, normal lymphocytes, which were not exposed to any damaging agent. The above DNA damaging effect of the conditioned media was not due to any residual presence of Polycyclic Aromatic Hydrocarbons, which were present in the smoke. The release of this factor was in correlation with the DNA damaging event, taking place in the cells. This secondary DNA damaging factor had a molecular weight less than 5 kd. The factor had the cell death inducing propensity when allowed to act on lymphocytes     

Pseudoreplication in experimental designs for the manipulation of seed germination treatments

In the published reports concerning the design of laboratory experiments examining signals that might trigger seed germination, there is currently a misunderstanding of what might constitute correct replication of the experimental treatment. This is particularly true for the study of dry heat, smoke and charcoal, where either an individual seed or a batch of seeds in a Petri dish or tray is being treated as the unit of replication of the experimental treatment, irrespective of whether or not those seeds were all subjected to the experimental manipulation simultaneously. Under these circumstances, the application of the treatment is unreplicated, while samples nested within that single application have been replicated, and the Petri dishes/trays are functioning solely as independent replicates of the variability in germination response within the seed batch used and variation within the pretreatment and post‐treatment environments. Thus any observed difference in germination may be due to the germination treatment but, potentially, it could also be due to any chance event affecting the treated sample. There are a number of alternative experimental designs that avoid this problem. The essential point with these designs is that the application of the experimental manipulation to each replicate should be separated in space by the use of separate experimental equipment and/or in time by the repeated use of the same experimental apparatus.     

Comparative seed germination ecology of Austrostipa compressa and Ehrharta calycina (Poaceae) in a Western Australian Banksia woodland

Austrostipa compressa, a native ephemeral of southwest Western Australia was stimulated to germinate under a range of temperatures, in the presence of light, and exposure to smoke-water. This combination of environmental cues results in winter-maximum germination in immediate postfire and disturbed-soil environments of this Mediterranean-type climate. In contrast, Ehrharta calycina, an introduced perennial grass from southern Africa that has invaded Banksia woodlands, germinated under a wide range of temperature and light conditions, but showed no promotive response to smoke-water. Although A. compressa seeds tolerated heat shock better than E. calycina, the self-burial mechanism of A. compressa seeds ensures protection from fire. High-intensity fire could have a greater impact on E. calycina, as the seeds of this species tend to accumulate in the top of the soil profile where they are more susceptible to high temperatures. Although seeds of E. calycina are more susceptible to high temperatures, survival of mature individuals by postfire resprouting ensures continued survival in native woodlands. Estimates of soil seed bank densities showed extreme variability, but some recently burnt areas of the Yule Brook Botany Reserve contained up to 8000 seed m^?2 of A. compressa and nearly 75 000 seeds m^?2 of E. calycina. Viable soil seed bank densities of A. compressa are reduced with time-since-last fire, but areas of greater than 45 years since the last fire, still contained up to 119 seeds m^?2. In both species, only about half their soil seed bank germinates following fire, thus ensuring the potential for later recruitment. Massive soil seed populations of E. calycina in native Banksia woodlands pose a major problem to management of this plant community type.     

Morpho‐functional modifications of human neutrophils induced by aqueous cigarette smoke extract: comparison with chemiluminescence activity

Cigarette smoking plays an important role as a cause of morbidity and mortality in humans, involving respiratory, cardiovascular, digestive and reproductive systems. Tobacco smoke contains a large number of molecules, some of which are proven carcinogens. Although not fully understood, polymorphonuclear leukocytes seem to play a crucial role in the mechanisms by which tobacco smoke compounds are implicated in smoke‐related diseases. In this paper the effects of an aqueous cigarette smoke extract on the expression of adhesion molecules of polymorphonuclear leukocytes together with the changes in the cell morphology have been related to the chemiluminescence activity. The results obtained show that polymorphonuclear leukocytes treated with aqueous cigarette smoke extract are significantly impaired, as suggested by the changes of chemiluminescence activity, of membrane receptors (CD18, CD62), myeloperoxidase expression and of cell morphology. Altogether the present data indicate that treated polymorphonuclear leukocytes are ineffectively activated and therefore unable to phagocytize zymosan particles. Copyright © 2010 John Wiley & Sons, Ltd.     

Switching from usual brand cigarettes to a tobacco-heating cigarette or snus: Part 2. Biomarkers of exposure

Abstract

A randomized, multi-center study of adult cigarette smokers switched to tobacco-heating cigarettes, snus or ultra-low machine yield tobacco-burning cigarettes (50/group) was conducted, and subjects’ experience with the products was followed for 24 weeks. Differences in biomarkers of tobacco exposure between smokers and never smokers at baseline and among groups relative to each other and over time were assessed. Results indicated reduced exposure to many potentially harmful constituents found in cigarette smoke following product switching. Findings support differences in exposure from the use of various tobacco products and are relevant to the understanding of a risk continuum among tobacco products.

Trial registration: ClinicalTrials.gov identifier: NCT02061917.     

Effect of bacoside A on membrane-bound ATPases in the brain of rats exposed to cigarette smoke

Membrane-bound enzymes play a vital role in neuronal function through maintenance of membrane potential and impulse propagation. We have evaluated the harmful effects of chronic cigarette smoking on membrane-bound ATPases and the protective effect of Bacoside A in rat brain. Adult male albino rats were exposed to cigarette smoke for a period of 12 weeks and simultaneously administered with Bacoside A (the active principle isolated from Bacopa monniera) at a dosage of 10 mg/kg b.w/day, p.o. The levels of lipid peroxides as marker for evaluating the extent of membrane damage, the activities of Na+/K+-ATPase, Ca2+-ATPase and Mg2+-ATPase, and associated cations sodium (Na+), potassium (K+), calcium (Ca2+), and magnesium (Mg2+) were investigated in the brain. Neuronal membrane damage was evident from the elevated levels of lipid peroxides and decreased activities of membrane-bound enzymes. Disturbances in the electrolyte balance with accumulation of Na+ and Ca2+ and depletion of K+ and Mg2+ were also observed. Administration of Bacoside A inhibited lipid peroxidation, improved the activities of ATPases, and maintained the ionic equilibrium. The results of our study indicate that Bacoside A protects the brain from cigarette smoking induced membrane damage.     

Smoke, Mulch, and Seed Broadcasting Effects on Woodland Restoration in Western Australia

Smoke, canopy‐derived mulch, and broadcast seeds were used to maximize the establishment of Banksia woodland species in sand quarries in Western Australia. Smoke, particularly aerosol smoke, had a positive effect on total seedling recruitment. Pre‐mined (woodland) sites showed a 42‐fold increase in total germinants and a 3‐fold increase in the number of species with aerosol smoke application. Post‐mined (restored) sites showed only a 3.6‐fold increase in total germinants and a 1.4‐fold increase in the number of species. Two water‐based smoke chemicals, DC10 (pH 4.5) and SC63 (pH 2.5), increased seedling recruitment at both sites but were not as effective in stimulating recruitment as aerosol smoke. Neither of the chemicals were effective in significantly increasing species richness. Application of aerosol smoke directly to seeds as a pretreatment before broadcasting had no effect on seedling recruitment. Broadcasting of seeds onto restoration sites significantly increased seedling abundance and richness. Application of a single layer of mulch from the canopy vegetation after seed broadcasting gave optimum seedling recruitment. Two layers of mulch significantly reduced recruitment, as did applying mulch before seed broadcasting. For broad‐scale restoration, the application of smoke on newly restored sites would be more effectively achieved using smoke water sprayed over the soil surface. Species that do not recruit from replaced topsoil could be effectively recovered from broadcast seed rather than from mulch.     

Dormancy break of celery (Apium graveolens L.) seeds by plant derived smoke extract

Seed dormancy of a highly-dormant cultivar of celery (Apium graveolens L.) was broken by combinations of plant-derived smoke extract or N⁶-benzyladenine (BA) and gibberellins A_4/7 (GA_4/7) in the dark at temperatures between 18 and 26°C. A less dormant cultivar which responded to GA_4/7 alone showed no additional response to smoke extract or BA. Neither smoke extract nor BA affected either cultivar in the dark in the absence of GA_4/7. The partial dormancy-breaking effect of short exposures to red-light was also enhanced by smoke extracts in this highly-dormant cultivar. The results suggest that smoke extracts act in a similar way to cytokinins, by enhancing gibberellin activity in the celery seed system.Abbreviations BA N⁶-benzyladenine - GA_4/7 A₄ and A₇ gibberellin mixture     

Combined genetic polymorphism and risk for development of lung cancer

Susceptibility to lung cancer has been shown to be modulated by host specific factors. Inheritance of different polymorphic cytochrome P450s (CYPs) and the glutathione S-transferases (GSTs) which affect metabolism of environmental toxicants may play a key role in individual susceptibility. Although individual polymorphic genes have been reported to be associated with development of lung cancer, little is known about the combined effects of several genes in carcinogenesis. From our study of 54 lung cancer patients and 50 matched controls, we observed that a combination of several versions of ‘unfavorable' metabolizing genes (CYP2D6, CYP2E1, GSTM1 and GSTT1) is strongly associated with lung cancer. The relative risk for the different combinations of these genotypes ranged between 1.3 and 14, with higher risk involving the activating genes. The duration and intensity of heavy smoking (expressed in pack-years) are the most important determinant for the development of lung cancer. For example, the estimated risk for development of lung cancer associated with smoking >30 pack-years is represented by an odds ratio=6.65; 95% CL=2.3–19.9 irrespective of an individual's genotype, whereas for smoking between >30 and <50 pack-years, odds ratio=4.5; 95% CL=1.37–15; and for smoking >50 pack-years, odds ratio=30; 95% CL=5.7–114. On the other hand, smoking of less than 30 pack-years is associated with an increased risk in the presence of the polymorphic genes (odds ratio=2.5; 95% CL=0.32–54). The results of our study indicate that the inheritance of multiple ‘unfavorable' genotypes, especially activating genes, is a crucial predisposing factor for the development of lung cancer from cigarette smoking. In addition, the genes may cause moderate smokers who would normally outlive the deleterious effects of smoking to develop lung cancer. The information can therefore be used to target individuals for prevention of health problems.