Peronia peronii as a bio-indicator to assess the toxicity of waterpipe tobacco leachates in aquatic and sediment media.

This study was aimed to survey toxicity of waterpipe wastes leachates on Peronia peronii in aquatic and sediment environments as two exposure media. For this, leachates of four tobacco types including burnt traditional tobacco (BTT), fresh traditional tobacco (FTT), burnt fruit-flavored tobacco (BFT) and fresh fruit-flavored tobacco (FFT)) were prepared and used to assess their toxic effects on P. peronei in two aquatic and sediment media. The in-vivo toxic effects of five different concentrations of waterpipe tobacco waste leachates on P. peronii were evaluated. The LC50 values of BTTs leachates to P. peronii were 17.50, 16.05, 11.31 and 9.38 g/L at exposure times of 24, 48, 72 and 96 h, respectively in aquatic media. These values for BFTs leachates were 14.86, 12.38, 9.53 and 7.46 g/L at exposure times of 24, 48, 72 and 96 h, respectively. In the case of sediment media, the LC50 values of BTTs leachates were 15.33, 13.70, 9.09 and 6.70 g/L at exposure times of 24, 48, 72 and 96 h, respectively while these values for BFTs leachates were 12.00, 10.32, 8.20 and 5.65 g/L. Fruit-flavored tobacco leachates had significantly higher toxicity than traditional tobacco leachates for P. peronii. The findings also showed significant differences between the LC50 values of different leachates in different media of water and sediment. The results demonstrated that even small amount of tobacco waste (~ 5 to 6 g/L) can lead to P. peronii mortality and may also pose a hazard to other aquatic and benthic organisms. The results obtained from the present study can be used as a baseline data to assess local effects causing from unsafe disposal of post-consumption tobacco waste in beach areas. In addition, these findings can lead to encouraging decision-makers to focus more on the types of tobacco waste in the municipal solid waste management system and to implement a source separation process for these wastes.

Maternal waterpipe tobacco smoke exposure during lactation induces hormonal and biochemical changes in rat dams and offspring.

Waterpipe smoking is common among pregnant and breastfeeding women. Herein, the effects of waterpipe tobacco smoke (WTS) exposure during lactation on milk composition, hormonal levels and biochemical profile in dams and pups were investigated. Lactating Wistar rats were randomly assigned to receive either WTS (2 hours per day) or fresh air (control group). Milk was collected on day 21 and analysed for protein, lactose and total fat. Blood, from dams and pups, was analysed for insulin, glucose, lipid profile, leptin, prolactin and corticosterone. WTS exposure during lactation increased the blood level of HDL and corticosterone in dams (P < .05). However, the level of milk lactose and blood glucose was reduced in dams after the exposure to WTS during lactation (P < .05). WTS during lactation significantly increased levels of triglycerides, LDL and leptin (P < .05), and a trend of increase in blood level of nicotine and prolactin in pups. Levels of other parameters were not affected by WTS exposure in dams and pups. In conclusion, WTS exposure during lactation altered the milk composition and altered lipid profile, glucose homeostasis and hormonal levels in dams and pups. It is necessary to adopt strategies to enhance tobacco cessation during breastfeeding.

Urinary metabolites of furan in waterpipe tobacco smokers compared to non-smokers in home settings in the US.

OBJECTIVES: Determine uptake of furan, a potential human carcinogen, in waterpipe tobacco (WPT) smokers in home settings. METHODS: We analysed data from a US convenience sample of 50 exclusive WPT smokers, mean age 25.3 years, and 25 non-smokers, mean age 25.5 years. For WPT smokers, data were collected at a home visit by research assistants during which participants smoked one WPT head of one brand for a mean of 33.1 min in their homes. Research assistants provided and prepared a WP for participants by weighing and loading 10 g of WPT in the WP head. At the completion of the smoking session, research assistants measured the remaining WPT. Cotinine and six furan metabolites were quantified in first morning urine samples provided on 2 consecutive days for non-smokers, and on the morning of a WPT smoking session and on the following morning for smokers. RESULTS: WPT smokers consumed a mean of 2.99 g WPT. In WPT smokers, urinary cotinine levels increased significantly 26.1 times the following morning; however, urinary metabolites of furan did not increase significantly. Compared to non-smokers, 2 furan metabolites, N-acetyl-S-[1-(5-acetylamino-5-carboxylpentyl)-1H-pyrrol-3-yl]-L-cysteine and N-acetyl-S-[1-(5-amino-5-carboxypentyl)-1H-pyrrol-3-yl]-L-cysteine sulfoxide, were significantly higher in WPT smokers in pre and in post WPT smoking levels. CONCLUSIONS: To enable a more rigorous assessment of furan exposure from WPT smoking, future research should determine furan concentrations in WPT smoke, quantify furan metabolites from users of various WPT brands; and extend the investigation to social settings where WPT smoking is habitually practiced.

Waterpipe Tobacco Smoke Inhalation Triggers Thrombogenicity, Cardiac Inflammation and Oxidative Stress in Mice: Effects of Flavouring.

The consumption of water-pipe smoking (WPS) has been promoted by the use of flavoured tobacco. However, little is known about the impact of flavouring on the cardiovascular toxicity induced by WPS inhalation. Here, we compared the cardiovascular effects and underlying mechanism of actions of plain (P) (unflavoured) versus apple-flavoured (AF) WPS (30 minutes/day, 5 days/week for 1 month) in mice. Control mice were exposed to air. Both P- and AF-WPS inhalation induced an increase in systolic blood pressure, thrombogenicity and plasma concentration of fibrinogen and von Willebrand factor. In heart homogenates, AF-WPS inhalation caused an increase of 8-isoprostane and a decrease in the levels of reduced glutathione (GSH) and superoxide dismutase (SOD). Nevertheless, P-WPS decreased only the activity of SOD. The concentrations of tumour necrosis factor α and interleukin 1ß were increased only in heart homogenates of mice exposed to AF-WPS. Although both P- and AF-WPS increased the concentration of troponin I in heart homogenates and induced DNA damage, the concentration of cleaved caspase 3 was only increased in mice exposed to AF-WPS. Immunohistochemical analysis of the hearts showed that both P- and AF- WPS inhalation decreased the expression of SOD. Moreover, the expression of nuclear factor erythroid-derived 2-like 2 at nuclear level in the heart was higher in both AF-WPS and P-WPS compared with control group, and the effect observed in AF-WPS group was more significant than that seen in P-WPS group. Likewise, the concentration of heme oxygenase-1 was significantly increased in both P-WPS and AF-WPS groups compared with control group, and the effect seen in AF-group was higher than that observed in P-WPS group. In conclusion, our findings showed that both P- and AF-WPS induce thrombogenicity and cardiac injury, and that this toxicity is potentiated by the presence of flavouring.

Waterpipe Tobacco Chemical Content, Microbial Contamination, and Genotoxic Effects: A Systematic Review.

The spread of tobacco smoking has increased over time at the global and national levels. One of the widely spread tobacco products is waterpipe. Recent studies showed that waterpipe tobacco smoke contains toxic substances, including carbon monoxide and nicotine. Some of them are genotoxic carcinogen, such as formaldehyde. This study aims to provide comprehensive insight into the types and depth of the scientific literature on waterpipe tobacco smoke chemical content, its genotoxic effects, and waterpipe device microbial contamination. We conducted a systematic comprehensive review of articles published between 1986 and December 2018. Primary research articles focusing on the content of waterpipe smoke, including chemical, genotoxic, and microbial contaminants, were eligible for inclusion. Of the 1,286 studies generated, 22 studies were included. Twenty-three chemical families were extracted from waterpipe smoke. Aldehydes were the most identified chemical family in 6 studies, and next is polycyclic hydrocarbons, found in 5 studies. About 206 chemical compounds were identified. Flavobacterium, Pseudomonas, coagulase-negative Staphylococci, and Streptococcus were the most abundant pathogen contaminants. Waterpipe smoke had elevated levels of many DNA damage markers (8-hydroxy-2'-deoxyguanosine and cytochrome P450 1A1) and inhibited levels of many DNA repair genes (OGG1 and XRCC1) in waterpipe smokers. Waterpipe smoke is associated with the genotoxic effect, which elevates the levels of many DNA damage markers and inhibits the levels of many DNA repair genes. In addition, waterpipe smoking can expose smokers to a range of pathogenic bacteria.

Contributions of charcoal, tobacco, and syrup to the toxicity and particle distribution of waterpipe tobacco smoke.

The use of waterpipes in the United States is increasing in a largely unregulated market. The shisha smoked in a waterpipe is a complex matrix of tobacco, flavorings, and humectant with smoke generated by an external heat source. This study explored the relationship between shisha components and the particulate matter size distributions and toxicity of smoke generated with heating. Standard waterpipe puff topography of charcoal- or electronic- heated whole shisha and shisha components generated smoke particulate matter that was characterized using a TSI Engine Exhaust Particle Sizer. Relative toxicity of the whole smoke was determined via measurement of lysosomal integrity and measures of membrane integrity following acute exposure of type II alveolar cells at the air-liquid interface. All waterpipe aerosols exhibited a unimodal particle size distribution, the peak and concentration of which varied depending upon the shisha components present. Acute exposure to charcoal-heated whole shisha, flavoring syrup, or humectant smoke, or electronic-heated whole shisha smoke caused significant alveolar cell damage and death, indicating neither tobacco nor charcoal are needed for these cytotoxic effects to occur.

Influence of prenatal waterpipe tobacco smoke exposure on reproductive hormones and oxidative stress of adult male offspring rats.

Male infertility is adversely affected by tobacco cigarette smoking. Herein, the effects of prenatal waterpipe tobacco smoke (WTS) exposure on reproductive hormones and oxidative stress of adult offspring rats were evaluated. Pregnant rats received either fresh air or mainstream WTS (2 hr daily). Pregnancy outcomes, circulatory levels of follicle stimulating hormone (FSH), luteinizing hormone (LH) and prolactin, testicular levels of oestrogen, testosterone and oxidative stress biomarkers [catalase, superoxide dismutase (SOD), glutathione peroxidase (GPx) and thiobarbituric acid reactive substances (TBARS)] were assessed in their adult male offspring rats. Prenatal WTS exposure reduced the number of born offspring, female to pups ratio and birthweight (p < 0.05). Prenatal WTS exposure increased the circulatory levels of FSH and the testicular levels of oestrogen, testosterone and TBARS and catalase activity compared with control group (p < 0.05). However, GPx activity was reduced by WTS exposure (p < 0.05). There appeared to be a trend of increased LH and prolactin levels with prenatal WTS exposure; however, it was not statistically significant compared with control group (p > 0.05). The activity of SOD was not affected by prenatal WTS exposure (p > 0.05). In conclusion, prenatal WTS exposure altered reproductive hormones as well as oxidative stress biomarkers in adult male offspring rats.

Secretome analysis of oral keratinocytes chronically exposed to shisha.

BACKGROUND: Shisha smoking has been associated with multiple diseases including oral cancer. However, a mechanistic study to investigate alteration of secreted proteins in oral cells due to shisha smoking is lacking. OBJECTIVES: Elucidation of differentially secreted proteins by immortalized human normal oral keratinocytes (OKF6/TERT1) upon chronic exposure to shisha. METHODS: OKF6/TERT1 was chronically treated with 0.5% shisha extract for 8 months. Conditioned media from shisha treated (OKF6/TERT1-Shisha) and untreated (OKF6/TERT1-Parental) cells were subjected to TMT-based quantitative proteomic analysis. Bioinformatics analysis of differentially secreted proteins was carried out using SignalP, SecretomeP and TMHMM. Immunoblot validation of selected proteins was carried out to confirm the proteomics results. RESULTS: Proteomic analysis of OKF6/TERT1-Parental and OKF6/TERT1-Shisha secretome resulted in the identification of 1,598 proteins, of which 218 proteins were found to be differentially secreted (⩾ 1.5-fold; p-value ⩽ 0.05) in shisha treated cells. Bioinformatics analysis using prediction tools showed secretory potential of differentially secreted proteins identified in OKF6/TERT1-Shisha. Western blotting validated the expression of AKR1C2, HSPH1 and MMP9 in OKF6/TERT1-Shisha secretome in agreement with proteomic data. CONCLUSION: This study serves as a useful resource to understand the effect of chronic shisha smoking on the milieu of secreted proteins of oral cells. In vivo studies are warranted to supplement our in vitro data to elucidate the role of these proteins as early diagnostic biomarkers for oral carcinogenesis among shisha smokers.

Strain- and sex-dependent pulmonary toxicity of waterpipe smoke in mouse.

Waterpipe smoking is emerging as a form of tobacco smoking, but its lung health/risks is not known. It has been shown that different mouse strains show differences in susceptibility to tobacco smoke. However, the effect of waterpipe smoke (WPS) exposure and strain differences in susceptibility to oxidative and inflammatory responses is not known. Here, we showed acute WPS exposure induced oxidative stress and inflammatory response in C57BL/6J and BALB/cJ mouse strains. WPS exposure induced inflammatory cell influx (neutrophils and T-lymphocytes) in bronchoalveolar lavage fluid (BAL fluid), which varied among mouse strains. Proinflammatory cytokines release differed among both the strains, but was significantly increased in C57BL/6J mice. Myeloperoxidase levels in BAL fluid were increased significantly in both the strains. Total reduced glutathione (GSH) level was decreased, whereas the level of oxidized or glutathione disulfide (GSSG) increased in lungs of both the strains. Similarly, the level of lipid peroxidation markers, 15-isoprostane (plasma), malondialdehyde and 4-hydroxy-2-nonenal (lung homogenates) were increased by WPS. Our data suggest that, oxidative stress and inflammatory responses are influenced by strain characteristics during acute WPS exposure. Overall, C57BL/6J mice showed more susceptibility to oxidative stress and inflammatory responses compared to BALB/cJ mice. Acute WPS mediated pulmonary toxicity is differentially regulated in different mouse strains.

Effect of prenatal waterpipe tobacco smoke on airway inflammation in murine model of asthma of adult offspring mice.

OBJECTIVE: Worldwide popularity of waterpipe tobacco smoking has increased, including in pregnant women. This study investigates the effect of prenatal waterpipe tobacco smoke (WTS) exposure on airway inflammation in a murine model of asthma of adult offspring mice. MATERIALS AND METHODS: Pregnant BALB/c mice were exposed to fresh air or WTS, using a whole-body exposure system that mimics human use during WTS. Adult male offspring mice were divided into; (1) control (prenatal fresh air, postnatal ovalbumin sensitization and saline challenge), (2) postnatal Ova S/C (prenatal fresh air, postnatal ovalbumin sensitization and challenge (Ova S/C)), (3) prenatal WTS (prenatal WTS, postnatal ovalbumin sensitization and saline challenge) and (4) prenatal WTS + postnatal Ova S/C. Cells from the bronchoalveolar lavage fluid, cytokines, and oxidative stress markers (superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx) and thiobarbituric acid reactive substances (TBARS)) from lung homogenates were evaluated. RESULTS: Prenatal WTS increased recruitment of cells in lungs and levels of SOD and catalase when compared to unexposed offspring's. The levels of cytokines, GPx and TBARS were not affected by prenatal WTS. Prenatal WTS exposure and postnatal Ova S/C increased airway inflammation and activity of SOD compared to control and Ova S/C mice and reduced IL-18 levels compared to Ova S/C mice. DISCUSSION AND CONCLUSIONS: Prenatal exposure to WTS induced airway inflammation, further enhanced by a murine model of asthma in adult offspring. Prenatal exposure to WTS adversely affects the lung function of the offspring and careful strategies for increasing public awareness regarding the harmful effects of WTS during pregnancy is important.