waterpipe tobacco smoke toxicant exposure and effects alan shihadeh american university of beirut 1...
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Waterpipe tobacco smoke toxicant exposure and effects
Alan ShihadehAmerican University of Beirut
1st International Conference on WTSOctober 20-23, 2013
Abu Dhabi
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Waterpipe toxicants & health effects
http://firefighterparamedicstories.blogspot.com/2011_10_01_archive.html
What happens to the user?• acute physiological effects• BP, HR, inflammatory responses,
lung function…
Does waterpipe smoking emit toxicants?
exhaled mainstream (EMS)
sidestream (SS)
SS + EMS
Does the smoke contain toxicants?• particle size• chemistry• biological activity
Does the user absorb toxicants?biomarkers in: blood, breath, urine
absorbed
exhaled
inhaled
What happened to the smoker?• long term mortality & morbidity
Molecule Particle Cell Animal Human individual Population health
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hose
mouthpiece
body
bowl
head
coal
tobacco
water
Narghile waterpipe
Photo credit: internet, source unknown
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hose
mouthpiece
body
bowl
head
coal
tobacco
water
Narghile waterpipe
Differences with cigarette
tobacco burn not self-sustaining charcoal needed
Tobacco temperature < 500 C (Cig < 900 C)
“molasses” flavorings
order of magnitude higher flow rates/puff volumes
long flow path with bubbler/humidifier (produces cool “smooth” smoke)
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Tobacco smoke constituents
Smoke aerosol
Particlephase10-1000 nm(visible)
Gas phase~ 90% wt(invisible)
“tar”
condensed water
nicotine
water vaporCON2, O2, CO2
C2-C6 hydrocarbons, aldehydes, nitriles …
Illustration from: N. deNevers Pollution Control
4000+ compounds PAH, nitrosamines, metals,… contains most important carcinogens
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disorder contributing agents possible enhancing agents
tobacco dependence major: nicotine
minor: secondary Nicotiana alkaloids, flavor components
acetaldehyde
cardiovascular disease major: carbon monoxide, nitrogen oxides, hydrogen cyanide, tar
minor: cadmium, zinc
nicotine, alkylating species
chronic obstructive pulmonary disease
hydrogen cyanide, volatile aldehydes, nitrogen oxides, carbon monoxide, tar
lung and larynx cancer major: PAH, NNK
minor: 210polonium, formaldehyde, acetaldehyde, butadiene, metals (Cr, Cd, Ni)
catechol, tumor promotersacetaldehydes, diet, alkylating species
oral cavity cancer major: NNN, NNK
minor: PAH
herpes simplex, irritation
esophageal cancer NNN ethanol, diet
pancreas cancer NNK, NNAL diet
Hoffmann et al, 2001; Hecht 1997
The “Hoffmann List“ of probable causative agents in cigarette smoke-related disorders
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disorder contributing agents possible enhancing agents
tobacco dependence major: nicotine
minor: secondary Nicotiana alkaloids, flavor components
acetaldehyde
cardiovascular disease major: carbon monoxide, nitrogen oxides, hydrogen cyanide, tar
minor: cadmium, zinc
nicotine, alkylating species
chronic obstructive pulmonary disease
hydrogen cyanide, volatile aldehydes, nitrogen oxides, carbon monoxide, tar
lung and larynx cancer major: PAH, NNK
minor: 210polonium, formaldehyde, acetaldehyde, butadiene, metals (Cr, Cd, Ni)
catechol, tumor promotersacetaldehydes, diet, alkylating species
oral cavity cancer major: NNN, NNK
minor: PAH
herpes simplex, irritation
esophageal cancer NNN ethanol, diet
pancreas cancer NNK, NNAL diet
*Hoffmann et al, 2001; Hecht 1997
The “Hoffmann List“ of probable causative agents in cigarette smoke-related disorders
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Particle size & lung dosimetry
“Ultrafine particles”Hinds, Aerosol Technology (1999)
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Tobacco smoke particle (100-200 nm)
Human hair
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Do waterpipes emit toxicants?
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Do waterpipes emit toxicants?
What is in the smoke?
1) Find out how people smoke waterpipes
2) Program a robot to smoke the same way
3) Analyze the smoke for toxicants
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Find out how people smokeShihadeh, Antonius, Azar, BRIMC,2005
Puff topography record
Average puff topography• Volume• Duration• Frequency• Number
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Field study of waterpipe users in a Beirut seaside café
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Results
Puff parameter NarghileShihadeh et al 2004
CigaretteDjordjevic et al
2000
Number of puffs 171 12.1
Session smoking time, min 61 3.7
Puff volume, ml/puff 530 44.1
Puff duration, s/puff 2.6 1.5
Interpuff interval, s/cycle 17 18.5
Total volume, ml 90,600 523
A. Shihadeh et al, Biochemistry, Pharmacology, and Behavior, 79(1),75-82, 2004
Field study of 52 café smokers in Beirut mean age 21 years, 14 f/38 m
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puff regimen• volume• duration• frequency• number
Ma’ssel quantity
charcoal type and application regimen
head preparation
“Beirut method” – a standard waterpipe smoking machine protocol for generating WTS
10 g ma’assel tobacco mixture load
1 easy-light charcoal disk + 0.5 added at 105th puff
18-hole aluminum perforation pattern
171 puffs, 530 ml, 2.6 s duration, 17 s IPI
the “Beirut method”
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Program smoking robot & sample/analyze smoke
Shihadeh & Azar, JAM, 2006
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Waterpipe smoke does contain toxicants (MS yields/unit)
NAT (ng)NNK (ng)NNN (ng)NAB (ng)
Lead, ng/10Chromium, ng/10
Arsenic, ng
Acrolein, ugAcetaldehyde, ug/10
Formaldehyde, ug
Indeno(1,2,3-cd)pyrene, ngDibenz(a,h)anthracene, ng
Benzo(a)pyrene, ng
Carbon monoxide, mgNicotine, ug/10
"Tar", mg
0 100 200 300 400 500 600 700 800 9001000
Cigarette Waterpipe (AUB-BfR)
PAHs
Aldehydes
Heavy metals
T/N/CO
TSNA
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narghile cigarette
Particle mass distribution mass median dia mm 0.61 0.33
Respirable mass
mg/puff 0.74 0.21Nanoparticles
#/puff 1.02x1011 1.11x1011
count median dia, nm 155 123
Waterpipe smoke particles are small
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Waterpipe smoke particles are biologically active
Rammah et al, 2013 Toxicology Letters Volume 219, Issue 2 2013 133 - 142
In vitro effects of WTS on HEACs• Oxidative stress• Inflammation• Cell cycle arrest • Impaired vasodialation• Impaired angiogenesis
Plausible cellular mechanism for vascular diseases
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Rammah et al, 2013 Toxicology Letters Volume 219, Issue 2 2013 133 - 142
Impaired angiogenesisCapillary tube formation of untreated and treated HAEC cells (mg/ml) WSC.
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Waterpipe smoke particles are biologically active
Rammah et al, 2012 Toxicology Letters
Effects on lung epithelial (A549) and endothelial cells and signaling mechanisms
Plausible cellular mechanism for COPD
Hoechst staining of cells 72 hours post-treatment with WSC. Pictures were taken using a 40x oil immersion lens.
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Does the user inhale toxicants?
• During a single WP use session the user inhales a large dose of toxicants known to cause tobacco-related diseases
• WTS particle size distribution is similar to cigarette smoke• WTS damages and interferes with repair mechanisms of lung
and vascular cells in culture
YES. Based on the Beirut Method:
Q. Does the Beirut Method provide a reasonable facsimile of real smoke?
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RESULTS charcoal electric charcoal contribution
CO, mg 57.2 5.7 90%
PAHs, ng/mg TPM
Benzo(a)pyrene 0.219 < 0.01 > 95%
∑ 2- and 3-ring PAH 5.262 1.444 73%
∑ 4- and 5-ring PAH 1.181 0.098-0.290a 75-92%
Charcoal(!)
Monzer, B., Sepetdjian, E., Saliba, N. and Shihadeh, A. Charcoal combustion as a source of CO and carcinogenic PAH in mainstream narghile waterpipe smok, Food and Chemical Toxicology, 2008
charcoal electrical heater temperature vs time
More charcoal than ma’ssel is consumed during a typical use session
Users continually “tune” it
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REALTIME sampling in the natural environment
Katurji et al, Inh Tox, 2010
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c) nicotine:tar
a) nicotine b) “tar”
d) CO
Nicotine “Tar”
CONicotine:Tar
Volume (l) Volume (l)
Volume (l)“tar” (mg)
Beirut Method
How does Beirut Method smoke compare to smoke made by people?
Katurji et al, Inh Tox, 2010
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Do waterpipe users absorb toxicants?
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Do waterpipe users absorb toxicants?
Contains “tar”, CO, Nicotine,PAHs, aldehydes….
What is in the smoker?
Measure toxicant levels in blood, breath, or urine.
Studies: placebo control, cigarette comparison, observational.
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CO and nicotine in blood
Carboxyhemoglobin (N=31)
0
1
2
3
4
5
0 5 15 30 45
Perc
ent
Time relative to smoking onset
Plasma nicotine (N=31)
0
2
4
6
8
10
12
14
0 5 15 30 45
Time relative to smoking onset
ng/m
l
Eissenberg & Shihadeh, American Journal of Preventive Medicine, 37, 518-523, 2009.
Clinical setting, one 45 min WP session
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CO and nicotine in blood: compared to cigarettes
Carboxyhemoglobin (N=31)
0
1
2
3
4
5
0 5 15 30 45
Perc
ent
Cigarette
Waterpipe
*
**
Time relative to smoking onset
*
Plasma nicotine (N=31)
0
2
4
6
8
10
12
14
0 5 15 30 45
Time relative to smoking onset
ng/m
l
*
Eissenberg & Shihadeh, American Journal of Preventive Medicine, 37, 518-523, 2009.
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CO and nicotine exposure over 24 hours
Jacob et al, 2013. Cancer Epidemiol Biomarkers Prev 2013;22:765-772.
Plasma nicotine (n=13) Exhaled breath CO (n=13)
• Hospital setting, cross-over design, N=13 dual users• All day ad libitim cigarette smoking (11 cpd mean) versus 3 WTS use sessions• Measurements on day 4 of 4-day protocol.
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Carcinogen exposure over 24 hours
Jacob et al, 2013. Cancer Epidemiol Biomarkers Prev 2013;22:765-772.
Urinary NNAL (TSNA biomarker) Urinary 1-HOP (PAH biomarker)
• Hospital setting, cross-over design, N=13 dual users• All day ad libitim cigarette smoking (11 cpd mean) versus 3 WTS use sessions• Measurements on day 4 of 4-day protocol.
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Carcinogens in populations of smokers
Al Ali R, et al. 2013 Tob Control doi:10.1136/
Urine samples collected from population in Aleppo. Mean cigarette use 27 cigs per day, mean WTS 2 per day.
CTRL (n=28) WP (n=24) CIG (n=23)0
20
40
60
80
100
Total NNAL ng/g creatinine*, **
*, **
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Do waterpipe users absorb toxicants?
YES!
• Clinical, hospital, natural environment findings consistent with one another
• Generally consistent with what we know about toxicants in smoke:
CO WP >> cigPAH WP > cigTSNA WP < cigNicotine WP ~ cig
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What happens to the smoker?
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Delivered nicotine is physiologically active
sympathetic activation reduced complexity
Cobb et al., Inhalation Toxicology 2012
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WP smoke suppresses “Urge to smoke” after 24 hour abstinence
Rastam et al., 2011
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WP smoke compromises cardiac autonomic regulation in human participants (with or without nicotine!)
0
1
2
3
4
baseline end 15 min
LF/HFTobacco
Herbal*
*
0
0.5
1
1.5
2
baseline end 15 min
SampEn Tobacco
Herbal
* *
baseline end 15 min baseline end 15 min
sympathetic activation reduced complexity
Cobb et al., Inhalation Toxicology 2012
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What happens to the smoker?
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WP smoke inhalation induces inflammation and oxidative stress in mice
Khabour et al 2012 Inh Tox
Inflammatory markers Oxidative stress
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WP smoke inhalation associated with:
• Genotoxicity (sister chromatid exchanges): WTS > CS > nonsmokers (Khabour et al, 2011)
• Reduction in exercise capacity : VO2 1.86 vs. 1.7 l/min, pre- post (Hawari et al., 2013)
• Reduction in lung function (Hakim et al., 2011; Mohammad et al., 2008; Koseoglu et al., 2006; Kiter et al., 2000; Aydin et al. 2004; Al Mutairi et al., 2006, Al Fayez et al., 1988):
WTS compared to:
Parameter a) non-smoker b) cigarette smokerFEV1 -4.04% nsFVC -1.38% nsFEV1/FVC -3.08% ns
“WPS negatively affects lung function and may be as harmful as cigarette smoking. WPS, therefore, is likely to be a cause of COPD.” – Raad et al., 2011
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What happened to the smoker?
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What happened to the smoker?
Waked, Khayat, Salameh, 2012
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What happened to the smoker?
(Jawad et al., 2013 based on Akl et al., 2010)
“A wide range of diseases have been associated with WTS, but research in this area is relatively underdeveloped and a better evidence base is needed.” - Akl et al., 2010
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Does waterpipe smoking emit toxicants into the environment?
http://firefighterparamedicstories.blogspot.com/2011_10_01_archive.html
What happens to the user?• acute physiological effects• BP, HR, inflammatory responses,
lung function…
Does waterpipe smoking emit toxicants?
exhaled mainstream (EMS)
sidestream (SS)
SS + EMS
Does the user inhale toxicants?• particle size• chemistry• biological activity
Does the user absorb toxicants?biomarkers in: blood, breath, urine
absorbed
exhaled
inhaled
What happened to the smoker?• long term mortality & morbidity
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Do waterpipe smoking emit toxicants?
• chamber based studies • observational studies of cafés
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Daher et. al, Atmospheric Environment, 44, 8-14, 2009.
Chamber study
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Waterpipe emits more nanoparticles
Daher et. al, Atmospheric Environment, 44, 8-14, 2009.
Particle concentration (particles/cm3)
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WP emits more of everything measured
Daher et. al, Atmospheric Environment, 44, 8-14, 2009.
mean±95% CI waterpipe SS cigarette SS N = 12 N=9Carbon monoxide, mg 2269 ± 108 65.5 ± 5.5
PAH, ng N = 11 N = 3Total PAH 1193 ± 226 305 ± 49 Particle number emissions N = 4 N = 4
ultrafine particles 5.6-99.5 nm, /1012 3.99 ± 0.60 0.639 ± 0.188total particles 5.6-560 nm, /1012 4.38 ± 0.66 1.68 ± 0.27count median diameter, nm 37.9 ± 4.1 130 ± 8 Volatile aldehydes, ug N = 6 N = 5Total aldehydes 12000 ± 1610 2954 ± 416
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• Waterpipe smokers release equivalent of 2-10 cigarette smokers per hour of smoking
What about per smoker-hour?
Daher et. al, Atmospheric Environment, 44, 8-14, 2009.
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Observational studies in WP cafés
* 1-Hour PM2.5 exposure Guideline-Alberta
*
Hammal et al., 2013
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Observational studies in WP cafés
Zhang et al., 2013
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Summary
• Delivers a large dose of toxicants
• Elicits inflammatory responses and dysfunction in human cells
• Elicits immediate physiological changes in users, including compromised lung function and autonomic function, subjective effects
• Is addictive
• Is associated with elevated markers of genetic damage and COPD
• Is associated with other long term health effects, but quality of evidence is very weak.
• Emits large quantities of toxicants in second-hand smoke
WTS…
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Thanks for listening
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RESULTS base case electrically heated charcoal contribution
CO, mg 57.2 5.7 90%
PAHs, ng/mg TPM
Benzo(a)pyrene 0.219 < 0.01 > 95%
∑ 2- and 3-ring PAH 5.262 1.444 73%
∑ 4- and 5-ring PAH 1.181 0.098-0.290a 75-92%
Monzer, B., Sepetdjian, E., Saliba, N. and Shihadeh, A. Charcoal combustion as a source of CO and carcinogenic PAH in mainstream narghile waterpipe smok, Food and Chemical Toxicology, 2008
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conc
entr
atio
n, a
rbitr
ary
units
electrically heated, R2 < 0.02
base case
charcoal extractR2 = 0.94
charcoal grilling (R2 = 0.96, Dyremark et al, 1995)
Relative PAH concentrations measured in the mainstream smoke of the base and electrically heated conditions, and in extracts of the unburned charcoal. PAH concentrations from smoke collected from a charcoal grill by Dyremark et al (1995) shown for comparison. Correlation coefficients are shown relative to the base condition. Unburned charcoal extract, charcoal grilling smoke, and base case narghile smoke exhibit similar PAH patterns. Electrically heated condition produces a different PAH pattern.
Monzer, B., Sepetdjian, E., Saliba, N. and Shihadeh, A. Charcoal combustion as a source of CO and carcinogenic PAH in mainstream narghile waterpipe smok, Food and Chemical Toxicology, 2008
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Tobacco Soex (N = 32) Baseline End 15 min Baseline End 15 minHR (1/min) 70.6 75.5* 72.2 72.8 71.5 68.1*LF (ms2) 1400 2420* 1190 1330 3570* 1610HF(ms2) 1270 1230 1140 1410 1690 1400LF/HF 1.67 2.84* 1.68 1.45 3.21* 1.55SampEn 1.63 1.48* 1.55 1.69 1.52* 1.65
Compromised cardiac autonomic regulation in human participants
Tobacco SoexTopography (N=32)
Total smoke volume (l) 31 (22) 57 (34)*Puffs drawn 95 (115) 95 (58)Mean puff volume (ml) 420 (250) 680 (330)*Mean interpuff interval (s) 45 (34) 35 (22)*Mean puff duration (s) 2.3 (0.9) 3.5 (2.0)*
Peak change from baseline plasma nicotine and CO
Nicotine (N=32) 7.8 (8.0) 0.0 (0.0)*CO% (N=29) 11.8 (7.7) 30.9 (19.0)*
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WP emits more of everything measured
Daher et. al, Atmospheric Environment, 44, 8-14, 2009.
mean±95% CI waterpipe SS cigarette SS N = 12 N=9Carbon monoxide, mg 2269 ± 108 65.5 ± 5.5
PAH, ng N = 11 N = 3Total PAH 1193 ± 226 305 ± 49 Particle number emissions N = 4 N = 4
ultrafine particles 5.6-99.5 nm, /1012 3.99 ± 0.60 0.639 ± 0.188total particles 5.6-560 nm, /1012 4.38 ± 0.66 1.68 ± 0.27count median diameter, nm 37.9 ± 4.1 130 ± 8 Volatile aldehydes, ug N = 6 N = 5Total aldehydes 12000 ± 1610 2954 ± 416
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Mass balance
𝑚𝑒=𝑉 [𝐶 ( 𝑡𝑠 )−𝐶 (0 ) ]+∫0
𝑡 𝑠
𝐶 (𝑡 )𝑄𝑑𝑡+𝑚𝑑
change of suspended mass within CV
For any toxicant…
advected mass
wall losses
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Sidestream emissions
Daher et. al, Atmospheric Environment, 44, 8-14, 2009.
mean±95% CI waterpipe SS cigarette SS N = 12 N=9Carbon monoxide, mg 2269 ± 108 65.5 ± 5.5
PAH, ng N = 11 N = 3Total PAH 1193 ± 226 305 ± 49
Particle number emissions N = 4 N = 4
ultrafine particles 5.6-99.5 nm, /1012 3.99 ± 0.60 0.639 ± 0.188
total particles 5.6-560 nm, /1012 4.38 ± 0.66 1.68 ± 0.27count median diameter, nm 37.9 ± 4.1 130 ± 8
Volatile aldehydes, ug N = 6 N = 5Total aldehydes 12000 ± 1610 2954 ± 416
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SS nanoparticle emissions: WP vs CIG
Daher et. al, Atmospheric Environment, 44, 8-14, 2009.
Particle concentration (particles/cm3)
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Is robot-generated smoke realistic?
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y = 0.018x + 1.470R² = 0.622
0
4
8
12
16
0 200 400 600 800
Bloo
d CO
(CO
Hb,
%)
Smoking machine CO (mg)
Toxicants in robot-generated smoke track blood level exposure in human participants
Carbon monoxide
Shihadeh & Eissenberg, CEBP, 2011
P<0.01R2>0.62
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Schubert et al., 2012
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Environ Mol Mutagen. 2011 Apr;52(3):224-8. doi: 10.1002/em.20601. Epub 2010 Aug 25. Assessment of genotoxicity of waterpipe and cigarette smoking in lymphocytes using the sister-chromatid exchange assay: a comparative study.Khabour OF, Alsatari ES, Azab M, Alzoubi KH, Sadiq MF.
AbstractTobacco smoking is a major world health problem. Recently, waterpipe smoking has become more popular in many countries. Although the genotoxicity associated with cigarette smoking has been extensively investigated, studies evaluating such toxicity in waterpipe users are still lacking. In this study, we examined the genotoxicity of waterpipe smoking in lymphocytes compared with the genotoxicity of cigarette smoking. Genotoxicity was evaluated using the sister chromatid exchanges (SCEs) assay. Fifty waterpipe smokers and 18 healthy nonsmokers participated in this study. Additionally, 18 heavy cigarette smokers (CS) were recruited for comparison. The results show that waterpipe smoking and cigarette smoking significantly increase the frequencies of SCEs (P < 0.01) compared with those of nonsmokers, indicating the genotoxic effect of tobacco smoking. In addition, frequencies of SCEs were significantly higher among waterpipe smokers compared with CS (P < 0.01), indicating that waterpipe smoking is more genotoxic than cigarette smoking. Moreover, the frequency of SCEs increased with the extent of waterpipe use. In conclusion, waterpipe smoking is genotoxic to lymphocytes and the magnitude of its genotoxicity is higher than that induced by regular cigarette smoking.