The Flemish biomonitoring study

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					                      The Flemish biomonitoring study

                                              Part 3
      Minimal group sizes yielding sufficient statistical power for particular
                               biomarker analysis




Interim report 3
Department of Health Risk Analysis and Toxicology
5 November 2003
De Jonge, J.D.
Kleinjans, J.C.




                   Universiteit Maastricht
                                         Table of contents

Table of contents ____________________________________________________ 2
Abstract ___________________________________________________________ 4
List of abbreviations _________________________________________________ 5
Introduction ________________________________________________________ 7
Approaches ________________________________________________________ 9
   Formula 1: Minimal group size calculations _________________________________________ 9
   Formula 2: calculation of the weighted means (the example used in this formula) and differences
   (method 1)__________________________________________________________________ 10
   Formula 3: calculation of the weighted SDs (method 2)_______________________________ 10
 In- and exclusion of biomarkers of effect __________________________________ 11
Results ___________________________________________________________ 13
 Genotoxicity markers___________________________________________________ 14
   Overview group sizes for TM (arb)* ______________________________________________ 14
                                                             -6
   Overview group sizes for HPRT mutation frequency (MF; 10 ), determined by the T-cell cloning
   assay______________________________________________________________________ 15
   Overview group sizes for HPRT variant frequency (VF; 10-6), determined by the BrdU assay _ 15
   Overview group sizes for oxidative damage expressed as 8-OHdG measured in lymphocytes 16
   Overview group sizes for oxidative damage expressed as 8-OHdG measured in leukocytes__ 16
   Overview group sizes for oxidative damage expressed as 8-OHdG measured in urine (µmol/mol
   creatine) ___________________________________________________________________ 17
   Overview group sizes for oxidative damage expressed as 8-OHdG measured in urine (µg/g
   creatine) ___________________________________________________________________ 17
   Overview group sizes for oxidative damage expressed as 8-OHdG measured in urine
   (pmol/kg/24h) _______________________________________________________________ 18
   Overview group sizes for oxidative damage expressed as 8-OHdG measured in urine (ng/BMI)18
   Overview group sizes for oxidative damage expressed as 8-OHdG measured in urine (nmol/24h)
    __________________________________________________________________________ 19
 Hormones ____________________________________________________________ 20
   Overview group sizes for serum testosterone levels in males (and females) ______________ 20
   Overview group sizes for serum oestradiol levels in healthy individuals __________________ 21
   Overview group sizes for serum sex hormon binding globulin (SHBG) levels in healthy individuals
    __________________________________________________________________________ 22
   Overview group sizes for serum FSH levels in healthy individuals ______________________ 23
   Overview group sizes for serum LH levels in healthy individuals ________________________ 24
   Overview group sizes for serum TSH levels in healthy individuals ______________________ 25
   Overview group sizes for serum inhibin B levels in healthy individuals ___________________ 25
 Proposed additional genotoxicity markers _________________________________ 26
   Overview group sizes for CA frequency (per 100 cells) in healthy individuals measured in PBL 26
   Overview group sizes for number of micronucleated cells per 1000 binucleated cells (MNCs) in
   healthy individuals____________________________________________________________ 27
   Overview group sizes for number of MN per 1000 binucleated cells (MNi) in healthy individuals28
                                                                                 8
   Overview group sizes for white blood cell PAH-DNA adduct levels (adducts/ 10 nucleotides) in
   healthy individuals____________________________________________________________ 29
Discussion ________________________________________________________ 32
 Genotoxicity markers___________________________________________________ 32
 Hormones ____________________________________________________________ 33
 Proposed additional genotoxicity markers _________________________________ 34
References ________________________________________________________ 37
Appendices _______________________________________________________ 50
Appendix 1: _______________________________________________________ 51
   DNA damage in healthy individuals measured by the Comet-assay, expressed in Tail Moment. 51
Appendix 2 ________________________________________________________ 53
   HPRT-mutation frequency in healthy individuals, determined by the T-cell cloning assay. ____ 53
   HPRT-mutation frequency in healthy individuals, determined by the BrdU assay. __________ 54
Appendix 3 ________________________________________________________ 56
   Oxidative damage measured in lymphocytes from healthy individuals, determined by HPLC-
   ECD. ______________________________________________________________________ 56
   Oxidative damage measured in leukocytes from healthy individuals, determined by HPLC-ECD.
    __________________________________________________________________________ 57
   Oxidative damage (8-OHdG) measured in urine from healthy individuals. ________________ 58
Appendix 4 ________________________________________________________ 63
   Total testosterone levels in serum of non-exposed healthy males (and females).___________ 63
Appendix 5 ________________________________________________________ 65
   Serum oestradiol levels in healthy individuals. ______________________________________ 65
Appendix 6 ________________________________________________________ 67
   Serum sex-hormone binding globulin (SHBG) levels in healthy individuals. _______________ 67
Appendix 7 ________________________________________________________ 69
   FSH serum levels in healthy individuals ___________________________________________ 69
Appendix 8 ________________________________________________________ 73
   LH serum levels in healthy males ________________________________________________ 73
Appendix 9 ________________________________________________________ 76
   Thyroid stimulating hormone (TSH) levels in healthy individuals ________________________ 76
Appendix 10 _______________________________________________________ 78
   Inhibin B levels in healthy individuals _____________________________________________ 78
Appendix 11: ______________________________________________________ 79
   CA frequency (per 100 cells) in healthy individuals measured in PBL by the CA assay ______ 79
Appendix 12: ______________________________________________________ 82
   Number of micronucleated cells per 1000 binucleated cells (MNCs) in healthy individuals ___ 82
   MN per 1000 binucleated cells (MNi) in healthy individuals ____________________________ 85
Appendix 13: ______________________________________________________ 94
   PAH-DNA adduct levels (adducts/ 108 nucleotides) in healthy individuals, as determined by
   ELISA and 32P-postlabelling methods_____________________________________________ 94
Abstract
The strategic program on Health and Environment in Flanders is performing a large-
scale biomonitoring study, known as the Flanders biomonitoring study. The Flanders
biomonitoring study explores multiple markers of adverse biological effects among
various environmentally exposed subpopulations. The Flanders biomonitoring study
has chosen a preliminary group size of both exposed and control groups of 200
individuals. The present project describes the smallest group sizes possible, still
yielding sufficient statistical power, of the exposed group for all foreseen biomarkers
of effect. Minimal group size calculations have been executed for genotoxicity
markers and hormones, and therefore assesses whether the foreseen group size of
n= 200 is sufficient for yielding statistical power for particular biomarker analysis.
Tumour markers have been excluded as a whole from this review for the following
reasons: (a) lack of reliable published data; in most studies tumour markers are
described for a variety of cancer patients, but not, or to a far less extent, for healthy
individuals (by default assumption no control values) and (b) comparisons with
healthy exposed persons are lacking for tumour markers. In conclusion: the intended
group size of 200 individuals in both exposed and control groups for the Flanders
biomonitoring study is sufficient for most biomarkers for effect described in this
review. Only the reported data on the hormone inhibin B did not satisfy the demands
needed for minimal group size calculations.
List of abbreviations
Σ                  sum of
@                  not significant
α                  the chance to detect a relation between determinant and
                   disease, whereas there is no actual relation
ß                  beta-fault: the chance to overlook an existing relation
                   between determinant and disease
δ                  standard deviation in the population of the continual
                   variable
AE                 accidental exposure
Arb                (visually scored) arbitrary units
BADGE              bisphenol A diglycidyl ether
BMI                body mass index
CBMN               cytochalasin B micronuclei assays
CBPI               cytokinesis-block proliferation index
CS                 current smoker
DAS                4,4’-diaminostilbene-2,2’ disulfonic acid
DBCP               dibromochloropropane
DE                 dietary exposure
DNT                dinitrotoluene
EE                 environmental exposure
ELISA              enzyme-linked immunosorbent assay
ExcE               exposure to physical exercise (regular exercisers as
                   controls)
F                  female
FS                 former smoker
FSH                follicle stimulating hormone
HCH                hexachlorocyclohexane
HPRT               hypoxanthine guanine phosphoribosyltransferase
HRT                hormone replacement therapy
IU                 international units
k                  amount of selected studies
LE                 lifestyle exposure
LH                 luteinizing hormone
M                  male
ME                 medical exposure
meanx              mean study x
MF                 mutation frequency
MP                 menopausal
n                  the number of exposed or unexposed individuals in one
                   study group of the research population
n1                 n (amount of individuals) in observed group
n2                 the calculated amount of exposed individuals in the
                   research population for that specific power: the minimal
                   group size
n2[0.95]           n2 with a power of 0.95
ND                 not defined; lack of relevant and/or significant values
NDI                nuclear division index
NS                         non-smoker
OE                         occupational exposure
8-OhdG                     8-hydroxy-2'-deoxyguanosine
32                         32
  P-postlabelling-HPLC       P-postlabelling followed by the HPLC method
32                         32
  P-postlabelling (n-Bu)     P-postlabelling with n-butanol extraction enrichment as
                           described by Gallagher et al. (1)
32                         32
  P-postlabelling (P1)       P-postlabelling with nuclease P1 enrichment
PAH                        polycyclic aromatic hydrocarbon
PBL                        peripheral blood lymphocytes
PS                         passive smoker
r                          ratio exposed/unexposed
SE                         smoking exposure (non-smokers vs. smokers)
SD                         standard deviation
TDA                        toluene diamine
TM                         tail moment (per scored cell)
TSH                        thyroid stimulating hormone
v                          the value of the difference to be detected or the required
                           difference between the proportions of the research groups
                           to be detected by the research
variance                   SD2
variancex                  variance study x
VF                         variant frequency
WBC                        white blood cells
wf                         weight factor: the proportion of the amount of individuals of
                           a particular study (n1study) as compared to the amount of
                           individuals in all selected studies (n1total).
wfx                        weight factor study x
wfSD                       weight factor for SD calculations: the proportion of the
                           amount of individuals of a particular study (n1study) minus 1
                           as compared to the amount of individuals in all selected
                           studies (n1total) minus the amount of selected studies (k).
wfSDx                      weight factor for SD calculations study x
wmav                       weighted mean as v; thus mean exposed is set at twice
                           the mean of the controls
x                          selected studies
Zα/2                       the value on the standard-normal distribution
Zß                         the value on the standard-normal distribution
Introduction
The strategic program on Health and Environment in Flanders is preparing a large-
scale biomonitoring study, known as the Flanders biomonitoring study (as described
in our earlier review). The Flanders biomonitoring study explores multiple markers of
adverse biological effects among various environmentally exposed subpopulations.
Group sizes for both exposed and control subjects have been set at 200 persons for
each group. The question that this review attempts to address is: is the intended
group size of 200 persons for the control and exposed groups sufficiently large for all
monitored biomarkers of effect to satisfy the need for statistical significance in order
to detect existing differences? The goal of this review is to describe for each
individual biomarker for effect the smallest group sizes possible, still yielding
sufficient statistical power, for the exposed group/ or the group at higher risk of
exposure(s).

A typical limitation of molecular epidemiology studies is the rather small size of study
groups in general, due to logistical difficulties. Solely for the purpose of illustration:
Gordon et al. (2) recently calculated that the statistical power of the published studies
on genetic susceptibility that they reviewed, ranges from 20 to 70%, and that only a
few of all the reviewed studies have an acceptable power to detect a real difference.
Since detecting a real difference, if present, is the practical goal of environmental
health studies in general, special care should be attributed to the design phase in
order to favour the circumstances that allow the detection of existing differences.
Sufficient group sizes lay the foundations on which field studies among humans can
be based. Based on the data on inter- and intra-variability, sensitivity and specificity,
power-calculations can be made, in order to determine the smallest group sizes
possible, still yielding sufficient statistical power. The statistical power regards the
discrimination of a marker between exposed and non-exposed groups, in other
words: the power is the chance that the study is able to detect an existing difference
between exposed and non-exposed groups, given the size of the studied groups.
The biomarkers of effect that have been described in this review, are genotoxicity
markers and hormones, but not tumor markers (as explained in our earlier review).
Additionally, validated genotoxicity markers (first introduced in our earlier review) that
were not yet selected for analysis in the Flanders biomonitoring study, are described
as well in the ‘proposed additional markers for effect’ subchapter.


General note:

Power calculations for some of the described biomarkers of effect are hampered by
limited published data. This study is designed to validate the biomarkers of effect in
healthy individuals; for this purpose, healthy control groups have been selected
(mean and standard deviation) from documented studies. Since the exposure source
in the studied populations of the Flanders biomonitoring study is expected to be of
environmental origin, the observed differences in biomarkers of effect between
environmentally exposed and control subjects have been extracted from earlier
studies. If the published data is inadequate in meeting the criterion of supplying the
studies that allow the extrapolation of such a preferred environmental exposure
based difference, the difference between non-smokers and smokers can be used as
an alternative because smoking has a significant effect on most of the biomarkers of
effect discussed in this review. Furthermore, smoking will be one of the relevant
exposures in the Flanders biomonitoring study.
Approaches

Minimal group size or statistical power can be calculated using the formulas,
described in this chapter. The formula as described by Bouter and van Dongen (3) by
which the smallest group size (n2) in a cause-effect research, given the preferred
power of the research, can be calculated as described in formula 1. The calculated
n2s are always rounded up to whole numbers, in order to gain minimal group size for
groups consisting of whole individuals.



Formula 1: Minimal group size calculations



              (Zα/2 + Zß)2 δ2 (r + 1)
       n2 =            v2r



With the variables:
ß       the chance to overlook an existing relation between determinant and disease;
Zß      the value on the standard-normal distribution;
α       the chance to detect a relation between determinant and disease, whereas there is no actual
        relation;
Zα/2    the value on the standard-normal distribution;
v       the value of the difference to be detected or the required difference between the proportions of
        the research groups to be detected by the research;
n2      the calculated number of exposed (or at higher risk) individuals or the number of patients with
        the disease in the research population;
r       ratio exposed/non-exposed;
δ       standard deviation of the continual variable in the population




An alpha fault (α) of 0.05 and a beta fault (β) of 0.05 or 0.10 (corresponding with
powers of 0.95 and 0.90 respectively) are commonly accepted and therefore also
used in the group size calculations for the Flanders biomonitoring study. For all
differences (v) an alpha fault (α) of 0.05 is considered significant. The differences
used for minimal group size calculations, are solely based on studies on respective
effect markers which reported significant differences between control and exposed
groups (unless specifically mentioned otherwise).
Since this biomonitoring study will use a ratio exposed/unexposed (r) of 1 this is also
applied at the group size of the unexposed group/ the group at lower risk
exposure(s). The power is 1-ß. The power and the sample size are further influenced
by among others (3):
    • Methodological faults: reduced power; more subjects needed;
    • Non-response: bigger sample population needed
    • Drop out (regarding cohort studies): bigger sample population needed;
      •   (individual) matching: increased power; smaller group size needed;
      •   Confounding (necessity to implement correctional methods during data
          analysis): decreased power; bigger group size needed.

Therefore it would be wise to use group sizes, somewhat larger than the minimal
group size, thereby buffering the above problems of limited power, and thus obviating
the possibly decreased ability to detect real differences.

If various studies have been used to calculate a mean value (of the value of a
specific biomarker, of the value of the standard deviation of this specific biomarker or
of the value of the difference between unexposed and exposed), weight factors had
to be applied when comparing unequally sized studies. Larger weight has been
applied to relative bigger studies. Different methods of weight factor determination
have been used for weighted means, weighted differences (formula 2) and weighted
SDs (formula 3). The formulas are described by Bouter and van Dongen (3).




Formula 2: calculation of the weighted means (the example used in this
formula) and differences (method 1)

      •   weighted mean (x) =            Σ(wfx*meanx)


With the variables:
Σ               sum of
meanx           mean study x
wf              weight factor: the proportion of the amount of individuals of a particular study (n1study)
                as compared to the amount of individuals in all selected studies (n1total).
wfx             weight factor study x
x               selected studies




Formula 3: calculation of the weighted SDs (method 2)

      •   weighted variance (x) =        Σ(wfSDx*variancex)

      •   weight factor (study a) =      n1study-1/n1total-k)

      •   weighted SD = square root of the weighted variance


With the variables:
Σ               sum of
k               amount of selected studies
SD              standard deviation
variance        SD2
variancex       variance study x
wfSD         weight factor for SD calculations: the proportion of the amount of individuals of a
             particular study (n1study) minus 1 as compared to the amount of individuals in all
             selected studies (n1total) minus the amount of selected studies.
wfSDx        weight factor for SD calculations study x
x            selected studies




Unless mentioned otherwise:
   • For weighted mean and SD calculations, only means with SDs (so only studies
      that offer both mean and SD) have been used.
   • For weighted differences only significant differences have been used (unless
      explicitly mentioned otherwise). For all differences (v) an alpha fault (α) of 0.05
      is considered significant.

If a discrimination between ‘male’, ‘female’ or ‘male and female’ classes has been
applied in the pooled/ weighted classes, different colourations have been appointed
to the characters describing these classes: black (‘male’), blue (‘female’) and green
(‘male and female’), respectively. For the ‘male and female’ weighted classes only
studies that have ‘male and female’ controls have been used, unless there is a fair
amount of ‘male’ and ‘female’ studies available as well (as can be seen in
Appendices), then the choice has been made to recruit these ‘male’ and ‘female’
studies into the ‘male and female’ pooled classes.
An appointed difference equal to the weighted mean (thus exposed with an estimated
biomarker of effect concentration of twice that of the controls) will also be described.
This has been done to allow alternative n2-calculations, additional to the study-based
n2-calculations, and as a crude hint when study-based n2-calculations could not be
calculated, due to the absence of essential parameters. All the differences used in
our minimal group size calculations are weighted differences, extrapolated from
earlier studies, and (unless mentioned otherwise) resemble significant (p ≤0.05)
differences in their original studies.
Microsoft Windows Office XP Excel has been used for all calculations.


In- and exclusion of biomarkers of effect
Regarding the COMET assay markers, tailmoment ((visually scored) arbitrary units)
is preferred over tailmoment (computer-generated linear scale), since the Flanders
biomonitoring study is using the first method. Tailmoment (TM) integrates tail length
(TL) and the percentage of DNA (%DNA) in the tail, thereby excusing TL and %DNA
from separate appearances as biomarkers in the Flanders biomonitoring study.
Regarding the HPRT assay, both T-cell cloning assay cloning efficiency and BrdU
labelling index merely reflect the yield of the respective procedures instead of
reflecting the value of a specific biomarker, hence their presence is also excused. 8-
hydroxy-2'-deoxyguanosine (8-OHdG) in urine (nmol/L) has been excluded since this
is not a normalized marker: no correction (e.g. expressed per amount of creatinine or
per 24 day) has been applied to correct for dilution rates. Tumour markers have been
excluded as a whole, because of lack of reliable published data; in most studies
tumour markers are described for a variety of cancer patients, but not, or to a far less
extent, for healthy individuals (by default assumption no control values). Furthermore,
comparisons with healthy exposed persons are lacking for tumour markers.
Regarding the CBMN assay, both the nuclear division index (NDI) and the
cytokinesis-block proliferation index (CBPI) merely reflect the yield of the specific
stages of the assay instead the value of a specific biomarker of effect, therefore their
presence is excused as well.
Results
The results for the pooled studies are displayed in the Tables, the Appendices add to
this information, the data derived from the specific studies. The numbers applied to
Tables and their respective Appendices correspond (e.g. Table 1 displays the same
data as Appendix 1).
For serum inhibin B (Table 10) no n2 could be calculated since no SDs were available
in any of the studies reviewed (Appendix 10).
Table 1 will be used to illustrate the process applied to derive the results as
described in this review. The studies used to obtain the weighted values as displayed
in Table 1 are shown in Appendix 1. The order of occurrence in the Appendices is
environmental exposure (EE) studies, followed by occupational exposure (EO)
studies and finally lifestyle exposure (LE) studies. The latter is subdivided in smoking
exposure (SE), dietary exposure (DE) and similar subgroups. The purpose of the
Tables is to give only the most relevant information, while the Appendices offer the
ability to take a more in depth view of the studies that have contributed to these
results. Weighted results display the weighted result of control groups derived from
separate studies that share similar characteristics (e.g. all non-smoking individuals).
For each class of pooled studies with similar characteristics, an attempt has been
made to derive a (multiple) study-based difference (v) between exposed and control
groups. The preferred exposure for such a v is of the environmental nature (EE), in
case a statistically significant EE based v could not be attained a SE based v has
been proposed (if possible). In the rare case that both EE- and SE-based differences
are not significant or not described, an alternatively based v has been used
(specifically mentioned) but this v is purely directional and the n2-calculations with
such a v are expected to be of little relevance for the Flanders biomonitoring study.
With this information a study-based minimal group size has been calculated.
Alternatively, a minimal group size based on a ratio exposed/control (or vice versa) of
2/1 has been calculated. In that case the weighted mean has been used as a v
(wmav).
For example: with respect to the TM (arb) of the COMET biomarker for the class of
‘NS+CS+ES’ (Table 1/Appendix 1) all studies in Appendix 1 have been used (4-12)
to determine a weighted mean (with a value of 0.62) and a weighted SD (with a value
of 0.26), while the v (with a value of 0.07) has been extrapolated from only one study
(4). Only one study had the preferred combination: significantly different from the
control group and EE. The symbol as displayed behind the weighted v ( ) indicates
that the v is derived from a study in which solely controls of the NS smoking class
were enrolled, while in this case the weighted mean and weighted SD is based on
controls of the NS, CS and ES smoking classes. Since no other v is available, the
option of accepting the NS smoking class-derived v for the NS+CS+ES smoking
class leaves open the possibility to determine an indication of the study-based
minimal group size. The ratio exposed/control (or vice versa) of 2/1, allows an
alternative calculation. For the class of ‘NS’ (Table 1/Appendix 1) only the studies
with NS (4-9) have been used (Appendix 1) to determine a weighted mean (with a
value of 0.76) and a weighted SD (with a value of 0.20), while the v (with a value of
0.07) again could only be extrapolated from one study (4), there is no symbol behind
the weighted v since both weighted SD and v are derived from studies with NS in this
case (class ‘NS’). The wmav based minimal group size calculation is again
substantially different from the study-based minimal group size calculation.
The other Tables have been attained in similar fashion as the above discussed
example (Table 1). It is clearly shown that the higher the statistical power one
pursues, the bigger the minimal group size has to be in order to meet that
requirement.



Genotoxicity markers


Table 1:

Overview group sizes for TM (arb)*
 Exposure     Smoking          Studies      TM (arb) weighted mean        Weighted v (n1)         n2
 type used    status                          ± weighted SD (n1)                                [power]
     to
determine v
EE            NS+CS+ES           (4-12)         0.62 ± 0.26 (375)           0.07 (20)         290 [0.90]
                                                                                              359 [0.95]
-             NS+CS+ES           (4-12)         0.62 ± 0.26 (375)           0.62 (wmav)         4 [0.90]
                                                                                                5 [0.95]
EE            NS                 (4-9)          0.76 ± 0.20 (167)            0.07 (20)        172 [0.90]
                                                                                              213 [0.95]
-             NS                 (4-9)          0.76 ± 0.20 (167)           0.76 (wmav)         2 [0.90]
                                                                                                2 [0.95]
EE            CS                (4, 5, 7)        0.48 ± 0.10 (47)           0.07 (20)          43 [0.90]
                                                                                               54 [0.95]
-             CS                (4, 5, 7)        0.48 ± 0.10 (47)           0.48 (wmav)         1 [0.90]
                                                                                                2 [0.95]
EE: environmetal exposure. NS: non-smoker. *: The principle of the COMET assay and tailmoments
(TMs) has been described in our earlier reviews. CS: current smoker. ES: ex-smoker. TM: tail moment
(per scored cell). Arb: (visually scored) arbitrary units. n1: n (amount of individuals) in observed group.
n2: the calculated amount of exposed individuals in the research population for that specific power. v:
difference in biomarker between exposed and control group. All detections were performed with the
alkaline COMET-assay. Biological medium in all the detections were leukocytes. wmav: weighted
mean as v; thus mean exposed is set at twice the mean of the controls. : the v is derived from a
study with NS instead of CS (or NS+CS) as in this case.
Table: 2

Overview group sizes for HPRT mutation frequency (MF; 10-6),
determined by the T-cell cloning assay
                                                    -6
 Exposure     Smoking         Studies         MF (10 ) weighted mean         Weighted v (n1)       n2
 type used    status                            ± weighted SD (n1)                               [power]
     to
determine v
EE            NS+CS+FS         (13-27)          11.05 ± 15.35 (836)             10.35 (9)        47 [0.90]
                                                                                                 58 [0.95]
-             NS+CS+FS         (13-27)          11.05 ± 15.35 (836)           11.05 (wmav)       41 [0.90]
                                                                                                 51 [0.95]
EE            NS               (13-19)           8.64 ± 5.88 (225)              10.35 (9)§       7 [0.90]
                                                                                                 9 [0.95]
-             NS               (13-19)           8.64 ± 5.88 (225)             8.64 (wmav)       10 [0.90]
                                                                                                 13 [0.95]
EE            CS               (13-15)            4.62 ± 3.25 (61)              10.35 (9)§       3 [0.90]
                                                                                                 3 [0.95]
-             CS               (13-15)            4.62 ± 3.25 (61)             4.62 (wmav)       11 [0.90]
                                                                                                 13 [0.95]
EE: environmetal exposure. NS: non-smoker. CS: current smoker. FS: former smoker. n1: n (amount
of individuals) in observed group. n2: the calculated amount of exposed individuals in the research
population for that specific power. ND: not defined; lack of relevant and/or significant values. v:
difference in biomarker between exposed and control group. wmav: weighted mean as v; thus mean
exposed is set at twice the mean of the controls. §: the v is derived from a study with a not defined
smoking status, instead of the defined smoking status as in this case.



Overview group sizes for HPRT variant frequency (VF; 10-6), determined
by the BrdU assay
 Exposure type     Smoking         Studies          VF (10-6) weighted         Weighted v (n1)      n2 [power]
     used to       status                          mean ± weighted SD
   determine v                                              (n1)
EE                 NS+CS            (28-30)         19.38 ± 30.50 (75)       240.00 (21)              1 [0.90]
                                                                                                      1 [0.95]
-                  NS+CS            (28-30)          19.38 ± 30.50 (75)      19.38 (wmav)            53 [0.90]
                                                                                                     65 [0.95]
EE                 NS              (28, 29)              5.51 ± 6.57 (30)    240.00 (21)§             1 [0.90]
                                                                                                      1 [0.95]
-                  NS              (28, 29)              5.51 ± 6.57 (30)    5.51 (wmav)             30 [0.90]
                                                                                                     37 [0.95]
EE                 CS                (28)                16.8 ± 24.9 (10)¶   240.00 (21)§             1 [0.90]
                                                                                                      1 [0.95]
-                  CS                (28)                16.8 ± 24.9 (10)¶   16.8 (wmav)             47 [0.90]
                                                                                                     58 [0.95]
EE: environmetal exposure. NS: non-smoker. CS: current smoker. n1: n (amount of individuals) in
observed group. n2: the calculated amount of exposed individuals in the research population for that
specific power. v: difference in biomarker between exposed and control group. wmav: weighted mean
as v; thus mean exposed is set at twice the mean of the controls. §: the v is derived from a study with
a NS+CS, instead of the only NS (or CS) as in this case. ¶: only based on one study, and a small one
at that (28).
Table 3:

Overview group sizes for oxidative damage expressed as 8-OHdG
measured in lymphocytes
                                                             -6
 Exposure type      Smoking        Studies       8-OHdG (10 ) weighted              Weighted v (n1)   n2 [power]
     used to        status                       mean ± weighted SD (n1)
   determine v
SE                  NS+CS         (27, 31-35)        4.61 ± 1.80 (615)          12.3 (21) ø             1 [0.90]
                                                                                                        1 [0.95]
-                   NS+CS         (27, 31-33)        4.61 ± 1.80 (615)          4.61 (wmav)             4 [0.90]
                                                                                                        4 [0.95]
SE                  NS            (27, 31-33)        7.69 ± 2.49 (229)          12.3 (21)ø              1 [0.90]
                                                                                                        2 [0.95]
-                   NS            (27, 31-33)        7.69 ± 2.49 (229)          7.69 (wmav)             3 [0.90]
                                                                                                        3 [0.95]
SE                  CS              (31, 32)          4.66 ± 2.24 (92)          12.3 (21) ø             1 [0.90]
                                                                                                        1 [0.95]
-                   CS              (31, 32)          4.66 ± 2.24 (92)          4.66 (wmav)             5 [0.90]
                                                                                                        7 [0.95]
SE: smoking exposure (non-smokers vs. smokers). NS: non-smoker. CS: current smoker. @: not
significant. ND: not defined; lack of relevant and/or significant values. n1: n (amount of individuals) in
observed group. n2: the calculated amount of exposed individuals in the research population for that
specific power. v: difference in biomarker between exposed and control group. wmav: weighted mean
as v; thus mean exposed is set at twice the mean of the controls. : the v is derived from a study with
NS (33) instead of CS (or CS+NS) as in this case. ø: since no EE based v could be extracted from
studies a SE based v has been applied.



Overview group sizes for oxidative damage expressed as 8-OHdG
measured in leukocytes
                                                              -6
 Exposure type      Smoking       Studies         8-OHdG (10 )           Weighted v (n1)         n2 [power]
     used to        status                      weighted mean ±
   determine v                                  weighted SD (n1)
EE                  NS+CS+F       (36-44)       13.61 ± 3.30 (236)       1.5 (24)                102 [0.90]
                    S                                                                            126 [0.95]
-                   NS+CS+F       (36-44)       13.61 ± 3.30 (236)       13.61 (wmav)             2 [0.90]
                    S                                                                             2 [0.95]
EE                  NS            (36-42)       11.44 ± 2.97 (160)       1.5 (24)                 83 [0.90]
                                                                                                 102 [0.95]
-                   NS            (36-42)       11.44 ± 2.97 (160)       11.44 (wmav)             2 [0.90]
                                                                                                  2 [0.95]
EE                  CS              (36)        33.30 ± 2.79 (30)¶       1.5 (24)                 73 [0.90]
                                                                                                  90 [0.95]
-                   CS              (36)        33.30 ± 2.79 (30)¶       33.30 (wmav)             1 [0.90]
                                                                                                  1 [0.95]
EE: environmental exposure. n1: n (amount of individuals) in observed group. n2: the calculated
amount of exposed individuals in the research population for that specific power. NS: non-smoker. CS:
current smoker. FS: former smoker. v: difference in biomarker between exposed and control group.
wmav: weighted mean as v; thus mean exposed is set at twice the mean of the controls. : the v is
derived from a study (37) with NS instead of CS (or CS+NS) as in this case. ¶: only based on one
study (36).
Overview group sizes for oxidative damage expressed as 8-OHdG
measured in urine (µmol/mol creatine)
 Exposure type     Smoking       Studies            8-OHdG (µmol/mol            Weighted v         n2 [power]
     used to       status                       creatine) weighted mean ±          (n1)
   determine v                                       weighted SD (n1)
EE                 NS+CS          (45-48)             1.60 ± 0.92 (130)        0.54 (89)           62 [0.90]
                                                                                                   76 [0.95]
-                  NS+CS          (45-48)            1.60 ± 0.92 (130)         1.60 (wmav)          7 [0.90]
                                                                                                    9 [0.95]
EE                 NS             (45-47)            1.64 ± 0.71 (103)         0.54 (89)           37 [0.90]
                                                                                                   45 [0.95]
-                  NS             (45-47)            1.64 ± 0.71 (103)         1.64 (wmav)          4 [0.90]
                                                                                                    5 [0.95]
EE                 CS              ND                      ND                  0.54 (89)               ND
-                  CS              ND                      ND                  ND                     ND
EE: environmental exposure. NS: non-smoker. CS: current smoker. n1: n (amount of individuals) in
observed group. n2: the calculated amount of exposed individuals in the research population for that
specific power. ND: not defined; lack of relevant and/or significant values. v: difference in biomarker
between exposed and control group. wmav: weighted mean as v; thus mean exposed is set at twice
the mean of the controls. : the v is derived from studies (45, 46) with NS instead of CS (or CS+NS)
as in this case.



Overview group sizes for oxidative damage expressed as 8-OHdG
measured in urine (µg/g creatine)
 Exposure type     Smoking          Studies           8-OHdG (µg/g creatine)        Weighted v (n1)      n2 [power]
     used to       status                                weighted mean ±
   determine v                                           weighted SD (n1)
EE                 NS+CS         (34, 44, 49, 50)        17.27 ± 4.16 (439)         1.97 (118)              94 [0.90]
                                                                                                           116 [0.95]
-                  NS+CS         (34, 44, 49, 50)         17.27 ± 4.16 (439)        17.27 (wmav)             2 [0.90]
                                                                                                             2 [0.95]
EE                 NS                (49, 50)             16.66 ± 1.88 (292)        2.1 (43)                17 [0.90]
                                                                                                            21 [0.95]
-                  NS                (49, 50)             16.66 ± 1.88 (292)        16.66 (wmav)             1 [0.90]
                                                                                                             1 [0.95]
EE                 CS                 (49)                 13.3 ± 4.5 (32)¶         1.97 (118)Ħ            110 [0.90]
                                                                                                           136 [0.95]
-                  CS                 (49)                 13.3 ± 4.5 (32)¶         13.3 (wmav)              3 [0.90]
                                                                                                             3 [0.95]
EE: environmental exposure. n1: n (amount of individuals) in observed group. n2: the calculated
amount of exposed individuals in the research population for that specific power. NS: non-smoker. CS:
current smoker. v: difference in biomarker between exposed and control group. wmav: weighted
mean as v; thus mean exposed is set at twice the mean of the controls. Ħ: since no significant v from
study groups with solely CS was available, the applied v was derived from studies with NS+CS or NS.
¶: only based on one study (49).
Overview group sizes for oxidative damage expressed as 8-OHdG
measured in urine (pmol/kg/24h)
 Exposure type     Smoking        Studies       8-OHdG (pmol/kg/24h)       Weighted v        n2 [power]
     used to       status                          weighted mean ±            (n1)
   determine v                                     weighted SD (n1)
EE                 NS+CS        (46, 51, 52)     212.84 ± 113.21 (124)   107 (53)             24 [0.90]
                                                                                              30 [0.95]
-                  NS+CS        (46, 51, 52)     212.84 ± 113.21 (124)   212.84 (wmav)        6 [0.90]
                                                                                              8 [0.95]
EE                 NS             (46, 51)       232.96 ± 113.97 (97)    107 (53)             24 [0.90]
                                                                                              30 [0.95]
-                  NS             (46, 51)       232.96 ± 113.97 (97)    232.96 (wmav)        6 [0.90]
                                                                                              7 [0.95]
EE                 CS               ND                    ND             107 (53)                ND
-                  CS               ND                    ND             ND                     ND
EE: environmental exposure. n1: n (amount of individuals) in observed group. n2: the calculated
amount of exposed individuals in the research population for that specific power. NS: non-smoker. CS:
current smoker. ND: not defined; lack of relevant and/or significant values. v: difference in biomarker
between exposed and control group. wmav: weighted mean as v; thus mean exposed is set at twice
the mean of the controls. : the v is derived from a study (51) with NS instead of CS (or CS+NS) as
in this case.



Overview group sizes for oxidative damage expressed as 8-OHdG
measured in urine (ng/BMI)
 Exposure type     Smoking       Studies       8-OHdG (ng/BMI)      Weighted v (n1)     n2 [power]
    used to        status                      weighted mean ±
  determine v                                  weighted SD (n1)
ND                 NS+CS                             ND             ND                     ND
-                  NS+CS                             ND             ND                     ND
SE                 NS              (33)         197 ± 31 (24)¶      43@ (24)ø           11 [0.90]ø
                                                                                        14 [0.95]ø
-                  NS              (33)         197 ± 31 (24)¶      197 (wmav)           1 [0.90]
                                                                                         1 [0.95]
ND                 CS                                ND             ND                      ND
-                  CS                                ND             ND                     ND
SE: smoking exposure (non-smokers vs. smokers). n1: n (amount of individuals) in observed group. n2:
the calculated amount of exposed individuals in the research population for that specific power. NS:
non-smoker. CS: current smoker. @: not significant. ND: not defined; lack of relevant and/or significant
values. v: difference in biomarker between exposed and control group. wmav: weighted mean as v;
thus mean exposed is set at twice the mean of the controls. ø: since no significant v was available, a
non-significant v has been chosen for this calculation. ¶: only based on one study (33).
Overview group sizes for oxidative damage expressed as 8-OHdG
measured in urine (nmol/24h)
 Exposure type     Smoking           Studies     8-OHdG (nmol/24h)      Weighted v (n1)    n2 [power]
     used to       status                         weighted mean ±
   determine v                                    weighted SD (n1)
EE                 NS+4 week         (46, 53)    21.94 ± 10.76 (102)    6.05 (102)          67 [0.90]
                   smoking                                                                  83 [0.95]
                   cessation CS
-                  NS+4 week           (46)       21.94 ± 10.76 (102)   21.94 (wmav)        6 [0.90]
                   smoking                                                                  7 [0.95]
                   cessation CS
EE                 NS                  (46)        19.1 ± 11.1 (44)¶    5.6 (44)           83 [0.90]
                                                                                           103 [0.95]
-                  NS                  (46)        19.1 ± 11.1 (44)¶    19.1 (wmav)         8 [0.90]
                                                                                            9 [0.95]
EE                 4 week              (53)        24.1 ± 10.5 (58)¶    6.4 (58)           57 [0.90]
                   smoking                                                                 70 [0.95]
                   cessation CS
-                  4 week              (53)        24.1 ± 10.5 (58)¶    24.1 (wmav)         4 [0.90]
                   smoking                                                                  5 [0.95]
                   cessation CS
EE: environmental exposure. n1: n (amount of individuals) in observed group. n2: the calculated
amount of exposed individuals in the research population for that specific power. NS: non-smoker. CS:
current smoker. @: not significant. ND: not defined; lack of relevant and/or significant values. v:
difference in biomarker between exposed and control group. wmav: weighted mean as v; thus mean
exposed is set at twice the mean of the controls. ¶: only based on one study: NS (46) or 4 week
smoking cessation CS (53).
Hormones


Table 4:

Overview group sizes for serum testosterone levels in males (and
females)
 Exposure type     Smoking status       Studies      Testosterone (ng/ml)     Weighted v       n2
     used to                                           weighted mean ±           (n1)        [power]
   determine v                                         weighted SD (n1)
EE                 M: NS+CS             (54-65)       3.72 ± 1.13 (1423)     0.44 (1158)    139 [0.90]
                                                                                            172 [0.95]
-                  M: NS+CS             (54-65)        3.72 ± 1.13 (1423)    3.72 (wmav)     2 [0.90]
                                                                                             3 [0.95]
EE                 M: NS+CS Ð          (54-58, 60,     3.54 ± 1.03 (1280)    0.44 (1158)    116 [0.90]
                                         61, 63)                                            143 [0.95]
-                  M: NS+CS Ð          (54-58, 60,     3.54 ± 1.03 (1280)    3.54 (wmav)     2 [0.90]
                                         61, 63)                                             3 [0.95]
EE                 M: NS (= M: NS        (54, 55)      2.87 ± 1.00 (959)     0.44 (1158)    109 [0.90]
                   Ð)                                                                       135 [0.95]
-                  M: NS (= M: NS       (54, 55)       2.87 ± 1.00 (959)     2.87 (wmav)     3 [0.90]
                   Ð)                                                                        4 [0.95]
EE                 F: NS                (66, 67)       0.21 ± 0.04 (392)     0.046 (377)     16 [0.90]
                                                                                             20 [0.95]
-                  F: NS                (66, 67)       0.21 ± 0.04 (392)     0.21 (wmav)     1 [0.90]
                                                                                             1 [0.95]
EE: environmental exposure. n1: n (amount of individuals) in observed group. n1: n (amount of
individuals) in observed group. n2: the calculated amount of exposed individuals in the research
population for that specific power. NS: non-smoker. CS: current smoker. FS: former smoker. v:
difference in biomarker between exposed and control group. wmav: weighted mean as v; thus mean
exposed is set at twice the mean of the controls. M: males. F: females. Ð: studies with potentially
‘compromised’ controls excluded (59, 62, 64, 65). : the v is derived from studies (54, 68) with NS
instead of CS+NS as in this case.
Table 5:

Overview group sizes for serum oestradiol levels in healthy individuals
 Exposure type     Smoking        Studies       Oestradiol (pg/ml)      Weighted v     n2 [power]
     used to       status                        weighted mean ±           (n1)
   determine v                                   weighted SD (n1)
EE                 M:            (59, 69-71)    29.18 ± 15.77 (162)    6.38 (23) ψ     129 [0.90]
                   NS+CS                                                               159 [0.95]
-                  M:            (59, 69-71)    29.18 ± 15.77 (162)    29.18 (wmav)     7 [0.90]
                   NS+CS                                                                8 [0.95]
ND                 M:              (70, 71)      38.54 ± 18.43 (59)    ND                 ND
                   NS+CS Ð
-                  M:              (70, 71)     38.54 ± 18.43 (59) ψ   38.54 (wmav)     5 [0.90]
                   NS+CS Ð                                                              6 [0.95]
EE                 M: NS ψ          (69)        26.65 ± 3.04 (23) ψ¶   6.38 (23) ψ      5 [0.90]
                                                                                        6 [0.95]
-                  M: NS ψ          (69)        26.65 ± 3.04 (23) ψ¶   26.65 (wmav)     1 [0.90]
                                                                                        1 [0.95]
EE                 Pre- and        (72-75)      8.63 ± 29.72 (3024)    11.69 (552)     136 [0.90]
                   post-MP                                                             169 [0.95]
                   F: NS+FS
-                  Pre- and        (72-75)      8.63 ± 29.72 (3024)    8.63 (wmav)     250 [0.90]
                   post-MP                                                             309 [0.95]
                   F: NS+FS
EE                 Post-MP         (72, 73)     7.14 ± 28.75 (2943)¶   3.7 (136)       1270 [0.90]
                   F: NS+FS                                                            1570 [0.95]
-                  Post-MP         (72, 73)     7.14 ± 28.75 (2943)¶   7.14 (wmav)      341 [0.90]
                   F: NS+FS                                                             422 [0.95]
EE                 Post-MP          (72)        7.14 ± 28.75 (2943)¶   ND                  ND
                   F: NS
-                  Post-MP          (72)        7.14 ± 28.75 (2943)¶   7.14 (wmav)     341 [0.90]
                   F: NS                                                               422 [0.95]
EE: environmental exposure. n1: n (amount of individuals) in observed group. n2: the calculated
amount of exposed individuals in the research population for that specific power. NS: non-smoker. CS:
current smoker. FS: former smoker. ND: not defined; lack of relevant and/or significant values. v:
difference in biomarker between exposed and control group. wmav: weighted mean as v; thus mean
exposed is set at twice the mean of the controls. M: males. F: females. MP: menopausal. Ψ: no real
controls, but patients with benign prostatic hyperplasia. Ð: excluding possibly ‘compromised’ control-
containing studies (59, 69). ¶: only based on one study: females (72) and males (69).
Table 6:

Overview group sizes for serum sex hormon binding globulin (SHBG)
levels in healthy individuals
    Exposure type used    Smoking        Studies     SHBG (nmol/L)       Weighted     n2 [power]
      to determine v      status                    weighted mean ±       v (n1)
                                                    weighted SD (n1)
   EE                     M: NS+CS        (76-78)   22.24 ± 4.85 (146)   6.55 (18)    12 [0.90]
                                                                                      15 [0.95]
   -                      M: NS+CS        (76-78)   22.24 ± 4.85 (146)   22.24        1 [0.90]
                                                                         (wmav)       2 [0.95]
   EE                     M: NS+CS       (76, 77)   22.66 ± 3.39 (109)   6.55 (18)    6 [0.90]
                          Ð                                                           7 [0.95]
   -                      M: NS+CS       (76, 77)   22.66 ± 3.39 (109)   22.66        1 [0.90]
                          Ð                                              (wmav)       1 [0.95]
   EE                     M: NS            (76)      28.6 ± 8.3 (18)¶    6.55 (18)    34 [0.90]
                                                                                      42 [0.95]
   -                      M: NS            (76)      28.6 ± 8.3 (18)¶    28.6         2 [0.90]
                                                                         (wmav)       3 [0.95]
   EE                     Pre- and        (79-81)          ND            8.95            ND
                          post-MP F:                                     (181)
                          NS
   -                      Pre- and        (79-81)          ND            ND              ND
                          post-MP F:
                          NS
   EE                     Pre-MP F:       (79-81)          ND            8.95 (181)      ND
                          NS
   -                      Pre-MP F:       (79-81)          ND            ND              ND
                          NS
EE: environmental exposure. n1: n (amount of individuals) in observed group. n2: the calculated
amount of exposed individuals in the research population for that specific power. NS: non-smoker. CS:
current smoker. FS: former smoker. @: not significant. ND: not defined; lack of relevant and/or
significant values. v: difference in biomarker between exposed and control group. wmav: weighted
mean as v; thus mean exposed is set at twice the mean of the controls. M: male. F: female. MP:
Menopausal. Ð: studies with potentially ‘compromised’ controls excluded (78, 82). ¶: only based on
one study (76). : the v is derived from a study (76) for males or studies for females (79-81) with NS
instead of CS+NS as in this case.
Table 7:

Overview group sizes for serum FSH levels in healthy individuals
Exposure type    Smoking status            Studies              FSH (IU/L)         Weighted v       n2
   used to                                                   weighted mean ±          (n1)        [power]
 determine v                                                 weighted SD (n1)
EE               M: NS+CS           (56-62, 78, 83-97)       6.21 ± 4.66 (1048)    0.81 (109)    696 [0.90]
                                                                                                 861 [0.95]
-                M: NS+CS           (56-62, 78, 83-97)       6.21 ± 4.66 (1047)    6.21 (wmav)    12 [0.90]
                                                                                                  15 [0.95]
EE               M: NS+CS Ð         (56-58, 60, 61, 83-94)    6.10 ± 1.96 (822)    0.81 (108)    124 [0.90]
                                                                                                 153 [0.95]
-                M: NS+CS Ð         (56-58, 60, 61, 83-94)    6.10 ± 1.96 (822)    6.10 (wmav)    3 [0.90]
                                                                                                  3 [0.95]
EE               M: NS              (83)                     12.22 ± 3.22 (20)¶    0.81 (108)†   333 [0.90]
                                                                                                 411 [0.95]
-                M: NS              (83)                     12.22 ± 3.22 (20)¶    12.22          2 [0.90]
                                                                                   (wmav)         2 [0.95]
EE               Pre- and post-     (74, 75, 98-103)         15.95 ± 15.36 (604)   2.9 (22)††    590 [0.90]
                 MP F: NS+CS                                                                     730 [0.95]
-                Pre- and post-     (74, 75, 98-103)         15.95 ± 15.36 (604)   15.95          20 [0.90]
                 MP F: NS+CS                                                       (wmav)         25 [0.95]
EE               Pre-MP F:          (74, 98-101)              9.20 ± 7.87 (492)    2.9 (22)††    155 [0.90]
                 NS+CS                                                                           192 [0.95]
-                Pre-MP F:          (74, 98-101)              9.20 ± 7.87 (492)    9.20 (wmav)    16 [0.90]
                 NS+CS                                                                            20 [0.95]
EE               Pre-MP F: NS       (99)                      9.5 ± 8.1 (447)¶     2.9 (22)††    164 [0.90]
                                                                                                 203 [0.95]
-                Pre-MP F: NS       (99)                      9.5 ± 8.1 (447)¶     9.5 (wmav)     16 [0.90]
                                                                                                  19 [0.95]
EE               Pre-MP F: CS       (98)                       6.0 ± 0.8 (11)¶     2.9 (22)††     2 [0.90]
                                                                                                  2 [0.95]
-                Pre-MP F: CS       (98)                       6.0 ± 0.8 (11)¶     6.0 (wmav)     1 [0.90]
                                                                                                  1 [0.95]
EE               Post-MP F:         (100-102)                63.98 ± 16.47 (31)    2.9 (22)      678 [0.90]
                 NS+CS                                                                           839 [0.95]
-                Post-MP F:         (100-102)                63.98 ± 16.47 (31)    63.98          2 [0.90]
                 NS+CS                                                             (wmav)         2 [0.95]
EE: environmental exposure. n1: n (amount of individuals) in observed group. n2: the calculated
amount of exposed individuals in the research population for that specific power. NS: non-smoker. CS:
current smoker. M= male. F=female. MP: menopausal. v: difference in biomarker between exposed
and control group. wmav: weighted mean as v; thus mean exposed is set at twice the mean of the
controls. †: the applied v is based on studies with NS+CS or ND smoking status, instead of NS as in
this case. ††: the applied v is based on studies (100, 101) with ND smoking status post-MP females
instead of the characteristics of the control group in this case. ¶: only based on one study: males (83)
and females (98) or (99). Ð: studies with potentially ‘compromised’ controls excluded (59, 62, 78, 95-
97, 104).
Table 8:

Overview group sizes for serum LH levels in healthy individuals
Exposure type    Smoking              Studies        LH (IU/L) weighted     Weighted v (n1)       n2
   used to       status                             mean ± weighted SD                          [power]
 determine v                                                 (n1)
EE               M:           (56-62, 78, 86-88,      8.34 ± 2.04 (944)     1.54 (5)†          37 [0.90]
                 NS+CS        90-93, 95, 96, 105)                                              46 [0.95]
-                M:           (56-58, 60, 61, 86-     8.34 ± 2.04 (944)     8.34 (wmav)        2 [0.90]
                 NS+CS        88, 90-93, 105)                                                  2 [0.95]
EE               M:           (56-62, 78, 86-88,      8.93 ± 1.51 (735)     1.54 (5)†          21 [0.90]
                 NS+CS Ð      90-93, 95, 96, 105)                                              25 [0.95]
-                M:           (56-58, 60, 61, 86-     8.93 ± 1.51 (735)     8.93 (wmav)        1 [0.90]
                 NS+CS Ð      88, 90-93, 105)                                                  1 [0.95]
EE               M: NS        (105)                    3.43 ± 0.77 (5)¶     1.54 (5)           6 [0.90]
                                                                                               7 [0.95]
-                M: NS        (105)                    3.43 ± 0.77 (5)¶     3.43 (wmav)        2 [0.90]
                                                                                               2 [0.95]
ND               Pre- and     (75, 99)               17.32 ± 16.08 (528)    ND                    ND
                 post-MP
                 F: NS
-                Pre- and     (75, 99)               17.32 ± 16.08 (528)    17.32 (wmav)       19 [0.90]
                 post-MP                                                                       23 [0.95]
                 F: NS
ND               Pre-MP F:    (99)                    14.9 ± 14.8 (447)¶    ND                    ND
                 NS
-                Pre-MP F:    (99)                    14.9 ± 14.8 (447)¶    14.9 (wmav)        21 [0.90]
                 NS                                                                            26 [0.95]
EE: environmental exposure. n1: n (amount of individuals) in observed group. n2: the calculated
amount of exposed individuals in the research population for that specific power. NS: non-smoker. CS:
current smoker. FS: former smoker. M= male. F=female. MP: menopausal. @: not significant. ND: not
defined; lack of relevant and/or significant values. v: difference in biomarker between exposed and
control group. ¥: men are recruited partners from couples on their first infertility consultation. §:
controls are greenhouse workers with 0-4 years OE in this occupation. Ф : male residents from Cairo,
Egypt (one of the most polluted cities in the world: especially EE to lead is very high) (62). Θ: no real
controls but group with lowest serum concentration of chromium (<1.07 nmol/ mmol creatinine): a
biomarker for exposure, thereby primarily consisting of controls but also a minority fraction consists of
‘exposed’ (metal welders). Ǿ: no real controls, since this group consists of pesticide applicators that
have not been exposed to DBCP in that year. DBCP: dibromochloropropane, a nematocide. HCH:
hexachlorocyclohexane. BADGE: bisphenol A diglycidyl ether. wmav: weighted mean as v; thus mean
exposed is set at twice the mean of the controls. †: the applied v is based on a study with NS (105),
instead of NS+CS as in this case. Ð: studies with potentially ‘compromised’ controls (59, 62, 78, 95,
96) are excluded. ¶: only based on one study: males (105) and females (99).
Table 9:

Overview group sizes for serum TSH levels in healthy individuals
 Exposure type      Smoking          Studies    TSH (mIU/L) weighted          Weighted v     n2 [power]
     used to        status                     mean ± weighted SD (n1)           (n1)
   determine v
EE                  M+pre- and        (106,           2.4 ± 0.1 (50)¶        0.14 (2800)      11 [0.90]
                    post-MP F:        107)                                                    14 [0.95]
                    NS+FS
-                   M+pre- and        (106,           2.4 ± 0.1 (50)¶        2.4 (wmav)       1 [0.90]
                    post-MP F:        107)                                                    1 [0.95]
                    NS+FS
ND                  M+pre-MP F:       (106)           2.4 ± 0.1 (50)¶        ND                 ND
                    NS
-                   M+pre-MP F:       (106)           2.4 ± 0.1 (50)¶        2.4 (wmav)       1 [0.90]
                    NS                                                                        1 [0.95]
EE                  M: NS+CS           (88,         1.86 ± 0.34 (49)         0.7 (4)          5 [0.90]
                                       108,                                                   7 [0.95]
                                      109)
-                   M: NS+CS           (88,         1.86 ± 0.34 (49)         1.86 (wmav)      1 [0.90]
                                       108,                                                   1 [0.95]
                                      109)
DE                  Pre- and post-     (77,         2.54 ± 1.54 (44)         0.17 (9)††      1725 [0.90]
                    MP F: NS+CS        103,                                                  2134 [0.95]
                                      110)
-                   Pre- and post-     (77,         2.54 ± 1.54 (44)         2.54 (wmav)      8 [0.90]
                    MP F: NS+CS        103,                                                   10 [0.95]
                                      110)
ND                  Pre-MP F:         (103,         2.54 ± 1.54 (44)         ND                 ND
                    NS+CS             110)
-                   Pre-MP F:         (103,         2.54 ± 1.54 (44)         2.54 (wmav)      8 [0.90]
                    NS+CS             110)                                                    10 [0.95]
ND                  Pre-MP F: CS      (110)         1.80 ± 0.33 (8)¶         ND                  ND
-                   Pre-MP F: CS      (110)         1.80 ± 0.33 (8)¶         1.80 (wmav)      1 [0.90]
                                                                                              1 [0.95]
EE: environmental exposure. DE: dietary exposure. NS: non-smoker. CS: current smoker. FS: former
smoker. n1: n (amount of individuals) in observed group. n2: the calculated amount of exposed
individuals in the research population for that specific power. M= male. F=female. MP: menopausal.
ND: not defined; lack of relevant and/or significant values. v: difference in biomarker between exposed
and control group. wmav: weighted mean as v; thus mean exposed is set at twice the mean of the
controls. ¶: only based on one study: M+F (106) and F (110). ††: this v is derived from a DE (to EE
fish exposed individuals) study (77), since no significant v could be derived from EE or SE studies, this
v has been used (the value of this v is in the same range as the value of the v found in a large study
by Knudsen et al. (107) with both males and females in the study population).


Table 10:

Overview group sizes for serum inhibin B levels in healthy individuals
 Exposure type         Studies        Inhibin B (ng/L) weighted            Weighted v (n1)      n2 [power]
     used to                           mean ± weighted SD (n1)
   determine v
EE                    (111, 112)                 ND                     20Ψ (85)                     ND
EE: environmental exposure. n1: n (amount of individuals) in observed group. n2: the calculated
amount of exposed individuals in the research population for that specific power. NS: non-smoker. CS:
current smoker. FS: former smoker. M= male. ND: not defined; lack of relevant and/or significant
values. v: difference in biomarker between exposed and control group. wmav: weighted mean as v;
thus mean exposed is set at twice the mean of the controls. Ψ: organic farmers serve as controls for
traditional farmers, but most organic farmers have also had traditional farmer exposure in the past.
Proposed additional genotoxicity markers

Table 11:

Overview group sizes for CA frequency (per 100 cells) in healthy
individuals measured in PBL
  Exposure     Smoking status        Studies     CA freq. (per 100cells)   Weighted v     n2 [power]
type used to                                       weighted mean ±            (n1)
 determine v                                       weighted SD (n1)
OE               ND: Total CAs      (113, 114)      2.02 ± 0.72 (86)        1.7 (86)†       4 [0.90]
                                                                                            5 [0.95]
-                ND: Total CAs      (113, 114)       2.02 ± 0.72 (86)      2.02 (wmav))     3 [0.90]
                                                                                            4 [0.95]
ND              ND: Chromatid-      (113, 114)       1.60 ± 0.70 (86)          ND              ND
                   subtotal
-               ND: Chromatid-      (113, 114)       1.60 ± 0.70 (86)      1.60 (wmav)      5 [0.90]
                   subtotal                                                                 5 [0.95]
ND             ND: Chomosome-       (113, 114)       0.51 ± 0.31 (86)          ND              ND
                   subtotal
-              ND: Chomosome-       (113, 114)       0.51 ± 0.31 (86)      0.51 (wmav)     8 [0.90]
                   subtotal                                                                10 [0.95]
ND                   ND:            (113, 114)       1.67 ± 0.37 (86)          ND             ND
                Chromosome-
                    break
-                    ND:            (113, 114)       1.67 ± 0.37 (86)      1.67 (wmav)      2 [0.90]
                Chromosome-                                                                 2 [0.95]
                    break
OE: occupational exposure. n1: n (amount of individuals) in observed group. n2: the calculated amount
of exposed individuals in the research population for that specific power. ND: not defined; lack of
relevant and/or significant values. v: difference in biomarker between exposed and control group.
wmav: weighted mean as v; thus mean exposed is set at twice the mean of the controls. †: since the
absence of a v derived from EE- and SE-based studies, an OE based v has been chosen as an
alternative. Since the results of Lander et al. (113) show lower values for CAs than those of
Kaiournova and Khabutdinova, (114) and only two studies (113, 114) were available to discuss, the
study of Lander et al. has not been excluded from the pooled classes.
Table 12:

Overview group sizes for number of micronucleated cells per 1000
binucleated cells (MNCs) in healthy individuals
                                                        a
  Exposure     Smoking status        Studies     MNCs weighted         Weighted v (n1)    n2 [power]
type used to                                     means ± weighted
 determine v                                         SDs (n1)
EE             NS+CS+FS+PS           (48, 115-   11.53 ± 5.24 (787)    9.58 (223)           7 [0.90]
                                       132)                                                 8 [0.95]
EE             NS+CS+FS+PS           (48, 115-    11.53 ± 5.24 (787)   8.37 (212)■          9 [0.90]
                                       132)                                                11 [0.95]
-              NS+CS+FS+PS           (48, 115-    11.53 ± 5.24 (787)   11.53 (wmav)         5 [0.90]
                                       132)                                                 6 [0.95]
EE             NS+CS+FS+PS Ħ         (48, 115-    10.93 ± 3.91 (604)   6.29 (106) (■)       9 [0.90]
                                       130)                                                11 [0.95]
-              NS+CS+FS+PS Ħ         (48, 115-    10.93 ± 3.91 (604)   10.93 (wmav)         3 [0.90]
                                       130)                                                 4 [0.95]
EE             NS                   (115, 116,     8.87 ± 3.91 (93)    9.58 (223)           4 [0.90]
                                       128)                                                 5 [0.95]
EE             NS                   (115, 116,     8.87 ± 3.91 (93)    8.37 (212) ■         5 [0.90]
                                       128)                                                 6 [0.95]
-              NS                   (115, 116,     8.87 ± 3.91 (93)    8.87 (wmav)          5 [0.90]
                                       128)                                                 6 [0.95]
EE             NS Ħ                 (115, 116,     8.87 ± 3.91 (93)    6.29 (106) (■)       9 [0.90]
                                       128)                                                11 [0.95]
-              NS Ħ                 (115, 116,     8.87 ± 3.91 (93)    8.87 (wmav)          5 [0.90]
                                       128)                                                 6 [0.95]
EE             CS                      (115)      22.19 ± 8.45 (16)¶   9.58 (223)          17 [0.90]
                                                                                           21 [0.95]
EE             CS                     (115)       22.19 ± 8.45 (16)¶   8.37 (212) ■        22 [0.90]
                                                                                           27 [0.95]
-              CS                     (115)       22.19 ± 8.45 (16)¶   22.19 (wmav)         4 [0.90]
                                                                                            4 [0.95]
                              a
EE: environmental exposure. Number of MNed cells per 1000 binucleated cells (MNCs). n1: n
(amount of individuals) in observed group. n2: the calculated amount of exposed individuals in the
research population for that specific power. NS: non-smoker. CS: current smoker. FS: former smoker.
PS: passive smoker. v: difference in biomarker between exposed and control group. wmav: weighted
mean as v; thus mean exposed is set at twice the mean of the controls. ■: for this weighted v the v of
post-Tsjernobyl radiation exposure (131) has been excluded from the weighted v calculation, due to
the extremity of this particular event. Ħ: (parts of) studies (130, 132) using assays other than the
standard assay and studies with children (131) instead of adults/ adolescents are excluded. : since
no smoking class specific EE based v was available the here applied v is based on studies without a
defined smoking status. (■): in this case ■ does also apply due to Ħ. ▲: since no EE based v for solely
M or solely F is available the v of M+F has been used in this case. ¶: only based on one study: M
(116) and F (115). Biological medium in all the detections were PBL. Unless mentioned otherwise:
data of cytochalasin B micronuclei assays (CBMN) have been described.
Overview group sizes for number of MN per 1000 binucleated cells (MNi)
in healthy individuals
                                                                   b
   Exposure     Smoking status            Studies             MNi weighted         Weighted        n2
 type used to                                                means ± weighted       v (n1)       [power]
 determine v                                                      SDs (n1)
EE              NS+CS+FS+PS        (11, 28, 116-121, 123,    6.38 ± 3.90 (4315)    3.04 (307)    35 [0.90]
                                  124, 129, 131, 133-163)                                        43 [0.95]
EE              NS+CS+FS+PS        (11, 28, 116-121, 123,    6.38 ± 3.90 (4315)    1.80          99 [0.90]
                                  124, 129, 131, 133-163)                          (296)♣       123 [0.95]
-               NS+CS+FS+PS        (11, 28, 116-121, 123,    6.38 ± 3.90 (4315)    6.38          8 [0.90]
                                  124, 129, 131, 133-163)                          (wmav)        10 [0.95]
EE              NS+CS+FS+PS        (11, 28, 116-121, 123,    6.34 ± 3.93 (4238)    1.80 (296)   101 [0.90]
                Ħ                 124, 129, 133-142, 144-                          (♣)          124 [0.95]
                                            163)
-               NS+CS+FS+PS        (11, 28, 116-121, 123,    6.34 ± 3.93 (4238)    6.34         9 [0.90]
                Ħ                 124, 129, 133-142, 144-                          (wmav)       10 [0.95]
                                            163)
EE              NS                (28, 116, 133-143, 162-    5.78 ± 3.12 (1235)    1.66 (70)     75 [0.90]
                                            164)                                   (♣)           92 [0.95]
-               NS                (28, 116, 133-143, 162-    5.78 ± 3.12 (1235)    5.78          7 [0.90]
                                            164)                                   (wmav)        8 [0.95]
EE              CS                (28, 135, 140-142, 164)    10.52 ± 10.98 (167)   3.06 (19)    242 [0.90]
                                                                                   (♣)          300 [0.95]
-               CS                 (28, 135, 140-142, 164)   10.52 ± 10.98 (167)   10.52         23 [0.90]
                                                                                   (wmav)        29 [0.95]
EE: environmental exposure. n1. n1: n (amount of individuals) in observed group. n2: the calculated
amount of exposed individuals in the research population for that specific power. NS: non-smoker. CS:
current smoker. FS: former smoker. PS: passive smoker. M: male. F: female. v: difference in
biomarker between exposed and control group. bNumber of MN per 1000 BN cells (MNi). Biological
medium in all the detections were PBL. Unless mentioned otherwise: data of cytochalasin B
micronuclei assays (CBMN) have been described. wmav: weighted mean as v; thus mean exposed is
set at twice the mean of the controls. ♣: for this weighted v the v of post-Tsjernobyl radiation exposure
(131) has been excluded from the weighted v calculation, due to the extremity of this particular event.
Ħ: (parts of) studies using assays other than the standard assay (134, 143) and studies with children
(131) instead of adults/ adolescents are excluded (those that apply to this specific class). (♣): in this
case ♣ does also apply due to Ħ or other class specific selection criteria.
Table 13:

Overview group sizes for white blood cell PAH-DNA adduct levels
(adducts/ 108 nucleotides) in healthy individuals
      Exposure     Smoking status               Studies     PAH-DNA adducts      Weighted     n2 [power]
                                                                 -8
    type used to                                              (10 ) weighted      v (n1)
    determine v                                             means ± weighted
                                                                 SDs (n1)
EE                 NS+FS+CS, ELISA+32P-        (165-178)     6.45 ± 7.22 (604)   14.83         5 [0.90]
                   postlabelling assays,                                         (121)         7 [0.95]
                   PBL+WBC
                                      32
-                  NS+FS+CS, ELISA+ P-         (165-178)     6.45 ± 7.22 (604)   6.45         27 [0.90]
                   postlabelling assays,                                         (wmav)       33 [0.95]
                   PBL+WBC
                                      32
EE                 NS+FS+CS, ELISA+ P-         (165-168,     5.34 ± 8.56 (147)   18.52 (63)    5 [0.90]
                   postlabelling assays, PBL    170, 175,                                      6 [0.95]
                                                  178)
-                  NS+FS+CS, ELISA+32P-        (165-168,     5.34 ± 8.56 (147)   5.34         55 [0.90]
                   postlabelling assays, PBL    170, 175,                        (wmav)       67 [0.95]
                                                  178)
                                      32
EE                 NS+FS+CS, ELISA+ P-         (168, 169,    6.81 ± 6.73 (457)   10.82 (58)    9 [0.90]
                   postlabelling assays, WBC    171-174,                                      11 [0.95]
                                               176, 177)
                                      32
-                  NS+FS+CS, ELISA+ P-         (168, 169,    6.81 ± 6.73 (457)   6.81         21 [0.90]
                   postlabelling assays, WBC    171-174,                         (wmav)       26 [0.95]
                                               176, 177)
EE                 NS+FS+CS, ELISA,            (168-170,     7.92 ± 9.37 (283)   10.09 (43)   19 [0.90]
                   PBL+WBC                      173-176,                                      23 [0.95]
                                                  178)
-                  NS+FS+CS, ELISA,            (168-170,     7.92 ± 9.37 (283)   7.92         30 [0.90]
                   PBL+WBC                      173-176,                         (wmav)       37 [0.95]
                                                  178)
EE                 NS+FS+CS, ELISA, PBL        (168, 170,    7.65 ± 9.61 (85)    10.09        20 [0.90]
                                               175, 178)                         (43)Ґ        24 [0.95]
-                  NS+FS+CS, ELISA, PBL        (168, 170,    7.65 ± 9.61 (85)    7.65         34 [0.90]
                                               175, 178)                         (wmav)       42 [0.95]
EE                 NS+FS+CS, ELISA, WBC        (168, 169,    8.04 ± 9.27 (198)   10.09 (43)   18 [0.90]
                                                173, 174,                                     22 [0.95]
                                                  176)
-                  NS+FS+CS, ELISA, WBC        (168, 169,    8.04 ± 9.27 (198)   8.04         28 [0.90]
                                                173, 174,                        (wmav)       35 [0.95]
                                                  176)
EE                 NS+CS, 32P-postlabelling    (165-168,     5.16 ± 4.53 (321)   17.44 (78)    2 [0.90]
                   assays, PBL+WBC              171, 172,                                      2 [0.95]
                                               176, 177)
                          32
-                  NS+CS, P-postlabelling      (165-168,     5.16 ± 4.53 (321)   5.16         17 [0.90]
                   assays, PBL+WBC              171, 172,                        (wmav)       21 [0.95]
                                               176, 177)
                          32
EE                 NS+CS, P-postlabelling      (165-168)     2.16 ± 6.81 (62)    18.52 (63)     3 [0.90]
                   assays, PBL                                                                  4 [0.95]
                           32
-                  NS+CS, P-postlabelling      (165-168)     2.16 ± 6.81 (62)    2.16         180 [0.90]
                   assays, PBL                                                   (wmav)       222 [0.95]
                           32
EE                 NS+CS, P-postlabelling      (171, 172,    5.87 ± 3.80 (259)   12.9 (15)      2 [0.90]
                   assays, WBC                 176, 177)                                        3 [0.95]
                           32
-                  NS+CS, P-postlabelling      (171, 172,    5.87 ± 3.80 (259)   5.87           9 [0.90]
                   assays, WBC                 176, 177)                         (wmav)        11 [0.95]
EE                 NS, ELISA+32P-              (165-167,     5.25 ± 5.49 (235)   18.98 (3)      2 [0.90]
                   postlabelling assays,       169, 171-                                        3 [0.95]
                   PBL+WBC                        175)
                               32
-                  NS, ELISA+ P-               (165-167,     5.25 ± 5.49 (235)   5.25         23 [0.90]
                   postlabelling assays,       169, 171-                         (wmav)       29 [0.95]
                   PBL+WBC                        175)
                32
EE   NS, ELISA+ P-               (165-167,    2.20 ± 6.54 (66)    18.98 (3)      3 [0.90]
     postlabelling assays, PBL      175)                                         4 [0.95]
                 32
-    NS, ELISA+ P-               (165-167,    2.20 ± 6.54 (66)    2.20         186 [0.90]
     postlabelling assays, PBL      175)                          (wmav)       230 [0.95]
                 32
EE   NS, ELISA+ P-               (169, 171-   6.44 ± 5.03 (169)   10.82          5 [0.90]
     postlabelling assays, WBC      174)                          (58)†          6 [0.95]
-    NS, ELISA+32P-              (169, 171-   6.44 ± 5.03 (169)   6.44          13 [0.90]
     postlabelling assays, WBC      174)                          (wmav)        16 [0.95]
EE   NS, ELISA, PBL+WBC          (169, 173-   4.70 ± 4.97 (90)    10.09          6 [0.90]
                                    175)                          (43)†          7 [0.95]
-    NS, ELISA, PBL+WBC          (169, 173-   4.70 ± 4.97 (90)    4.70          24 [0.90]
                                    175)                          (wmav)        30 [0.95]
EE   NS, ELISA, PBL                (175)      1.35 ± 0.78 (27)¶   10.09          1 [0.90]
                                                                  (43)‡          1 [0.95]
-    NS, ELISA, PBL                (175)      1.35 ± 0.78 (27)¶   1.35           8 [0.90]
                                                                  (wmav)         9 [0.95]
EE   NS, ELISA, WBC              (169, 173,   6.13 ± 5.93 (63)    10.09          8 [0.90]
                                    174)                          (43)†          9 [0.95]
-    NS, ELISA, WBC              (169, 173,   6.13 ± 5.93 (63)    6.13          20 [0.90]
                                    174)                          (wmav)        25 [0.95]
         32
EE   NS, P-postlabelling         (165-167,    5.60 ± 5.79 (145)   18.98 (3)      2 [0.90]
     assays, PBL+WBC             171, 172)                                       3 [0.95]
         32
-    NS, P-postlabelling         (165-167,    5.60 ± 5.79 (145)   5.60          23 [0.90]
     assays, PBL+WBC             171, 172)                        (wmav)        28 [0.95]
         32
EE   NS, P-postlabelling         (165-167)    2.79 ± 8.56 (39)    18.98 (3)      5 [0.90]
     assays, PBL                                                                 6 [0.95]
-    NS, 32P-postlabelling       (165-167)    2.79 ± 8.56 (39)    2.79         198 [0.90]
     assays, PBL                                                  (wmav)       245 [0.95]
EE   NS, 32P-postlabelling       (171, 172)   6.63 ± 4.41 (106)   18.98 (3)Γ     1 [0.90]
     assays, WBC                                                                 1 [0.95]
         32
-    NS, P-postlabelling         (171, 172)   6.63 ± 4.41 (106)   6.63          10 [0.90]
     assays, WBC                                                  (wmav)        12 [0.95]
                 32
EE   CS, ELISA+ P-               (165, 172-   6.26 ± 3.97 (94)    14.72          2 [0.90]
     postlabelling assays,          174)                          (118)†         2 [0.95]
     PBL+WBC
                 32
-    CS, ELISA+ P-               (165, 172-   6.26 ± 3.97 (94)    6.26          9 [0.90]
     postlabelling assays,          174)                          (wmav)       11 [0.95]
     PBL+WBC
                 32
EE   CS, ELISA+ P-                 (165)       1.1 ± 1.6 (23)¶    18.5 (60)†    1 [0.90]
     postlabelling assays, PBL                                                  1 [0.95]
-    CS, ELISA+32P-                (165)       1.1 ± 1.6 (23)¶    1.1          45 [0.90]
     postlabelling assays, PBL                                    (wmav)       56 [0.95]
EE   CS, ELISA+32P-              (172-174)    7.93 ± 4.48 (71)    10.82         4 [0.90]
     postlabelling assays, WBC                                    (58)†         5 [0.95]
-    CS, ELISA+32P-              (172-174)    7.93 ± 4.48 (71)    7.93          7 [0.90]
     postlabelling assays, WBC                                    (wmav)        9 [0.95]
EE   CS, ELISA, WBC              (173, 174)   10.02 ± 6.07 (35)   10.09         8 [0.90]
                                                                  (43)†        10 [0.95]
-    CS, ELISA, WBC              (173, 174)   10.02 ± 6.07 (35)   10.02         8 [0.90]
                                                                  (wmav)       10 [0.95]
         32
EE   CS, P-postlabelling         (165, 172)   4.03 ± 1.81 (59)    17.38         1 [0.90]
     assays, PBL+WBC                                              (75)†         1 [0.95]
         32
-    CS, P-postlabelling         (165, 172)   4.03 ± 1.81 (59)    4.03          5 [0.90]
     assays, PBL+WBC                                              (wmav)        6 [0.95]
         32
EE   CS, P-postlabelling           (165)       1.1 ± 1.6 (23)¶    18.5 (60)†    1 [0.90]
     assays, PBL                                                                1 [0.95]
         32
-    CS, P-postlabelling           (165)       1.1 ± 1.6 (23)¶    1.1          45 [0.90]
     assays, PBL                                                  (wmav)       56 [0.95]
         32
EE   CS, P-postlabelling           (172)      5.90 ± 1.93 (36)¶   12.9 (15)†    1 [0.90]
     assays, WBC                                                                1 [0.95]
-    CS, 32P-postlabelling         (172)      5.90 ± 1.93 (36)¶   5.90          2 [0.90]
     assays, WBC                                                  (wmav)        2 [0.95]
EE: environmental exposure. OE: occupational exposure. SE: smoking exposure (non-smokers vs.
smokers). ND: not detectable. NS: never smoker. CS: current smoker. FS: former smoker. wmav:
weighted mean as v; thus mean exposed is set at twice the mean of the controls. : this v is based on
                      32
a study, that uses a P-postlabelling assay (168), while this class (also) consists of ELISA-based
studies. ¶: solely based on one study: (175), (165) or (172). †: since no ‘NS’- or ‘CS’-specific v was
available, a ‘NS+CS’-derived v has been applied. ‡: since no ‘NS, ELISA, PBL’-specific v was
available, a ‘NS+CS, ELISA, WBC’-derived v has been applied. Γ: since no WBC-derived v could be
attained for this class a PBL-derived v has been used. Ґ: since no PBL-derived v was available a
WBC-derived v ahs been used for this class.
Discussion
Considering the intention of using a group size of 200 individuals for both exposed
and control groups (r=1), we can ascertain that this group size offers sufficient power
for most markers. For most biomarkers of effect a group size of 200 individuals per
group is adequate to satisfy the calculated minimal group sizes (n2) yielding the
preferred statistical power, as shown in the Tables and Appendices. However, it
should be noted that, as discussed earlier, these are minimal group sizes and that
other factors could decrease the power. A further remark is that the study-based and
wmav-based minimal group sizes tend to differ substantially. In most cases wmav-
based minimal group sizes are (very) low compared to study-based minimal group
sizes. This is mostly due to the fact that weighted means (and thus the v used for
wmav-based n2 calculations) are substantially bigger than their weighted SDs,
resulting in lower n2s.
The most noteworthy deviations will be elucidated in the subchapters for all
biomarkers of effect. With noteworthy deviations those that either abundantly exceed
(n2[0.95]s exceeding 400 (two times the intended group size of 200)) the intended
group size of 200 individuals or are excessively small (n2[0.95]s less than 20 (one-
tenth of the intended group size of 200)). Furthermore, only those calculations that
noteworthy deviate from the intended group size that are calculated using a v that is
derived from actual studies will be discussed, for the following rationale: (I) that in
general wmav-based n2s are substantially lower than those based on actual studies
and (II) that n2s based on actual studies have more relevance for the Flanders
biomonitoring study.



Genotoxicity markers


TM (arb). No noteworthy deviations: the intended group size is adequate for all
weighted calculation for this biomarker of effect.
HPRT MF determined by the T-cell cloning assay. In Table 2 (MF; T-cell cloning
assay) the separate ‘NS’ and ‘CS’ classes, which have lower SDs than the
‘NS+CS+FS’ class (Table 2), also had to apply an on ‘NS+CS+FS’ studies based v,
thus decreasing the SD with an unchanged v (based on a higher variance based
smoking status influenced class) as compared to the ‘NS+CS+FS’ class, what will
logically result in smaller n2s than those displayed in the ‘NS+CS+FS’ class (Table 2).
HPRT VF determined by the BrdU assay. The small n2s found in Table 2 (VF; BrdU
assay) all share the same founding factor: the v is solely based on one study (30),
this v is based on a significant v of exactly ten times the value of the SD of the control
group SD (30). The weighted SD is based on three studies (28-30), of which the
studies that don’t contribute to the v have a net weighted mean- and weighted SD-
decreasing effect (Appendix 2; MF; BrdU assay), thereby increasing the ratio v:SD,
thus further decreasing the size of the pooled n2s (Table 2; MF; BrdU assay).
8-OHdG in lymphocytes. The small n2s as seen in Table 3 (in lymphocytes), can be
reduced to the relative (compared to the weighted SDs) size of the SE-based v (one
study; the only significant v) (33). The n2s calculated for this specific study (Appendix
3; lymphocytes) are already larger than those of the pooled classes (Table 3;
lymphocytes), but an additional factor resulting in the smaller n2s for the pooled
classes is that the weighted means and weighted SDs are based on a relative large
amount of studies (27, 31-35), of which all have a weighted mean- and weighted SD-
decreasing effect (Appendix 3; lymphocytes); further decreasing the size of the
pooled n2s (Table 3; lymphocytes) as compared to the n2s of the specific study (33)
(Appendix 3; lymphocytes).
8-OHdG in leukocytes. No noteworthy deviations: the intended group size is
adequate for all weighted calculation for this biomarker of effect.
8-OHdG in urine (µmol/mol creatine). No noteworthy deviations: the intended
group size is adequate for all weighted calculation for this biomarker of effect.
8-OHdG in urine (µg/g creatine). No noteworthy deviations: the intended group size
is adequate for all weighted calculation for this biomarker of effect.
8-OHdG in urine (pmol/kg/24h). No noteworthy deviations: the intended group size
is adequate for all weighted calculation for this biomarker of effect.
8-OHdG in urine (ng/BMI). The small n2s shown in Table 3 (ng/BMI) are founded on
only one study (33), the used v is not even significant (Appendix 3; ng/BMI), but has
been used since a significant v is lacking. Larger n2s than those calculated with this
not significant v are therefore expected in the Flanders biomonitoring study.
8-OHdG in urine (nmol/24h). No noteworthy deviations: the intended group size is
adequate for all weighted calculation for this biomarker of effect.



Hormones


Serum testosterone. No noteworthy deviations: the intended group size is adequate
for all weighted calculation for this biomarker of effect.
Serum oestradiol. The class ‘post-MP F:NS+FS’ abundantly exceeds the intended
group size. The difference between control and exposed classes tends to be
relatively smaller in females of the post-menopausal type for hormonal biomarkers of
effect. In the Flanders biomonitoring study these minimal group size-related problems
in post-MP females do not occur since all hormones (including serum oestradiol
(Table 5), serum FSH (Table 7), and serum TSH (Table 9)) will only be monitored in
adolescents (thus pre-MP females and males). In Table 5, the small n2s (M:NS) are
solely based on one study (69), which results suggested that cigarette smoking has a
highly significant (p<0.01) effect on serum oestradiol levels in males (69), allowing
the use of relative small group sizes, while retaining the ability to detect real
differences. Mechanistic support for this finding is discussed by Kupeli et al. (69).
Serum SHBG. In Table 6 small n2s have been found with the ‘M:NS+CS’ class and
the same class without potentially ‘compromised’ controls. The weighted means and
SDs have been derived from several studies, the v however has only been derived
from one study (76) and this study was solely conducted with NS controls instead of
‘NS+CS’, the class ‘M:NS’ (with NS-based SD and NS-based v) has larger n2s (a
n2[0.90] of 34 and a n2[0.95] of 42), as shown in Appendix 6. Furthermore, the
studies that don’t contribute to the applied v have a weighted mean- and weighted
SD-decreasing effect, thus lowering pooled n2s in these classes (Table 6).
Serum FSH. Classes that contain post-menopausal female (Table 7) abundantly
exceed the intended group size, due to relatively smaller differences between
controls and exposed. This problem is not expected in the Flanders biomonitoring
study for this marker, since it is not monitored in post-menopausal female (as
discussed earlier). The pooled comparison for serum FSH in males (Table 7) shows
a relatively small v as compared to the SD, thus resulting in big n2s. The separate
studies however, the only two on which the weighted v has been based (56, 94),
show that if a minimal group size still yielding the preferred power would be
calculated for this separate studies this would result in n2s smaller than 200 (n2[0.90]
and n2[0.95] of 2 and 2 (56) and 31 and 38 (94), respectively), as shown in Appendix
7. This can be reduced to relative smaller SDs in these separate studies (0.2 (56)
and 0.6 (94)) (Appendix 7) as compared to the pooled SDs of 4.66 (M: NS+CS) and
3.22 (M: NS) (Table 7). The class ‘pre-MP F:CS’ in Table 7 shows small n2s and is
solely based on one study (98), the v is based on two other studies (100, 101) that
have been conducted on ND smoking status post-MP females, thereby explaining the
relative high ratio of v:SD (Appendix 7) that is responsible for low n2s for this class as
seen in Table 7. The post-MP based v has also been applied for the class of ‘pre-MP
F:CS’ (Table 7), since no pre-MP based v was available.
Serum LH. The class ‘M:NS’ in Table 8 has small n2s and is based on one small
study (105) which results suggest that acute toluene exposure has a significant
(p<0.05) effect on serum LH levels in males (105), the small group size able to
discriminate existing differences could be due to the potentially high impact factor of
the acute toluene exposure intervention on this specific biomarker of effect in males.
Serum TSH. As with serum FSH, classes that contain post-menopausal female
(Table 9) abundantly exceed the intended group size, due to relatively smaller
differences between controls and exposed, but this is not a problem since the
Flanders biomonitoring study does not monitor this biomarker of effect in post-
menopausal females. The class ‘M+pre- and post-MP F: NS+FS’ in Table 9 is based
on one study (106) for weighted means and weighted SDs, while the v is derived
from another study (107). Knudsen et al. found a highly significant difference
(p<0.001) between ‘NS+FS’ and ‘CS’ in his large study (107) for serum TSH levels,
thus allowing the use of small studies while retaining the ability to monitor real
differences. A possible underlying mechanism of tobacco smoke on TSH levels has
also been suggested by Knudsen et al. (107). The class ‘M: NS+CS’ (Table 9) has
weighted means and weighted SDs derived from three studies (88, 108, 109), while
the v is only based on one small study (109). This v is based on EE to extreme cold
(109), thus it is logical that a relative large difference between exposed and
unexposed individuals could manifest if TSH levels are sensitive to this sort of
environmental influence. These sorts of extremities (109) in exposure as compared to
the exposure expected among the general population are not to be expected in the
Flemish populations monitored in the Flanders biomonitoring study, thus bigger
groups are needed in order to sustain the ability to discriminate real differences.
Serum inhibin B. For serum inhibin B (Table 10) no n2 could be calculated since no
SDs were available in any of the studies reviewed (Appendix 10).



Proposed additional genotoxicity markers



CA frequency in PBL. The small calculated n2s of the class ‘ND: Total CAs’ can be
deduced to the origin of the v, which is derived from a study on OE to pesticides
(113, 114). OE-based differences, tend to be larger than those based on EE or SE,
due to higher levels of exposure, thus with unchanged SDs of unexposed classes,
smaller groups are sufficient to discriminate real effects. The n2-calculations of the
separate studies show considerably higher n2s (n2[0.90] and n2[0.95] of 73 and 90,
respectively) for the study by Lander et al. (113) and even lower n2s (n2[0.90] and
n2[0.95] of 1 and 1, respectively) for the study by Kaiournova and Khabutdinova
(114).
MNCs. All the pooled classes of Table 12 (MNCs) have small n2s. The individual
studies that allow calculations for those specific studies show that the study of Chang
et al. (130) has n2s larger than 20 (n2[0.90] and n2[0.95] of 38 and 47, respectively),
but for the same study specified to compare only matched group A controls and
exposed showed n2s smaller than 20 (n2[0.90] and n2[0.95] of 7 and 9, respectively)
(Appendix 12, MNCs). Group A matches consist of close relatives, siblings, brother-
in-laws, or sister-in-laws matched for age and sex. Futhermore, this same study (130)
showed that the alternative assay resulted for both total controls with total exposed
comparisons and for group A matched controls exposed comparisons to have
specific n2s larger than 20 (n2[0.90] and n2[0.95] of 22 and 26, respectively and for
both groups). The study by Fenech et al. (131), was not only conducted with children
(expected to have higher susceptibility to such exposures than adults) but also with
an extreme event (post-Chernobyl radiation nuclear radiation exposed residents),
seen in this light it seems logical that the low SD:v ratio results in the possibility to
use extremely small n2s (Appendix 12, MNCs) in that specific study cohort (Appendix
12, MNCs). The EE in the Flanders biomonitoring study is not expected to be of
similar intensity to that of post-Chernobyl radiation.
MNi. No noteworthy deviations: the intended group size is adequate for all weighted
calculation for this biomarker of effect.
PAH-DNA adduct levels. All the pooled classes of Table 13 have small n2s, the only
exceptions are the classes which only consist of ELISA based data. All other classes
have relatively lower SDs due to the relatively lower values in the 32P-postlabelling
assays as compared to the ELISA values (table 13) (this reason only applies for
classes with both ELISA and 32P-postlabelling assay data) and the fact that the
difference of the 32P-postlabelling studies is solely based on (one of the) two studies
(166, 168) and is relatively big (table 13). The fact that both the calculated n2s of the
ELISA and the 32P-postlabelling assay classes is relatively low can be reduced to the
nature of the used differences. The differences are based on EE, but their nature is
more extreme than that expected in the Flanders biomonitoring study: the ambient air
pollution of bus-drivers (168) is of a higher degree than that expected in the
‘exposed’ Flemish population (one of the two studies that offer a 32P-postlabelling v,
appendix 13) and the all the other differences (for both ELISA and 32P-postlabelling
assays) are based on EE as residents of the Silesian region of Poland (appenix 13).
Silesia is one of the most heavily polluted areas in the world (especially during
winter), among the most prevalent air pollutants in Silesia are PAHs (176). The
Flemish populations are probably not exposed to such extremities in environmental
exposure, thus smaller differences and higher n2s are expected to be relevant for the
Flanders biomonitoring study.

In conclusion: the intended group size of 200 individuals in both exposed and control
groups for the Flanders biomonitoring study is sufficient for all the biomarkers for
effect described in this review with the exception of inhibin B. As pointed out before:
the reported data on the hormone inhibin B did not satisfy the demands needed for
minimal group size calculations. Small minimal group sizes are the result of an
unsatisfactory amount of reported data on these biomarkers of effect (especially the
lack of reported studies that are big enough to discriminate significant differences,
and to a lesserer extent: the lack of reported SDs in studies for some biomarkers of
effect), of heterogenecity in study populations, of relative big differences between
control group and exposed group and/or of (smoking) exposure-sensitive biomarkers.
All factors, which are not expected to play major roles in the Flanders biomonitoring
study.




.
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198.   Swennen B, Buchet JP, Stanescu D, Lison D, Lauwerys R. Epidemiological
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       J. Ind. Med. 1993;50:835-42.
199.   Muller B, Zulewski H, Huber P, Ratcliffe JG, Staub JJ. Impaired action of
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200.   Bolognesi C, Parrini M, Bonassi S, Ianello G, Salanitto A. Cytogenetic analysis
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       1993;285:239-49.
201.   Bolognesi C, Parrini M, Merlo F, Bonassi S. Frequency of micronuclei in
       lymphocytes from a group of floriculturists exposed to pesticides. J. Toxicol.
       Environ. Health 1993;40:405-11.
202.   Gattas GJ, Cardoso Lde A, Medrado-Faria Mde A, Saldanha PH. Frequency
       of oral mucosa micronuclei in gas station operators after introducing methanol.
       Occup. Med. (Lond.) 2001;51:107-13.
203.   Nielsen PS, Andreassen A, Farmer PB, Ovrebo S, Autrup H. Biomonitoring of
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       hydroxypyrene as markers of exposure. Toxicol. Lett. 1996;86:27-37.
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Appendices
Appendix 1:
DNA damage in healthy individuals measured by the Comet-assay,
expressed in Tail Moment.

Studies and their exposure types
Exposure        Smoking                      n1    TM (arb) mean ±       v           n2 [power]
type            status                                   SD
EE (rural vs    NS        Piperakis et       20     0.710 ± 0.078    0.070    (4)     27 [0.90]
urban                     al., 2000                                                   33 [0.95]
inhabitants)
EE (rural vs    CS        Piperakis et       20     0.980 ± 0.125    0.010@   (4)       ND
urban                     al., 2000
inhabitants)

OE (rubber      ND        Collins et al.,    22      0.64 ± 0.42              (10)      ND
tire factory              1997
workers)
OE              ND        Maluf and          12     0.162 ± 0.103             (11)      ND
(antineoplast             Erdtmann,
ic drugs)                 2000
OE (airport     NS        Pitarque et        8       0.80 ± 0.15              (5)       ND
personnel)                al., 1999
OE (airport     CS        Pitarque et        3       0.69 ± 0.23              (5)       ND
personnel)                al., 1999
OE (airport     CS+NS     Pitarque et        11      0.77 ± 0.12              (5)       ND
personnel)                al., 1999
OE              NS        Piperakis et      28 M    0.823 ± 0.141             (6)       ND
(greenhouse               al., 2003
(pesticides))
OE              ES        Piperakis et      13 M    0.803 ± 0.161             (6)       ND
(greenhouse               al., 2003
(pesticides))
OE              NS        Piperakis et      17 F    0.811 ± 0.161             (6)       ND
(greenhouse               al., 2003
(pesticides))
OE              ES        Piperakis et      8F      0.841 ± 0.171             (6)       ND
(greenhouse               al., 2003
(pesticides))
OE (waste       NS        Hartmann et        23      0.05 ± 0.02              (7)       ND
disposal                  al., 1998
workers)
OE (waste       CS        Hartmann et        24      0.03 ± 0.03              (7)       ND
disposal                  al., 1998
workers)
OE (waste       NS+CS     Hartmann et        47      0.041 ± 0.03             (7)       ND
disposal                  al., 1998
workers)
OE (X-ray       NS+CS     Maluf et al.,      22     0.171 ± 0.142             (12)      ND
exposed                   2001
workers)
                                                                                        ND
SE              NS        Dhawan et          31       1.3 ± 0.4               (8)       ND
                          al., 2001
SE (20-25       NS        Piperakis et       20      0.63 ± 0.02              (9)       ND
years old                 al., 1998
males)
SE (55-60       NS        Piperakis et       20      0.77 ± 0.02              (9)       ND
years old                 al., 1998
males)
DE              NS+CS     Dhawan et          26       1.4 ± 0.7               (8)       ND
(vegetarians               al., 2001
vs non-
vegetarians)

Weighted       NS+CS+E                            0.62 ± 0.26 (375)   0.07 (20)            290 [0.90]
(n1)           S                                                                           359 [0.95]
Weighted       NS+CS+E                            0.62 ± 0.26 (375)     0.62                4 [0.90]
(n1)           S                                                      (wmav)                5 [0.95]
Weighted       NS                                 0.76 ± 0.20 (167)   0.07 (20)            172 [0.90]
(n1)                                                                                       213 [0.95]
Weighted       NS                                 0.76 ± 0.20 (167)      0.76               2 [0.90]
(n1)                                                                   (wmav)               2 [0.95]
Weighted       CS                                 0.48 ± 0.10 (47)    0.07 (20)            43 [0.90]
(n1)                                                                                       54 [0.95]
Weighted       CS                                 0.48 ± 0.10 (47)       0.48               1 [0.90]
(n1)                                                                   (wmav)               2 [0.95]
EE: environmental exposure. OE: occupational exposure. SE: smoking exposure (non-smokers vs.
smokers). DE: dietary exposure. NS: non-smoker. CS: current smoker. ES: ex-smoker. TM: tail
moment (per scored cell). Arb: (visually scored) arbitrary units. n1: n (amount of individuals) in
observed group. n2: the calculated amount of exposed individuals in the research population for that
specific power. @: not significant. ND: not defined; lack of significant and or relevant data. v:
difference in biomarker between exposed and control group. All detections were performed with the
alkaline COMET-assay. Biological medium in all the detections were leukocytes. M: male. F: female.
wmav: weighted mean as v; thus mean exposed is set at twice the mean of the controls. : the v is
derived from a study with NS instead of CS (or NS+CS) as in this case.
Appendix 2
HPRT-mutation frequency in healthy individuals, determined by the T-cell
cloning assay.

Studies and their exposure types
Exposure        Smoking                    n1        Mutation           v                 n2 [power]
type            status                           Frequency (10-6)
                                                    mean ± SD
EE (vinyl       NS        Becker et        46       3.15 ± 2.62     0.33@         (13)       ND
chloride                  al., 2001
momomer)
EE (vinyl       CS        Becker et        41      2.72 ± 1.87      0.19@         (13)       ND
chloride                  al., 2001
momomer)
AE              ND        da Cruz et        9      14.65 ± 8.20     23.15*/4.3    (20)     14 [0.90]
(radiological             al., 1996                                 5**/3.55***            17 [0.95]
exposure)

OE (vinyl       NS        Huttner and      8       10.24 ± 6.33                   (14)       ND
chloride                  Holzapfel,
exposed)                  1996
OE (vinyl       CS        Huttner and      15      6.02 ± 4.15                    (14)       ND
chloride                  Holzapfel,
exposed)                  1996
OE (diesel      NS        Hou et al.,      22          8.4                        (179)      ND
exhaust                   1995
exposed)
OE              NS+CS     Jones et al.,    117     15.4 ± 14.2                    (21)       ND
(Chernobyl                   1999
liquidator)
OE              NS+CS+F   Thomas et        66      16.1 ± 14.1                    (22)       ND
(Chernobyl      S         al., 1999
liquidator)
OE              NS+CS+F   Thomas et        231     11.1 ± 24.1                    (22)       ND
(Chernobyl      S         al., 1999
liquidator)
OE (coke-       NS+CS     Zanesi et al.,   26      15.8 ± 14.6                    (23)       ND
oven                      1998
workers)
OE (arsenic     ND        Harrington-       4        9.0 ± 2.9                    (24)       ND
exposed                   Brock et al.,
copper plant              1999
workers)
OE (1,3-        NS+CS     Tates et al.,    31     14.71 ± 11.00                   (25)       ND
butadiene                 1996
exposed)
OE (1,3-        NS+CS     Tates et al.,    26      13.93 ± 9.04                   (25)       ND
butadiene                 1996
exposed)
OE (styrene     NS        Vodicka et        8       13.2 ± 4.6                    (15)       ND
exposure)                 al., 1999
OE (styrene     CS        Vodicka et        5       16.0 ± 7.6                    (15)       ND
exposure)                 al., 1999
OE (styrene     NS+CS     Vodicka et       13       14.2 ± 6.5                    (15)       ND
exposure)                 al., 1999
OE (styrene     NS+CS     Vodicka et       19       13.3 ± 6.3                    (26)       ND
exposure)                 al., 2001
                                                                                             ND
SE              NS        Huttner and      24      5.88 ± 3.53                    (14)       ND
                          Holzapfel,
                            1996
SE              NS          Duthie et al.,    25           16.6 ± 1.7                    (16)       ND
                            1995
SE              NS          Hou et al.,       76              17.4                       (180)      ND
                            1999
SE              NS          Podlutsky et      38          18.7 ± 12.0                    (17)       ND
                            al., 1999
SE              NS          Burkhart-         34            8.0 ± 5.7                    (18)       ND
                            Schultz et
                            al., 1996
SE              NS          Ammenheus         42          1.74 ± 0.13                    (19)       ND
                            er et al.,
                            1997
ME (HBO         ND          Dennog et         8            18.6 ± 12.3                   (27)       ND
treatment)                  al., 1999

Weighted        NS+CS+F                                   11.05 ± 15.35     10.35 (9)             47 [0.90]
(n1)            S                                             (836)                               58 [0.95]
Weighted        NS+CS+F                                   11.05 ± 15.35       11.05               41 [0.90]
(n1)            S                                             (836)          (wmav)               51 [0.95]
Weighted        NS                                      8.64 ± 5.88 (225)   10.35 (9)§            7 [0.90]
(n1)                                                                                              9 [0.95]
Weighted        NS                                      8.64 ± 5.88 (225)      8.64               10 [0.90]
(n1)                                                                         (wmav)               13 [0.95]
Weighted        CS                                      4.62 ± 3.25 (61)    10.35 (9)§            3 [0.90]
(n1)                                                                                              3 [0.95]
Weighted        CS                                      4.62 ± 3.25 (61)       4.62               11 [0.90]
(n1)                                                                         (wmav)               13 [0.95]
EE: environmental exposure. AE: accidental exposure. OE: occupational exposure. SE: smoking
exposure (non-smokers vs. smokers). ME: medical exposure. NS: non-smoker. CS: current smoker.
FS: former smoker. n1: n (amount of individuals) in observed group. n2: the calculated amount of
exposed individuals in the research population for that specific power. *: 3 years after accidental
(radiation) exposure. **: 4 years after accidental (radiation) exposure. ***: 5 years after accidental
(radiation) exposure. @: not significant. ND: not defined; lack of relevant and/or significant values. v:
difference in biomarker between exposed and control group. HBO treatment: hyperbaric oxygen
treatment. wmav: weighted mean as v; thus mean exposed is set at twice the mean of the controls. §:
the v is derived from a study with a not defined smoking status, instead of the defined smoking status
as in this case.




HPRT-mutation frequency in healthy individuals, determined by the BrdU
assay.

Studies and their exposure types
Exposure      Smoking status                       n1         Variant             v               n2 [power]
                                                                       -6
type                                                      Frequency (10 )
                                                             mean ± SD
EE (nitrate   NS+CS            van Maanen          14         44 ± 60        20@¥/220      (30)      ND
contamina                      et al., 1996                                  @¥¥/104
ted                                                                          @¥¥¥
drinking
water)
EE (nitrate   NS+CS            van Maanen          21          24 ± 24       20@ф/24       (30)    1 [0.90]
contamina                      et al., 1996                                  0фф/124               1 [0.95]
ted                                                                          @ффф
drinking
water)

OE (coal      NS               Stierum et          7          3.9 ± 3.7                    (28)      ND
fly ash                        al., 1993
processin
g workers)
OE (coal     CS               Stierum et       10       16.8 ± 24.9                 (28)         ND
fly ash                       al., 1993
processin
g workers)

SE           NS                Hageman et      23       6.00 ± 7.15       6.03@     (29)         ND
                                al., 1993

Weighted     NS+CS                                     19.38 ± 30.50     240.00               1 [0.90]
(n1)                                                        (75)         (21)                 1 [0.95]
Weighted     NS+CS                                     19.38 ± 30.50     19.38                53 [0.90]
(n1)                                                        (75)         (wmav)               65 [0.95]
Weighted     NS                                       5.51 ± 6.57 (30)   240.00               1 [0.90]
(n1)                                                                     (21)§                1 [0.95]
Weighted     NS                                       5.51 ± 6.57 (30)   5.51                 30 [0.90]
(n1)                                                                     (wmav)               37 [0.95]
Weighted     CS                                      16.8 ± 24.9 (10)¶   240.00               1 [0.90]
(n1)                                                                     (21)§                1 [0.95]
Weighted     CS                                      16.8 ± 24.9 (10)¶   16.8                 47 [0.90]
(n1)                                                                     (wmav)               58 [0.95]
EE: environmental exposure. OE: occupational exposure. SE: smoking exposure (non-smokers vs.
smokers). NS: non-smoker. CS: current smoker. n1: n (amount of individuals) in observed group. n2:
the calculated amount of exposed individuals in the research population for that specific power. ¥: 0.02
mg nitrate/L drinking water group compared with 17.5 mg nitrate/L drinking water group. ¥¥: 0.02 mg
nitrate/L drinking water group compared with 25 ± 15.5 mg nitrate/L drinking water group. ¥¥¥: 0.02
mg nitrate/L drinking water group compared with 135 ± 76 mg nitrate/L drinking water group.
Ф: 17.5 mg nitrate/L drinking water group compared with 0.02 mg nitrate/L drinking water group. ФФ:
17.5 mg nitrate/L drinking water group compared with 25 ± 15.5 mg nitrate/L drinking water group.
ФФФ: 17.5 mg nitrate/L drinking water group compared with 135 ± 76 mg nitrate/L drinking water
group. @: not significant. ND: not defined; lack of relevant and/or significant values. v: difference in
biomarker between exposed and control group. wmav: weighted mean as v; thus mean exposed is set
at twice the mean of the controls. §: the v is derived from a study with a NS+CS, instead of the only
NS (or CS) as in this case. ¶: only based on one study, and a small one at that (28).
Appendix 3
Oxidative damage measured in lymphocytes from healthy individuals,
determined by HPLC-ECD.

Studies and their exposure types
                                                                6
Exposure       Smoking status                 n1    8-OHdG / 10 dG      v               n2
type                                                   mean ± SD                      [power]
OE                   NS         Marczynsk     143      1.55 ± 0.38            (31)      ND
(asbestos                       i et al.,
fiber                           2000
exposed
workers)
OE                   CS         Marczynsk     71      1.45 ± 0.40             (31)      ND
(asbestos                       i et al.,
fiber                           2000
exposed
workers)
OE                 NS+CS        Marczynsk     214     1.52 ± 0.39             (31)      ND
(asbestos                       i et al.,
fiber                           2000
exposed
workers)
OE                   NS         Marczynsk     46       15.1 ± 4.3              (32)     ND
(styrene                        i et al.,
exposed                         1997
workers)
OE                   CS         Marczynsk     21       15.5 ± 4.7              (32)     ND
(styrene                        i et al.,
exposed                         1997
workers)

SE                   NS         Nia et al.,   21       38.6 ± 5.2      12.3    (33)   4 [0.90]
                                  2001                                                5 [0.95]
DE                   NS         Collins et    4           5.8                 (181)     ND
(carotenes)                     al., 1998
DE                   NS         Collins et    4           9.0                 (181)     ND
(carotenes)                     al., 1998
DE                   NS         Collins et    4           3.4                 (181)     ND
(carotenes)                     al., 1998
DE                   NS         Collins et    4           5.1                 (181)     ND
(carotenes)                     al., 1998
DE                   NS         Collins et    4           4.6                 (181)     ND
(carotenes)                     al., 1998
DE                   NS         Collins et    4           3.3                 (181)     ND
(carotenes)                     al., 1998
DE                  ND          Thompso       28       7.9 ± 1.2               (34)     ND
(vegetable                       n et al.,
s and fruit)                       1999
ND                  ND          Bianchini     24      1.59 ± 1.01              (35)     ND
                                  et al.,
                                   2000
ND                  ND          Bianchini     28      2.30 ± 0.78              (35)     ND
                                  et al.,
                                   2000
DE                  ND          Bianchini     28    2.17 (1.27-4.40)          (182)     ND
(alcohol                          et al.,
intake)                            2001
ME (HBO              NS          Dennog       19      1.75 ± 0.27              (27)     ND
treatment)                        et al.,
                                   1999
Weighted      NS+CS                                    4.61 ± 1.80 (615)         12.3                 1 [0.90]
(n1)                                                                             (21) ø               1 [0.95]
Weighted      NS+CS                                    4.61 ± 1.80 (615)         4.61                 4 [0.90]
(n1)                                                                             (wmav)               4 [0.95]
Weighted      NS                                       7.69 ± 2.49 (229)         12.3 (21)ø           1 [0.90]
(n1)                                                                                                  2 [0.95]

Weighted      NS                                       7.69 ± 2.49 (229)         7.69                 3 [0.90]
(n1)                                                                             (wmav)               3 [0.95]
Weighted      CS                                          4.66 ± 2.24 (92)       12.3                 1 [0.90]
(n1)                                                                             (21) ø               1 [0.95]
Weighted      CS                                          4.66 ± 2.24 (92)       4.66                 5 [0.90]
(n1)                                                                             (wmav)               7 [0.95]
OE: occupational exposure. SE: smoking exposure (non-smokers vs. smokers). ME: medical
exposure. NS: non-smoker. CS: current smoker. @: not significant. ND: not defined; lack of relevant
and/or significant values. n1: n (amount of individuals) in observed group. n2: the calculated amount of
exposed individuals in the research population for that specific power. v: difference in biomarker
between exposed and control group. wmav: weighted mean as v; thus mean exposed is set at twice
the mean of the controls. : the v is derived from a study with NS (33) instead of CS (or CS+NS) as in
this case. ø: since no EE based v could be extracted from studies a SE based v has been applied.



Oxidative damage measured in leukocytes from healthy individuals,
determined by HPLC-ECD.
Studies and their exposure types
                                                                             6
Exposure      Smoking status                          N1      8-OHdG / 10             v              n2 [power]
type                                                         dG mean ± SD
EE (2h              NS          Bergamaschi et al.,   8        4.53 ± 0.37          2.94†     (37)    1 [0.90]
traffic                               2001                                                            1 [0.95]
exposure
on
motorcycle
O3
concentrati
ons >80
ppb)
EE (2h              NS          Bergamaschi et al.,   16       3.87 ± 0.61         0.78††     (37)    13 [0.90]
traffic                               2001                                                            16 [0.95]
exposure
on
motorcycle
O3
concentrati
ons >80
ppb)

OE                  NS            Ivancsits et al.,   12       0.72 ± 0.70                    (38)      ND
(vanadium                              2002
exposed
workers)
OE (coal           NS+CS          Toraason et al.,    15        19.6 ± 8.3                    (44)      ND
tar and/or                             2001
asphalt
fume
exposed
workers)
OE                  NS          Irie et al., 2001     10         3.1 ± 0.7                    (39)      ND
(perceived
workload,
slight vs.
great)
OE                  NS         Irie et al., 2001      10     2.8 ± 1.3                   (39)      ND
(perceived
stress,
slight vs.
great)
OE                  NS         Irie et al., 2001      7      2.9 ± 0.3                   (39)      ND
(possibility
to alleviate
stress,
possible
vs.
impossible
)

SE                  NS           Lee et al., 1998     5     22.1 ± 1.0                   (36)      ND
SE                  NS          van Zeeland et al.,   29    34.0 ± 6.5                   (40)      ND
                                      1999
SE                  NS            Lodovici et al.,    31    15.3 ± 1.8                   (41)      ND
                                      2000
DE                  NS           Chen et al., 1999    32    2.28 ± 0.38                  (42)      ND
DE                  CS           Lee et al., 1998     15    29.5 ± 1.4                   (36)      ND
(vitamin E
intake)
DE (red             CS           Lee et al., 1998     15    37.1 ± 1.9                   (36)      ND
ginseng
intake)
DE              NS+CS+FS         Nakajima et al.,     31    2.89 ± 1.37                  (43)      ND
(alcohol)                            1996

Weighted       NS+CS+FS                                    13.61 ± 3.30   1.5 (24)              102 [0.90]
(n1)                                                          (236)                             126 [0.95]
Weighted       NS+CS+FS                                    13.61 ± 3.30   13.61                  2 [0.90]
(n1)                                                          (236)       (wmav)                 2 [0.95]
Weighted       NS                                          11.44 ± 2.97   1.5 (24)               83 [0.90]
(n1)                                                          (160)                             102 [0.95]
Weighted       NS                                          11.44 ± 2.97   11.44                  2 [0.90]
(n1)                                                          (160)       (wmav)                 2 [0.95]
Weighted       CS                                          33.30 ± 2.79   1.5 (24)               73 [0.90]
(n1)                                                          (30)¶                              90 [0.95]
Weighted       CS                                          33.30 ± 2.79   33.30                  1 [0.90]
(n1)                                                          (30)¶       (wmav)                 1 [0.95]
EE: environmental exposure. OE: occupational exposure. SE: smoking exposure (non-smokers vs.
smokers). n1: n (amount of individuals) in observed group. n2: the calculated amount of exposed
individuals in the research population for that specific power. NS: non-smoker. CS: current smoker.
FS: former smoker. @: not significant. ND: not defined; lack of relevant and/or significant values. v:
difference in biomarker between exposed and control group. wmav: weighted mean as v; thus mean
exposed is set at twice the mean of the controls. †: comparison of persons with NQO1 wild type and
GSTM1null genotypes pre- and post-ride of 2 hour on a motorcycle exposed to O3 exceeding 80 parts
per billion (ppb). ††: comparison of persons with other than † genotypes pre- and post-ride of 2 hour
on a motorcycle exposed to O3 exceeding 80 parts per billion (ppb). : the v is derived from a study
(37) with NS instead of CS (or CS+NS) as in this case. ¶: only based on one study (36).



Oxidative damage (8-OHdG) measured in urine from healthy individuals.

Expressed as µmol/mol creatine:

Studies and their exposure types
Exposure       Smoking status                          n1     8-OHdG              v               n2 [power]
type                                                         (µmol/mol
                                                              creatine)
                                                             mean ± SD
OE (cobalt-         NS+CS         De Boeck et al.,     27    1.46 ± 1.48                   (48)      ND
containing                            2000
dust
exposed
workers)
OE                  NS+CS       Nilsson et al., 1996   32        0.85                      (18       ND
(gasoline                                                                                   3)
exposed
workers)

SE                   NS          Pilger et al., 2001   45    1.62 ± 0.50         0.33      (45)    49 [0.90]
                                                                                                   60 [0.95]
SE                   NS          Pilger et al., 2002   44    1.61 ± 0.95         0.75      (46)    34 [0.90]
                                                                                                   42 [0.95]
DE (β-               NS         Sumida et al., 1997    8     1.70 ± 0.11                   (47       ND
carotene)                                                                                   )
ExcE (a              NS         Sumida et al., 1997    6     1.94 ± 0.10                   (47       ND
single bout                                                                                 )
of
exhaustive
exercise)

Weighted       NS+CS                                         1.60 ± 0.92    0.54 (89)              62 [0.90]
(n1)                                                            (130)                              76 [0.95]
Weighted       NS+CS                                         1.60 ± 0.92    1.60 (wmav)            7 [0.90]
(n1)                                                            (130)                              9 [0.95]
Weighted       NS                                            1.64 ± 0.71    0.54 (89)              37 [0.90]
(n1)                                                            (103)                              45 [0.95]
Weighted       NS                                            1.64 ± 0.71    1.64 (wmav)            4 [0.90]
(n1)                                                            (103)                              5 [0.95]
Weighted       CS                                                ND         0.54 (89)                 ND
(n1)
Weighted       CS                                                ND         ND                       ND
(n1)
OE: occupational exposure. SE: smoking exposure (non-smokers vs. smokers). DE: dietary exposure.
ExcE: exposure to physical exercise (regular exercisers as controls). NS: non-smoker. CS: current
smoker. n1: n (amount of individuals) in observed group. n2: the calculated amount of exposed
individuals in the research population for that specific power. @: not significant. ND: not defined; lack
of relevant and/or significant values. v: difference in biomarker between exposed and control group.
wmav: weighted mean as v; thus mean exposed is set at twice the mean of the controls. : the v is
derived from studies (45, 46) with NS instead of CS (or CS+NS) as in this case.


Expressed as µg/g creatine:

Studies and their exposure types
Exposure       Smoking status                          n1   8-OHdG (µg/g          v               n2 [power]
type                                                          creatine)
                                                             mean ± SD
EE (long             NS            Chuang et al.,      43     10.1 ± 4.5         2.1       (49)    97 [0.90]
term traffic                          2003                                                        120 [0.95]
exposure
(taxi-
drivers))
EE (long             CS            Chuang et al.,      32     13.3 ± 4.5         1.0@      (49)      ND
term traffic                          2003
exposure
(taxi-
drivers))
EE (long            NS+CS         Chuang et al.,     75      11.5 ± 4.7          1.9      (49)   129 [0.90]
term traffic                         2003                                                        160 [0.95]
exposure
(taxi-
drivers))

OE (coal            NS+CS         Toraason et al.,   12      3.3 ± 1.9                    (44)      ND
tar and/or                             2001
asphalt
fume
exposed
workers)

ExcE (not            NS         Huang et al., 2000   101     16.6 ± 0.9                   (50)      ND
regular
exercise)
DE                   NS         Huang et al., 2000   148     18.6 ± 0.8                   (50)      ND
(antioxidan
t
supplemen
tuse)
DE (fruit            NS         Huang et al., 2000   35        20.5‫٭‬                      (50)      ND
and
vegetable
intake)
DE (fruit            ND          Thompson et al.,    28     49.6 ± 12.4                   (34)      ND
and                                  1999
vegetable
intake)

Weighted       NS+CS                                       17.27 ± 4.16     1.97 (118)            94 [0.90]
(n1)                                                          (439)                              116 [0.95]
Weighted       NS+CS                                       17.27 ± 4.16     17.27                  2 [0.90]
(n1)                                                          (439)         (wmav)                 2 [0.95]
Weighted       NS                                          16.66 ± 1.88     2.1 (43)              17 [0.90]
(n1)                                                          (292)                               21 [0.95]
Weighted       NS                                          16.66 ± 1.88     16.66                  1 [0.90]
(n1)                                                          (292)         (wmav)                 1 [0.95]
Weighted       CS                                           13.3 ± 4.5      1.97 (118)Ħ          110 [0.90]
(n1)                                                          (32)¶                              136 [0.95]
Weighted       CS                                           13.3 ± 4.5      13.3 (wmav)            3 [0.90]
(n1)                                                          (32)¶                                3 [0.95]
EE: environmental exposure. OE: occupational exposure. ExcE: exposure to physical exercise
(regular exercisers as controls). n1: n (amount of individuals) in observed group. n2: the calculated
amount of exposed individuals in the research population for that specific power. NS: non-smoker. CS:
current smoker. @: not significant. ND: not defined; lack of relevant and/or significant values. v:
difference in biomarker between exposed and control group. ‫ 2 :٭‬or less servings of fruit or vegetables
a day. wmav: weighted mean as v; thus mean exposed is set at twice the mean of the controls. Ħ:
since no significant v from study groups with solely CS was available, the applied v was derived from
studies with NS+CS or NS. ¶: only based on one study (49).



Expressed as pmol/kg/24h:

Studies and their exposure types
Exposure       Smoking status                        n1       8-OHdG              v                 n2
type                                                       (pmol/kg/24h)                          [power]
                                                             mean ± SD
OE (diesel          NS+CS       Loft et al., 1999    23    145.0 ± 115.0^                 (52)      ND
exposed
workers)
OE (diesel        NS+CS         Loft et al., 1999      4    115.0 ± 66.0^                 (52)      ND
exposed
workers)

SE                  NS            Loft et al., 1992    53      213 ± 84           107     (51)   13 [0.90]
                                                                                                 17 [0.95]
SE                  NS           Pilger et al., 2002   44     257 ± 142                   (46)     ND


Weighted     NS+CS                                             212.84 ±      107 (53)            24 [0.90]
(n1)                                                         113.21 (124)                        30 [0.95]
Weighted     NS+CS                                             212.84 ±      212.84              6 [0.90]
(n1)                                                         113.21 (124)    (wmav)              8 [0.95]
Weighted     NS                                                232.96 ±      107 (53)            24 [0.90]
(n1)                                                          113.97 (97)                        30 [0.95]
Weighted     NS                                                232.96 ±      232.96              6 [0.90]
(n1)                                                          113.97 (97)    (wmav)              7 [0.95]
Weighted     CS                                                   ND         107 (53)               ND
(n1)
Weighted     CS                                                  ND          ND                     ND
(n1)
OE: occupational exposure. SE: smoking exposure (non-smokers vs. smokers). n1: n (amount of
individuals) in observed group. n2: the calculated amount of exposed individuals in the research
population for that specific power. NS: non-smoker. CS: current smoker. @: not significant. ND: not
defined; lack of relevant and/or significant values. v: difference in biomarker between exposed and
control group. wmav: weighted mean as v; thus mean exposed is set at twice the mean of the controls.
^: no real ‘controls’, but rural bus drivers on a day off. Ф: rural/ suburban bus-drivers on a day off
compared with city center drivers. : the v is derived from a study (51) with NS instead of CS (or
CS+NS) as in this case.


Expressed as ng/BMI:

Studies and their exposure types
Exposure     Smoking status                            n1     8-OHdG               v             n2 [power]
type                                                          (ng/BMI)
                                                             mean ± SD
SE                  NS            Nia et al., 2001     24     197 ± 31            43@     (33)      ND


Weighted     NS+CS                                               ND          ND                     ND
(n1)
Weighted     NS+CS                                               ND          ND                     ND
(n1)
Weighted     NS                                             197 ± 31 (24)¶   43@ (24)ø           11 [0.90]ø
(n1)                                                                                             14 [0.95]ø
Weighted     NS                                             197 ± 31 (24)¶   197 (wmav)           1 [0.90]
(n1)                                                                                              1 [0.95]
Weighted     CS                                                  ND          ND                      ND
(n1)
Weighted     CS                                                  ND          ND                     ND
(n1)
SE: smoking exposure (non-smokers vs. smokers). n1: n (amount of individuals) in observed group. n2:
the calculated amount of exposed individuals in the research population for that specific power. NS:
non-smoker. CS: current smoker. @: not significant. ND: not defined; lack of relevant and/or significant
values. v: difference in biomarker between exposed and control group. wmav: weighted mean as v;
thus mean exposed is set at twice the mean of the controls. ø: since no significant v was available, a
non-significant v has been chosen for this calculation. ¶: only based on one study (33).


Expressed as nmol/24h:

Studies and their exposure types
Exposure     Smoking status                            n1     8-OHdG              v              n2 [power]
type                                                         (nmol/24h)
                                                             mean ± SD
SE                  NS           Pilger et al., 2002   44    19.1 ± 11.1         5.6      (46)    83 [0.90]
                                                                                                 103 [0.95]
SE            4 week smoking    Prieme et al., 1998    58    24.1 ± 10.5       6.4Ю       (53)    57 [0.90]
               cessation CS                                                                       70 [0.95]

Weighted     NS+4 week                                      21.94 ± 10.76   6.05 (102)           67 [0.90]
(n1)         smoking                                            (102)                            83 [0.95]
             cessation CS
Weighted     NS+4 week                                      21.94 ± 10.76   21.94                 6 [0.90]
(n1)         smoking                                            (102)       (wmav)                7 [0.95]
             cessation CS
Weighted     NS                                              19.1 ± 11.1    5.6 (44)              83 [0.90]
(n1)                                                            (44)¶                            103 [0.95]
Weighted     NS                                              19.1 ± 11.1    19.1 (wmav)           8 [0.90]
(n1)                                                            (44)¶                             9 [0.95]
Weighted     4 week smoking                                  24.1 ± 10.5    6.4 (58)              57 [0.90]
(n1)         cessation CS                                       (58)¶                             70 [0.95]
Weighted     4 week smoking                                  24.1 ± 10.5    24.1 (wmav)           4 [0.90]
(n1)         cessation CS                                       (58)¶                             5 [0.95]
SE: smoking exposure (non-smokers vs. smokers). n1: n (amount of individuals) in observed group. n2:
the calculated amount of exposed individuals in the research population for that specific power. NS:
non-smoker. CS: current smoker. @: not significant. ND: not defined; lack of relevant and/or significant
values. v: difference in biomarker between exposed and control group. wmav: weighted mean as v;
thus mean exposed is set at twice the mean of the controls. Ю: CS before and after 4 week smoking
cessation. ¶: only based on one study: NS (46) or 4 week smoking cessation CS (53).
Appendix 4
Total testosterone levels in serum of non-exposed healthy males (and
females).

Studies and their exposure types
Exposure      Smoking status                            n1   Testosterone          v               n2 [power]
type                                                         (ng/mL) mean
                                                                  ± SD
EE (traffic        ND           Rosa et al., 2003       85 M    4.7 ± 0.2        0.1@       (56)      ND
exposure
(motorway
tollgate
employees
))

OE (DAS           NS+CS         Grajewski et al.,       22 M       5.56                     (18       ND
exposed                              1996                                                    4)
workers)
OE (lead          NS+CS          Ng et al., 1991        49 M    7.39 ± 2.21                 (57)      ND
exposed
workers)
OE                NS+CS         Grajewski et al.,       34 M   5.238 ± 1.390                (58)      ND
(radiofrequ                          2000
ency
exposed
workers)
OE                 ND           Oliva et al., 2001      80 M     5.3 ± 2.3                  (59)      ND
(various                                                             ψ
exposures)
OE                 ND           Padungtod et al.,       38 M    4.72 ± 1.43                 (60)      ND
(pesticde                            1998
factory
workers)
OE                 ND           Garry et al., 2001      15       5.4 ± 0.9                  (61)      ND
(chlorophe                                              M
noxy
herbicides
applicators
)
OE (lead          NS+CS         El-Zohairy et al.,      10      6.66 ± 0.46                 (62)      ND
exposed                              1996               M            Ŧ
workers)

SE                  NS          Attia et al., 1989      35 M    4.55 ± 0.24      0.02@      (55)      ND
SE                  NS          Freedman et al.,        234        6.23        1.19Φ/1.20   (68)      ND
                                        1991             M                         ΦΦ
SE                  NS         Sofikitis et al., 1995   28 M        9.2           0.9@       (18      ND
                                                                                              5)
SE                  NS          Field et al., 1994      924     2.81 ± 1.02       0.25      (54)   350 [0.90]
                                                         M                                         433 [0.95]
DE                NS+CS         Freedman et al.,        109        6.48                     (68)      ND
(alcohol)                            1991                M
DE (alpha         NS+CS          Hartman et al.,        100     5.74 ± 0.14                 (63)      ND
tocopherol                           2001                M
supplemen
tation)
DE (PCB            ND          Persky et al., 2001      26 M      3.905                     (77)      ND
polluted
fish)
ExcE Ǽ             ND             Roberts et al.,       5M      8.68 ± 0.93                 (64)      ND
                                        1993
ExcE Œ                ND          Friedl et al., 2000   48 M    4.70 ± 0.46                  (65)      ND
(multistres
sor
environme
nt)
SE                    NS         Ortego-Centeno et      15 F    0.45 ± 0.19       0.06@      (67)      ND
                                      al., 1994
SE                    NS         Sowers et al., 2001    377    0.196 ± 0.025   0.021¥/0.07   (66)    4 [0.90]
                                                         F                         1¥¥               5 [0.95]

Weighted      M: NS+CS                                          3.72 ± 1.13    0.44 (1158)          139 [0.90]
(n1)                                                              (1423)                            172 [0.95]
Weighted      M: NS+CS                                          3.72 ± 1.13    3.72 (wmav)           2 [0.90]
(n1)                                                              (1423)                             3 [0.95]
Weighted      M: NS+CS Ð                                        3.54 ± 1.03    0.44 (1158)          116 [0.90]
(n1)                                                              (1280)                            143 [0.95]
Weighted      M: NS+CS Ð                                        3.54 ± 1.03    3.54 (wmav)           2 [0.90]
(n1)                                                              (1280)                             3 [0.95]
Weighted      M: NS (= M: NS                                    2.87 ± 1.00    0.44 (1158)          109 [0.90]
(n1)          Ð)                                                   (959)                            135 [0.95]
Weighted      M: NS (= M: NS                                    2.87 ± 1.00    2.87 (wmav)           3 [0.90]
(n1)          Ð)                                                   (959)                             4 [0.95]
Weighted      F: NS                                             0.21 ± 0.04    0.046 (377)           16 [0.90]
(n1)                                                               (392)                             20 [0.95]
Weighted      F: NS                                             0.21 ± 0.04    0.21 (wmav)           1 [0.90]
(n1)                                                               (392)                             1 [0.95]
EE: environmental exposure. OE: occupational exposure. SE: smoking exposure (non-smokers vs.
smokers). DE: dietary exposure. ExcE: exposure to physical exercise (regular exercisers as controls).
n1: n (amount of individuals) in observed group. n2: the calculated amount of exposed individuals in the
research population for that specific power. NS: non-smoker. CS: current smoker. FS: former smoker.
@: not significant. ND: not defined; lack of relevant and/or significant values. v: difference in biomarker
between exposed and control group. wmav: weighted mean as v; thus mean exposed is set at twice
the mean of the controls. M: males. F: females. ψ: men are recruited partners from couples on their
first infertility consultation. Ŧ : male residents from Cairo, Egypt (one of the most polluted cities in the
world: especially EE to lead is very high) (62). Ǽ: in this specific case controls are endurance-trained
men, and ‘exposed’ are these men after overtraining (twice the previous average weekly training
volume with unchanged intensity). Œ: in this specific case controls are militia, and ‘exposed’ are these
men after finishing a rigorous 8 week during US army Ranger course. Multistressor environment in the
study by Friedl et al. (65) includes: semi starvation, sustained workload, inadequate sleep, and thermal
strain. Φ: 1-20 cigarettes a day. ΦΦ: more than 20 cigarettes a day. ¥: compared with FS. ¥¥:
compared with CS. Ð: studies with potentially ‘compromised’ controls excluded (59, 62, 64, 65). : the
v is derived from studies (54, 68) with NS instead of CS+NS as in this case.
Appendix 5
Serum oestradiol levels in healthy individuals.

Studies and their exposure types
Exposure     Smoking status                          n1     Oestradiol ±         v                n2 [power]
type                                                        SD (pg/mL)
OE                 ND         Oliva et al., 2001    80 M    23.0 ± 15.8 ¥                  (59)      ND
(various
exposures)
OE                 ND          Aw et al., 1989      8M         41 ± 13                     (70)      ND
(zeranol
exposed
workers)
OE (lead           ND          Telisman et al.,     51 M    38.15 ± 19.07                  (71)      ND
exposed                             2000
workers)

SE                 NS           Bancroft and          81     62.7 ± 54.5    2.7@٨/8.1@     (75)      ND
                               Cawood., 1996        pre-                        ٨٨
                                                     and
                                                    post-
                                                    MP F
SE                 NS         Austin et al., 1993     84         8.9        1.7@٨/0.1@     (18       ND
                                                    pre-                        ٨٨          6)
                                                     and
                                                    post-
                                                    MP F
SE                 NS         Verkasalo et al.,      416       96.73        22.07^/6.54^   (74)      ND
                                   2001             pre-                         ^
                                                    MP F
SE                 NS         Verkasalo et al.,      299        4.96        0.41@^/0.05    (74)      ND
                                   2001             post-                       @^^
                                                    MP F
SE                 NS          Law et al., 1997      154        12.3           0.1@        (18       ND
                                                    post-                                   7)
                                                    MP F
SE               NS+FS         Wu et al., 2002       136        13.0            3.7        (73)      ND
                                                    post-
                                                    MP F
SE                 NS          Chapurlat et al.,    2943    7.14 ± 28.75       0.87@       (72)      ND
                                   2001             post-
                                                    MP F
SE                 NS         Kupeli et al., 1997   23 M    26.65 ± 3.04Ψ   7.18^/5.58^^   (69)    5 [0.90]
                                                                                                   6 [0.95]

Weighted     M: NS+CS                                       29.18 ± 15.77   6.38 (23) ψ           129 [0.90]
(n1)                                                             (162)                            159 [0.95]
Weighted     M: NS+CS                                       29.18 ± 15.77   29.18                  7 [0.90]
(n1)                                                             (162)      (wmav)                 8 [0.95]
Weighted     M: NS+CS Ð                                     38.54 ± 18.43   ND                        ND
(n1)                                                              (59)
Weighted     M: NS+CS Ð                                     38.54 ± 18.43   38.54                  5 [0.90]
(n1)                                                            (59) ψ      (wmav)                 6 [0.95]
Weighted     M: NS ψ                                         26.65 ± 3.04   6.38 (23) ψ            5 [0.90]
(n1)                                                           (23) ψ¶                             6 [0.95]
Weighted     M: NS ψ                                         26.65 ± 3.04   26.65                  1 [0.90]
(n1)                                                           (23) ψ¶      (wmav)                 1 [0.95]
Weighted     Pre- and post-                                  8.63 ± 29.72   11.69 (552)           136 [0.90]
(n1)         MP F: NS+FS                                        (3024)                            169 [0.95]
Weighted     Pre- and post-                                  8.63 ± 29.72   8.63 (wmav)           250 [0.90]
(n1)         MP F: NS+FS                                       (3024)                            309 [0.95]
Weighted     Post-MP F:                                     7.14 ± 28.75   3.7 (136)            1270 [0.90]
(n1)         NS+FS                                            (2943)¶                           1570 [0.95]
Weighted     Post-MP F:                                     7.14 ± 28.75   7.14 (wmav)          341 [0.90]
(n1)         NS+FS                                            (2943)¶                           422 [0.95]
Weighted     Post-MP F: NS                                  7.14 ± 28.75   ND                      ND
(n1)                                                          (2943)¶
Weighted     Post-MP F: NS                                  7.14 ± 28.75   7.14 (wmav)           341 [0.90]
(n1)                                                          (2943)¶                            422 [0.95]
OE: occupational exposure. SE: smoking exposure (non-smokers vs. smokers). n1: n (amount of
individuals) in observed group. n2: the calculated amount of exposed individuals in the research
population for that specific power. NS: non-smoker. CS: current smoker. FS: former smoker. @: not
significant. ND: not defined; lack of relevant and/or significant values. v: difference in biomarker
between exposed and control group. wmav: weighted mean as v; thus mean exposed is set at twice
the mean of the controls. M: males. F: females. MP: menopausal. ¥: men are recruited partners from
couples on their first infertility consultation. ٨: compared with CS. ٨٨: compared with FS. Ψ: no real
controls, but patients with benign prostatic hyperplasia. Ð: excluding possibly ‘compromised’ control-
containing studies (59, 69). ¶: only based on one study: females (72) and males (69). : the v is
derived from a study (69) with NS instead of CS+NS as in this case.
Appendix 6
Serum sex-hormone binding globulin (SHBG) levels in healthy
individuals.

Studies and their exposure types
Exposure     Smoking status                          n1     SHBG ± SD           v               n2 [power]
type                                                         (nmol/L)
OE (Agent            ND         Michalek et al.,    996        28.5¥                     (82)      ND
Orange                              1999             M
exposed
veterans)
OE (Agent            ND         Michalek et al.,    125       24.0¥■                     (82)      ND
Orange                              1999             M
exposed
veterans)
OE                   ND        Abell et al., 2000   37      21.0 ± 7.7§                  (78)      ND
(pesticide                                          M
exposed
greenhous
e workers)
OE (carbon           ND       Wagar et al., 1983    22          53                       (18       ND
disulfide                                           M                                     8)
exposed
workers)

SE                   NS       Daniel et al., 1992     29       57.85           5.53      (79,      ND
                                                    pre-                                 80)
                                                    MP F
SE                   NS       Moore et al., 1987     152        68.4            9.6      (81)      ND
                                                    pre-
                                                    MP F
                                                      Φ
SE                   NS       Moore et al., 1987     134        62.6          3.3@       (81)      ND
                                                    post-
                                                    MP F
                                                      Φ
SE                   NS        Cassidenti et al.,     17        158            35@       (18       ND
                                    1992            post-                                 9)
                                                    MP F
SE                   NS         Eliasson et al.,     18      28.6 ± 8.3      5.7/7.4Ħ    (76)    34 [0.90]
(smoking                             1993            M                                           42 [0.95]
and snuff
using)
DE (dioxin           ND       Persky et al., 2001   28         36.1                      (77)      ND
polluted                                            M
fish)
ND                   ND       Mifsud et al., 2001   91 M    21.48 ± 0.80                 (19       ND
                                                                                          0)

Weighted     M: NS+CS                                       22.24 ± 4.85   6.55 (18)            12 [0.90]
(n1)                                                           (146)                            15 [0.95]
Weighted     M: NS+CS                                       22.24 ± 4.85   22.24                1 [0.90]
(n1)                                                           (146)       (wmav)               2 [0.95]
Weighted     M: NS+CS Ð                                     22.66 ± 3.39   6.55 (18)            6 [0.90]
(n1)                                                           (109)                            7 [0.95]
Weighted     M: NS+CS Ð                                     22.66 ± 3.39   22.66                1 [0.90]
(n1)                                                           (109)       (wmav)               1 [0.95]
Weighted     M: NS                                           28.6 ± 8.3    6.55 (18)            34 [0.90]
(n1)                                                           (18)¶                            42 [0.95]
Weighted     M: NS                                           28.6 ± 8.3    28.6 (wmav)          2 [0.90]
(n1)                                                             (18)¶                              3 [0.95]
Weighted      Pre- and post-                                      ND         8.95 (181)                ND
(n1)          MP F: NS
Weighted      Pre- and post-                                      ND         ND                        ND
(n1)          MP F: NS
Weighted      Pre-MP F: NS                                        ND         8.95 (181)                ND
(n1)
Weighted      Pre-MP F: NS                                        ND         ND                        ND
(n1)
OE: occupational exposure. SE: smoking exposure (non-smokers vs. smokers). DE: dietary exposure.
n1: n (amount of individuals) in observed group. n2: the calculated amount of exposed individuals in the
research population for that specific power. NS: non-smoker. CS: current smoker. FS: former smoker.
@: not significant. ND: not defined; lack of relevant and/or significant values. v: difference in biomarker
between exposed and control group. M: male. F: female. MP: Menopausal. ¥: men (including
‘controls’) are veterans from the war in Vietnam. ■: diabetic controls (also compared with diabetic
exposed groups). §: controls are greenhouse workers with 0-4 years OE in this occupation.
Ħ: as compared with snuff users. : compared with CS smoking less than 10 cigarettes a day. :
compared with CS smoking 11-20 cigarettes a day. : compared with CS smoking more than 20
cigarettes a day. : compared with FS. Φ: never used oral contraceptives or hormone replacement
therapy. wmav: weighted mean as v; thus mean exposed is set at twice the mean of the controls. Ð:
studies with potentially ‘compromised’ controls excluded (78, 82). ¶: only based on one study (76). :
the v is derived from a study (76) for males or studies for females (79-81) with NS instead of CS+NS
as in this case.
Appendix 7
FSH serum levels in healthy individuals

Studies and their exposure types
Exposure      Smoking status                         n1      FSH ± SD        v           n2 [power]
type                                                           (IU/L)
EE (light           CS         Martikainen et al.,    11      6.0 ± 0.8           (98)      ND
vs. dark                             1996            pre-
season in                                             MP
Finland)                                               F
EE (high           ND           Gonzales and          34      6.3 ± 5.5            (10      ND
altitude)Ж                     Gones or Villena,     pre-                           0,
                                    2000              MP                          101
                                                       F                            )
EE (high           ND           Gonzales and          22    64.8 ± 18.0     2.9    (10    810 [0.90]
altitude)Ж                     Gones or Villena,     post                           0,   1002 [0.95]
                                    2000             -MP                          101
                                                       F                            )
EE (light          ND          Martikainen et al.,    24      5.2 ± 0.6     0.5   (94)    31 [0.90]
vs. dark                             1985             M                                   38 [0.95]
season in
Finland)
EE (traffic        ND          Rosa et al., 2003     85 M     3.2 ± 0.2     0.9   (56)    2 [0.90]
exposure                                                                                  2 [0.95]
(motorway
tollgate
employees
))

OE                 ND          Zaire et al. 1997     29     4.56 ± 3.31           (93)      ND
(Namibian                                            M
uranium
miners)
OE                NS+CS        Grajewski et al.,     34 M     5.8 ± 1.4           (58)      ND
(radiofrequ                         2000
ency
exposed
workers)
OE                NS+CS        Roels et al. 1992     101    4.76 ± 3.67           (92)      ND
(manganes                                             M
e dioxide
dust
exposed
workers)
OE (HCH           NS+CS         Tomczak et al.       20       4.0 ± 1.8           (91)      ND
exposed                            1981              M
workers)
OE                 ND          Oliva et al., 2001    80     10.6 ± 15.3 ¥         (59)      ND
(various                                             M
exposures)
OE                NS+CS        Padungtod et al.,     34       3.4 ± 2.2           (60)      ND
(pesticide                          1998             M
factory
workers)
OE                 ND          Abell et al., 2000    44      4.2 ± 1.5 §          (78)      ND
(pesticide                                           M
exposed
greenhous
e workers)
OE                 ND          Garry et al., 2001    15       5.0 ± 0.9           (61)      ND
(chlorophe                                           M
noxy
herbicides
applicators
)
OE (DBCP-       ND       Glass et al., 1979     22     3.1 ± 0.3 Ǿ    (95)   ND
exposed                                         M
pesticide
applicators
)
OE (dioxin     NS+CS       Egeland et al.,      231    10.9 ± 1.07    (90)   ND
exposed                        1994              M
production
workers)
OE (TDA        NS+CS     Hamill et al., 1982    103      4.8 Ю        (10    ND
and DNT                                          M                    4)
exposed
workers)
OE              ND        Hanaoka et al.,       42         7.6        (19    ND
(BADGE                        2002              M                      1)
exposed
sprayers)
OE            NS+CS+FS   Welch et al., 1988     40      5.5 ± 3.9     (89)   ND
(ethylene                                       M
glycol
esters
exposed
painters)
OE (lead        ND        Gustafson et al.,     25     4.5 ± 0.37     (88)   ND
exposed                        1989             M
workers)
OE (lead       NS+CS      El-Zohairy et al.,    10    5.48 ± 0.58 Ф   (62)   ND
exposed                        1996             M
workers)
OE (lead        ND         Ng et al., 1991      49     1.92 ± 1.20    (57,   ND
exposed                                         M                     62)
workers)
OE (metal      NS+CS      Hjollund et al.,      200        3.3        (19    ND
welders)                       1998              M                    2)
OE             NS+CS     Bonde and Ernst,        60    4.7 ± 2.9 Θ    (96)   ND
(hexavalen                     1992              M
t
chromium
exposed
welders)
OE              ND         Li et al., 2001      21     2.41 ± 0.08    (87)   ND
(hexavalen                                      M
t
chromium
exposed
welders)
OE              ND       Erfurth et al., 1990   10         3.6        (19    ND
(mercury                                        M                     3)
vapour
exposed
plant
workers)
OE              ND       Erfurth et al., 1990   11         5.8        (19    ND
(mercury                                        M                     3)
vapour
exposed
dentists)
OE              ND         Mason, 1990          83      4.4 ± 1.7     (86)   ND
(cadmium                                        M
exposed
workers)
OE              ND       Zeng et al., 2002      27        5.26        (19    ND
(cadmium                                               M                                    4)
exposed
workers)
OE                    ND        Mutti et al., 1984     30          8.8                      (10       ND
(styrene                                              pre-                                  3)
exposed                                               MP
workers)                                               F

SE                    NS        Berta et al. 1992      447      9.5 ± 8.1                   (99)      ND
                                                      pre-
                                                      MP F
SE                    NS       Cooper et al., 1995     232         8.4                      (19       ND
                                                      pre-                                   5)
                                                      MP F
SE                    NS        Verkasalo et al.,      416         7.0                      (74)      ND
                                     2001             pre-
                                                      MP F
SE                    NS        Law et al., 1997       288        70.1                      (18       ND
                                                      post-                                  7)
                                                      MP F
SE                    NS          Bancroft and          81     38.6 ± 35.9                  (75)      ND
                                 Cawood, 1996         pre-
                                                       and
                                                      post-
                                                      MP F
DE              NS+CS+FS        Verkasalo et al.,      132         7.8                      (74)      ND
(alcohol                             2001             pre-
intake)                                               MP F
ME (HRT               ND         Johnson et al.,        9     61.99 ± 11.5                  (10       ND
treatment)                           1997             post-                                  2)
                                                      MP F
SE                    NS        Shaarawy and          20 M    12.22 ± 3.22                  (83)      ND
                                Mahmoud, 1982
DE (PCB               ND       Persky et al., 2001    28          3.00                      (77)      ND
polluted                                              M
fish)
DE (long-             ND       Luboshitzky et al.,    6M        2.0 ± 0.6                   (85)      ND
term                                2000
melatonin
administrat
ion)
ME                    ND       Gillman et al., 1988   6M      1.267 ± 1.325                 (84)      ND
(analgesic
nitrous
oxide)
ExcE (ski-            ND        Vasankari et al.,     9M       5.9 ± 1.9■                   (97)      ND
racing)                             1993

Weighted      M: NS+CS                                          6.21 ± 4.66   0.81 (109)           696 [0.90]
(n1)                                                              (1048)                           861 [0.95]
Weighted      M: NS+CS                                          6.21 ± 4.66   6.21 (wmav)           12 [0.90]
(n1)                                                              (1047)                            15 [0.95]
Weighted      M: NS+CS Ð                                        6.10 ± 1.96   0.81 (108)           124 [0.90]
(n1)                                                               (822)                           153 [0.95]
Weighted      M: NS+CS Ð                                        6.10 ± 1.96   6.10 (wmav)           3 [0.90]
(n1)                                                               (822)                            3 [0.95]
Weighted      M: NS                                            12.22 ± 3.22   0.81 (108)†          333 [0.90]
(n1)                                                               (20)¶                           411 [0.95]
Weighted      M: NS                                            12.22 ± 3.22   12.22                 2 [0.90]
(n1)                                                               (20)¶      (wmav)                2 [0.95]
Weighted      Pre- and post-                                  15.95 ± 15.36   2.9 (22)††           590 [0.90]
(n1)          MP F: NS+CS                                          (604)                           730 [0.95]
Weighted      Pre- and post-                                  15.95 ± 15.36   15.95                 20 [0.90]
(n1)          MP F: NS+CS                                          (604)      (wmav)                25 [0.95]
Weighted      Pre-MP F:                                         9.20 ± 7.87   2.9 (22)††           155 [0.90]
(n1)         NS+CS                                              (492)                             192 [0.95]
Weighted     Pre-MP F:                                        9.20 ± 7.87     9.20 (wmav)          16 [0.90]
(n1)         NS+CS                                               (492)                             20 [0.95]
Weighted     Pre-MP F: NS                                      9.5 ± 8.1      2.9 (22)††          164 [0.90]
(n1)                                                            (447)¶                            203 [0.95]
Weighted     Pre-MP F: NS                                      9.5 ± 8.1      9.5 (wmav)           16 [0.90]
(n1)                                                            (447)¶                             19 [0.95]
Weighted     Pre-MP F: CS                                   6.0 ± 0.8 (11)¶   2.9 (22)††           2 [0.90]
(n1)                                                                                               2 [0.95]
Weighted     Pre-MP F: CS                                   6.0 ± 0.8 (11)¶   6.0 (wmav)           1 [0.90]
(n1)                                                                                               1 [0.95]
Weighted     Post-MP F:                                     63.98 ± 16.47     2.9 (22)            678 [0.90]
(n1)         NS+CS                                              (31)                              839 [0.95]
Weighted     Post-MP F:                                     63.98 ± 16.47     63.98                2 [0.90]
(n1)         NS+CS                                              (31)          (wmav)               2 [0.95]
EE: environmental exposure. OE: occupational exposure. SE: smoking exposure (non-smokers vs.
smokers). DE: dietary exposure. ME: medical exposure. ExcE: exposure to physical exercise (regular
exercisers as controls). n1: n (amount of individuals) in observed group. n2: the calculated amount of
exposed individuals in the research population for that specific power. NS: non-smoker. CS: current
smoker. FS: former smoker. PS: passive smoker. M= male. F=female. MP: menopausal. @: not
significant. ND: not defined; lack of relevant and/or significant values. v: difference in biomarker
between exposed and control group. : the month May compared to the average of all other months
(12 in total) of that year (from October to October). ¥: men are recruited partners from couples on their
first infertility consultation. §: controls are greenhouse workers with 0-4 years OE in this occupation.
Ф : male residents from Cairo, Egypt (one of the most polluted cities in the world: especially EE to lead
is very high) (62). Θ: no real controls but group with lowest serum concentration of chromium (<1.07
nmol/ mmol creatinine): a biomarker for exposure, thereby primarily consisting of controls but also a
minority fraction consists of ‘exposed’ (metal welders). Ǿ: no real controls, since this group consists of
pesticide applicators that have not been exposed to DBCP in that year. Ю: ‘control’ group consists of
not and slightly exposed individuals. ■: no real controls but professional ski-athletes before the race.
DNT: dinitrotoluene. TDA: toluene diamine. BADGE: bisphenol A diglycidyl ether. HRT: hormone
replacement therapy. wmav: weighted mean as v; thus mean exposed is set at twice the mean of the
controls. †: the applied v is based on studies with NS+CS or ND smoking status, instead of NS as in
this case. ††: the applied v is based on studies (100, 101) with ND smoking status post-MP females
instead of the characteristics of the control group in this case. ¶: only based on one study: males (83)
and females (98) or (99). Ð: studies with potentially ‘compromised’ controls excluded (59, 62, 78, 95-
97, 104).
Appendix 8
LH serum levels in healthy males

Studies and their exposure types
Exposure      Smoking status                          n1     LH ± SD (IU/L)    v            n2 [power]
type
EE (acute           NS         Luderer et al., 1999   5M      3.43 ± 0.77     1.54   (10     6 [0.90]
toluene                                                                               5)     7 [0.95]
exposure)
EE (traffic        ND           Rosa et al., 2003     85 M      2.8 ± 0.1            (56)      ND
exposure
(motorway
tollgate
employees
))

OE                 ND           Oliva et al., 2001    80       5.1 ± 4.3 ¥           (59)      ND
(various                                              M
exposures)
OE                NS+CS         Grajewski et al.,     34 M     10.3 ± 1.5            (58)      ND
(radiofrequ                          2000
ency
exposed
workers)
OE                 ND           Zaire et al. 1997     29      3.86 ± 2.10            (93)      ND
(Namibian                                             M
uranium
miners)
OE                 ND            Hanaoka et al.,      42          4.0                (19       ND
(BADGE                               2002             M                               1)
exposed
sprayers)
OE (HCH           NS+CS          Tomczak et al.       20       6.1 ± 2.1             (91)      ND
exposed                             1981              M
workers)
OE                 ND           Padungtod et al.,     38       1.8 ± 1.0             (60)      ND
(pesticde                            1998             M
factory
workers)
OE                 ND           Abell et al., 2000    37       4.3 ± 2.0 §           (78)      ND
(pesticide                                            M
exposed
greenhous
e workers)
OE                 ND           Garry et al., 2001    15       4.7 ± 0.6             (61)      ND
(chlorophe                                            M
noxy
herbicides
applicators
)
OE (dioxin        NS+CS          Egeland et al.,      231     16.5 ± 1.05            (90)      ND
exposed                              1994              M
production
workers)
OE (DBCP-          ND           Glass et al., 1979    22      12.1 ± 1.4 Ǿ           (95)      ND
exposed                                               M
pesticide
applicators
)
OE (metal         NS+CS          Hjollund et al.,     200         3.3                (19       ND
welders)                              1998             M                             2)
OE (lead     NS+CS    El-Zohairy et al.,    10      6.90 ± 0.55 Ф                (62)   ND
exposed                    1996             M
workers)
OE (lead      ND     Erfurth et al., 2001   11           4.3                     (19    ND
exposed                                     M                                     6)
workers)
OE (lead      ND      Gustafson et al.,     25       9.0 ± 0.34                  (88)   ND
exposed                    1989             M
workers)
OE (lead     NS+CS     Ng et al., 1991      49 M     3.24 ± 1.61                 (57)   ND
exposed
workers)
OE            ND       Li et al., 2001      21       6.85 ± 0.30                 (87)   ND
(hexavalen                                  M
t
chromium
exposed
welders)
OE           NS+CS   Bonde and Ernst,       60       6.8 ± 3.0 Θ                 (96)   ND
(hexavalen                1992              M
t
chromium
exposed
welders)
OE            ND       Mason, 1990          82        7.8 ± 1.9                  (86)   ND
(cadmium                                    M
exposed
workers)
OE            ND     Zeng et al., 2002      27          5.26                     (19    ND
(cadmium                                    M                                    4)
exposed
workers)
OE           NS+CS   Roels et al. 1992      101      6.04 ± 2.56                 (92)   ND
(manganes                                    M
e dioxide
dust
exposed
workers)
OE            ND     Erfurth et al., 1990   10           6.9                     (19    ND
(mercury                                    M                                    3)
vapour
exposed
plant
workers)
OE            ND     Erfurth et al., 1990   11           8.0                     (19    ND
(mercury                                    M                                    3)
vapour
exposed
dentists)
OE            ND     Mutti et al., 1984      30         11.7                     (10    ND
(styrene                                    pre-                                 3)
exposed                                     MP
workers)                                     F

SE            NS      Berta et al. 1992      447     14.9 ± 14.8      0.1@       (99)   ND
                                            pre-
                                            MP F
SE            NS      Verkasalo et al.,      416        11.9        1.6@^/1.5@   (74)   ND
                           2001             pre-                        ^^
                                            MP F
SE            NS      Law et al., 1997        61        38.2          1.1@       (18    ND
                                            post-                                 7)
                                            MP F
SE            NS        Bancroft and          81     30.7 ± 21.9     3.4@٨/      (75)   ND
                       Cawood, 1996         pre-                     1.2@٨٨
                                             and
                                                      post-
                                                      MP F
DE              NS+CS+FS          Verkasalo et al.,    132        13.5                      (74)     ND
(alcohol                               2001           pre-
intake)                                               MP F
SE                   NS         Allen et al., 2002    372         5.22                       (19     ND
                                                       M                                      7)
DE (PCB              ND         Persky et al., 2001    27         2.29                      (77)     ND
polluted                                               M
fish)

Weighted     M: NS+CS                                          8.34 ± 2.04    1.54 (5)†            37 [0.90]
(n1)                                                              (944)                            46 [0.95]
Weighted     M: NS+CS                                          8.34 ± 2.04    8.34 (wmav)          2 [0.90]
(n1)                                                              (944)                            2 [0.95]
Weighted     M: NS+CS Ð                                        8.93 ± 1.51    1.54 (5)†            21 [0.90]
(n1)                                                              (735)                            25 [0.95]
Weighted     M: NS+CS Ð                                        8.93 ± 1.51    8.93 (wmav)          1 [0.90]
(n1)                                                              (735)                            1 [0.95]
Weighted     M: NS                                             3.43 ± 0.77    1.54 (5)             6 [0.90]
(n1)                                                               (5)¶                            7 [0.95]
Weighted     M: NS                                             3.43 ± 0.77    3.43 (wmav)          2 [0.90]
(n1)                                                               (5)¶                            2 [0.95]
Weighted     Pre- and post-                                   17.32 ± 16.08   ND                      ND
(n1)         MP F: NS                                             (528)
Weighted     Pre- and post-                                   17.32 ± 16.08   17.32                19 [0.90]
(n1)         MP F: NS                                             (528)       (wmav)               23 [0.95]
Weighted     Pre-MP F: NS                                      14.9 ± 14.8    ND                      ND
(n1)                                                             (447)¶
Weighted     Pre-MP F: NS                                      14.9 ± 14.8    14.9 (wmav)          21 [0.90]
(n1)                                                             (447)¶                            26 [0.95]
EE: environmental exposure. OE: occupational exposure. SE: smoking exposure (non-smokers vs.
smokers). DE: dietary exposure. n1: n (amount of individuals) in observed group. n2: the calculated
amount of exposed individuals in the research population for that specific power. NS: non-smoker. CS:
current smoker. FS: former smoker. M= male. F=female. MP: menopausal. @: not significant. ND: not
defined; lack of relevant and/or significant values. v: difference in biomarker between exposed and
control group. ¥: men are recruited partners from couples on their first infertility consultation. §:
controls are greenhouse workers with 0-4 years OE in this occupation. Ф : male residents from Cairo,
Egypt (one of the most polluted cities in the world: especially EE to lead is very high) (62). Θ: no real
controls but group with lowest serum concentration of chromium (<1.07 nmol/ mmol creatinine): a
biomarker for exposure, thereby primarily consisting of controls but also a minority fraction consists of
‘exposed’ (metal welders). Ǿ: no real controls, since this group consists of pesticide applicators that
have not been exposed to DBCP in that year. DBCP: dibromochloropropane, a nematocide. HCH:
hexachlorocyclohexane. BADGE: bisphenol A diglycidyl ether. ^: compared to CS. ^^: compared to
FS. wmav: weighted mean as v; thus mean exposed is set at twice the mean of the controls. †: the
applied v is based on a study with NS (105), instead of NS+CS as in this case. Ð: studies with
potentially ‘compromised’ controls (59, 62, 78, 95, 96) are excluded. ¶: only based on one study:
males (105) and females (99).
Appendix 9
Thyroid stimulating hormone (TSH) levels in healthy individuals

Studies and their exposure types
Exposure      Smoking status                           n1     TSH (mIU/L)       v              n2 [power]
type
EE                 ND           Solter et al., 1989   4M       2.0 ± 0.1       0.7      (10     1 [0.90]
(extreme                                                                                 9)     1 [0.95]
cold)ω
EE                 ND           Solter et al., 1989    6       2.7 ± 0.7      0.1@      (10       ND
(extreme                                              pre-                               9)
cold)ω                                                MP F

OE                 ND          Zaidi et al., 2000     20      2.88 ± 0.51               (10       ND
(pesticide                                            M                                  8)
formulator
s)
OE                 ND          Erfurth et al., 1990   10          1.2                   (19       ND
(mercury                                              M                                 3)
vapour
exposed
plant
workers)
OE                 ND          Erfurth et al., 1990   11          1.2                   (19       ND
(mercury                                              M                                 3)
vapour
exposed
dentists)
OE (lead           ND           Gustafson et al.,     25      1.02 ± 0.13               (88)      ND
exposed                              1989             M
workers)
OE (cobalt        NS+CS          Swennen et al.,      82         4.72                   (19       ND
(-oxides, -                          1993             M                                  8)
salts and -
metal)
exposed
refinery
workers)
OE                 ND           Mutti et al., 1984     30      2.7 ± 1.8                (10       ND
(styrene                                              pre-                              3)
exposed                                               MP
workers)                                               F

LE                  CS         Szostak-Wegierek         8     1.80 ± 0.33               (11       ND
(obesity)                         et al., 1996        pre-                               0)
                                                      MP F
SE                  NS         Muller et al., 1995     109        0.9         0.0@      (19       ND
                                                      pre-                               9)
                                                       and
                                                      post-
                                                      MP F
DE (PCB            ND          Persky et al., 2001      9        1.64         0.17      (77)      ND
polluted                                              pre-
fish)                                                  and
                                                      post-
                                                      MP F
DE (PCB            ND          Persky et al., 2001    26 M       1.82                   (77)      ND
polluted
fish)
SE                  NS           Sepkovic et al.,      50      2.4 ± 0.1    0.2@▼/0.4   (10       ND
                                     1984             M+pr                  @▼▼/0.5@     6)
                                                    e-MP                         ▼▼▼
                                                      F
SE                NS+FS           Knudsen et al.,   2800         1.28              0.14     (10      ND
                                      2002          M+pr                                     7)
                                                      e-
                                                     and
                                                    post-
                                                    MP F

Weighted     M+pre- and                                     2.4 ± 0.1 (50)¶   0.14 (2800)          11 [0.90]
(n1)         post-MP F:                                                                            14 [0.95]
             NS+FS
Weighted     M+pre- and                                     2.4 ± 0.1 (50)¶   2.4 (wmav)           1 [0.90]
(n1)         post-MP F:                                                                            1 [0.95]
             NS+FS
Weighted     M+pre-MP F: NS                                 2.4 ± 0.1 (50)¶   ND                     ND
(n1)
Weighted     M+pre-MP F: NS                                 2.4 ± 0.1 (50)¶   2.4 (wmav)           1 [0.90]
(n1)                                                                                               1 [0.95]
Weighted     M: NS+CS                                        1.86 ± 0.34      0.7 (4)              5 [0.90]
(n1)                                                            (49)                               7 [0.95]
Weighted     M: NS+CS                                        1.86 ± 0.34      1.86 (wmav)          1 [0.90]
(n1)                                                            (49)                               1 [0.95]
Weighted     Pre- and post-                                  2.54 ± 1.54      0.17 (9)††          1725 [0.90]
(n1)         MP F: NS+CS                                        (44)                              2134 [0.95]
Weighted     Pre- and post-                                  2.54 ± 1.54      2.54 (wmav)          8 [0.90]
(n1)         MP F: NS+CS                                        (44)                               10 [0.95]
Weighted     Pre-MP F:                                       2.54 ± 1.54      ND                      ND
(n1)         NS+CS                                              (44)
Weighted     Pre-MP F:                                       2.54 ± 1.54      2.54 (wmav)          8 [0.90]
(n1)         NS+CS                                              (44)                               10 [0.95]
Weighted     Pre-MP F: CS                                    1.80 ± 0.33      ND                      ND
(n1)                                                            (8)¶
Weighted     Pre-MP F: CS                                    1.80 ± 0.33      1.80 (wmav)          1 [0.90]
(n1)                                                            (8)¶                               1 [0.95]
EE: environmental exposure. OE: occupational exposure. SE: smoking exposure (non-smokers vs.
smokers). DE: dietary exposure. NS: non-smoker. CS: current smoker. FS: former smoker. n1: n
(amount of individuals) in observed group. n2: the calculated amount of exposed individuals in the
research population for that specific power. M= male. F=female. MP: menopausal. @: not significant.
ND: not defined; lack of relevant and/or significant values. v: difference in biomarker between exposed
and control group. ω: after work shift extreme cold factory workers (meat cutters) and normal
temperature factory workers. ▼: compared with light smokers. ▼▼: compared with moderate
smokers. ▼▼▼: compared with heavy smokers. wmav: weighted mean as v; thus mean exposed is
set at twice the mean of the controls. ¶: only based on one study: M+F (106) and F (110). ††: this v is
derived from a DE (to EE fish exposed individuals) study (77), since no significant v could be derived
from EE or SE studies, this v has been used (the value of this v is in the same range as the value of
the v found in a large study by Knudsen et al. (107) with both males and females in the study
population).
Appendix 10
Inhibin B levels in healthy individuals

Studies and their exposure types
Exposure     Smoking status                           n1     Inhibin B          V               n2 [power]
type                                                           (ng/L)
OE              NS+CS+FS        Ishoy et al., 2001    215       173            1@         (11      ND
(Persian                                               M                                   1)
Gulf War
exposed
veterans)
OE                  ND          Larsen et al., 1999   85       164Ψ            20Ψ       (11       ND
(organic                                              M                                  2)
farmers)

Weighted      M: NS+CS+FS                                       ND           20Ψ (85)              ND
(n1)
OE: occupational exposure. n1: n (amount of individuals) in observed group. n2: the calculated amount
of exposed individuals in the research population for that specific power. NS: non-smoker. CS: current
smoker. FS: former smoker. M= male. @: not significant. ND: not defined; lack of relevant and/or
significant values. v: difference in biomarker between exposed and control group. wmav: weighted
mean as v; thus mean exposed is set at twice the mean of the controls. Ψ: organic farmers serve as
controls for traditional farmers, but most organic farmers have also had traditional farmer exposure in
the past.
Appendix 11:

CA frequency (per 100 cells) in healthy individuals measured in PBL by
the CA assay

Studies and their exposure types
Exposure     Smoking   CA-type                       n1    Mean CA freq.     v               n2
type          status                                       per 100 cells                   [power]
                                                               ± SD
OE           NS+FS     Chromoso    Lander et al.,   29 Ψ         *           *      (11      ND
(pesticide              me: gap       2000                                           3)
exposed
greenhou
se
workers)
OE           NS+FS     Chromoso    Lander et al.,   29 Ψ    0.17 ± 0.38    0.01@    (11      ND
(pesticide             me: break      2000                                 &/0.11    3)
exposed                                                                     @&&
greenhou
se
workers)
OE           NS+FS     Chromoso    Lander et al.,   29 Ψ    0.10 ± 0.31    0.18@    (11      ND
(pesticide                me:         2000                                 &/0.01    3)
exposed                exchange                                             @&&
greenhou
se
workers)
OE           NS+FS     Chromoso    Lander et al.,   29 Ψ    0.28 ± 0.45    0.16@    (11      ND
(pesticide             me: break      2000                                 &/0.07    3)
exposed                    +                                                @&&
greenhou               exchange
se
workers)
OE           NS+FS     Chromoso    Lander et al.,   29 Ψ    0.28 ± 0.45    0.19@    (11      ND
(pesticide                me:         2000                                 &/0.10    3)
exposed                 subtotal                                            @&&
greenhou
se
workers)
OE           NS+FS     Chromatid   Lander et al.,   29 Ψ     0.31 ± 1.6    0.25@    (11   150 [0.90]
(pesticide               : gap        2000                                 &/0.60    3)   185 [0.95]
exposed                                                                      &&
greenhou
se
workers)
OE           NS+FS     Chromatid   Lander et al.,   29 Ψ    1.00 ± 0.80    0.10@    (11      ND
(pesticide              : break       2000                                 &/0.03    3)
exposed                                                                     @&&
greenhou
se
workers)
OE           NS+FS     Chromatid   Lander et al.,   29 Ψ         *           *      (11      ND
(pesticide                 :          2000                                           3)
exposed                exchange
greenhou
se
workers)
OE           NS+FS     Chromatid   Lander et al.,   29 Ψ    1.03 ± 0.82    0.12@    (11      ND
(pesticide             : break +      2000                                 &/0.03    3)
exposed                exchange                                             @&&
greenhou
se
workers)
OE           NS+FS   Chromatid    Lander et al.,   29 Ψ   1.34 ± 1.14   0.12@    (11     ND
(pesticide           : subtotal      2000                               &/0.56    3)
exposed                                                                  @&&
greenhou
se
workers)
OE           NS+FS   Total CAs    Lander et al.,   29 Ψ   1.62 ± 1.15   0.29@    (11   73 [0.90]
(pesticide                           2000                               &/0.62    3)   90 [0.95]
exposed                                                                   &&
greenhou
se
workers)
OE           NS+FS     Total      Lander et al.,   29 Ψ   1.31 ± 0.85   0.03@    (11     ND
(pesticide           breaks +        2000                               &/0.02    3)
exposed              exchange                                            @&&
greenhou                 s
se
workers)
OE            ND       Mean        Kaiournova       36    2.18 ± 0.32    2.29    (11   1 [0.90]
(herbicide           incidence         and                                        4)   1 [0.95]
productio              of CAs     Khabutdinova,
n                                     1998
workers)
OE            ND     Chromoso      Kaiournova       36    2.42 ± 0.36   2.11@    (11     ND
(herbicide           me: break         and                                        4)
productio                         Khabutdinova,
n                                     1998
workers)
OE            ND     CAs single    Kaiournova       36    1.72 ± 0.28   0.81@    (11     ND
(herbicide           fragments         and                                        4)
productio                         Khabutdinova,
n                                     1998
workers)
OE            ND      CAs pair     Kaiournova       36    0.61 ± 0.15   1.39@    (11     ND
(herbicide           fragments         and                                        4)
productio                         Khabutdinova,
n                                     1998
workers)
OE            ND        Other      Kaiournova       36    0.11 ± 0.05     #      (11     ND
(herbicide           endoredup         and                                        4)
productio             lications   Khabutdinova,
n                                     1998
workers)
OE            ND        4N         Kaiournova       36    0.08 ± 0.05     #      (11     ND
(herbicide                             and                                        4)
productio                         Khabutdinova,
n                                     1998
workers)
OE            ND       Mean        Kaiournova      21Ж    2.29 ± 0.34    2.18    (11   1 [0.90]
(herbicide           incidence         and                                        4)   1 [0.95]
productio              of CAs     Khabutdinova,
n                                     1998
workers)
OE            ND     Chromoso      Kaiournova      21Ж    2.43 ± 0.37   2.10@    (11     ND
(herbicide           me: break         and                                        4)
productio                         Khabutdinova,
n                                     1998
workers)
OE            ND     CAs single    Kaiournova      21Ж    1.76 ± 0.32   0.77@    (11     ND
(herbicide           fragments         and                                        4)
productio                         Khabutdinova,
n                                     1998
workers)
OE            ND      CAs pair     Kaiournova      21Ж    0.67 ± 0.28   1.33@    (11     ND
(herbicide                   fragments        and                                           4)
productio                                Khabutdinova,
n                                            1998
workers)
OE               ND            Other      Kaiournova      21Ж          ф            #      (11      ND
(herbicide                  endoredup         and                                           4)
productio                    lications   Khabutdinova,
n                                            1998
workers)
OE               ND             4N        Kaiournova      21Ж          ф            #      (11      ND
(herbicide                                    and                                           4)
productio                                Khabutdinova,
n                                            1998
workers)

Weighted         ND         Total CAs                              2.02 ± 0.72      1.7           4 [0.90]
(n1)                                                                  (86)        (86)†           5 [0.95]
Weighted         ND         Total CAs                              2.02 ± 0.72     2.02           3 [0.90]
(n1)                                                                  (86)       (wmav)           4 [0.95]
                                                                                     )
Weighted         ND         Chromati                               1.60 ± 0.70     ND               ND
(n1)                           d:                                     (86)
                            subtotal
Weighted         ND         Chromati                               1.60 ± 0.70     1.60           5 [0.90]
(n1)                           d:                                     (86)       (wmav)           5 [0.95]
                            subtotal
Weighted         ND         Chomoso                                0.51 ± 0.31     ND               ND
(n1)                           me:                                    (86)
                            subtotal
Weighted         ND         Chomoso                                0.51 ± 0.31     0.51           8 [0.90]
(n1)                           me:                                    (86)       (wmav)           10 [0.95]
                            subtotal
Weighted         ND         Chromos                                1.67 ± 0.37     ND               ND
(n1)                          ome:                                    (86)
                             break
Weighted         ND         Chromos                                1.67 ± 0.37     1.67           2 [0.90]
(n1)                          ome:                                    (86)       (wmav)           2 [0.95]
                             break
OE: occupational exposure. n1: n (amount of individuals) in observed group. n2: the calculated amount
of exposed individuals in the research population for that specific power. @: not significant. ND: not
defined; lack of relevant and/or significant values. v: difference in biomarker between exposed and
control group. * Chromatid-type exchanges and chromosome-type gaps were rare and have not been
shown separately. &: preseason greenhouse workers. &&: postseason greenhouse workers. Ψ:
organic farmers serve as controls for traditional farmers, but most organic farmers have also had
traditional farmer exposure in the past. #: no comparison with ’exposed’ class. Ж: controls working in
the close vicinity of the herbicide plant. ф: not detected. wmav: weighted mean as v; thus mean
exposed is set at twice the mean of the controls. †: since the absence of a v derived from EE- and SE-
based studies, an OE based v has been chosen as an alternative. Since the results of Lander et al.
(113) show lower values for CAs than those of Kaiournova and Khabutdinova, (114) and only two
studies (113, 114) were available to discuss, the study of Lander et al. (Ψ) has not been excluded from
the pooled classes.
Appendix 12:

Number of micronucleated cells per 1000 binucleated cells (MNCs) in
healthy individuals

Studies and their exposure types
Exposure       Smoking status                           n1   MNCsa ± SD       v              n2 [power]
type
EE                  ND          Chang et al., 1997      75     11 ± 8         6        (13    38 [0.90]
(chronic                                                                               0)     47 [0.95]
low-dose,
low-dose-
rate
gamma
radiation
exposed
irradiated
building
residents)
EE                  ND          Chang et al., 1997      31      9±4           7        (13    7 [0.90]
(chronic                                                Ǽ                              0)     9 [0.95]
low-dose,
low-dose-
rate
gamma
radiation
exposed
irradiated
building
residents)
EE                  ND          Fenech et al.,          11    9.1 ± 1.0   32.9 /4.2@   (13    1 [0.90]
(Nuclear                        1997                                        /2.1@      1)     1 [0.95]
radiation
exposed
residents
in the post-
chernobyl
period)
EE                  ND          Chang et al., 1997      77    19 ± 11Ǿ       11Ǿ       (13    22 [0.90]
(chronic                                                                               0)     26 [0.95]
low-dose,
low-dose-
rate
gamma
radiation
exposed
irradiated
building
residents)
EE                  ND          Chang et al., 1997      29    16 ± 9Ǿ         9Ǿ       (13    22 [0.90]
(chronic                                                Ǽ                              0)     26 [0.95]
low-dose,
low-dose-
rate
gamma
radiation
exposed
irradiated
building
residents)

OE                 NS+CS        Sari-Minodier et al.,   17    6.5 ± 2.9                (12      ND
(ionizing                 2002                                        6)
radiation
exposed
industrial
radiograph
ers)
OE               ND       Cardoso et al.,         8     23.8 ± 2.8    (12    ND
(chronicall               2001                                        9)
y radiation
exposed
hospital
workers)
OE               NS       Testa et al., 2002      30    4.7 ± 3.0     (11    ND
(various                                                              6)
chemicals
exposed
laborants)
OE               NS       Testa et al., 2002      23    3.0 ± 1.2     (11    ND
(various                                                              6)
chemicals
exposed
laborants)
OE             NS+FS+CS   Carere et al., 1998     12   16.96 ± 1.19   (12    ND
(gasoline                                                             5)
station
attendants)
OE               NS       Pitarque et al.,        20   24.15 ± 7.49   (11    ND
(solvent                  2002                                        5)
exposed
shoe
factory
workers)
OE               CS       Pitarque et al.,        16   22.19 ± 8.45   (11    ND
(solvent                  2002                                        5)
exposed
shoe
factory
workers)
OE              NS+CS     Crebelli et al., 2002   16    9.7 ± 4.9     (12    ND
(primary                                                              4)
aluminium
industry
workers)
OE (cobalt-     NS+CS       De Boeck et al.,      27    3.9 ± 1.7     (48)   ND
containing                      2000
dust
exposed
workers)
OE              NS+CS     Vaglenov et al.,        18   19.83 ± 1.97   (12    ND
(chromium                 1999                                        3)
exposed
electroplati
ng
workers)
OE               ND         Medeiros et al.,      27   3.58 ± 1.69    (12    ND
(trivalent                      2003                                   2)
chromium
exposed
tannery
workers)
OE             NS+CS Ш     Falck et al., 1999     33   6.8 ± 2.7 ╬    (13    ND
(pesticide                                                             2)
exposed
greenhous
e workers)
OE             NS+CS Ш     Falck et al., 1999     33   6.8 ± 2.6 ╬╬   (13    ND
(pesticide                                                                         2)
exposed
greenhous
e workers)
OE              NS+FS+PS    Pastor et al., 2002   22   8.45 ± 1.04                 (11     ND
(pesticide                                                                         8)
exposed
Spanish
greenhous
e workers
during
high
exposure
season)
OE              NS+FS+PS    Pastor et al., 2002   22   11.59 ± 1.07                (11     ND
(pesticide                                                                         8)
exposed
Spanish
greenhous
e workers
during
lower
exposure
season)
OE             NS+FS+CS     Pastor et al., 2002   41   8.07 ± 0.96                 (11     ND
(pesticide                                                                         7)
exposed
Hungarian
agricultura
l workers)
OE             NS+FS+CS     Pastor et al., 2002   12   12.83 ± 2.09                (11     ND
(pesticide                                                                         7)
exposed
Hungarian
agricultura
l workers)
OE             NS+FS+CS     Pastor et al., 2001   50   17.67 ± 1.14                (12     ND
(pesticide                                                                         0)
exposed
Polish
farmers)
OE              NS+FS+PS    Pastor et al., 2001   41   12.90 ± 1.20                (11     ND
(pesticide                                                                         9)
exposed
Greek
farmers)
OE              NS+FS+PS    Pastor et al., 2001   25   16.92 ± 2.77                (11     ND
(pesticide                                                                         9)
exposed
Greek
farmers)
OE (MOCA         NS+CS      Murray et al., 1999   18   5.93 ± 0.13                 (12     ND
exposed                                                                             1)
workers)

SE                NS        Burgaz et al., 1995   20    6.6 ± 0.5                  (12     ND
                                                                                    8)
DE (folic        NS+CS      Fenech et al., 1998   33   6.92 ± 0.58                 (12     ND
acid and                                                                            7)
vitamin
B12
supplemen
tation)

Weighted      NS+CS+FS+PS                              11.53 ± 5.24   9.58 (223)         7 [0.90]
(n1)                                                      (787)                          8 [0.95]
Weighted       NS+CS+FS+PS                                  11.53 ± 5.24   8.37 (212)■           9 [0.90]
(n1)                                                           (787)                             11 [0.95]
Weighted       NS+CS+FS+PS                                  11.53 ± 5.24   11.53                 5 [0.90]
(n1)                                                           (787)       (wmav)                6 [0.95]
Weighted       NS+CS+FS+PS                                  10.93 ± 3.91   6.29 (106)            9 [0.90]
(n1)           Ħ                                               (604)       (■)                   11 [0.95]

Weighted       NS+CS+FS+PS                                  10.93 ± 3.91   10.93                 3 [0.90]
(n1)           Ħ                                               (604)       (wmav)                4 [0.95]

Weighted       NS                                            8.87 ± 3.91   9.58 (223)            4 [0.90]
(n1)                                                             (93)                            5 [0.95]
Weighted       NS                                            8.87 ± 3.91   8.37                  5 [0.90]
(n1)                                                             (93)      (212) ■               6 [0.95]
Weighted       NS                                            8.87 ± 3.91   8.87 (wmav)           5 [0.90]
(n1)                                                             (93)                            6 [0.95]
Weighted       NS Ħ                                          8.87 ± 3.91   6.29 (106)            9 [0.90]
(n1)                                                             (93)      (■)                   11 [0.95]
Weighted       NS Ħ                                          8.87 ± 3.91   8.87 (wmav)           5 [0.90]
(n1)                                                             (93)                            6 [0.95]
Weighted       CS                                           22.19 ± 8.45   9.58 (223)            17 [0.90]
(n1)                                                            (16)¶                            21 [0.95]
Weighted       CS                                           22.19 ± 8.45   8.37 (212) ■          22 [0.90]
(n1)                                                            (16)¶                            27 [0.95]
Weighted       CS                                           22.19 ± 8.45   22.19                 4 [0.90]
(n1)                                                            (16)¶      (wmav)                4 [0.95]
EE: environmental exposure. OE: occupational exposure. SE: smoking exposure (non-smokers vs.
                                   a
smokers). DE: dietary exposure. Number of MNed cells per 1000 binucleated cells (MNCs). n1: n
(amount of individuals) in observed group. n2: the calculated amount of exposed individuals in the
research population for that specific power. NS: non-smoker. CS: current smoker. FS: former smoker.
PS: passive smoker. @: not significant. ND: not defined; lack of relevant and/or significant values. v:
difference in biomarker between exposed and control group. Ǿ: using the alternative micronucleus
assay: a CBMN test involving incubation with cytarabine (CBMNA). Ǽ: comparison of matched group
A (close relatives, siblings, brothers-in-law, or sisters-in-law matched for age and sex) controls with
exposed. : children aged 12-15 years (131). : residents of Minsk (controls) compared with residents
of Yazvenki. : residents of Minsk (controls) compared with residents of Vetka.         : residents of
Minsk (controls) compared with residents of Kalinkovichi. ╬: evaluated by the anti-bromodeoxyuridine
(anti-BrdU) assay, with 0.5 µg/ml BrdU. ╬╬: evaluated by the anti-bromodeoxyuridine (anti-BrdU)
assay, with 1 µg/ml BrdU. MOCA: 4,4’-methylenebis-(2-chloroaniline). Ш: in multiple regression
analysis controlled for age, sex, smoking and genotypes (132). wmav: weighted mean as v; thus mean
exposed is set at twice the mean of the controls. ■: for this weighted v the v of post-Tsjernobyl
radiation exposure (131) has been excluded from the weighted v calculation, due to the extremity of
this particular event. : since no smoking class specific EE based v was available the here applied v is
based on studies without a defined smoking status. Ħ: (parts of) studies (130, 132) using assays other
than the standard assay and studies with children (131) instead of adults/ adolescents are excluded.
(■): in this case ■ does also apply due to Ħ. ¶: only based on one study (115). Biological medium in all
the detections were PBL. Unless mentioned otherwise: data of cytochalasin B micronuclei assays
(CBMN) have been described.




MN per 1000 binucleated cells (MNi) in healthy individuals

Studies and their exposure types
                                                                b
Exposure       Smoking status                         n1     MNi ± SD           v               n2 [power]
type
EE               NS+FS+CS       Barale et al., 1998   439   4.13 ± 3.20      0.94@▼       (15       ND
(residential                                                                               1)
exposure)
EE               NS+FS+CS       Barale et al., 1998   341   4.10 ± 3.55     0.91@▼▼       (15       ND
(residential                                                                               1)
exposure)
EE              NS+CS     Martelli et al., 2000   103   4.93 ± 4.04      1.47      (13   159 [0.90]
(residential                                                                        5)   197 [0.95]
exposure
to an
urban
industrializ
ed area
(coke
factory and
metallurgic
al plantŦ)
EE              NS+CS     Martelli et al., 2000   43    4.08 ± 3.80     1.13@      (13      ND
(residential                                                                        5)
exposure
to an
urban
industrializ
ed area
(coke
factory and
metallurgic
al plantŦ)
EE              NS+CS     Martelli et al., 2000   60    5.55 ± 4.13      1.67      (13   129 [0.90]
(residential                                                                        5)   160 [0.95]
exposure
to an
urban
industrializ
ed area
(coke
factory and
metallurgic
al plantŦ)
EE               NS       Martelli et al., 2000   70    5.33 ± 4.17      1.66      (13   133 [0.90]
(residential                                                                        5)   165 [0.95]
exposure
to an
urban
industrializ
ed area
(coke
factory and
metallurgic
al plantŦ)
EE               CS       Martelli et al., 2000   33    4.10 ± 3.68     1.49@      (13      ND
(residential                                                                        5)
exposure
to an
urban
industrializ
ed area
(coke
factory and
metallurgic
al plantŦ)
EE             NS+FS+CS   Jen et al., 2002        10    2.73 ± 0.59      2.06      (15    2 [0.90]
(residents                                                                          2)    3 [0.95]
of
radionuclid
e-
contaminat
ed regions)
EE               ND       Fenech et al.,          11     9.8 ± 1.0    36.6 /4.5@   (13    1 [0.90]
(nuclear                  1997                                          /2.3 @     1)     1 [0.95]
radiation
exposed
residents
in the post-
chernobyl
period)
EE (air         NS     Bolognesi et al.,      23      3.89 ± 1.30   0.13@   (14     ND
pollution              1997                                                  1)
exposed
traffic
police
workers)
EE (air         CS     Bolognesi et al.,      11      4.31 ± 2.18   0.61@   (14     ND
pollution              1997                                                  1)
exposed
traffic
police
workers)
EE (air        NS+CS   Zhao et al., 1998      34      3.22 ± 1.31   2.50    (14   6 [0.90]
pollution                                                                    0)   8 [0.95]
exposed
traffic
police
workers)
EE (air         NS     Zhao et al., 1998      15      2.26 ± 1.14   1.24@   (14     ND
pollution                                                                    0)
exposed
traffic
police
workers)
EE (air         CS     Zhao et al., 1998      19      3.89 ± 2.40   3.06    (14   13 [0.90]
pollution                                                                    0)   16 [0.95]
exposed
traffic
police
workers)
EE (air         NS     Zwingmann et al.,      13       6.0 ± 1.1    1.4@    (13     ND
pollution              1998                                                  9)
exposed
traffic
police
workers)
EE (air        NS+CS   Zwingmann et al.,      23       7.1 ± 1.6    0.4@    (13     ND
pollution              1998                                                  9)
exposed
traffic
police
workers)

OE             NS+CS   Somorovská et al.,      22     1.05 ± 0.20           (15     ND
(multiple                   1999             (facto                          3)
compound                                       ry
s exposed                                    contr
rubber                                        ols)
factory
workers)
OE             NS+CS   Somorovská et al.,      17     1.23 ± 0.26           (15     ND
(multiple                   1999             (labo                           3)
compound                                     rator
s exposed                                       y
rubber                                       contr
factory                                       ols)
workers)
OE (MOCA       NS+CS   Murray et al., 1999    18      9.24 ± 0.29           (12     ND
exposed                                                                      1)
workers)
OE              NS     Anwar et al., 1995     18      6.00 ± 2.83           (13     ND
(styrene                                                        8)
exposed
workers)
OE (low        ND     Laffon et al., 2002   30   13.91 ± 0.81   (14    ND
levels of                                                       4)
styrene
exposed
workers)
OE             NS     Chang et al., 1996    18    4.3 ± 1.0     (13    ND
(excessive                                                      4)
and
chronic
nitrous
oxide
exposed
nurses)
OE             NS     Testa et al., 2002    30    5.0 ± 3.5     (11    ND
(various                                                        6)
chemicals
exposed
laborants)
OE             NS     Testa et al., 2002    23    3.1 ± 1.2     (11    ND
(various                                                        6)
chemicals
exposed
laborants)
OE            NS+CS   Maluf et al., 2000    10   5.75 ± 2.61    (11)   ND
(antineopla
stic drugs
exposed
hospital
workers)
OE            NS+CS   Maluf et al., 2000    12   5.54 ± 2.98    (11)   ND
(antineopla
stic drugs
exposed
hospital
workers)
OE            NS+CS   Pilger et al., 2000   39   11.65 ± 3.75   (15    ND
(cytostatic                                                     4)
drugs
exposed
pharmacy
personnel)
OE             ND     Cardoso et al.,       8      26 ± 3       (12    ND
(chronicall           2001                                      9)
y radiation
exposed
hospital
workers)
OE             ND     Thierens et al.,      60   18.63 ± 7.53   (15    ND
(chronicall           2000                                      5)
y low level
ionizing
radiation
exposed
hospital
workers)
OE (low        NS     Maffei et al., 2002   20   5.15 ± 3.67    (16    ND
level                                                           4)
ionizing
radiation
exposed
hospital
workers)
OE (low        CS     Maffei et al., 2002   17   6.00 ± 1.94    (16    ND
level                                                                4)
ionizing
radiation
exposed
hospital
workers)
OE              NS     Garaj-Vrhovac and       20    5.10 ± 1.38     (13    ND
(Doppler               Kopjar, 2000                                  7)
ultrasound
exposed
hospital
workers)
OE              ND     Hessel et al., 2001     60         9          (15    ND
(Doppler                                                             6)
ultrasound
exposed
hospital
workers)
OE              ND     Hoerauf et al.,         10    4.00 ± 0.36     (15    ND
(isoflurane            1999                                          7)
and
nitrous
oxide
exposed
operating
room
personnel)
OE (radon-     NS+CS   Bilban, 1998            61   12.800 ± 4.232   (14    ND
and heavy                                                            5)
metal-
exposed
lead-zinc
mine
miners)
OE             NS+CS   Crebelli et al., 2002   16     11.1 ± 6.7     (12    ND
(primary                                                             4)
aluminium
industry
workers)
OE             NS+CS   Vaglenov et al.,        18    21.05 ± 2.07    (12    ND
(chromium              1999                                          3)
exposed
electroplati
ng
workers)
OE             NS+CS   Cavallo et al., 2002    23    4.00 ± 2.96     (14    ND
(antimony                                                            6)
trioxide
exposed
workers)
OE (coal        NS     Stierum et al., 1993    6      4.4 ± 2.6      (28)   ND
fly ash
exposed
workers)
OE (coal        CS     Stierum et al., 1993    11     7.1 ± 4.4      (28)   ND
fly ash
exposed
workers)
OE (diesel      ND     Schoket et al.,         55     19.9 ± 6.5     (14    ND
exposed                1999                                          7)
mechanics
)
OE              NS     Carere et al., 1995     24    5.50 ± 0.60◘    (14    ND
(gasoline                                                            3)
station
attendants)
OE               NS       Carere et al., 1995   24    7.70 ± 0.90◘◘   (14   ND
(gasoline                                                             3)
station
attendants)
OE (low-        NS+CS     Lemasters et al.,      8     11.8 ± 3.4     (14   ND
level                     1997                                        8)
solvent
and fuel
exposed
aircraft
sheet
metal
workers)
OE (oil         NS+CS     Lillienberg et al.,   31         3.6        (14   ND
tank                      1992                                        9)
cleaners)
OE             NS+FS+CS   Lucero et al., 2000   50    7.900 ± 0.125   (15   ND
(pesticide                                                            0)
exposed
greenhous
e workers)
OE             NS+FS+PS   Pastor et al., 2002   22     8.95 ± 1.15    (11   ND
(pesticide                                                            8)
exposed
Spanish
greenhous
e workers
during
high
exposure
season)
OE             NS+FS+PS   Pastor et al., 2002   22    11.81 ± 1.07    (11   ND
(pesticide                                                            8)
exposed
Spanish
greenhous
e workers
during
lower
exposure
season)
OE              NS+CS     Bolognesi et al.,     75     6.67 ± 0.36    (15   ND
(pesticide                1993                                        8,
exposed                                                               200
floriculturi                                                          ,
sts)                                                                  201
                                                                      )
OE              NS+CS     Bolognesi et al.,     92     2.64 ± 1.28    (13   ND
(pesticide                2002                                        6)
exposed
floriculturi
sts)
OE              NS+CS     Bolognesi et al.,     15     3.92 ± 1.52    (13   ND
(pesticide                2002                                        6)
exposed
floriculturi
sts)
OE              NS+CS     Bolognesi et al.,     107    3.04 ± 2.14    (13   ND
(pesticide                2002                                        6)
exposed
floriculturi
sts)
OE              NS+CS     Scarpato et al.,      22      4.1 ± 2.1     (15   ND
(pesticide                1996                                        9)
exposed
floriculturi
sts during
high
exposure
period)
OE              NS+CS     Scarpato et al.,      22     7.4 ± 3.0     (15    ND
(pesticide                1996                                       9)
exposed
floriculturi
sts during
lower
exposure
period)
OE             NS+FS+CS   Pastor et al., 2002   41    9.07 ± 1.01    (11    ND
(pesticide                                                           7)
exposed
Hungarian
agricultura
l workers)
OE             NS+FS+CS   Pastor et al., 2002   12    14.50 ± 2.19   (11    ND
(pesticide                                                           7)
exposed
Hungarian
agricultura
l workers)
OE             NS+FS+CS   Pastor et al., 2001   50    20.10 ± 1.34   (12    ND
(pesticide                                                           0)
exposed
Polish
farmers)
OE             NS+FS+PS   Pastor et al., 2001   41    14.68 ± 1.44   (11    ND
(pesticide                                                           9)
exposed
Greek
farmers)
OE             NS+FS+PS   Pastor et al., 2001   25    19.16 ± 3.16   (11    ND
(pesticide                                                           9)
exposed
Greek
farmers)
OE              NS+CS     Garaj-Vrhovac and     20        154        (16    ND
(pesticide                Zeljezic, 2001                             0)
exposed
Croatian
workers)
OE              NS+CS     Garaj-Vrhovac and     20    7.70 ± 3.20    (16    ND
(Croatian                 Zeljezic, 2002                             1)
pesticide
production
workers)
OE               NS       Chang et al., 1996    18    12.4 ± 1.9Ǿ    (13    ND
(excessive                                                           4)
and
chronic
nitrous
oxide
exposed
nurses)

SE               NS       Xue et al., 1992      183    1.6 ± 3.5     (14    ND
                                                                     2)
SE               NS       Barale et al., 1998   275   4.82 ± 3.50     (16   ND
                                                  †                    2)
SE               NS       Barale et al., 1998   276   3.66 ± 3.21     (16   ND
                                                 ††                    2)
SE               NS       Bolognesi et al.,      22   3.36 ± 1.33    (13    ND
                          2002                                       6)
SE                  NS         Cheng et al., 1996    18      10.2 ± 6.3                    (16       ND
                                                                                           3)
DE                  CS         Xue et al., 1992      41      10.5 ± 9.1                    (14       ND
(alcohol■)                                                                                 2)
DE              NS+FS+CS       Barale et al., 1998   209    4.20 ± 3.36                     (16      ND
(alcohol■)                                             †                                     2)
DE              NS+FS+CS       Barale et al., 1998   229    3.49 ± 3.23                     (16      ND
(alcohol■)                                            ††                                     2)
DE              NS+FS+CS       Cheng et al., 1996     50     9.3 ± 6.5                     (16       ND
(alcohol♦)                                                                                 3)
DE                  CS         Xue et al., 1992      35     25.4 ± 21.1                    (14       ND
(tea■■■■)                                                                                  2)
DE              NS+FS+CS       Barale et al., 1998   84     4.59 ± 3.49                     (16      ND
(coffee■■)                                            †                                      2)
DE              NS+FS+CS       Barale et al., 1998   81     3.53 ± 3.13                     (16      ND
(coffee■■)                                           ††                                      2)
DE (broiled        NS+CS       Bolognesi et al.,     22     4.27 ± 1.80                     (14      ND
beef■■■)                       1997                                                          1)
DE              NS+FS+CS       Cheng et al., 1996    32      9.5 ± 6.7                     (16       ND
(vitamin                                                                                   3)
A♦♦)
DE              NS+FS+CS       Cheng et al., 1996    27      8.1 ± 4.1                     (16       ND
(folate♦♦♦)                                                                                3)
DE                  NS         Fenech and            18     7.16 ± 1.19                    (13       ND
(vegetarian                    Rinaldi, 1995                                               3)
ю)
DE                  NS         Fenech and            24    20.18 ± 1.79                    (13       ND
(vegetarian                    Rinaldi, 1995                                               3)
ю)
DE                  NS         Fenech and            24    22.57 ± 1.79                    (13       ND
(vegetarian                    Rinaldi, 1995                                               3)
ю)
DE                  NS         Fenech and            20     8.36 ± 1.31                    (13       ND
(vegetarian                    Rinaldi, 1995                                               3)
ю)
DE                  NS         Fenech and            23    19.46 ± 2.15                    (13       ND
(vegetarian                    Rinaldi, 1995                                               3)
ю)
DE                  NS         Fenech and            30    24.48 ± 1.91                    (13       ND
(vegetarian                    Rinaldi, 1995                                               3)
ю)

Weighted      NS+CS+FS+PS                                   6.38 ± 3.90    3.04 (307)              35 [0.90]
(n1)                                                          (4315)                               43 [0.95]
Weighted      NS+CS+FS+PS                                   6.38 ± 3.90    1.80 (296)♣             99 [0.90]
(n1)                                                          (4315)                              123 [0.95]
Weighted      NS+CS+FS+PS                                   6.38 ± 3.90    6.38 (wmav)             8 [0.90]
(n1)                                                          (4315)                               10 [0.95]
Weighted      NS+CS+FS+PS                                   6.34 ± 3.93    1.80 (296)             101 [0.90]
(n1)          Ħ                                               (4238)       (♣)                    124 [0.95]
Weighted      NS+CS+FS+PS                                   6.34 ± 3.93    6.34 (wmav)             9 [0.90]
(n1)          Ħ                                               (4238)                               10 [0.95]
Weighted      NS                                            5.78 ± 3.12    1.66 (70) (♣)           75 [0.90]
(n1)                                                          (1235)                               92 [0.95]
Weighted      NS                                            5.78 ± 3.12    5.78 (wmav)             7 [0.90]
(n1)                                                          (1235)                               8 [0.95]
Weighted      CS                                           10.52 ± 10.98   3.06 (19) (♣)          242 [0.90]
(n1)                                                           (167)                              300 [0.95]
Weighted      CS                                           10.52 ± 10.98   10.52                   23 [0.90]
(n1)                                                           (167)       (wmav)                  29 [0.95]
EE: environmental exposure. OE: occupational exposure. SE: smoking exposure (non-smokers vs.
smokers). DE: dietary exposure. n1. n1: n (amount of individuals) in observed group. n2: the calculated
amount of exposed individuals in the research population for that specific power. NS: non-smoker. CS:
current smoker. FS: former smoker. PS: passive smoker. M: male. F: female. @: not significant. ND:
not defined; lack of relevant and/or significant values. v: difference in biomarker between exposed and
control group. bNumber of MN per 1000 BN cells (MNi). ▼: the small city Cascina compared with the
city Pisa (higher degree of air pollution) (151). ▼▼: the small city Navacchio compared with the city
Pisa (higher degree of air pollution). : children aged 12-15 years (131). : residents of Minsk
(controls) compared with residents of Yazvenki. : residents of Minsk (controls) compared with
residents of Vetka.      : residents of Minsk (controls) compared with residents of Kalinkovichi. Ŧ:
residential suburb: Genova-Nervi (controls) compared to urban industrialized area near a metallurgical
plant and a coke factory: Genova-Cornigliano (exposed). ◘: from phytohemagglutinin (PHA)-stimulated
cultures. ◘◘: from pokeweed (PKW)-stimulated cultures. Ǿ: using the alternative micronucleus assay:
a CBMN test involving incubation with cytarabine (CBMNA). MEG: a mixture of 33% methanol, 60%
ethanol and 7% gasoline (202). MOCA: 4,4’-methylenebis-(2-chloroaniline). †: population of the small
cities: Cascina and Navacchio (162). ††: general population (Pisa); in Pisa there is a higher degree of
air pollution than in the small cities (162). ■: non-alcohol drinkers compared with alcohol drinkers. ♦:
less than 30 grams alcohol a day drinkers (controls) compared to more than 30 grams of alcohol a day
drinkers (exposed). ■■: non-coffee drinkers compared with coffee drinkers. ■■■: less than once a
month compared with more than two times a month. ■■■■non-tea drinkers compared with tea
drinkers. ♦♦: less than 15,000 IU vitamin A per day consumers (controls) compared with more than
15,000 IU vitamin A per day consumers (exposed). ♦♦♦: less than 400 grams of folate per day
consumers (controls) compared with more than 400 grams of folate per day consumers (exposed). Ю:
non vegetarians (controls) compared with vegetarians (‘exposed’). Biological medium in all the
detections were PBL. Unless mentioned otherwise: data of cytochalasin B micronuclei assays (CBMN)
have been described. wmav: weighted mean as v; thus mean exposed is set at twice the mean of the
controls. ♣: for this weighted v the v of post-Tsjernobyl radiation exposure (131) has been excluded
from the weighted v calculation, due to the extremity of this particular event. Ħ: (parts of) studies using
assays other than the standard assay (134, 143) and studies with children (131) instead of adults/
adolescents are excluded (those that apply to this specific class). (♣): in this case ♣ does also apply
due to Ħ or other class specific selection criteria.
Appendix 13:

PAH-DNA adduct levels (adducts/ 108 nucleotides) in healthy individuals,
as determined by ELISA and 32P-postlabelling methods

Studies and their exposure types
Exposure       Smoking                Additional      n1    PAH-DNA adducts        v                n2
                                                               -8
type           status                information            (10 ) ± SD (range)                    [power]
EE             NS+CS     Perera et   ELISA,          21 M   11.0 ± 22.6■■        19.4    (176)   29 [0.90]
(inhabitant              al., 1992   WBC, winter                                                 36 [0.95]
s of the
highly
industrializ
ed Silesian
region of
Poland■)
EE             NS+CS     Perera et   ELISA,          22 M   3.0 ± 3.3■■          1.2     (176)   156 [0.90]
(inhabitant              al., 1992   WBC,                                                        197 [0.95]
s of the                             summer
highly
industrializ
ed Silesian
region of
Poland■)
                                     32
EE             NS+CS     Perera et    P-             21 M   3.1 ± 1.7■■          2.4@    (176)      ND
(inhabitant              al., 1992   postlabeling,
s of the                             WBC, winter
highly
industrializ
ed Silesian
region of
Poland■)
                                     32
EE             NS+CS     Perera et    P-             22 M   2.9 ± 2.8■■          0.7@    (176)      ND
(inhabitant              al., 1992   postlabeling,
s of the                             WBC,
highly                               summer
industrializ
ed Silesian
region of
Poland■)
                                     32
EE             ND        Hemminki       P-           15 M   8.2■■                12.9    (177)      ND
(inhabitant              et al.,     postlabelling
s of the                 1990        , WBC,
highly                               laboratory 1
industrializ
ed Silesian
region of
Poland■)
                                     32
EE             ND        Hemminki      P-            15 M   4.4■■                5.8@    (177)      ND
(inhabitant              et al.,     postlabelling
s of the                 1990        , WBC,
highly                               laboratory 2
industrializ
ed Silesian
region of
Poland■)
                                     32
EE             NS        Möller et     P-            3M     2.42 ± 3.45■■        0.46    (166)    1 [0.90]
(inhabitant              al., 1996   postlabelling                               @▲/              1 [0.95]
s of the                             -HPLC, PBL,                                 18.98
highly                               summer                                      ▲▲
industrializ
ed Silesian
region of
Poland■)
                                      32
EE             NS+CS   Nielsen et        P-           60     2.2 (0.1-266)   18.5   (168)   ND
(ambient               al., 1996      postlabelling   M+F
air                                   (n-Bu), PBL
exposed
bus
drivers)

OE (PAH        NS+CS   Perera et      ELISA, PBL      10 M   2.2 ± 0.9              (178)   ND
exposed                al., 1988
iron
foundry
workers)
 OE            NS      Liou et al.,   ELISA, WBC      8M     28.8 ± 13.8            (173)   ND
(pyrolysis             1989
products
exposed
firefighters
)
 OE            CS      Liou et al.,   ELISA, WBC      7M     38.1 ± 13.2            (173)   ND
(pyrolysis             1989
products
exposed
firefighters
)
 OE            NS+CS   Liou et al.,   ELISA, WBC      13 M   34.5 ± 13.8            (173)   ND
(pyrolysis             1989
products
exposed
firefighters
)
OE (PAH        CS      van            ELISA, WBC      28 M   3.00 ± 2.52            (174)   ND
exposed                Schooten
coke oven              et al.,
workers)               1990
OE (PAH        NS      van            ELISA, WBC      16 M   1.71 ± 0.72            (174)   ND
exposed                Schooten
coke oven              et al.,
workers)               1990
OE (PAH        NS+CS   van            ELISA, WBC      44 M   2.49 ± 2.07            (174)   ND
exposed                Schooten
coke oven              et al.,
workers)               1990
                                      32
OE (PAH-       CS      Rojas et         P-            23 M   1.1 ± 1.6              (165)   ND
exposed                al., 1995      postlabelling
coke oven                             , PBL
workers)
                                      32
OE (PAH-       NS      Rojas et         P-            16 M   3.7 ± 13.1             (165)   ND
exposed                al., 1995      postlabelling
coke oven                             , PBL
workers)
                                      32
OE             ND      Hemminki         P-            15 M   8.2■■                  (177)   ND
(Silesian              et al.,        postlabelling
region of              1990           , WBC,
Poland                                laboratory 1
coke oven
workers■)
                                      32
OE             ND      Hemminki         P-            15 M   4.4■■                  (177)   ND
(Silesian              et al.,        postlabelling
region of              1990           , WBC,
Poland                                laboratory 2
coke oven
workers■)
                                      32
OE (PAH      NS         van Delft        P-           19 M   10.9 ± 7.2                   (172)     ND
exposed                 et al.,       postlabelling
coke oven               2001          -HPLC,
workers)                              WBC
                                      32
OE (PAH      CS         van Delft        P-           18 M   8.2 ± 2.5                    (172)     ND
exposed                 et al.,       postlabelling
coke oven               2001          -HPLC,
workers)                              WBC
                                      32
OE (PAH      NS+CS      van Delft        P-           37 M   9.7 ± 5.8                    (172)     ND
exposed                 et al.,       postlabelling
coke oven               2001          -HPLC,
workers)                              WBC
                                      32
OE (diesel   NS         Nielsen et       P-           12 M   7.8 (3.9-42.0)               (203)     ND
exhaust                 al., 1996     postlabelling
exposed                               (n-Bu), PBL
workers)
                                      32
OE (PAH      NS         van Delft       P-            19 M   2.7 ± 1.2                    (172)     ND
exposed                 et al.,       postlabelling
coke oven               2001          (n-Bu), WBC
workers)
                                      32
OE (PAH      CS         van Delft       P-            18 M   3.6 ± 1.1                    (172)     ND
exposed                 et al.,       postlabelling
coke oven               2001          (n-Bu), WBC
workers)
                                      32
OE (PAH      NS+CS      van Delft       P-            37 M   3.2 ± 1.2                    (172)     ND
exposed                 et al.,       postlabelling
coke oven               2001          (n-Bu), WBC
workers)
                                      32
OE (diesel   NS         Nielsen et      P-            12 M   2.4 (0.9-8.7)                (203)     ND
exhaust                 al., 1996     postlabelling
exposed                               (P1), PBL
workers)
                                      32
OE (PAH      CS         Kuljukka        P-            3M     1.1 (ND-1.9)                 (204)     ND
exposed                 et al.,       postlabelling
coke oven               1998          (P1), WBC
workers)
                                      32
OE (PAH      NS         Kuljukka        P-            6M     0.9 (ND-1.9)                 (204)     ND
exposed                 et al.,       postlabelling
coke oven               1998          (P1), WBC
workers)

SE           FS+NS      Perera et     ELISA, PBL      48     12.33 ± 12.67                (170)     ND
                        a., 1989                      M+F
SE           NS         Tang et       ELISA, WBC      39     3.3 ± 4.5                    (169)     ND
                        al., 1995                     M+F
                                      32
SE           NS         Phillips et     P-            20     2.12 ± 1.44                  (167)     ND
                        al., 1990     postlabelling   M+F
                                      (P1), PBL
SE           NS         Santella et   ELISA, PBL      27 M   1.35 ± 0.78                  (175)     ND
                        al., 1992
                                      32
SE           NS         van Delft        P-           19 M   10.9 ± 7.2                   (172)     ND
                        et al.,       postlabelling
                        2001          -HPLC,
                                      WBC
                                      32
SE           NS         van Delft        P-           19 M   2.7 ± 1.2                    (172)     ND
                        et al.,       postlabelling
                        2001          (n-Bu), WBC
                                      32
SE           NS         Tang et          P-           30 M   6.2 ± 1.2                    (171)     ND
                        al., 2001     postlabelling
                                      (P1), WBC

Weighted     NS+FS+C                                          6.45 ± 7.22 (604)   14.83           5 [0.90]
(n1)         S,                                                                   (121)           7 [0.95]
             ELISA+32
             P-
           postlabell
           ing
           assays,
           PBL+WB
           C
Weighted   NS+FS+C      6.45 ± 7.22 (604)   6.45    27 [0.90]
(n1)       S,                               (wma    33 [0.95]
                  32
           ELISA+                           v)
           P-
           postlabell
           ing
           assays,
           PBL+WB
           C
Weighted   NS+FS+C      5.34 ± 8.56 (147)   18.52   5 [0.90]
(n1)       S,                               (63)    6 [0.95]
                  32
           ELISA+
           P-
           postlabell
           ing
           assays,
           PBL
Weighted   NS+FS+C      5.34 ± 8.56 (147)   5.34    55 [0.90]
(n1)       S,                               (wma    67 [0.95]
                  32
           ELISA+                           v)
           P-
           postlabell
           ing
           assays,
           PBL
Weighted   NS+FS+C      6.81 ± 6.73 (457)   10.82   9 [0.90]
(n1)       S,                               (58)    11 [0.95]
                  32
           ELISA+
           P-
           postlabell
           ing
           assays,
           WBC
Weighted   NS+FS+C      6.81 ± 6.73 (457)   6.81    21 [0.90]
(n1)       S,                               (wma    26 [0.95]
           ELISA+32                         v)
           P-
           postlabell
           ing
           assays,
           WBC
Weighted   NS+FS+C      7.92 ± 9.37 (283)   10.09   19 [0.90]
(n1)       S, ELISA,                        (43)    23 [0.95]
           PBL+WB
           C
Weighted   NS+FS+C      7.92 ± 9.37 (283)   7.92    30 [0.90]
(n1)       S, ELISA,                        (wma    37 [0.95]
           PBL+WB                           v)
           C
Weighted   NS+FS+C      7.65 ± 9.61 (85)    10.09   20 [0.90]
(n1)       S, ELISA,                        (43)Ґ   24 [0.95]
           PBL
Weighted   NS+FS+C      7.65 ± 9.61 (85)    7.65    34 [0.90]
(n1)       S, ELISA,                        (wma    42 [0.95]
           PBL                              v)
Weighted   NS+FS+C      8.04 ± 9.27 (198)   10.09   18 [0.90]
(n1)       S, ELISA,                        (43)    22 [0.95]
           WBC
Weighted   NS+FS+C      8.04 ± 9.27 (198)   8.04    28 [0.90]
(n1)       S, ELISA,                        (wma    35 [0.95]
           WBC                              v)
Weighted   NS+CS,       5.16 ± 4.53 (321)   17.44    2 [0.90]
           32
(n1)          P-                            (78)     2 [0.95]
           postlabell
           ing
           assays,
           PBL+WB
           C
Weighted   NS+CS,       5.16 ± 4.53 (321)   5.16    17 [0.90]
           32
(n1)          P-                            (wma    21 [0.95]
           postlabell                       v)
           ing
           assays,
           PBL+WB
           C
Weighted   NS+CS,       2.16 ± 6.81 (62)    18.52    3 [0.90]
           32
(n1)          P-                            (63)     4 [0.95]
           postlabell
           ing
           assays,
           PBL
Weighted   NS+CS,       2.16 ± 6.81 (62)    2.16    180 [0.90]
           32
(n1)          P-                            (wma    222 [0.95]
           postlabell                       v)
           ing
           assays,
           PBL
Weighted   NS+CS,       5.87 ± 3.80 (259)   12.9     2 [0.90]
           32
(n1)          P-                            (15)     3 [0.95]
           postlabell
           ing
           assays,
           WBC
Weighted   NS+CS,       5.87 ± 3.80 (259)   5.87    9 [0.90]
           32
(n1)          P-                            (wma    11 [0.95]
           postlabell                       v)
           ing
           assays,
           WBC
Weighted   NS,          5.25 ± 5.49 (235)   18.98    2 [0.90]
                  32
(n1)       ELISA+                           (3)      3 [0.95]
           P-
           postlabell
           ing
           assays,
           PBL+WB
           C
Weighted   NS,          5.25 ± 5.49 (235)   5.25    23 [0.90]
(n1)       ELISA+32                         (wma    29 [0.95]
           P-                               v)
           postlabell
           ing
           assays,
           PBL+WB
           C
Weighted   NS,          2.20 ± 6.54 (66)    18.98    3 [0.90]
(n1)       ELISA+32                         (3)      4 [0.95]
           P-
           postlabell
           ing
           assays,
           PBL
Weighted   NS,          2.20 ± 6.54 (66)    2.20    186 [0.90]
                  32
(n1)       ELISA+                           (wma    230 [0.95]
           P-                               v)
           postlabell
           ing
           assays,
           PBL
Weighted   NS,          6.44 ± 5.03 (169)   10.82    5 [0.90]
(n1)       ELISA+32                         (58)†    6 [0.95]
           P-
           postlabell
           ing
           assays,
           WBC
Weighted   NS,          6.44 ± 5.03 (169)   6.44    13 [0.90]
                   32
(n1)       ELISA+                           (wma    16 [0.95]
           P-                               v)
           postlabell
           ing
           assays,
           WBC
Weighted   NS,          4.70 ± 4.97 (90)    10.09    6 [0.90]
(n1)       ELISA,                           (43)†    7 [0.95]
           PBL+WB
           C
Weighted   NS,          4.70 ± 4.97 (90)    4.70    24 [0.90]
(n1)       ELISA,                           (wma    30 [0.95]
           PBL+WB                           v)
           C
Weighted   NS,          1.35 ± 0.78 (27)¶   10.09    1 [0.90]
(n1)       ELISA,                           (43)‡    1 [0.95]
           PBL
Weighted   NS,          1.35 ± 0.78 (27)¶   1.35     8 [0.90]
(n1)       ELISA,                           (wma     9 [0.95]
           PBL                              v)
Weighted   NS,          6.13 ± 5.93 (63)    10.09    8 [0.90]
(n1)       ELISA,                           (43)†    9 [0.95]
           WBC
Weighted   NS,          6.13 ± 5.93 (63)    6.13    20 [0.90]
(n1)       ELISA,                           (wma    25 [0.95]
           WBC                              v)
Weighted   NS, 32P-     5.60 ± 5.79 (145)   18.98    2 [0.90]
(n1)       postlabell                       (3)      3 [0.95]
           ing
           assays,
           PBL+WB
           C
               32
Weighted   NS, P-       5.60 ± 5.79 (145)   5.60    23 [0.90]
(n1)       postlabell                       (wma    28 [0.95]
           ing                              v)
           assays,
           PBL+WB
           C
               32
Weighted   NS, P-       2.79 ± 8.56 (39)    18.98    5 [0.90]
(n1)       postlabell                       (3)      6 [0.95]
           ing
           assays,
           PBL
Weighted   NS, 32P-     2.79 ± 8.56 (39)    2.79    198 [0.90]
(n1)       postlabell                       (wma    245 [0.95]
           ing                              v)
           assays,
           PBL
               32
Weighted   NS, P-       6.63 ± 4.41 (106)   18.98    1 [0.90]
(n1)       postlabell                       (3)Γ     1 [0.95]
           ing
           assays,
           WBC
               32
Weighted   NS, P-       6.63 ± 4.41 (106)   6.63    10 [0.90]
(n1)       postlabell                       (wma    12 [0.95]
           ing                              v)
           assays,
           WBC
Weighted   CS,          6.26 ± 3.97 (94)    14.72   2 [0.90]
(n1)       ELISA+32                         (118)   2 [0.95]
           P-                               †
           postlabell
           ing
           assays,
           PBL+WB
           C
Weighted   CS,          6.26 ± 3.97 (94)    6.26    9 [0.90]
(n1)       ELISA+32                         (wma    11 [0.95]
           P-                               v)
           postlabell
           ing
           assays,
           PBL+WB
           C
Weighted   CS,           1.1 ± 1.6 (23)¶    18.5    1 [0.90]
                   32
(n1)       ELISA+                           (60)†   1 [0.95]
           P-
           postlabell
           ing
           assays,
           PBL
Weighted   CS,           1.1 ± 1.6 (23)¶    1.1     45 [0.90]
                   32
(n1)       ELISA+                           (wma    56 [0.95]
           P-                               v)
           postlabell
           ing
           assays,
           PBL
Weighted   CS,          7.93 ± 4.48 (71)    10.82   4 [0.90]
(n1)       ELISA+32                         (58)†   5 [0.95]
           P-
           postlabell
           ing
           assays,
           WBC
Weighted   CS,          7.93 ± 4.48 (71)    7.93    7 [0.90]
                   32
(n1)       ELISA+                           (wma    9 [0.95]
           P-                               v)
           postlabell
           ing
           assays,
           WBC
Weighted   CS,          10.02 ± 6.07 (35)   10.09   8 [0.90]
(n1)       ELISA,                           (43)†   10 [0.95]
           WBC
Weighted   CS,          10.02 ± 6.07 (35)   10.02   8 [0.90]
(n1)       ELISA,                           (wma    10 [0.95]
           WBC                              v)
               32
Weighted   CS, P-       4.03 ± 1.81 (59)    17.38   1 [0.90]
(n1)       postlabell                       (75)†   1 [0.95]
           ing
           assays,
           PBL+WB
           C
Weighted   CS, 32P-     4.03 ± 1.81 (59)    4.03    5 [0.90]
(n1)       postlabell                       (wma    6 [0.95]
           ing                              v)
           assays,
           PBL+WB
           C
               32
Weighted   CS, P-        1.1 ± 1.6 (23)¶    18.5    1 [0.90]
(n1)       postlabell                       (60)†   1 [0.95]
             ing
             assays,
             PBL
Weighted     CS, 32P-                                           1.1 ± 1.6 (23)¶   1.1               45 [0.90]
(n1)         postlabell                                                           (wma              56 [0.95]
             ing                                                                  v)
             assays,
             PBL
                 32
Weighted     CS, P-                                           5.90 ± 1.93 (36)¶   12.9              1 [0.90]
(n1)         postlabell                                                           (15)†             1 [0.95]
             ing
             assays,
             WBC
                 32
Weighted     CS, P-                                           5.90 ± 1.93 (36)¶   5.90              2 [0.90]
(n1)         postlabell                                                           (wma              2 [0.95]
             ing                                                                  v)
             assays,
             WBC
EE: environmental exposure. OE: occupational exposure. SE: smoking exposure (non-smokers vs.
smokers). ■: Silesia is one of the most heavily polluted areas in the world and is characterized by high
cancer mortality and high infant mortality, among the most prevalent carcinogenic and mutagenic air
pollutants in Silesia are PAHs (176). ■■: controls are inhabitants of Biala Podlaska, a rural province in
northeastern Poland where winter atmospheric levels of B(a)P are estimated to be 10-fold lower than
in Silesia (176). ▲: compared to Silesian residents in the summer. ▲▲: compared to Silesia residents
                                                                             32                       32
in the winter. PBL: peripheral blood lymphocytes. WBC: white blood cells. P-postlabelling (n-Bu): P-
                                                                                         32
postlabelling with n-butanol extraction enrichment as described by Gallagher et al (1). P-postlabelling
(P1): 32P-postlabelling with nuclease P1 enrichment. 32P-postlabelling-HPLC: 32P-postlabelling
followed by the HPLC method. ND: not detectable. CS: current smoker. FS: former smoker. NS: never
smoker. wmav: weighted mean as v; thus mean exposed is set at twice the mean of the controls. ¶:
solely based on one study: (175), (165) or (172). †: since no ‘NS’- or ‘CS’-specific v was available, a
‘NS+CS’-derived v has been applied. ‡: since no ‘NS, ELISA, PBL’-specific v was available, a
‘NS+CS, ELISA, WBC’-derived v has been applied. Γ: since no WBC-derived v could be attained for
this class a PBL-derived v has been used. Ґ: since no PBL-derived v was available a WBC-derived v
ahs been used for this class.

				
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