Articles

Effect of NOX on the Somatic Chromosomes of Goldsmiths

Joginder S. Yadav and Neena Seth
Human Genetics Unit, Department of Zoology, Kurukshetra University, Kurukshetra, India

The genotoxic effc of NO2 was investigated on somatic human chromosomes obtained from
lymphocytes of 45 goldsmiths exposed to 1770.5 mg/m3 NO1 in ambient air at normal temperatre and pressure and compared to an equalnumber of matched control breadting air containing 50 pg/m3 NOr. Short-term lymphocyte culres were set up from blood collected from both
exposed and control individuals by venipuncue in heparinized sterile syringes. Mitotic index
(MI), chromosome aberrations (CAs), sister chromatid
(SCEs), and satellite associations (SAs) were analyzed. All the parameters showed a significant increase (pO.01 and pr.05)
in the exposed individuals as compard to the controls: MI (9.57 vs. 5.01), CAM (3.48 vs. 0.711),
SCEs (10.56 vs. 7.02), and SM (25.97 vs. 12.84), respecvly. Occurrence of DG-type SM (one
D-group chromosome and one G-group chromosome) was highest and 3 D-type (three D-group
chromosomes) lowest. NO2 was thus found to be genotoxic for humans. Key work chromosome
aberrations, genetic hazards, genotoxicity, mitotic index, NO., satellite association, sister chromatid xchng. Environ Heakh Perpect 106:643-647 (1998). [Online 4 September 1998]
hbnp://ebpnetl. niebs. nibgov.docs/d9,98i106p643-647yadavlabstrahbml

NOX (a mixture of NO2 and NO gas) is a

red brown toxic gas. When inhaled through

the respiratory tract and absorbed through
the skin, it causes various skin diseases and
respiratory problems. Based on in vitro
studies, Beckett et al. (1) postulated that the
environmental concentration of HNO2 is
formed within the respiratory system predominantly by hydrogen abstraction, with
subsequent conversion of HNO2 at physiologic pH, to H+ and NO2. Victorin (X) proposed that HNO2 formed in this way may
contribute to the bronchoconstricting
effects of NO2 seen in normal subjects and
asthmatics. Victorin (2) also observed eye
irritation just before, during, and immediately after exposure of asthmatic patients to
Eye irritation was reported to be sigNO..
nificantly higher in individuals with HNO2
exposure than in controls. Moreover NO2
levels as low as 0.5 ppm increase susceptibility to bacterial infection of lungs (4.
In India, laborers are rarely aware of the
potential damage that can accumulate in
their genomes due to pollutants in workplaces. NO. gas, evolved from aqua regia
used by goldsmiths, is such an example.
When gold or silver is added to aqua regia
(a mixture of HNO3 and HCR), copper and
other metal impurities present in these metals are reduced with the evolution of brown
fumes of NO. gas. This prompted us to
screen goldsmiths for genetic damage.

Materials and Methods

This study included 45 goldsmiths exposed
to NOX and 45 controls matched with
respect to age, sex, smoking habits, drinking
habits, and social status. Through spectrophotomety the average concentration of
NOX in the workplace was found to be

1770.05 mg/m3 compared to 50 pg/m3 at

normal temperature and pressure.
We conducted an epidemiological survey using a proforma especially designed for
this purpose. After obtaining consent, we
performed subsequent medical examinations of subjects suffering from asthma.
Short-term lymphocyte cultures were
prepared from heparinized blood (heparin
500 IU/ml without preservative) according
to the method of Moorhead et al. (3), with
slight modifications. Blood (0.5 ml) was
added to 5 ml RPMI 1640 medium containing 20% fetal calf serum, 100 IU/ml
penicillin, 100 pg/ml streptomycin, and
0.1 ml phytohemagglutinin (Sigma, St.
Louis, MO). Colchicine (10 pg/ml) was
added to the culture 2 hr before harvesting.
Lymphocytes were harvested after 48
hr for determination of mitotic index
(MI), chromosomal aberrations (CAs), and
satellite associations (SAs). Slides were air
dried and stained with 4% Giemsa. As
many as 100 good metaphases (well-spread
metaphases showing 2N-46 and proper
morphology of chromosomes) per individual were screened for CAs.
To determine sister chromatid exchanges
(SCEs), we added 5-bromodeoxyuridine (10
pg/ml/culture; Sigma) 24 hr after preparing
cultures. We used the method of Perry and
Wolff (4) with the following modifications: 1) the cells on slides were stained
directly in excess solution of Hoechst
33258 (50 pg/ml in H20; Sigma); 2) the
slides were exposed to long-wave UV light
of about 320-400 mm intensity; 3) the
slides were rinsed with water and incubated for 15-30 min in 2X SSC at 65C; and
4) the slides were rinsed with water and
stained with 2-3% Giemsa in phosphate

Environmental Health Perspectives * Volume 106, Number 10, October 1998

buffer (pH 6.8). For calculating the frequency of SCEs per cell, 25 metaphases per
individual were analyzed as per international practice.
For calculation of MI, 5,000 cells per
individual were scanned from Giemsastained slides. The mitotic index was calculated using the formula
MI =

No. of dividing cells x 100.

Total no. of cells scored

For evaluating the frequency of SAs, 100

good second division metaphases per individual were scanned. The following criteria
given by Hansson (5) were applied: satellite
ends of the associating chromosomes had to
be directed towards each other with their
longitudinal axes meeting between their
short arms, and the distance between the
centromeres of associated chromosomes
could not exceed the total length of one G
chromosome, its satellite excluded.
The samples were analyzed blindly by
two observers to remove possible laboratory
scoring bias. For statistical analysis of
results, the Student's t-test was applied.

Results
The epidemiological survey showed that 6
individuals out of the sample of 45 goldsmiths were asthmatic. As many as 30 persons also complained of irritation in the eyes.
The data obtained during this study are
presented in Tables 1-8. It is evident from
Tables 1 and 2 that mean MI in exposed
workers (9.57) was significantly higher
(p<0.01) than in matched controls (5.01).
It was highest in workers with an exposure
period of 0-5 years. Thereafter, the MI
showed a gradual decline.
We found chromosomal aberrations to
be elevated in the group exposed to NOX.
Yields of aberrations in goldsmiths are presented in Table 3, and raw data are given in
Table 4. Frequencies of all types of CAs
such as dicentrics, rings, acentric fragments,
chromatid gaps, breaks, and isochromatid
aberrations were significantly higher in the
Address correspondence to J.S. Yadav, Human
Genetics Unit, Department of Zoology, Kurukshetra
University, Kurukshetra-136 119 India.
We thank Kurukshetra University for use of laboratory facilities and A.S. Yadav for assistance. N.S.
received a Senior Research Fellowship from the
University Grants Commission, New Delhi, India.
Received 16 January 1998; accepted 28 April 1998.

643

Articles * Yadav and Seth

Table 1. Mitotic index (Ml) in goldsmiths exposed to NO5

Duration of
No. of cells
No.of
exposure
screened (nl)
samples
(years)
45
220,500
Control
45
219,150
Exposed
6
0-5
29,220
18
6-10
87,660
14
68,180
11-15
7
34,090
16-20
ap<0.01; tvalue = 20.72.

Table 2. Raw data showing mitotic index of goldsmiths exposed to NOX

No. of cells Duration of Mitotic
Case no. calculated exposure (years) index
9.824
0-5
4,856
1
9.810
0-5
2
4,659
9.820
0-5
4,820
3
9.760
0-5
4
5,123
9.793
0-5
5
4,689
9.783
0-5
6
5,073
9.713
6-10
7
5,069
9.668
6-10
8
4,650
9.682
6-10
9
4,832
9.677
6-10
10
4,739
9.623
6-10
11
5,068
9.655
6-10
12
4,875
6-10
9.629
13
4,865
9.680
6-10
14
4,733
9.720
6-10
4,987
15
9.648
6-10
16
4,678
9.733
6-10
17
4,859
9.723
6-10
18
4,963
9.670
6-10
19
4,578
9.678
6-10
20
5,088
9.657
6-10
21
4,992
9.681
6-10
22
5,000
9.700
6-10
23
4,698
6-10
9.710
24
4,986
11-15
10.375
25
4,822
11-15
9.625
26
4,299
11-15
9.462
27
4,939
11-15
9.538
28
5,000
11-15
9.600
29
5,023
11-15
9.571
30
4,995
9.629
11-15
31
4,923
11-15
9.620
32
4,977
9.638
11-15
33
4,633
11-15
9.422
34
5,000
9.450
11-15
35
4,698
9.548
11-15
36
4,699
11-15
9.617
37
5,000
11-15
9.583
38
5,172
16-20
8.98
39
4,666
8.96
16-20
40
4,996
41
16-20
8.92
5,000
16-20
9.05
42
5,000
16-20
9.08
43
4,987
44
16-20
8.99
4,962
45
16-20
9.01
4,479
-

Table 4. Raw data showing chromosomal aberrations in goldsmiths exposed NOX

No. of
metaphases (n2)
11,069
20,977
2,864
8,486
6,559
3,068

Ml SD
(n2In1 x 100)
5.01 0.05
9.57 0.128
9.80 0.02
9.68 0.03
9.61 0.22
8.99 0.04

Table 3. Frequency of chromosomal aberrations

in lymphocytes of goldsmiths and controls
Control Exposed tValues
Group
45
45
No. of persons
4,500
4,500
No. of metaphases
Chromosome type
aberrations
20 (0.444)
0
Dicentrics
2 (0.044)
0
Rings
Acentric fragments 7 (0.155) 39 (0.866)
7 (0.155) 61(1.35) 10.4
Total
Chromatid type
aberrations
56 (1.244) 179 (3.977)
Gaps
25 (0.555) 93. (2.06)
Breaks
lsochromatid
0
13 (0.288)
aberrations
25 (0.555) 106 (2.355) 9.79
Total
Total chromosomal
aberrations
32(0.711) 157(3.488) 24.01
(except gaps)
Values in parentheses indicate aberrations per 100
metaphases. All values are significant at p<O.01.

Range of cells calculated, 4,299-5,172.

p<0.01) than in controls (7.02). Both

among exposed workers and controls, individuals who drink alcohol showed higher
SCE frequency (10.30) than controls
(7.31). Similarly exposed tobacco smokers
showed a higher frequency of SCEs (10.31)
than control smokers (7.37). The highest
frequency of SCEs (11.02) was observed in
an exposed worker who was both a drinker
and a smoker (see Table 6). SCEs showed
an elevation in frequencies with increase in
the duration of exposure (Table 7).
Frequencies of SAs in both exposed and
control groups are shown in Table 8. The
exposed group showed more than a twofold
increase in the frequency of SAs (25.97)
compared to controls (12.84). DG association (one D-group chromosome and one
G-group chromosome) was found to be
highest (7.428%), while 3D association
(three D-group chromosomes) showed the
lowest frequency (1%).

exposed sample than in controls. The frequency of total CAs per 100 metaphases
was 3.48 in goldsmiths and 0.711 in controls; the difference was significant (p <0.01).
Frequency of SCEs (Table 5) in goldsmiths was significantly higher (10.56;

In the present study, 30 out of 45 goldsmiths complained of eye irritation. These

30 persons include 6 asthmatic workers.
We concluded that fumes of NOX cause eye
irritation in goldsmiths, and the effect is

644

Discussion

Case no.
-

2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45

na DC
100
100 2
100
100 2
100 2
100
100 2
100
100
100
100
100
100 2
100 1
100
100 2
100
100
100 2
100
100 2
100
100 1
100
100 1
100
100
100
100
100
100
100 1
100
100
100
100 1
100
100
100
100
100
100
100
100
100

Rg

AC
21
2
3
2

2
3
3
2
2
3

2
2
2

Br Iso ab

2
2
2
2
2

2
2
l
l
2
2
2
l

4
2
1
1
2

3
3
4
2
3
2
1
3
2
2
3
1
2
1
3
1
2
2
4
3
4
3
3
1
2
3
2

Abbreviations: DC, dicentrics; Rg, ring; AC, acentric fragments; Br, breaks; Iso ab, isochromatid aberrations.
aNumber of cells screened per individual.

more pronounced in asthmatic goldsmiths.

However, genotoxicity per se has no direct
relationship either with asthma or eye irritation. These abnormalities only depict general deterioration in health due to NOX
exposure.
There was a significant increase in total
CAs in the exposed group as compared to
the controls. Chromatid gaps were not taken
into account because their significance in
cytological monitoring of populations is still
a matter of discussion (6). The total chromatid aberrations were more than the total
chromosome aberrations in both goldsmiths
and controls. A similar elevation in CAs was
reported earlier in human populations
exposed to polycyclic aromatic hydrocarbons

Volume 106, Number 10, October 1998 * Environmental Health Perspectives

Articles * Effect of NO, on chromosomes

Table 5. Frequency of sister chromatid exchanges

(SCEs) (25 cells per individual) in goldsmiths and
controls as shown by alcohol drinking and tobacco
smoking status
Exposed
Control
Parameters
n Mean SE n Mean SE
Total SCEs
45 10.56 0.09 45 7.02 + 0.088
Drinker
2 10.30 0.45 1 7.31 0.54
Drinker/smoker 1 11.02 0.66 1 8.08 0.56
Smoker
6 10.31 0.26 8 7.37 + 0.19
Nondrinker 36 10.29 0.10 35 6.90 0.08
/nonsmoker
SE, standard error.
stValue = 13.1; significant at p50.01.

(PAHs), nitrosamines, carbon monoxide, and

sulfur dioxide (7). A significant elevation in
the frequency of CAs in peripheral blood
lymphocytes among persons belonging to a
given group may indicate an increased cancer
risk (8,9). Au et al. (10) and Santos-Mello et
al. (11) reported that CAs are initial events in
carcinogenesis and constitute an early warning signal for development ofcancer. Lu et al.
(12 treated mice and rats twice intragastrically with nitrite in doses of 1.7-47 mg/kg body
weight and rabbits were given the same dose
in drinking water for 3 months; CAs in bone
marrow were induced in all three species.
In this study, we also found a high frequency of dicentrics. Dicentrics are typical
and are normally used as a dosimeter for
ionizing irradiation. The present situation,
however, reveals that nitric oxides are one
of the few NOxs that may confound biological radiation dosimetry. Moreover,
dicentrics are known to be lethal in cell
proliferation, implying that carcinogenic
risk, as discussed by Awa (9), originates
mainly from CAs that survive cell division,
the so-called stable aberrations.
The frequency of SCEs in goldsmiths
was higher than in controls. Both among
exposed workers and controls, alcohol
drinkers and tobacco smokers showed
higher frequencies. A 1-8 ppm concentration of NO2 in culture bottles for a duration of 2 hr induced SCEs (13). Elevation
in the level of SCEs in the human population exposed to PAHs, nitrosamines, carbon monoxide, and sulfur dioxide as compared to controls has also been reported
(7). The elevated SCE level in operating
room personnel exposed to waste anesthetic
gases (mostly halothane, nitrous oxide, and
isoflurane) indicates that these gases could
be possible genotoxic hazards (14). In
human populations exposed to nitrate concentrations in the range of 50-300 mg/l,
no increase in SCE frequency was observed
(15). However, CAs (12,16-19), SCEs
(18), and gene mutations (16,20) were
induced in cell cultures after treatment
with high doses of sodium nitrite.

Table 6. Raw data showing frequency of sister chromatid exchanges (SCEs) in goldsmiths and controls as
shown by alcohol drinking and tobacco smoking status
Duration of
Dri/Sm
Duration of
Dri/Sm
Case no. exposure (years) status
SCE
Case no. exposure (years) status
SCE
Dri
6-10
10.65
46
Sm
7.22
6-10
2
11-15
47
9.95
7.42
6-10
3
0-5
Sm
9.68
48
6-10
8.00
4
11-15
Sm
10.99
49
Dr
7.19
6-10
5
0-5
Sm
9.86
50
Sm
7.00
6-10
6
11-15
Sm
10.39
51
Sm
6-10
8.00
7
11-15
Sm
10.58
52
7.13
6-10
8
11-15
Sm
10.38
53
7.00
6-10
9
11-15
11.02
54
Dri
7.31
6-10
10
16-20
10.98
55
6.75
6-10
11
16-20
10.86
56
6.85
6-10
12
16-20
10.57
57
6.15
6-10
13
16-20
11.06
58
6.85
6-10
14
16-20
11.68
59
6.03
6-10
15
16-20
11.62
60
6-10
6.58
16
16-20
10.23
61
8.08
6-10
Dri/Sm
17
11-15
11.25
62
6-10
6.45
18
11-15
11.06
63
6.88
6-10
19
11-15
11.14
64
6-10
6.97
20
11-15
11.56
65
6-10
6.82
21
11-15
11.76
66
6-10
6.33
22
11-15
10.46
67
6-10
6.32
23
11-15
11.94
68
6-10
6.52
24
11-15
11.38
69
6-10
6.35
25
0-5
8.08
70
6-10
6.91
26
0-5
8.32
71
6-10
6.58
Sm
27
6-10
10.60
72
6-10
6.35
28
6-10
10.59
73
7.39
6-10
29
6-10
10.58
74
Sm
6-10
7.05
30
6-10
10.50
75
6-10
7.31
31
6-10
10.00
76
7.34
6-10
32
6-10
10.61
77
Sm
6-10
7.02
33
6-10
10.51
78
6-10
7.31
34
6-10
Dri/Sm
11.02
79
6-10
7.36
35
6-10
10.50
80
6-10
7.31
36
6-10
10.00
81
6-10
7.05
37
6-10
10.89
82
6-10
7.01
38
6-10
10.80
83
6-10
6.98
39
6-10
10.80
84
Sm
6-10
7.25
40
6-10
Sm
10.69
85
6-10
7.35
41
6-10
10.58
86
6-10
7.10
42
6-10
10.80
87
6-10
7.50
43
6-10
10.25
88
6-10
7.54
44
6-10
Dri
9.95
89
6-10
7.05
45
6-10
8.11
90
6-10
7.00
-

Abbreviations: Dri, drinker; Sm, smoker; -, nondrinker/nonsmoker. Cases 1-45 are indviduals exposed to NOx and cases
46-90 are controls.

Beyond 5 years, the MI showed a slight

dedine. Similar declines in MI were reported in workers exposed to S02 over 5 years
(21) and in workers exposed to NH3 over
10 years (22). Rupa et al. (23) observed a
decrease in MI in smokers exposed to pesticides for longer periods. The hypothesis
that the increased amount of pollutants in
exposed individuals could destroy the lymphocytes, resulting in decline of MI (22),

Table 7. Frequency of sister chromatid exchanges

(SCEs) in goldsmiths with exposure to NO,
Duration of
No. of
exposure (years)
individuals
SCEs SE
0-5
6
9.00 0.24
6-10
18
10.57 0.15
11-15
14
10.99 0.17
16-20
7
11.00 0.25
SE, standard error.

Table 8. Frequency of satellite association pattern observed in goldsmiths and controls

No. of
Individuals

Exposed(45)

Control (45)

Total cells
scanned D-D
225
4,500
90
4,500

Environmental Health Perspectives * Volume 106, Number 10, October 1998

D-G
405
180

Type of satellite association

G-G 2D-G 2G-D 2G-2D
180 135
90
89
89 59
45
79

3D Total
45 1,169
36 578

Associations
per cell SE
25.97 0.71
12.8 0.4

645

Articles * Yadav and Seth

may be applicable in the present case, albeit

at a shorter exposure period. A decline in
MI with increasing dose of extract of particulate matter has also been reported (24).
Human cancers are reflections of sustained cell proliferative factors (consisting of
chemical agents, hormones, etc.) because
nondividing cells in adults, such as nerve cells
and cardiomyocytes, never develop tumors
(25). It has also been reported that some
PAHs, which are genotoxic carcinogens, have
the capability to increase cell proliferation
(26,24). Earlier, Cohen and Ellwein (28) suggested that potential to cause proliferation is
common to carcinogens, regardless of
whether they have genotoxic potency.
As such, an increased proliferative/mitotic index does indicate genotoxicity/carcinogenicity of the chemical to which the cells
were exposed.
We observed a twofold increase in the
frequency of SAs per cell. The difference is
significant and may lead to increased probability of chromosomal translocations. The
frequency of DG-type associations was
highest, whereas the frequency of 3D-type
associations was lowest. Similar results were
reported earlier for workers exposed to S02
(21) and NH3 (22) and for grape garden
workers occupationally exposed to different
pesticides (29). Trezepizur et al. (30) found
that chromosomes 21 and 22, which are
smaller than chromosomes 13, 14, and 15,
possess more extended nucleolus organizer
regions; consequently, acrocentric chromosomes 21 and 22 enter into associations
more frequendy than chromosomes 13, 14,
and 15.
A high incidence of SAs has often been
considered as a predisposition to an
increased tendency of nondisjunction in
satellite chromosomes and thus to the
induction of D and G trisomies (5).
Klossoglu et al. (31) reported cases of
increased SAs in families in which such trisomies occur. Among four families in which
new cases of translocation trisomics were
observed, the trisomics and their parents had
an unusually high incidence of SAs (32.
Hansson (5) suggested that the tendency
of SA is genetically controlled and specific
environments may influence the SA. In the
present study, NOX in ambient air increased
SA in exposed goldsmiths to twice that of
controls. The controls were exposed to a
background frequency of 0.266 ppb NOX,
which is near international standards: World
Health Organization, 0.25 ppb (33) and
EPA, 0.23 ppb (34).
Nitrous acid (HNO2) deaminates bases
in DNA; this is one mechanism for direct
mutagenic action. Alkylating agents are
formed by nitrosation of primary amines.
The formation of N-nitroso compounds
646

from secondary amines and amides is another way for indirect mutagenic activity (35).
Nitrite and nitrate are found in blood
after inhalation of NO (36), and nitrosated
compounds are formed after inhalation of
NO2 (37). Nitrosating agents are formed
in the skin when mice are exposed to 50
ppm NO2 for 4 hr; most of these nitrosating agents were derived from cholesterol
and some from triglycerides via a peroxidized product (38).
Nitrite derived from nitrate may react
in vivo with amines and amides to form Nnitroso compounds, which may have carcinogenic properties. Analysis of urine samples of subjects exposed to nitrates in
drinking water revealed the presence of Nnitroso compounds (39). NO2 is also
known for its potential to produce free radicals and other potent oxidizing species that
may oxidize tissue unsaturated fatty acids.
Tabacova (40) suspected that peroxidation
of membrane lipid is a primary mechanism
for the toxicity of NO2-induced lipid peroxidation in maternal tissues and is related
to its embryonic effect.
NO may react intracellularly, or in the
medium, with any dissolved oxygen or oxygen-derived radicals that may be generated
during exposure to NOX, resulting in the
formation of NO2 (41). The combination
of NO with NO2 yields a potent nitrosating agent, which has been shown to react
with secondary amines to yield carcinogenic N-nitrosamines both in aqueous and
lipid phase (42). Two mechanisms for this
reaction have been postulated involving
either nucleophilic attack by an amine on
N203 or a two-step process of radical reaction with the amine (43). While secondary
N-nitrosamines generally require metabolic
activation for mutagenicity (44), nitrosation of primary amines, as found on DNA
bases, results in immediate deamination
and DNA codon alteration (45). This latter
mechanism may, at least in part, be responsible for the observed mutagenic effects of
NO in our system.
Deamination of cytosine leads to the
formation of uracil, which, if not repaired,
causes a base substitution mutation.
Likewise, the deamination of methylcytosine
results in the formation of a thymine residue
in the DNA chain, which is repairable. NO
has recently been shown to alter bases in
both nucleosides and DNA under aerobic
conditions and at physiological pH (46,47).
Accordingly, exposure to NO could result in
the expected deamination of cytosine to
uracil and account for the observed mutagenicity of NO.
These results have significant implications for evaluating the genotoxic potential
of NOR, especially that of NO, because in

most situations involving exposure to high

concentrations of NOX, nitric oxide is
present in greater concentration relative to
NO2. The ability of NO to traverse the cell
and sequester in the lipophilic environment
within the DNA chain may be crucial to its
mutagenic activity (41).

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Faculty Position in Environmental Risk Analysis

The University of Californa at Santa Barbara (UCSB) Donald Bren School of Environmental Science and Management
i.nvites applications for a tenure-track position at the assistant professor level. We seek an interdisciplinary environmental
scientist who conducts research in environmental risks that result from human activities, with a focus on the quantification
of probabilities and expected consequences for biological communities, ecosystem processes, or society. Candidates must
be able to teach graduate-level courses in all aspects of environmental nrsk ainalysis, including risk identification, estimation,
evaluation, and management. Research foci could include pollution, natural resource exploitation, climate change, land use
and habitat conversion, or other areas. Postdoctoral or professional experience will be regarded favorably. The Donald Bren
School of Environmental Science and Management is a graduate school offering a two-year professional master's degree and
an interdisciplinary Ph.D. The curriciulum focuses on climatic, hydrological, ecological, and human systems at local, regional,
and global levels, and addresses the consequences of governmental policies, regulations of industrial practice, and
corporate strategies. Applicants for faculty positions must have a strong interest in collaborations with faculty and students
who conduct interdisciplinary research in environmental science, economics, and policy with implications for environmental
problem solving. Our Web page (http',/www.bren.ucsb.edu9) contains more information about the School.

Applications should be sent to Environmental Risk Search Committee, Donald Bren School of Environmental Science and
Management, University of California, Santa Barbara, CA 93106-5131 USA, tel: 805-893-7363, fax: 805-893-7612, e-mail:
erisk@bren.ucsb.edu. The Ph.D. is normally required at the time of appointment. Applications must include a curriculum
vitae, a statement of research interests and teaching philosophy, and copies of three refereed publications. The applicant
should arrange for three letters of reference to be sent in support of the application. For fullest consideration, all material
should be received by 7 December 1998, although the position will remain open until filled. The start date for the position
is 1 July 1999. The University of California is an equal opportunity/affirmative action employer. We encourage all qualified
applicants to apply, including minorities, women, and persons with disabilities.

Environmental Health Perspectives * Volume 106, Number 10, October 1998

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