Indoor Air 2009; 19: 6874  2008 The Authors

www.blackwellpublishing.com/ina Journal compilation  Blackwell Munksgaard 2008

Printed in Singapore. All rights reserved
INDOOR AIR
doi:10.1111/j.1600-0668.2008.00562.x

Eects on human eyes caused by experimental exposures to oce

dust with and without addition of aldehydes or glucan

Abstract Thirty-six volunteers (in three susceptibility groups: 11 subjects were L. Mlhave1, Z. Pan2, S. K.
non-allergic with nasal histamine hypersensitivity, 13 were non-allergic with Kjrgaard1, J. H. Bnlkke1, J.-E.
normal sensitivity, and 12 were pollen allergic with or without nasal hypersen- Juto3, K. Andersson4, G. Stridh4, H.
sitivity) were exposed for three and a half hours in a climate chamber. Each Lfstedt4, L. Bodin5, T. Sigsgaard1
subject was exposed to clean air (dust 45 38 lg/m3 total suspended particle,
TSP), house dust at 357 180 lg/m3 TSP, house dust 382 175 lg/m3 TSP 1
Department of Environmental and Occupational
with added glucan (50 ng/m3) and house dust 394 168 lg/m3 TSP with Medicine, Institute of Public Health, Aarhus University,
added aldehydes corresponding to a gaseous phase of 300 lg/m3 in the air. Aarhus, Denmark, 2Wuhan Centres for Disease
The study was explorative by nature. No signicant eects of exposures as such Prevention and Control, Wuhan, China, 3Department of
were seen on break-up time, conjunctival epithelial damage score and Trolox Otorhinolaryngology, Huddinge University Hospital,
Huddinge, Sweden, 4Department of Occupational and
Equivalent Antioxidant Capacity (TEAC) in tear lm and subjective ratings.
Environmental Medicine, rebro University Hospital,
However, in TEAC a signicant dierent time course was seen during exposures rebro, Sweden, 5Unit of Biostatistics, rebro
to aldehyde-containing dust indicating a subacute and late response to the University Hospital, rebro, Sweden
exposures. Perceived eye irritation increased signicantly during exposures to
normal dust. The perception ratings were highly correlated, whereas no Key words: Ocular effects; Humans; Eyes; Experimental
correlation was found between the subjective responses and the objective exposure; House dust; Glucan; Volatile organic
measurements. compounds; Aldehydes.
L. Mlhave
Department of Environmental and Occupational
Medicine
Institute of Public Health
Aarhus University
Vennelyst Boulevard 6
Building 260
DK-8000 Aarhus
Denmark
Tel.: +45 8942 6181
Fax: +45 8942 6199
e-mail: lm@mil.au.dk

Received for review 20 August 2007. Accepted for

publication 10 July 2008.
 Indoor Air (2008)

Practical Implications
The ndings indicate that measurement eects on the eyes are rather insensitive measures of short time eects of oce
dust exposures.

This type of air pollution has been associated with

Introduction
In the past decades, eld investigations in non-indus-
trial buildings (e.g. in day care centres, oces or
schools) and homes have demonstrated air pollution
with airborne particulate matter in the range of 20
200 lg/m3 (total suspended particles, TSP) with
extreme values up to 1000 lg/m3 (Gyntelberg et al.,
1994; Rudblad et al., 2002; Skov et al. 1989, 1990).
health eects. The Danish Town hall study (Gyntel-
berg et al., 1994) showed a 50% increased frequency of
symptoms associated with sedimented dust but no
association with the concentrations of airborne dust.
The investigations show that exposure to dust also
causes increased prevalence of complaints about air
quality, discomfort and symptoms related to irritation
of the eyes, nose, or throat. Several of these symptoms

68

have been related to objective eects, e.g. changed
numbers of inammatory cells in the eyes and
decreased tear lm stability (Kjrgaard et al., 1989;
Mlhave et al., 2000; Pan et al., 2000). Five human
exposure studies of the eects of dust (a review is found
in Mlhave, 2008) give strong evidence that dust
exposures cause decreased tear lm stability or break-
up time (BUT) and increased number of eosinophil
cells in nasal lavages. There is also strong evidence that
general well-being decreases during exposures. This is
also reected in increasing general irritation in the eyes,
nose, and throat. No signs of eects in non-exposed
organs or tissues were seen, not even in sensitive
persons.
The composition of the dust seems to matter. During
the past 10 years, employees in the occupational
environment reported symptoms associated with expo-
sures to airborne organic dust containing microbes or
chemicals (Eduard et al., 2001). Candidate compounds
are endotoxin (lipopolysaccharides, LPS from Gram-
negative bacteria, b-(1,3)-D-glucan from mold and
other compounds of microbiological origin (Hauswirth
and Sundy, 2004). Microorganisms and their endotox-
ins also caused adverse respiratory and systemic eects
in the indoor environment (Douwes et al., 2000).
Furthermore, the severity of asthma was associated
with the LPS concentration in house dust (Michel
et al., 1991). Epidemiological studies showed that
exposure to organic dust contaminated with micro-
organisms and their endotoxins may cause a wide
range of symptoms (Hagmar et al., 1990; Hollander
et al., 1994; Milton et al., 1995). b-Glucan is an
inammatory agent recognized by receptors on cell
surfaces. Dectin-1 has been identied as the major
cellular receptor (Brown and Gordon, 2001). The
cytokine release after glucan stimulation on cells is
usually less than that after LPS stimulation. However,
IL8 release from blood cells is signicantly higher after
glucan compared with LPS exposure in whole blood
assays (Kruger et al., 2004). Human studies with
inhalation of pure Curdlan also showed, that glucan
is an inammatory agent, however, weaker than LPS
(Fogelmark et al., 1992). In addition, volatile organic
compounds (VOCs) were reported to be associated
with health eects (Mlhave, 2001; Mlhave et al.,
1986). The adsorption of VOCs (here exemplied by
aldehydes) on dust is also of interest.
The present experimental study tests the hypothesis
that human exposures to oce dust at levels normally
occurring indoors with or without added glucan and
aldehydes cause eects on the eyes. These are present in
the form of weak inammatory or allergic objective
responses or subjective symptoms. The main hypoth-
eses of this study were: (1) dust exposure as such causes
weak inammatory or allergic reactions on human
eyes; (2) adsorption of VOCs (exemplied by alde-
hydes) on dust increases the toxic potential of the dust;
Eects of oce dust on human eyes
(3) contamination of dust with b-(1,3)-D-glucan
increases the toxic potential of the dust; and
(4) individuals respond dierently to these exposures
depending on personal factors. (5) The study also
investigates possible associations between objective
measures and the subjective responses related to the
eyes.
Materials and methods
This publication is one in a series of publications from
a larger study on house dust composition, human
sensitivity, and dierent types of health eects. More
details about the study including subject selection,
measuring methods, biomarkers and mediators in nasal
lavages and tear uids, rhinostereometry, and acoustic
rhinometry, as well as the 29 symptoms and percep-
tions as a whole can be found in Bnlkke et al. (2006)
and Bodin et al. (2008).
Thirty-six subjects were selected from about 150
potential volunteers and assigned to one of three
subgroups (GR1: non-allergic with nasal histamine
hypersensitivity, GR2: non-allergic with normal sensi-
tivity and GR3: pollen allergic with or without nasal
hypersensitivity). Histamine sensitivity was dened as
an increase in the swelling of the inferior turbinates of
at least 6 units (0.6 mm) measured with rhinostereo-
metry after 2 and 5 min (Hallen and Juto, 1993). The
healthy and non-smoking subjects were selected after
an investigation months prior to the exposures. The
male/female ratio was 14:22 and the average age was
30 years (range 2259).
The study included three replications of the same
basic design. For each replication, 12 subjects from the
36 subjects available were divided into four exposure
subgroups. These four subgroups (each of three
persons) included one person from each of the three
susceptibility groups mentioned above. Each exposure
subgroup was exposed to four types of exposures in a
balanced 4 4 Latin square design. Due to practical
diculties to obtain the same number of subjects in the
susceptibility groups, we had to introduce a minor
imbalance in the Latin square design.
The dust for the exposures was extracted from
vacuum cleaner bags from oce buildings and institu-
tions without reported indoor air quality problems.
The bulk dust was collected during a period without
pollen production according to the local Allergen
Monitoring Station. The collection and preparation
of dust followed a standardized procedure including
sieving (Mlhave et al., 2000). No homogenization was
made. Parts of the resulting processed dust were
supplemented with aldehydes or glucan. The aldehyde
addition was made by injecting the liquid compounds
into the bulk dust in a closed container and then
stirring the mixture at room air temperature for 12 h to
equilibrate the amount of adsorbed gasses with the air
69
Mlhave et al.
concentration of vapours of the three aldehydes:
n-hexanal, n-nonanal, and n-decanal respectively (ratio
1:10:100 volume). The glucan addition was made using
b-(1,3)-D-glucan (Curdland; WAKO Chemicals, Neuss,
Germany). Aldehyde mixture (0.1 ll) was added to 1 g
of processed dust. One gram of glucan was added to
100 g of bulk dust, and the mixture was mixed in a
rotating drum for 12 h.
Exposures were: (1) clean air (actual background
concentration of dust 45 38 lg/m3 TSP); (2) house
dust 357 180 lg/m3 TSP; (3) house dust
382 175 lg/m3 TSP with added glucan (equivalent
to 50 ng/m3); and (4) house dust 394 168 lg/m3 TSP
with added aldehydes. The amount of aldehydes added
to the dust corresponded to a gaseous concentration of
300 lg/m3 and was calculated from available adsorp-
tion isotherms. The amount of glucan and aldehyde
reected what had been reported from eld studies.
The exposures were performed in a 32-m3 stainless
climate chamber using a dust generator, placed outside
the chamber. The dust generator was slightly modied
from the previously developed acoustic generator
(Mlhave et al., 2000). The outlet from the generator
was connected to the chamber in the inlet of fresh
ventilation air. The climatic exposure conditions were
air temperature 22.923.0C and RH 26.834.2%.
The exposures were arranged in daily sessions
covering 1 h of clean air for acclimatization, 4 h in
the chamber, and 1 h post-exposure measurements in
clean air. The 4-h climate chamber exposure included
30 min of clean air exposure, 30 min of increasing
exposure, and 3 h of full exposure. Each time, the
subjects wore new protective clean room clothing
(Kimberly-Clark Kleengard T65xp, Bagsvaerd, Den-
mark) to avoid unintended contamination of the
chamber air.
This study was performed according to the Helsinki
declarations and was approved by the local research
ethical committee for ethics use of human subjects in
experimentation (reference number 1999/4486). The
participants gave their informed written consent prior
to the exposures.
Eect measures included subjective symptoms and
perceptions registered in a computer-based question-
naire, biosampling for analyses of mediators in nasal
lavage, and tear uids, rhinostereometry, acoustic
rhinometry and several ocular measures. This paper
reports only the ocular measurements which are tear
lm stability or break up time (BUT), the Epithelium
Damage (ED) Score and Trolox Equivalent Antioxi-
dant Capacity (TEAC) in the tear uid. Nasal mea-
surements have been published earlier (Bnlkke et al.,
2006). The subjective symptoms and perceptions were
registered in a questionnaire of 29-items of which 10
related to the eyes are reported here.
Tear lm stability or BUT was measured using a slit
lamp (model Top Con SL 75; Norn, 1983). Tear lm
70
stability was measured as the time (s) from an eye blink
to the appearance of visible break-up of the tear lm.
No blinking was allowed in this period. The pre-
corneal area of the left eye was tested. The eye was
stained using 1% Na-uoescein and then observed in
Cobalt-blue illumination. The measurements were
made before the registration of epithelial defects (see
below) to avoid interference. Three repeated measure-
ments of the same eye were made and the average value
was reported.
The Conjunctival ED Score was measured as the
number of dots in the intact epithelium using a vital
stain (Frank, 1986; Kjrgaard et al., 2004a). Erosion
of the conjunctival epithelia was measured as the
number of dead epithelial cells visible in a slit lamp
after staining with 10:l, 1% Lissamin green in the lower
conjunctival sack. After a blinking period, the number
of dots was counted and categorized as four groups: A:
010; B: 1150; C: 51100; D: >100.
The TEAC in the tear uid was measured in 10 ll of
the tear uid sampled from the outer eye cantus using a
calibrated capillary tube. An enhanced chemilumine-
scent method (Kjrgaard et al., 2004b; Whitehead
et al., 1992) was used. The addition of solutions of
known antioxidants such as ascorbate, urate or vitamin
E (or biological uids containing them) to a glowing
chemiluminescent reaction temporarily interrupts light
output. Light emission is restored after an interval that
is linearly related to the molar quantity of antioxidant
added. In this way, it is possible to qualify the total
antioxidant capacity in a variety of biological uids.
The capacity is expressed in Trolox equivalents (lM/l)
as the standard is made using Trolox, an articial
vitamin E.
The Tear lm stability/BUT and the measurements
of the Conjunctival ED Score were performed together
once per exposure session at the day after each
exposure session to reduce the potential impact of
colour staining on other measurements such as nasal
lavage measurements. Tear uids for TEAC measure-
ments were sampled three times during each exposure
session of 2 days, i.e. before exposure, after exposure,
and on the next day.
Subjective evaluations were made using a computer-
based questionnaire with 29 questions. The subjects
scored the symptom intensity on a PC screen by
placing a marker on a 130-mm-long Visual Analogue
Scale. The intensity was registered as the length in mm
from the left end of the scale to the marker (Mlhave
et al., 1986). For this eye-related publication, only the
following 10 of 29 questions were included in the
analyses: air quality, need of ventilation, eye irritation,
dry eyes, runny eyes, sleepiness, headache, concentra-
tion diculty, general well-being, and feeling stressed
by chamber occupancy. The questionnaire ratings were
made four times, i.e. before the exposure as a baseline
and three times during the exposure: just at the start, in
 Eects of oce dust on human eyes

the middle, and just before the termination of the
exposure.
Variables related to personal factors and other
response modiers were also measured but are pub-
lished elsewhere. They included age, gender, smoking
habits, bronchial sensitivity, allergy, nasal responses to
histamine measured with rhinostereometry, and blood
pressure (Bnlkke et al., 2006).
As no specic biological model for the responses
existed before the experiment, the statistical analysis
should be considered exploratory rather than conr-
matory. To reduce the between-subject variation, all
variables (except BUT and ED) were subtracted a
baseline or pre-exposure value. The rst four hypo-
theses mentioned above were tested for each eect
variable as an independent analysis. Initially, data were
explored by general descriptive statistics both as a
whole and split into exposure categories and suscepti-
bility groups. Based on the results of descriptive
analyses, data were transformed into a normal distri-
bution to meet the needs for the further analyses
(general linear model GLM for repeated measurements
and Pearson correlation). In GLM analyses, we
focused on the eects caused by the exposure factor
(four levels), the susceptibility group factor (three
levels), and the time course factor (three or four levels,
i.e. before exposure and dierent phases of the expo-
sure). Some co-factors (such as age, gender, and the
dierent replications of the design) were also included.
To explore a potential interaction between the expo-
sure factor and the susceptibility group factor, we
performed analyses of exposure also separately for the
three susceptibility groups, i.e. a subgroup analysis.
The day after the exposures, data for the BUT, ED,
and tear TEAC were used for analyses of correlation
between the objective measurements. Baseline cor-
rected ratings in the PC questionnaire at the end of the
exposure were used for correlation of the subjective
measurements. Baseline-corrected ratings in the PC
questionnaire and tear TEAC measures at the end of
the exposure together with BUT and ED measure-
ments on the day after the exposure were used for the
analyses of correlations between the objective and
subjective measurements. SPSS 10. was used for all
analyses. A P-value of 5% was set as statistical
signicance level.
Results
This publication deals with the objective eects on
human eyes and include tear lm stability/BUT, ED
score, and TEAC in tear uids. In addition, eects on
10 of 29 registered subjective symptoms and percep-
tions related to the eyes are described. When entered
individually, the personal or co-factors (e.g. age,
gender, and sensitivity group) did not show statistically
signicant eects on the outcome and further statistical
testing was performed without these co-factors.
Mean values and standard deviations for the BUT
are shown in Table 1. An eect of exposures to dust
with added glucan was seen in the initial analyses but
no signicant eects of exposures as such were seen in
the nal analyses. Mean values and standard devia-
tions for ED were all between 8 and 9 with standard

Table 1 Effects of dust exposures on the Trolox Equivalent Antioxidant Capacity (TEAC) and break-up time (BUT)

Group 1 Group 2 Group 3 Total

Measurements
and schedule Exposures Mean s.d. Mean s.d. Mean s.d. Mean s.d.

Before exposure
TEAC (lM/l) Clean air 642 205 716 417 620 156 661 285
Glucan 663 207 618 297 598 189 625 233
Aldehydes 632 186 771 321 558 121 658* 243
Normal dust 702 224 770 259 653 208 710 231
At end of exposure
TEAC (lM/l) Clean air 585 148 668 280 555 168 605 211
Glucan 674 248 570 233 654 230 632 234
Aldehydes 669 189 739 258 632 312 682* 258
Normal dust 632 183 680 395 649 251 655 289
The day after exposure
TEAC (lM/l) Clean air 596 194 677 245 536 130 605 200
Glucan 603 185 641 259 698 311 649 254
Aldehydes 613 184 607 245 522 181 581* 206
Normal dust 609 220 767 413 673 407 687 359
BUT (s) Clean air 7.9 3.0 9.6 4.1 11.2 10.5 9.6 6.7
Glucan 8.3 5.3 6.7 2.9 7.1 3.6 7.3** 3.9
Aldehydes 9.6 5.2 7.6 2.7 10.0 4.2 9.0 4.2
Normal dust 8.5 2.9 8.5 4.9 7.3 3.5 8.1 3.8

TEAC indicates different change trends *(P = 0.012). Paired t-test Glucan versus clean air (delay effect) **(P = 0.039).
Group 1: non-allergic with nasal histamine hypersensitivity; Group 2: non-allergic with normal sensitivity; Group3: pollen-allergic with or without nasal hypersensitivity.

71
Mlhave et al.

Table 2 The correlation (Pearson r and P-value) of the eye-related subjective responses
and symptoms

Q7 Q9 Q10 Q11 Q12 Q19 Q20 Q21 Q28 Q29

Q7 0.488 0.286 0.221 0.092 0.350 0.192 0.364 0.385 0.372

Q9 ** 0.404 0.300 0.150 0.163 0.208 0.319 0.307 0.440
Q10 ** ** 0.543 0.418 0.139 0.137 0.248 0.400 0.318
Q11 ** ** ** 0.241 0.132 0.109 0.117 0.152 0.166
Q12 ** ** )0.112 00.085 )0.115 0.148 0.123
Q19 ** 0.114 0.607 0.521 0.269
Q20 * * 0.258 0.300 0.403
Q21 ** ** ** ** ** 0.562 0.448
Q28 ** ** ** ** ** ** 0.589
Q29 ** ** ** * ** ** ** **

Q7, air quality; Q9, need for more ventilation; Q10, eye irritation; Q11, dry eyes; Q12,
runny eyes; Q19, sleepiness; Q20, headache; Q21, concentration difficulty; Q28, gen-
eral well-being; Q29, feeling stressed by chamber occupancy.
*P < or = 0.05, **P < or = 0.01, : not significant.

Fig. 1 The time course of the tear TEAC equivalent values

(lM/l; mean and 95% CI) for the four exposure types. Each
cluster represents before, end of exposure, and the day after
deviations between 2 and 3 and no signicant eects of
exposures as such were seen. Mean values and their
standard deviations for Tear TEAC are shown in
Table 1 and Figure 1. No signicant eects were found
for the exposure as such, although an indication of
changes was found after exposures to dust with added
aldehydes. However, the baseline-corrected data
showed signicantly dierent time courses in
(P = 0.012) during exposure to dust containing alde-
hydes. These time courses (Figure 1) from baseline,
subacute, and to measurements next day showed that
when the dust contained aldehydes and compared with
the other exposures, TEAC had increased signicantly
shortly after the start of the exposure and decreased the
next day. The GLM analysis of 10 of 29 questionnaire
responses to exposures showed that eye irritation
intensity signicantly increased during exposure
(P = 0.016) to untreated dust.
No correlation was found between the BUT and the
two other objective measurements, whereas TEAC was
signicantly correlated with ED during the glucan
exposure (R = 0.407, P = 0.014). However, this sig-
nicance disappeared when an outlying data point was
excluded. The subjective responses used in the correla-
tion analysis were the data registered just before the end
of the exposure, because these ratings were the closest in
time to the BUT and ED measurements. The perception
ratings were highly correlated (Table 2), whereas no
correlation was found between the subjective responses
and the objective measurements.
Discussion and conclusion
It should be underlined that because of its size and a
slight imbalance in the composition of the subject
exposure groups, this study should be considered
explorative rather than conclusive and denitive con-
clusions about the presence or absence of eects on the
eyes of the exposures must await future investigations.
Basically, our results seem to show no major
dierences neither between subgroups of subject sen-
sitivity nor between clean air and the dust exposures.
Frank (1986) showed in a eld study a signicant
correlation between the prevalence of sensory irritation
and the BUT and a similar correlation for epithelium
damage in the eyes. Kjrgaard et al. (1989) showed in
an experimental study the same association between
the BUT and acute sensory irritation by n-decane
exposure but no eect on epithelium damage. Pan et al.
(2000) showed a signicantly decreased BUT by
exposure to normal oce dust but no eect on the ED.
The discrepancy between the present study and
previous studies may be due to dierent timings of
the measurements. The insignicant results could also
be caused by the lack of baseline measurements. The
objective measures of the BUT and ED were measured
only once on the day after exposure, because the vital
staining would otherwise aect the other eect mea-
surements, especially the nasal lavage. These missing
baseline measurements prevented us from comparing
changes in the BUT and ED and limited the analyses to
comparisons of these eye-related endpoint variables the
mornings after the exposure. However, the observed
indication of a decreased BUT on the day after glucan
exposure suggests that glucan absorbed in dust may
trigger eects, although it requires a relatively long time
to reach full eect. It may also be a chance nding.
Glucan has been reported to be present in indoor air
and associated with symptoms such as nasal irritation,
itchy eyes, cough, etc. (Rylander, 1998), but its action
and function linking it to the symptoms are unknown.
Glucan may, in epidemiological studies, be an indicator
of other mold-related exposures, but glucan may also
by itself stimulate humoral and cell-mediated reactions
(Bnlkke et al., 2004; Kruger et al., 2004).

72
 Eects of oce dust on human eyes

A statistically signicant time course was found as
the TEAC increased at the end of the exposure to dust
with aldehydes and decreased the day after (Table 1
and Figure 1). During exposure to dust with glucan,
the TEAC increased with increased epithelium damage.
It should be noticed that these ndings appeared only
in the sensitivity subgroup analyses. The positive
correlation between the TEAC and ED after exposure
to dust with glucan might be attributed to a single
outlying observation.
The TEAC represents the overall antioxidative
capacity and thereby the resistance to oxidative
impairments. Rahman et al. (2000) suggested that a
decreased level of plasma antioxidants and increased
markers of oxidative stress may be related to injury and
inammation, although there are conicting reports
about the association between the oxidant/antioxidant
imbalances.
The indication that the BUT decreased after glucan
exposure (Table 1) may possibly be related to the
beginning of the tissue damage. In addition, the time
course of TEAC after aldehyde exposure indicate that
TEAC concentrations might at rst slightly increase
and then decrease till the next day. The decrease in the
TEAC the day after the exposure to aldehydes may be
explained by a depletion of antioxidants due to
inammation or oxidative stress induced by aldehyde
exposures. However, there were no signs of such
inammation.
From the same study, we previously reported that
IL-8 concentrations in the nasal lavage increased after
the aldehyde and glucan exposures a sign of nasal
inammation (Bnlkke et al., 2006). Nasal inamma-
tion was conrmed by nasal mucosal swelling and
decreasing nasal volumes after these exposures. Ten-
dencies for dierences in the kinetics of these inam-
matory changes between glucan and aldehydes were
also observed a parallel to the tendencies for the BUT
and TEAC in the eyes. Nasal eects were small despite
exposures to dust and it may not be surprising that due
to limited absorbed dose, the particle-bound irritants
used in this study provoked no or only very mild
reactions in the eyes.
In summary, inconclusive signs of the eects
expected according to the main hypothesis were found.
Inconclusive evidences indicate that an eect may be
present. The hypothesis could therefore neither be
rejected nor accepted at the exposure levels used.
However, in the TEAC indications of dierent time
courses were seen during exposures to aldehyde-
containing dust indicating a subacute and late response
to the exposures. Perceived eye irritation increased
signicantly during exposures to normal dust. The
perception ratings were highly correlated, whereas no
correlation was found between the subjective responses
and the objective measurements. The missing inuence
of sensitivity subgroup assignment is not surprising
and given the missing eects, but it might also provide
clues for further studies, i.e. that contents of glucan
and aldehydes change the nature of dust and modulate
the health eects on human eyes caused by indoor
exposures to dust.
Acknowledgements
This study was supported by the Building Research
Foundation (BFR #G1970187), Sweden; FORMAS,
Sweden, (FORMAS #21.2/2001-01.7) Statens
Miljforsknings Projekter, Denmark (SMP-98-2001)
and The National Research Foundation for Health
Sciences (SSVF #22-00-0191). Special thanks are due
to Gert Doekes, Institute for Risk Assessment Sciences
(IRAS), University of Utrecht, PO Box 80176, 3508
TD, Utrecht, The Netherlands, for glucan analyses, the
Danish Meteorological Institute (DMI) for outdoor
weather data, ISS A/S for assistance in dust sampling
in buildings, and Lisbeth Larsen, Danish Institute of
Technology (TI) for fungi analyses of the dust.

References
Bodin, L., Andersson, K., Bnlkke, J.,
Mlhave, L., Sigsgaard, T., Kjrgaard,
S.K. and Juto, J.E. (2008).Nasal hyper
responders and atopic subjects report
different symptoms intensity related to
IAQ; a climate chamber experiment,
Indoor Air, INA 07 02 016.
Bnlkke, J.H., Thomassen, M., Viskum, S.,
Omland, O., Bonefeld-Jorgensen, E.C.
and Sigsgaard, T. (2004) Respiratory
symptoms and ex vivo cytokine release
are associated in workers processing her-
ring, Int. Arch. Occup. Environ. Health,
77, 136141.
Bnlkke, J., Stridh, G., Sigsgaard, T.,
Kjrgaard, S.K., Lofsted, H., Andersson,
K., Bonefeld-Jrgensen, E.C., Jayatissa,
M.N., Bodin, L., Juto, J.E. and Mlhave,
L. (2006) Upper airway inflammation in
relation to dust spiked with aldehydes or
glucan, Scand. J. Work Environ. Health,
32, 374382.
Brown, G.D. and Gordon, S. (2001) Immune
recognition. A new receptor for beta-
Glucan, Nature, 413, 3637.
Douwes, J., Zuidhof, A., Doekes, G., van der
Zee, S.C., Wouters, I., Boezen, M.H. and
Brunekreef, B. (2000) (1 3)-beta-D-
glucan and endotoxin in house dust and
peak flow variability in children, Am. J.
Respir. Crit. Care Med., 162, 13481354.
Eduard, W., Douwes, J., Mehl, R., Heederik,
D. and Melbostad, E. (2001) Short term
exposure to airborne microbial agents
during farm work: exposureresponse
relations with eye and respiratory symp-
toms, Occup. Environ. Med., 8, 113
118.
Fogelmark, B., Goto, H., Yuasa, K., Mar-
chat, B. and Rylander, R. (1992) Acute
pulmonary toxicity of inhaled b-1, 3-glu-
can and Endotoxin, Agents Actions, 35,
5056.
Frank, C. (1986) Eye symptoms and signs in
buildings with indoor climate problems
(office eye syndrome), Acta Ophthalmol.,
64, 306311.
Gyntelberg, F., Suadicani, P., Nielsen, J.W.,
Skov, P., Valbjrn, O., Nielsen, P.A.,
Schneider, T., Jrgensen, O., Wolkoff, P.,
Wilkins, C.K., Gravesen, S. and Norn, S.

73
Mlhave et al.
(1994) Dust and sick building syndrome,
Indoor Air, 4, 223238.
Hagmar, L., Schutz, A., Hallberg, T. and
Sjoholm, A. (1990) Health effects of
exposure to endotoxins and organic dust
in poultry slaughter-house workers, Int.
Arch. Occup. Environ. Health, 62, 159
164.
Hallen, H. and Juto, J.E. (1993) A test for
objective diagnosis of nasal hyperreactiv-
ity, Rhinology, 31, 2325.
Hauswirth, D.W. and Sundy, J.S. (2004)
Bioaerosols and innate immune responses
in airway diseases, Curr. Opin. Allergy
Clin. Immunol., 4, 361366.
Hollander, A., Heederik, D. and Kauffman,
H. (1994) Acute respiratory effects in the
potato processing industry due to a bio-
aerosol exposure, Occup. Environ. Med.,
153, 7378.
Kjrgaard, S., Mlhave, L. and Pedersen,
O.F. (1989) Human exposure to indoor
air pollutants: n-decane, Environ. Int., 15,
473482.
Kjrgaard, S.K., Hempel-Jrgensen, A.,
Mlhave, L., Andersson, K., Juto, J.-E.
and Stridh, G. (2004a) Eye trigeminal
sensitivity, tear film stability and con-
junctival epithelium damage in 182 non-
allergic, non-smoking Danes, Indoor Air,
14, 200207.
Kjrgaard, S.K., Pedersen, O.F., Miller,
M.R., Rasmussen, T.R., Hansen, J.C.
and Mlhave, L. (2004b) Ozone exposure
decreases the effect of deep inhalation on
forced expiratory flow in normal subjects,
J. Appl. Physiol., 96, 16511657.
Kruger, T., Sigsgaard, T. and Bonefeld-
Jrgensen, E.C. (2004) Ex vivo induction
74
of cytokines by mould components in
whole blood of atopic and non-atopic
volunteers, Cytokine, 25, 7384.
Michel, O., Ginanni, R., Duchateau, J.,
Vertongen, F., Le Bon, B. and Sergysels,
R. (1991) Domestic endotoxin exposure
and clinical severity of asthma, Clin. Exp.
Allergy, 21, 441448.
Milton, D.K., Amsel, J., Reed, C.E.,
Enright, P.L., Brown, L.R., Aughenb-
augh, G.L. and Morey, P.R. (1995)
Cross-sectional follow-up of a flu-like
respiratory illness among fiber glass
manufacturing employees: endotoxin
exposure associated with two distinct
sequelae, Am. J. Ind. Med., 28,
469488.
Mlhave, L. (2001) Sensory irritation in
humans caused by volatile organic
compounds (VOCs) as indoor air pol-
lutants: a summary of 12 exposure
experiments. In: Spengler, J.D., Samet,
J.M. and McCarthy, J.F. (eds) Indoor
Air Quality Handbook, New York, The
McGraw-Hill Companies,
25.125.28.
Mlhave, L. (2008) Inflammatory and
allergic responses to airborne office dust
in five human provocation experiments,
Indoor Air, 18, 261270.
Mlhave, L., Bach, B. and Pedersen, O.F.
(1986) Human reactions to low concen-
trations of volatile organic compounds,
Environ. Int., 12, 167175.
Mlhave, L., Schneider, T., Kjrgaard,
S.K., Larsen, L., Norn, S. and Jrgen-
sen, O. (2000) House dust in seven
Danish buildings, Atmos. Environ., 34,
47674779.
Norn, M.S. (1983). The External Eye.
Methods of Examination, Copenhagen,
Denmark, Scriptor.
Pan, Z., Mlhave, L. and Kjrgaard, S.K.
(2000) Effects on eyes and noise in
humans after experimental exposure to
airborne office dust, Indoor Air, 10, 237
245.
Rahman, I., Swarska, E., Henry, M., Stolk,
J. and MacNee, W. (2000) Is there any
relationship between plasma antioxidant
capacity and lung function in smokers
and in patients with chronic obstructive
pulmonary disease? Thorax, 55, 189193.
Rudblad, S., Andersson, K., Stridh, G.,
Bodin, L. and Juto, J.E. (2002) Slowly
decreasing mucosal hyperreactivity years
after working in a school with moisture
problems, Indoor Air, 12, 138144.
Rylander, R. (1998) Microbial cell wall
constituents in indoor air and their rela-
tion to disease, Indoor Air, (Suppl. 4), 59
65.
Skov, P., Valbjrn, O., Pedersen, B.V. and
DISG (1989) Influence of personal char-
acteristics, job related factors and psy-
chological factors on the Sick Building
Syndrome, Scand. J. Work Environ.
Health, 15, 286295.
Skov, P., Valbjrn, O., Pedersen, B.V. and
DISG (1990) Influence of indoor climate
on the Sick Building Syndrome in an
office environment, Scand. J. Work
Environ. Health, 16, 363371.
Whitehead, T.P., Thorpe, G.H.G. and
Maxwell, S.R.J. (1992) Enhanced chemi-
luminescent assay for antioxidant capa-
city in biological fluid, Anal. Chim. Acta,
266, 265277.