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The prevalence of allergic disease has increased world wide during the last decades. Pollen allergy is the most typical form of allergic disease. The increase in its frequency during recent years is the most evident. Environmental factors play an important role in the problem of pollen allergy in large cities. The aim of this research is determination of allergenicity of Canna pollen in polluted and non-polluted conditions, detection of their allergenic proteins and also elucidation of some microscopic effects of air pollutants on pollen structure and proteins. Mature and immature pollen grains of Canna indica were collected from polluted and non-polluted areas. Pollen grains were studied by scanning electron microscopy. Mice were sensitized by injection of pollen extract and an adjuvant for five times. Allergy potency of different pollen extracts were compared by means of skin test, blood eosinophills number and IgE levels in sensitized and treated animals. Pollen proteins were studied by SDS-PGE and allergenic proteins were detected by immunoblotting techniques. Scanning electron microscope study of the pollen grains showed that in polluted areas, air born particles accumulated on the surface of pollen and changed both pollen's shape and pollen's tectum. Also many vesicles were released out of polluted pollen and the pollen material agglomerated on the surface of pollen. SDS-PAGE showed that different proteins exist in mature and immature pollen. In pollen collected from polluted area, some of protein bands between 22 and 45 kDa were disappeared . Also in all polluted pollen grains, protein content of pollen decreased in response to air pollution causing the release of pollen proteins. According to our experiments and regarding induction of allergic symptoms, the polluted pollen is more effective than non-polluted one, and mature pollen has more allergy potency than immature one.
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The effects of air pollution on structures, proteins and allergenicity of pollen
Ahmad Majd
, Abdolkarim Chehregani
*, Mostafa Moin
, Mansour Gholami
Shigekatsu Kohno
, Takeshi Nabe
& M.A. Shariatzade
Laboratory of Cytology, Department of Biology, Teacher Training University, Tehran, Iran;
Department of
Biology, Bu-Ali Sina University, Hamadan, Iran;
Asthma and allergy research center, Tehran, Iran;
Department of Pharmacology, Kyoto Pharmaceutical University, Kyoto, Japan;
Faculty of Health, Urmia
University. Iran. (*Author for correspondence: E-mail:
Received 21 February 2003; accepted 6 February 2004
Key words: air pollution, allergen, allergy, electron microscopy, pollen, proteins
The prevalence of allergic disease has increased world wide during the last decades. Pollen allergy is the
most typical form of allergic disease. The increase in its frequency during recent years is the most evident.
Environmental factors play an important role in the problem of pollen allergy in large cities. The aim of this
research is determination of allergenicity of Canna pollen in polluted and non-polluted conditions,
detection of their allergenic proteins and also elucidation of some microscopic effects of air pollutants on
pollen structure and proteins. Mature and immature pollen grains of Canna indica were collected from
polluted and non-polluted areas. Pollen grains were studied by scanning electron microscopy. Mice were
sensitized by injection of pollen extract and an adjuvant for five times. Allergy potency of different pollen
extracts were compared by means of skin test, blood eosinophills number and IgE levels in sensitized and
treated animals. Pollen proteins were studied by SDS-PGE and allergenic proteins were detected by
immunoblotting techniques. Scanning electron microscope study of the pollen grains showed that in pol-
luted areas, air born particles accumulated on the surface of pollen and changed both pollen’s shape and
pollen’s tectum. Also many vesicles were released out of polluted pollen and the pollen material agglom-
erated on the surface of pollen. SDS-PAGE showed that different proteins exist in mature and immature
pollen. In pollen collected from polluted area, some of protein bands between 22 and 45 kDa were dis-
appeared . Also in all polluted pollen grains, protein content of pollen decreased in response to air pollution
causing the release of pollen proteins. According to our experiments and regarding induction of allergic
symptoms, the polluted pollen is more effective than non-polluted one, and mature pollen has more allergy
potency than immature one.
1. Introduction
Over the centuries, pollen allergy has increased
from a local nuisance to a global problem (Em-
berlin, 1998). The reasons for this increase are
unknown. Among many hypotheses, the idea that
environmental pollutants may play a role has
gained substantial public and scientific attention
(Ishizaki et al.1987; Braun-Fahrlander et al., 1992;
Ring and Behrendt, 1993; Rusznak et al., 1994;
Behrendt et al., 1997). Air pollutants have detri-
mental effects on organisms and many researchers
are working on the effects of air pollution on
plants, animals and people. Air pollution may af-
fect pollen grains indirectly via stress on the
growth on the plant or directly either through
contamination of the anthers on the plant or
during the flight of pollen grains through the air
Aerobiologia 20: 111–118, 2004.
2004 Kluwer Academic Publishers. Printed in the Netherlands. 111
when it dispersed (Emberlin, 1998; Chehregani et
al., 2004). Airborne pollen grains can be attacked
directly by air pollutants and changes in pollen
shape and tectum occur under polluted condition
(Behrendt et al., 1992; Knox and Suphioglu 1996;
Knox et al., 1997). Changes of the structure, ultra-
structure, chemical compound and allergenicity of
some pollen were reported in polluted areas
(Ghanati and Majd 1995; Majd and Kiabi 1997).
Laboratory experiments have shown that NO
and CO treatments can cause changes in the
soluble protein composition of pollen grains. Some
studies showed that air pollutants could induce the
breakage of proteins or the formation of new
proteins in the pollen that exposed to pollutants
(Ruffin et al., 1983; Chakraborty et al., 1996). A
decrease of total pollen protein in polluted areas
was reported by Behrendt et al.(1997).
There are some evidences today that air pollu-
tion may be responsible for the increase in pollen-
induced allergies and asthma in highly polluted
areas (Emberlin, 1998; Behrendt et al., 1997;
Ruffin et al., 1983; Behrendt and Becker, 2001).
Aberg (1989) had reported increasing of asthma in
children due to SO
, cigarette smoke and air pol-
lution, while Riedel and Kramer (1988) and Jae-
Kyoung et al.(2001) had suggested that air con-
taminants such as SO
and other chemical pollu-
tants are responsible for developing asthma in
industrial regions. Additionally, irritant gases and/
or diesel exhaust particles have been shown to
express both adjuvant activity for sensitization
against common allergens and enhancing effects
on allergic symptoms in sensitized individuals
(Matsuramara, 1970; Holt et al., 1986; Riedel and
Kramer, 1988; Kainka-Staniche et al., 1989; Ito
et al., 1996; Helander et al., 1997).
In this report allergenicity of Canna pollen
grains was examined in different condition. We
tried detecting of the allergenic proteins. Also the
complex interaction between allergy potency of
pollen and air pollutants is investigated.
2. Materials and methods
2.1 Sampling
We chose Tehran as a highly-polluted city and as a
real condition, for our studies. In Tehran, the
amount of air pollutants is several times more than
the standard amounts (Table 1). Canna indica
belonging to Cannaceae family is one of the pop-
ular plants which use for landscaping in Tehran.
We selected samples planted in the central region
of Tehran as a polluted region and also out of
Tehran as a non-polluted region. Passing through
mesh with pores 30–40 lm in diameter, the fresh
pollen was being purified. We collected immature
pollen from immature anthers before releasing
their pollen grains.
2.2 Scanning electron microscopy
Polluted and non-polluted pollen were studied by
the scanning electron microscope. After coating
with gold, samples were analyzed using a scanning
electron microscope model SEM-EDS, EX 300
Link, England. The purified pollen was fixed by
glutaraldehyde, post fixed with Osmium, dehy-
drated and dried prior to gold coating.
2.3 Protein studies
Pollen protein extraction of non-polluted and pol-
luted pollen grains was carried out separately at
4C in Tris-HCl buffer, pH 7.6. Finally, 12%SDS-
polyacrylamid gel electrophoresis was performed
for the total soluble protein according to the
Table 1. Comparison of average amount of air pollutants in Tehran and standard amounts
Pollutants Carbon monoxide
Sulfur dioxide
Nitrogen dioxide
Hydro carbons
particle (lg/m
Average in 2000 5.89 56 81 3.6 135
Average in 2001 4.39 72 78 2.47 112
Standard amount 9.5 35 54 – <10 lm51
>1 0 lm15
Data evaluated at during spring and summer in 2 years, 2000 and 2001. Data obtained from 6 to 8 air pollution measurement stations.
Standard amount of air pollutants adopted from Nation Ambient Air Quality Standards, USA. This table shows that the amount of
some pollutants are several times more than standard in Tehran.
method of Laemmli (1970). The extraction of sol-
uble proteins was made in sample buffer (0.125 M
Tris-HCl, pH 6.8, 4%SDS, 20%glycerol, 10%b-
mercaptoethanol, 0.1%bromophenol blue dye)
with heating for 3–4 minutes at 100 C before
loading. The amount of protein was 10 lg per
channel and the total current was 14 mA. The gel
run in Tris-glycine buffer (pH 8.3) with 0.1%SDS
and calibrated with a marker protein was obtained
from ‘Sigma Company’. The evaluation of protein
concentration of the pollen extract was accom-
plished by using the method of Lowry et al.(1951).
2.4 Animals
We used sensitized Mice as experimental models.
Male Balb/C Mice (8–10 weeks of age) were sen-
sitized and treated with Canna indica pollen. The
animals were housed in an air-conditioned room at
a temperature of 25 ± 2 C; they were fed a
standard laboratory diet and given with water
added Vitamins. The experimental animal research
section at Razi Institute provided these animals.
Animals were sensitized and treated by intraperi-
toneal injection of 50 ll pollen extract (containing
about 20 lg protein in phosphate saline buffer) of
immature, non-polluted and polluted pollen sepa-
rately, with added the same dose AL(OH)
as an
adjuvant. Treatments were repeated five times, for
5 weeks, one in each week.
2.5 Skin test
We used sensitized Mice for skin tests. Pollen ex-
tracts were prepared from all samples (including
immature, mature and polluted pollen) by using
the method of Sheldon et al.(1967), and the skin
prick tests performed on Mice. Each animal was
tested with 50 ll of the total extract diluted with
0.05 M phosphate buffered saline, pH 7.4, and
containing 25 lg protein without adjuvant. The
negative control was buffered saline and the posi-
tive control was histamine acid phosphate. The
skin reactions were read after 4, 8, and 24 hours
from the start of the test and quantified on the
basis of wheal diameter.
2.6 Clinical and serological tests
Sensitized Mice were used in serological tests.
Blood samples were directly drawn from the heart,
one week after skin test (Aberg, 1989). At the end
of each experiment, the animals were anaesthetized
with Ether and blood samples were directly ob-
tained from the heart. Up to the time they were
needed, the sera of all samples were stored at
)76 C. Serums of 10 Canna-allergic sensitized
animals were individually tested for IgE reaction
to Canna pollen grains. Serums of 10 sensitized
animals were tested for polluted pollen, and 10
animals were tested for immature pollen. Sera
from 10 animals, which were not sensitized to
Canna pollen, were tested as the control sera poll.
The total serum IgE was evaluated by using ELI-
SA for the blood of mice. The standard Pharm-
ingen mouse IgE protocol for sandwich ELISA
was used to quantify the total amount of IgE. The
serum IgE level was expressed in nonogram per
milliliter after comparing with mouse IgE stan-
dards (Pharmingen, Calif, USA). Peripheral blood
smears of healthy controls were compared to
experimental animals (Mondal et al., 1997). We
determined blood cells in each group and com-
pared them by using statistical analysis.
2.7 IgE-specific immunoblotting
Pollen samples were subjected to SDS-PAGE and
then electerophoretically transferred to PVDF
membrane (Robtom et al., 2002). After SDS-
PAGE, proteins were transferred to a 0.2 lm
PVDF membrane (Bio Rad, USA) 1 hour at 100V
through use of an Electro-Transfer unit (Bio Rad).
The membranes were blocked over night at 4 Cin
PBS/5%non-fat dry milk/0.05%NaN
. Diluted
sensitized and treated mice sera, 1:5 (v:v) in PBS/
1%non-fat dry milk/0.01%NaN
, were added to
membrane and incubated over night at 4 C. The
membrane were then washed for 20 minutes 3
times in PBS/0.1%Tween 20 and incubated over
night at 4 C with polycolonal Goat anti-Mouse
IgE (Abcam, Cambridge, UK), diluted 1:400 in
non-fat milk buffer. The membranes were washed
for 20 minutes 3 times and incubated 2 hour at RT
with peroxidase-conjugated polycolonal Rabbit
anti-Goat IgG(H+L)(Jackson Immunol research,
USA) diluted 1:2000 in non-fat milk buffer. The
membrane were washed for 20 minutes 5 times and
incubated 1 minutes in enhanced chemilumines-
cence plus (Amersham Bioscience). The membrane
were exposed to Kodak medical X-ray film 3
minutes and developed by Fuji X-ray film
processor model FPM 700. The prestained protein
ladder for western blot was purchased from New
England Biolabs.
3. Results
3.1 SEM analysis
Ultra-structural studies of pollen grains of Canna
indica showed that they were spherical apolar, 30–
35 lm in diameter, which had a thick exin with
one visible pore. SEM analysis showed that tectum
on pollen grains was small mast (Figure 1). On the
surface of non-polluted pollens neither atmo-
spheric fine dust nor agglomeration was shown.
After contamination by polluted air, the color of
the pollen grains changed dramatically and dark-
ened considerably. Pollen grains became folded
when they exposed to polluted air and vermicular
particles accumulated on the surface of pollen
grains (Figure 2). Pollen grains collected from
polluted regions were covered by high amounts of
pollutants too; the tectum of polluted pollen grains
was disrupted. The interaction of pollen and par-
ticulate pollutants caused the pollen constituents
to be released and agglomeration of organic par-
ticles on to pollen surface to be formed (Figure 3).
3.2 Protein analysis
The SDS-PAGE protein profiles of different pollen
samples (mature, immature, polluted and non-
polluted) are shown in Figure 4. The bands are
present in the molecular weight range between 12
and 67 kDa. SDS-PAGE showed that several
proteins exist in immature pollen grains, which are
different from bands observed in mature pollen
grains. Some bands between 22 and 45 kDa dis-
appeared and there was an increased quantity of
another protein band with 22 kDa weight in pollen
grains which were collected from polluted areas.
Also in all polluted pollen, protein content was
Figure 1. Scanning micrograph of Canna pollen grains that
collected from non-polluted regions. Pollen grains are spherical
having small mast as the tectum (1700 X).
Figure 2. Scanning micrograph of Canna pollen grains that
collected from polluted regions. Pollen grains be came folded,
the tactum disrupted and air born particles accumulated in the
surface of pollen grains. app, air pollutant particles (1700 X).
Figure 3. Scanning micrograph of Canna pollen grains that
were exposed to Tehran polluted air for 20 days. Pollen grains
became folded; air born particles accumulated on the surface of
pollen and pollen material are released out of the pollen and
agglomerated on the surface of pollen. P, pore; amp, agglom-
erated pollen’s material (2000 X).
decreased in response to air pollution. The protein
content of extracts from pollen collected on the
same day from plants which were growing in non-
polluted and polluted area are shown in Table 2.
Significant changes were found in concentration of
total soluble protein per gram of pollen, between
polluted and non-polluted pollens.
3.3 Skin tests
Results of skin tests for different samples are
shown in Table 3. Pollen extract had allergenic
effect. While performing the skin-prick test, the
maximum allergenic sensitivity was seen for the
pollen which were collected from polluted areas,
with an average wheal diameter about 6 cm. Al-
lergy reactions were relatively large in normal
pollens which were collected from non-polluted
regions, with average 3 cm wheal diameters. Sta-
tistical analysis showed that the effect of polluted
pollen is about two times more than normal pollen
extract. Injection of these extracts caused the for-
mation of deep wound in the place of injection
after 24 hours whereas normal pollen caused only
small and superficial wound. Mature non-polluted
pollens were also observed to be more effective in
causing allergic symptoms than immature pollens.
3.4 Clinical and serological tests
Results of blood smears from different groups are
illustrated in Table 3. Study of blood smears
showed that the amount of eosinophiles and neo-
trophils were increased in animals which were
treated with pollen extracts. The quantity of eo-
sinophiles in animals which were treated with
polluted pollen was about two times more than
those treated with non-polluted pollens and ten
times more than the control group (Figure 5). The
Figure 4. Protein profile of different pollen grains. Immature
pollen grains have several different bands from mature ones.
Some bands disappeared and increased the quantity of band
22 kDa() in plants which were grown in polluted area. M,
markers; Pl, the bands of pollen grains that collected from
polluted area; N, the bands of normal pollen grains that col-
lected from non-polluted area; Im, the bands of immature
Table 2. Total protein content of polluted and non-polluted pollen grains of Canna indica
Specimen Pollen grains collected from
non-polluted area
Pollen grains collected from
polluted area
Pollen grains that 20 days
exposed to air pollution
Protein content mg/g 6.9 ± 2.6 4.6 ± 0.8 2.8 ± 0.7
Data shows that pollen protein content are decreased in polluted condition.
Table 3. Comparison of allergic reaction by means skin test (wheal diameter), blood eosinophil, neotrophil and IgE levels in control
and treated Mice
Samples C Im N Pl
Wheal diameter (cm) 0.4 ± 0.2 1.6 ± 0.9 3.1 ± 0.7 5.6 ± 1.03
Eosinophil number (·10
cells/ml blood) 5.5 ± 1.3 9.8 ± 2.2 38.6 ± 3.9 57.7 ± 5.6
Neotrophil number(·10
cells/ml blood) 35 ± 1.9 68.5 ± 3.4 84.2 ± 2.8 93.4 ± 3.7
IgE (ng/ml) 0.56 ± 0.2 2.32 ± 0.47 7.82 ± 1.2 11.7 ± 0.9
Data shows in all groups that have treated by pollen extracts, skin reaction, amount of eosinophils and IgE increased considerably. C,
control group that treated by salin; Im, the group that treated by immature pollen extract; N, the group that treated by mature non-
polluted pollen extract; Pl, the group that treated by mature polluted pollen extract.
data of the determination of total IgE in different
groups are shown in Table 3. All pollen extracts
caused an increase in IgE levels in the blood of
treated animals. Serum of animals, which were
treated with non-polluted pollen extracts, have
more IgE than the control animals. The results
indicate that the amount of IgE increased signifi-
cantly in animals which were treated by polluted
pollen extract. The amount of IgE in serum of
animals which were treated by immature pollen
extracts were more than the control group, but less
than other experimental groups. Statistical analy-
sis indicates that differences between polluted and
non-polluted groups are significant (Figure 6).
3.5 IgE-Specific Immunoblotting
To detect the allergen by allergen-sensitive sera,
immunoblotting was carried out on the pollen
extracts. Figure 4 shows the different bands of
pollen proteins. About 15 distinct protein bands
were detected by Comassi blue staining. To iden-
tify bands corresponding to the allergen(s), IgE-
binding bands were visualized by immunoblotting
which were using sera from animals that sensitized
and treated with Canna pollen extracts. After
immunoblotting, two different bands with molec-
ular mass of about 22 and 50 kDa were demon-
strated (Figure 7). Intense reactivity was seen with
the sera of 17 animals from a total of 20 sensitized
Eosiophilia in the peripheral blood of different
experimental groups
Eosinophil number (X104
cell/ml blood)
Figure 5. Eosinophilia in the peripheral blood of Mice that
were treated by different pollen extract or saline injection.
Polluted and non-polluted pollen extracts cause to increase in
eosinophil cells as allergic characteristics, but polluted pollen
grains were more effective. C, control group that treated by
saline; Im, the group that treated by immature pollen; N, the
group that treated by non-polluted mature pollen; Pl, the group
that treated by polluted mature pollen. Each column represent
the mean ± SE of 8–10 animals p< 0.05.
Increasing of total IgE in
animals that treated by pollen
Amount of total IgE (ng/ml)
Figure 6. Amount of IgE in blood of Mice that were treated
with different pollen extract or saline. Result shows that mature
non-polluted and mature polluted pollen extracts induce
increasing of blood total IgE as an allergy inducer. Effect of
polluted and non-polluted pollen grains was significantly dif-
ferent. C, control group that treated by saline; Im, the group
that treated by immature pollen grains; N, the group that
treated by non-polluted mature pollen grains; Pl, the group that
treated by polluted mature pollen grains. Each column repre-
sent the mean ± SE of 8–10 animals p< 0.05.
Figure 7. Results of immunoblotting by the using sensitized
and treated Mice sera and pollen proteins. Results shows that
Canna have two allergenic proteins with molecular mass of 22
and 50 kDa. M, pre-stained molecular mass markers; line 1,
protein bands of pollen extract that stained by Comassi Bril-
liant blue; line 2, protein bands that probed with pooled posi-
tive sera. The strong bands indicate reactivity of IgE antibodies
with corresponding protein bands that act as allergens.
animals. There was not any difference between
polluted and non-polluted animals. We could not
detect any allergen bands in immature pollen ex-
4. Discussion
Air pollution has different detrimental effects on
organisms. Ultra-structural studies (Figures 1–3)
showed that air pollution has affected pollen
grains (Figures 1–3). On the surface of non-pol-
luted pollens neither atmospheric fine dust nor
agglomeration was shown (Figure 1). Pollen
grains became folded when they exposed to pol-
luted air and vermicular particles accumulated on
the surface of pollen grains. Pollen grains collected
from polluted regions were covered by high
amounts of pollutants (Figure 2). Agglomeration
of air borne particles were seem on the surface of
pollen collected from polluted regions, this is in
agreement with observations which were reported
by some researchers (Behrendt et al., 1992; Beh-
rent et al., 1997; Knox and Suphioglu, 1996).
The results of 12%SDS-PAGE analysis for
soluble proteins (Figure 4), demonstrated the dif-
ferent bands in polluted and non-polluted pollen
grains. Some bands between 22 and 45 kDa dis-
appeared and there was an increased quantity of
another protein band with 22 kDa weight in pollen
grains which were collected from polluted areas
(Figure 4). It seems that air pollution could affect
the developmental process of pollen that is accor-
dance with finding of previous reports (Rusznak
et al., 1994; Majd and Kiabi, 1997; Ghanati and
Majd, 1995; Ruffin et al., 1983), but that is dif-
ferent than the reports of Behrendt et al.(1992,
1997). According to our results, total protein of
pollen grains decreased significantly in all polluted
pollen (Table 2). This is in accordance with the
findings of Behrendt et al.(1997), but not with the
findings of Ruffin et al.(1983). The releasing of
pollen material (Figure 3), culminate to decrease
the pollen protein content.
Allergenic potential of polluted and non-pol-
luted pollens were measured as different size of
wheal, leukocyte number changes and amount of
total IgE. Our clinical and serological results
showed that abundance of eosinophils, average
wheal diameter and amount of total IgE in animals
that were treated with extracts of fresh pollen are
more than the control group (Table 3). Statistical
analysis indicates that Canna indica is an allergenic
plant and immunoblotting studies showed that
Canna have two allergenic proteins with molecular
mass of about 22 and 50 kDa that reacted with
serum IgE strongly (Figure 7). These bands were
seemed in both polluted and non-polluted pollen
grains. It means that air pollutants could not affect
pollen allergenic proteins. We did not find any
reports regarding to this observation, therefore we
will try to get separation and more analysis of
these allergenic proteins. Also the allergenic po-
tential of polluted pollen is more than that of non-
polluted pollen grains (Figures 5 and 6). Our re-
sults indicated that mature pollens are more
allergenic than immature pollens. We could not
detect any allergenic bands in the immature pollen
proteins by using immunoblotting. It means that
most of the allergenic proteins were produced in
the developmental process of pollen formation and
were accumulated in mature pollen grains that
finished their development.
Our different results including skin tests, eval-
uation IgE and determination blood cells showed
that allergic potential of polluted pollen is more
than of non-polluted pollen grains. That is accor-
dance with some prior researches (Rusznak et al.,
1994; Ito et al., 1996; Chakraborty et al., 1996;
Behrendt et al., 1997; Knox et al., 1997; Majd and
Kiabi, 1997; Emberlin, 1998; Jae-Kyoung et al.,
2001). According to our observations organic
substances absorbed to air pollutants mediate the
agglomeration of particles on to the pollen surface
followed by local preactivation of the coated pol-
len grains. Under appropriate conditions, i.e.,
humidity, water-soluble compounds of air pollu-
tants may induce local allergen release and
agglomeration of pollen material including pollen
proteins on the surface of pollens (Figure 3). A
consequence of the release of allergen molecules as
aerosols is that the molecules are free to interact
with other types of air born particles, such as diesel
exhaust particles, that are associated with air
pollution. Findings of several researchers support
this idea (Knox and Suphioglu 1996; Behrendt
et al., 1997). In this condition, sensitive individuals
are in more contact with pollen proteins (allergens)
than in the non-polluted condition. This phe-
nomenon is one of the reasons of increase allergy
and asthma frequency in polluted cities. On the
other hand, one has to take into consideration that
in regions with high pollution, air pollutant par-
ticles are not only a carrier of pollutants, but also a
carrier of allergens and probably altered allergens,
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... Inorganic pollution and the degree of maturity for pollen grains have been reported as other factors to be considered, as pollen grains from polluted areas and from longer matured pollen grains showed a higher allergenicity (Buters et al., 2010), even changing their shape, colour and composition for tectum Majd et al., 2004). The fragility of the exine when pollen is exposed to pollutants and the consequent release of intracellular content could influence pollen allergenicity. ...
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The drivers affecting the Pollen Allergen Potency (PAP, amount of allergen released per pollen) are sparsely known. Betula and Poaceae airborne pollen are the two main allergenic pollen in the World. Airborne pollen and their allergens Bet v 1 and Phl p 5 were simultaneously measured from 2010 to 2015 in Davos (Switzerland) and Munich (Germany) by using volumetric traps and ChemVol cascade impactors. Daily variations in PAP were analysed in PM>10 and PM2.5-10 air fractions and generalized additive models were created to explain which factors determine PAP, including meteorological parameters and inorganic pollutants. 87.1 ± 13.9% of Bet v 1 and 88.8 ± 15.5% of Phl p 5 was detected in the fraction PM>10 where most pollen grains were collected. Significantly higher PAP for grasses (3.5 ± 1.9 pg Phl p 5/pollen grain) were observed in Munich than in Davos (2.4 ± 1.5 pg/pollen grain, P < 0.001), but not for Betula (2.5 ± 1.6 pg Bet v 1/pollen grain in Munich and 2.3 ± 1.7 in Davos, N.S.). PAP varied between days, years and location, and increased along the pollen season for Poaceae, but remaining constant for Betula. Free allergens (allergens observed in the fraction with limited pollen, PM2.5- 10) were recorded mostly at the beginning or at the end of the pollen season, being linked to higher humidity and rainy days. Also, PAP was higher when the airborne pollen concentrations increased rapidly after one day of low/moderate levels. Our findings show that pollen exposure explains allergen exposure only to a limited extend, and that day in the season, geographic location and some weather conditions need to be considered also to explain symptoms of allergic individuals.
... The effect of different air pollutants and vehicle exhaust particles on pollen viability and germination has been documented in earlier studies, such as reduction in pollen viability in Cannabis sativa, Cassia fistula and Thevetia peruviana growing along the roadsides of Amritsar city in India (Kaur et al., 2015); significant reduction in pollen viability in eight different species due to intense road traffic of Perugia (Iannotti et al., 2000); and significant decrease in pollen viability and germination in Betula pendula, Ostrya carpinifolia and Carpinus betulus pollen after exposure to NO 2 (Cuinica et al., 2014). Many authors have claimed that air pollution changes pollen protein content, structure and biochemical content such as reduction in pollen protein and lipids content in Artemisia vulgaris L. collected from the heavy-traffic site of Rzeszow, Poland (Depciuch et al., 2016); decrease in protein content and changed pollen shape and tectum of Canna indica pollen collected from polluted site of Tehran city (Majd et al., 2004); increase in phenol content in pollen of Thuja orientalis L. collected from the polluted environment of Tehran city (Rezanejad, 2009); and increase in allergenic protein in Platanus pollen after exposure to pollutant gases and vehicle exhaust particles (Lu et al., 2014). These biochemical changes in pollen alter its allergenicity (Ackaert et al., 2014;Karle et al., 2012;Lu et al., 2014;Sénéchal et al., 2015). ...
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Plants have been exposed to the urban environment for many years, and in response to air pollution, they have adopted selective and adaptive changes. In this study, we examined Datura pollen deposition on the stigma for germination and also assessed the viability of the pollen along with its element and protein content. According to the hypothesis that pollen physiology is negatively impacted by air pollutants, we expected a highly polluted area to have a high degree of pollen abortion with low amount of total protein content with accumulation of different elements because the high amount of particulate pollutants deposited on pollen should alter its physiology. We found that pollen viability at all three different locations is significantly similar, while pollen germination is significantly affected by pollution in Amravati City. The protein content in pollen and its shape is also affected. Correlation analysis reveals the interrelationship between pollen viability, germination, elements and protein content with respect to the polluted area. Principal component analysis was used to determine pollen characteristics contributing to discriminate at the three locations studied. Results revealed that Datura is adaptive in nature. Further study is needed to evaluate the adaptive evolution of Datura with respect to pollen tube sensitivity and tolerance to environmental pollution.
... Los espacios verdes urbanos, especialmente los ocupados por árboles, aportan importantes beneficios a los habitantes de las ciudades (Nowak et al. 2007, Hartig et al. 2011, Dadvand et al. 2016, Carrus et al. 2017, Vaz et al. 2017, pero también algunos inconvenientes (Von Döhren & Haase 2015, Cariñanos et al. 2017a, Vaz et al. 2017, entre los que queremos destacar los problemas de salud ocasionados por la alergia al polen (Thompson & Thompson 2003, Cariñanos et al. 2011. Esta situación suscita cada vez más interés, ya que las particularidades del entorno urbano, especialmente la contaminación, hace que los efectos sobre la población sean más notables (Majd et al. 2004, Heinrich & Wichmann 2004, Sénéchal et al. 2015 incluso cuando las concentraciones polínicas sean menores que en entornos rurales (Nico-lau et al. 2005). ...
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Los bosques poseen un rol clave en la mitigación del cambio climático ya que contribuyen a reducir los gases del efecto invernadero. Este trabajo tiene como objetivo conocer la diversidad, la biomasa y la captura de carbono de un bosque secundario de caldén (Prosopis caldenia) de la Argentina. Su importancia radica en ser una formación vegetal nativa y endémica que ha sido modificado a través de la historia por la valoración de sus recursos naturales. Para cumplir con el objetivo, se aplicó la metodología de REDD+ que consta de un inventario aleatorio de 10 parcelas circulares cuya superficie total es de 1 ha. Se seleccionó como área de estudio la Reserva Provincial Laguna Guatraché, provincia de La Pampa. Los resultados determinaron que el bosque de caldén contiene 156,4 Mg/ha de biomasa y la captura de carbono que realiza es de 78,2 t/ha. Conocer este valor contribuye a comprender los servicios ecosistémicos de este bosque y la importancia de su conservación.
... Changes in the protein patterns of Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and formation of the new bands have also been reported in the plants affected by the environmental pollutants (Majd et al. 2004). Also, it has been shown that these new detoxifying proteins were produced in response to the effects of diesel exhaust particles as an environmental pollutant (Chehregani and Kouhkan 2008). ...
Phytoremediation is a novel and cost-effective technology for removing contaminants from soil. This study investigated the effect of Chitosan (CS) and Chitosan nanoparticles (CSNPs) on phytoremediation ability of Datura stramonium L. for Cadmium (Cd) polluted soils. The results showed that D. stramonium is an accumulator plant for Cd, with the translocation factor (TF) of 1.12. Its accumulation ability was increased by the application of CS and CSNPs considerably. The concentration of Cd, in the soil, was decreased by increasing application of CS and CSNPs; also CSNPs were more effective in enhancing phytoremediation of Cd, in compared to CS. The application of CS and CSNPs significantly increased the bioconcentration factor (BCF) and TF. The highest BCF (1.85) and TF (1.65) were observed in the group treated by 5% CSNPs, showing the best function for phytoremediation of Cd. Increasing the uptake of Cd, after the CSNPs treatment, could be described as an effect of the small size, higher surface area and the low crystallinity of the nanoparticles. In the leaves of the plants treated with Cd + CS and Cd + CSNPs, the total protein content decreased in comparison to Cd- treated plants, that is the sign of Cd resistance in D. stramonium. Application of CS and CSNPs caused the increase in the peroxidase and polyphenol oxidase activities as compared to the group treated just by Cd. The results, therefore, demonstrated that D. stramonium could be suitable for phytoremediation of Cd-contaminated soils and application of CS and CSNPs could enhance BCF and TF.
... In particular, particulate matter deposited onto allergenic PGs could play a proinflammatory role contributing to the enhancement of pollinosis symptoms (D'Amato 2002). For instance, PGs with atmospheric particles accumulated on their surface are more effective at eliciting allergic symptoms in mice (Majd et al. 2004). Furthermore, a high exposure to PM 10 (particulate matter \ 10 lm) and PM 2.5 (particulate matter \ 2.5 lm) is associated with an increase in rhinitis severity (Burte et al. 2020). ...
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Hirst-type sampler (HTS) is the standard equipment for pollen sampling while cascade impactors are commonly used for particulate matter (PM) samplings. In this work, we investigated the feasibility of using such devices to reliably assess the atmospheric PM adhesion onto pollen grains (PGs). Birch PGs were deposited on sampling substrates and then exposed to ambient PM in each type of sampling device operating simultaneously. In the HTS, the surface of PGs was significantly polluted with PM during sampling (93.5% of polluted PGs with on average 4 particles per PG with a mean diameter of 2.2 µm). A modified entrance slit was tested and proved to significantly reduce the PM sampling artifact (28% of polluted PGs), although not completely canceling it. In contrast, in the PM10 impactor, all PGs remained free from particulate pollutants. We concluded that the PM pollution of PGs reported in previous studies using a HTS may be overestimated. While PG samplings using a HTS might provide insights into the co-exposure to allergenic pollen and other ambient PM, a size-selective aerosol sampler proves to be more appropriate for a reliable assessment of the extent of PM pollution of airborne pollen in order to give a realistic depiction of the physical and chemical state of ambient PGs that could be inhaled.
Introduction: Air pollution has steadily increased for several decades, with widely studied effects on human health, including increased mortality, incidence of stroke, respiratory and allergic disease. However, the effects of pollution on the ocular surface, in direct contact with the outside world, have been less precisely studied. Materials and methods: We conducted a literature review of articles on the subject published from 1966 to October 2020. Among the 661 articles identified, 33 were retained. Ocular surface disease associated with pollution included non-specific conjunctivitis, dry eye disease, blepharitis, and allergic conjunctivitis. The studied pollutants were particulate matter less than 2.5μm and 10μm (PM2.5, PM10), ozone (O3), nitrogen dioxide (NO2), carbon monoxide (CO) and sulfur dioxide (SO2). Certain air quality parameters such as temperature and relative humidity were also studied. Results: Among the markers of air pollution possibly associated with ophthalmic disease, NO2 and SO2 appear to be the most frequent and highly correlated. High temperatures and low humidity levels also appear to be aggravating factors for the ocular surface. However, due to the heterogeneity of the studies, the results must be interpreted with caution. Indeed, the methodology and the results of the various studies are sometimes contradictory. The inclusion of patients, the analysis of environmental data, and the correlation between these two elements indeed raise numerous methodological questions. Conclusion: Air pollution control would appear essential, as well as the development of new studies based on reliable methods of studying the environmental and its clinical effects.
The content for this preprint is already published as peer reviewed article with this DOI: 10.1016/j.envres.2022.113987
Pollen, climatic variables and air pollutants coexist in nature with the potential to interact with one another and play a crucial role in increasing allergic diseases. The current study evaluates the influence of meteorological parameters and air pollutants on the airborne pollen in an urban city, Chandigarh, situated in the Indo-Gangetic Plains. Airborne pollen monitoring was done following Spanish Aerobiological Network guidelines and dynamics of daily total pollen and six most abundant taxa were studied from June 2018 to June 2020. Among meteorological parameters, temperature and wind were the most correlated and influential parameters to airborne pollen concentration. Annual Pollen Integral (APIn) of Cannabis sativa (r = 0.52), Parthenium hysterophorus (r = 0.27), Poaceae (r = 0.32) and total pollen concentration (r = 0.30) showed a statistically significant positive correlation with temperature. In contrast, precipitation and relative humidity negatively correlated with APIn of total pollen concentration, Eucalyptus sp. and Poaceae except for Parthenium hysterophorus and Celtis occidentalis. Similar results were found with Seasonal Pollen Integral (SPIn) of total pollen concentration, six major taxa and meteorological variables. Spearman correlation performed for NOx showed a significant positive correlation among APIn and SPIn of Celtis occidentalis and insignificant among APIn and SPIn of Eucalyptus sp. and Morus alba. In contrast, except for Eucalyptus sp., PM10 and PM2.5 were negatively correlated among APIn and SPIn of total pollen concentration and other major taxa. Spearman's correlation of APIn and SPIn for each pollen taxon, meteorological parameters and air pollutants suggests that each taxon has a different pattern in response to all parameters. The study findings suggest that pollen response must be examined at the taxon level, not the assemblage level, having long time-series data. This will help to compute future scenarios of changing environmental factors and comprehend the relationships and trends among meteorology, air pollutants and aerobiology.
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Environmental factors such as air pollution are known to exacerbate respiratory illness and increase the overall health risk. However, on a daily or seasonal basis, the relation between air pollutants, weather and a disease such as asthma is not clear. When combined with aeroallergens such as birch pollen and under specific weather conditions, synergistic effects may increase symptoms of respiratory illness and morbidity and then reveal interesting links with environmental factors. Hence, it is important to improve the understanding of pollution-pollen-weather and broaden the public health message. Combined analysis and model simulation of aeroallergens, air pollution and weather as presented here is important to correctly evaluate health burdens and allow a better forecast of the potential health risk. However, analyzing the combined effects of several environmental factors is not well understood and represents a challenging task. This paper shows: (1) the results of data analysis performed in Montreal for asthma hospitalization in relation to complex synergistic environmental factors, and (2) model simulation of birch pollen using a coupled weather-air quality model (GEM-MACH) compared with model-data fusion of classical chemical species (e.g., near-surface ozone, nitrogen dioxide and fine particulate matter) in order to evaluate spatiotemporal vulnerable zone for asthma health risk.
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A diary study on a random sample of 1,063 Swiss children aged 0-5 y was conducted in four different areas of Switzerland (two urban, one suburban, and one rural area) to investigate the association between air pollution and respiratory symptoms. Passive samplers inside and outside the home measured NOâ concentration during the 6 wk each child was studied. Diaries were completed by parents, and 20% of them were validated with the attending pediatrician's case notes. Other pollutants were measured by city monitor in the two urban locations. Incidence and duration of symptom episodes were examined separately. The study included any episode: episodes of coughing without runny nose, upper respiratory episodes, and episodes of breathing difficulty. Annual average NOâ was associated with the duration of any episode and of upper respiratory episodes in Poisson regressions that controlled for medical history. Nitrogen dioxide (NOâ) was not associated with the incidence of episodes. In regressions using 6-wk average pollution, NOâ measured outdoors, but not indoors, was associated with the duration of any symptom. Total suspended particulates were a more significant predictor of duration in the two urban locations where it was available. Six-week average TSP was also associated with the incidence of coughing episodes. We conclude that duration of respiratory symptom episodes are associated with particulate concentrations, and possibly with NOâ.
Although in this new edition the format is very similar to that of the first and the number of chapters remains at 20, several improvements have been made and much information added. The entire book has been revised and some chapters completely rewritten. For example, chapter 1, "Immunology and Immunochemistry of Allergy," discusses the immediate and delayed types of hypersensitivity, the chemical mediators involved in the former and the transfer factor in the latter, and describes the immunoglobulins IgA, IgG, and IgM together with their physical properties. Other valuable additions include chapter 2, "Non Immunologic Factors in Allergic Disease," in which the role of infection and nonspecific influence such as irritants, air pollution, and weather are mentioned; chapter 15, "Management of Hypersensitization to Stinging Insects," in which the prevention and treatment of allergic reactions to antigens of the Hymenoptera group of insects are presented; chapter 19, "Immunologic Aspects of Some
Many people suffer from allergic diseases and much research has illustrated that pollen can play an important role in inducing such diseases. The fact that allergic symptoms are more prevalent in larger cities may be related to the increase in air pollution resulting from the mineral or synthetic materials released into the atmosphere by cars and factories. On the other hand, recent investigations have considered that it is the exinic mineral elements which induce or control pollen allergenicity. Different degrees of Pinus elderica allergenicity in guinea pigs were studied along with the ultrastructural changes and variation in the mineral element concentration of the pollen exine after exposure of the pollen to the atmosphere of Tehran for certain periods. The results were analyzed qualitively and also statistically by the unpaired T test, whenever necessary. P values of < 0.005 were considered significant.