The Effects of Air Pollution on Structures, Proteins and Allergenicity of Pollen Grains

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.     

A 2-year aeropalynological survey of allergenic pollen in the atmosphere of Kastamonu, Turkey

Knowledge of airborne pollen concentrations and the weather conditions influencing them is important for air quality forecasters, allergists and allergy sufferers. For this reason, a 7-day recording volumetric spore trap of the Hirst design was used for pollen monitoring between January 2006 and December 2007 in Kastamonu, Turkey. A total of 293,427 pollen grains belonging to 51 taxa were recorded during the study period. In the 2?years of study, the period March–August was identified as the main pollination season for Kastamonu. The highest monthly pollen counts were observed in May in both years. Six taxa made up 86.5% of the total amount of pollen recorded in the atmosphere of Kastamonu. These were as follows: Pinaceae (42.9%), Cupressaceae (20.6%), Poaceae (9.7%), Quercus (5.5%) Betula (5.3%) and Carpinus (2.6%). Four of these are considered to be highly allergenic (Betula, Carpinus, Cupressaceae and Poaceae). There were also a greater percentage of highly allergenic taxa found within the city, including Betula pendula that is not part of the local flora. This shows that through urban planting, the public and municipalities can unconsciously create a high risk for allergy sufferers. Daily average pollen counts from the six most frequently recorded pollen types were entered into Spearman’s correlation analysis with meteorological data. Mean daily temperature, relative humidity, daily rainfall and wind speed were found to significantly (p?<?0.05) affect atmospheric pollen concentrations, but the relationships between pollen concentrations and meteorological variables can vary and so there is a need for more local studies of this nature.     

Cloning and expression of cyclophilin from Platanus orientalis pollens in Escherichia coli

Background:

Allergy is a clinical disorder affecting the human population with wide geographical distribution. Platanus orientalis (P. orientalis) trees are planted in many countries and their pollen causes allergic reactions.Cyclophilin has recently been identified as one of the most important allergens of P. orientalis pollen. We aimed to clone and purify this allergen in Escherichia coli for further studies and therapeutic and diagnostic purposes for allergy to P. orientalis.

Methods:

RNA was extracted from P. orientalis. A full-length fragment encoding cyclophilin was prepared by polymerase chain reaction amplification of the first-strand cDNA synthesized from P. orientalis RNA. The cDNA was inserted into the pET32b (+) vector, and the construct transformed into E. coli Top10 and BL21 cells. The expressed protein was purified by the CuSO4 method.

Results:

The cDNA for the cyclophilin of P. orientalis pollen was cloned, and a specific reactivity of recombinant cyclophin was confirmed by immunoblotting using sera from patients allergic to P. orientalis pollen.

Conclusion:

The recombinant cyclophilin has a potential for immunologic assays for evaluation of allergy to P. orientalis pollen.Key Words: Allergy; Recombinant allergen; Cyclophilin, Escherichia coli, Platanus orientalis, Pollen, Cloning     

A study on the allergenicity and ontogeny of Acacia farnesiana pollen grains in guinea pigs

Pollen grains as the angiosperm male gametophytes transfer male genetic material during sexual reproduction. Pollen grains are among the important plant allergens, such that almost 80–90 % of plant allergens are of pollen origin. Acacia farnesiana is a plant with economical values, and due to its resistance to dry climates, it has had a widespread distribution in Southern Iran. This study was aimed at investigating the allergenicity and the ontogeny of pollen grains of A. farnesiana. Pollen grains were collected from the blossoms and flowers of A. farnesiana in the suburbs of Ahvaz-Iran. Pollen extracts (15 %) were prepared in PBS (pH 7.2). 4–6-week-old male guinea pigs (Hartly) were used for allergenicity tests. Skin tests showed a significant increase in flare diameter. Clinical tests also showed a significant change in the levels of eosinophils, neutrophils, and IgE. Histotechnical analysis was performed on male gametophytes, and photographs were taken using a camera-equipped light microscope. Pollen characteristics were identified using both light and electron microscopy. Sporoderm ultrastructure and pollen morphology were studied using Scanning Electron Microscopy. PAGE analysis of the total protein content of mature pollen grains showed seven clear bands of 10–83 kDa.     

Allergenicity of Acroptilon repens and Juglans regia pollen in rats

Pollen proteins that are located in the cytoplasm or on the surface of the exine can function as allergens and evoke immune system responses in sensitive patients, leading to allergic rhinitis and asthma. In this research, the pollen allergenicity and ability to induce IgE response of the pollen of two plant species were studied in rats. Acroptilon repens is an herbaceous, invasive plant with entomophilous pollen, while Juglans regia which is a tree crop produces anemophilous pollen. Immunoblot analysis using sera of sensitised rats revealed IgE reactivity to three protein bands including the 70, 41 and 25.12 kDa bands present in the A. repens pollen extract, while only one single immunogenic band of 11 kDa was detected in J. regia pollen extract. Both pollen extracts increased the eosinophil content and caused some clinical signs of allergy in treated rats. The results showed that both entomophilous and anemophilous pollen can be allergenic.     

Trends in the frequency of different pollen types in Vienna from 1976 to 1989

Summary For allergological purposes it is of interest whether the number of allergenic pollen will change in a significant order. Linear regression and correlation of different pollen types show that the annual totals of most of the allergenic pollen remain constant. None of the pollen grains show distinct and significant disappearance, but four types show significant increase of annual totals: Varia, Sambucus, Cupressaceae and Rubicaea. Pollen of Cupressaceae and Castanea fairly will become of more allergological interest within the next years, if the upward-trend should prove to be continous. No significant change was found in the number of the investigated spores.     

Characterization of protein expression of Platanus pollen following exposure to gaseous pollutants and vehicle exhaust particles

Being major ornamental street trees, species of Platanus are widely planted in the Shanghai urban area. A great deal of allergenic Platanus pollen is released from the trees and suspended in the atmosphere during its flowering season, ultimately causing allergic respiratory diseases. Few papers have focused on the distribution of this type of pollen and its expression of allergenic proteins. In order to investigate any differences in protein expression in Platanus pollen following exposure to gaseous and particulate pollutants, a special apparatus was designed. Exposure condition (such as temperature, humidity, and exposure time) of Platanus pollen and gaseous pollutants can be simulated using of this apparatus. Fresh Platanus orientalis pollen, pollutant gases (NO₂, SO₂, NH₃), and typical urban ambient particles (vehicle exhaust particles, VEPs) were mixed in this device to examine possible changes that might occur in ambient airborne urban pollen following exposure to such pollutants. Our results showed that the fresh P. orientalis pollen became swollen, and new kinds of particles could be found on the surface of the pollen grains after exposure to the pollutants. The results of SDS-PAGE showed that five protein bands with molecular weights of 17–19, 34, 61, 82, and 144 kDa, respectively, were detected and gray scale of these brands increased after the pollen exposure to gaseous pollutants. The two-dimensional gel electrophoresis analysis demonstrated that a Platanus pollen allergenic protein (Pla a1, with a molecular weight of 18 kDa) increased in abundance following exposure to pollutant gases and VEPs, implying that air pollutants may exacerbate the allergenicity of pollen.     

Pollen Lipidomics: Lipid Profiling Exposes a Notable Diversity in 22 Allergenic Pollen and Potential Biomarkers of the Allergic Immune Response

Background/Aim

Pollen grains are the male gametophytes that deliver sperm cells to female gametophytes during sexual reproduction of higher plants. Pollen is a major source of aeroallergens and environmental antigens. The pollen coat harbors a plethora of lipids that are required for pollen hydration, germination, and penetration of the stigma by pollen tubes. In addition to proteins, pollen displays a wide array of lipids that interact with the human immune system. Prior searches for pollen allergens have focused on the identification of intracellular allergenic proteins, but have largely overlooked much of the extracellular pollen matrix, a region where the majority of lipid molecules reside. Lipid antigens have attracted attention for their potent immunoregulatory effects. By being in close proximity to allergenic proteins on the pollen surface when they interact with host cells, lipids could modify the antigenic properties of proteins.

Methodology/Principal Findings

We performed a comparative pollen lipid profiling of 22 commonly allergenic plant species by the use of gas chromatography-mass spectroscopy, followed by detailed data mining and statistical analysis. Three experiments compared pollen lipid profiles. We built a database library of the pollen lipids by matching acquired pollen-lipid mass spectra and retention times with the NIST/EPA/NIH mass-spectral library. We detected, identified, and relatively quantified more than 106 lipid molecular species including fatty acids, n-alkanes, fatty alcohols, and sterols. Pollen-derived lipids stimulation up-regulate cytokines expression of dendritic and natural killer T cells co-culture.

Conclusions/Significance

Here we report on a lipidomic analysis of pollen lipids that can serve as a database for identifying potential lipid antigens and/or novel candidate molecules involved in allergy. The database provides a resource that facilitates studies on the role of lipids in the immunopathogenesis of allergy. Pollen lipids vary greatly among allergenic species and contain many molecules that have stimulatory or regulatory effects on immune responses.     

Twenty years of aerobiological monitoring in Trentino (Italy): assessment and evaluation of airborne pollen variability

The aim of the study is to report a reliable airborne pollen spectrum composition and seasonal timings for the monitored area as a basis for allergy management and to ascertain possible modifications through the detection of trends during the 20-year time series (1989–2008). Pollen was collected at San Michele all’Adige (Trento, Italy) by means of a Hirst-type spore trap. Sampling and counting of airborne pollen grains were carried out according to a national standard. Pollen concentration data for the period were processed in order to characterize the main pollen seasons for a subset of taxa, selected on the basis of their allergenicity and local representativeness. Variations in the pollen data over the years surveyed were analyzed using non-parametric tests. The results showed the presence of 63 pollen taxa, 40 of which belonged to tree and shrub species and 23 to herbaceous species. The local pollen spectrum was characterized by the presence of highly allergenic taxa, such as Urticaceae, Graminaceae, Ostrya sp., and Cupressaceae, in terms of percentage contribution as well as mean daily pollen count or peak values over the years surveyed. A significant upward trend was observed for daily mean pollen amount, mainly due to pollen from woody species and probably ascribable to a temperature-driven increase in pollen production. Evaluation of the results presented will form the basis of further research focussed on the climate change-related causes of modifications to vegetational dynamics as well as on the phenology of flowering and on pollen production.     

A 10-year survey of allergenic airborne pollen in the city of Porto (Portugal)

Airborne pollen calendars are useful to estimate the flowering season of the different plants as well as to indicate the allergenic potential present in the atmosphere at a given time. In this study, it is presented a 10-year survey of the atmospheric concentration of allergenic pollen types. Airborne pollen was performed, from 2003 to 2012, using a 7-day Hirst-type volumetric trap. The interannual variation of the daily mean concentration of the number of pollen grains and the main pollen season was determined as well as the hourly variations and correlation with meteorological parameters. During the study period, 18 different allergenic pollen types were considered based on its representativeness on the total annual airborne pollen concentration. The lowest annual concentrations were sampled in 2006 and the highest in 2007. The highest airborne pollen concentration was found during early spring and early summer. On the contrary, December was the month with the lowest pollen concentration. The major pollen sampled belongs to trees followed by weeds and grasses, being the most representative pollen types in the atmosphere: Urticaceae, Platanus, Poaceae, Pinaceae, Cupressaceae, Acer, Quercus, Castanea, Plantago, Alnus, Olea europaea, Betula, Myrtaceae and Populus. Intradiurnal distribution patterns of the pollen types studied presented differences with some taxa being predominantly sampled in the morning (9–11 a.m.) while others in first night hours (between 9 and 12 p.m.). Significantly correlations were found between the airborne pollen concentration and meteorological parameters.     

A comparative study on the allergenicity, structure, and ultrastructure of two Acacia pollen grains in guinea pigs

Acacia saligna and Acacia victoria are native to Australia brought to and grown in Southern Iran. They have since had a widespread distribution and become native in Iran. As people of the region are exposed to the pollen grains from these plants during a 5 months period, this study aimed at investigating the allergenicity of their mature pollen grains. In addition, the structural and ultrastructural as well as the total protein content of the mature pollen grains were analyzed. Pollen grains of A. saligna and A. victoria were collected from the suburbs of Ahvaz, Iran. Pollen extracts (15 % concentration) were prepared in PBS (pH 7.2), and 4- to 6-week-old male guinea pigs (Hartly) were used for allergenicity tests. Skin tests showed a significant increase in flare diameter. Clinical tests also showed a significant change in the levels of eosinophils, neutrophils, and IgE. Comparative analysis of the allergenicity of A. saligna and A. victoria pollen grains revealed a higher IgE level in the latter. Comparative analysis of pollen characteristics was performed using both light and electron microscopy for sporoderm structure and ultrastructure, respectively. Twelve percent PAGE analysis of the total protein content of mature pollen grains showed a greater number of bands in A. victoria, while four bands were common in both species. In conclusion, this study demonstrates that both A. saligna and A. victoria pollen grains are allergenic, with a greater allergenicity of A. victoria pollen grains. In addition, the structural and ultrastructural as well as the total protein content of the mature pollen grains are revealed, and a potential protein allergen is proposed.     

Pectate Lyase Pollen Allergens: Sensitization Profiles and Cross-Reactivity Pattern

BackgroundPollen released by allergenic members of the botanically unrelated families of Asteraceae and Cupressaceae represent potent elicitors of respiratory allergies in regions where these plants are present. As main allergen sources the Asteraceae species ragweed and mugwort, as well as the Cupressaceae species, cypress, mountain cedar, and Japanese cedar have been identified. The major allergens of all species belong to the pectate lyase enzyme family. Thus, we thought to investigate cross-reactivity pattern as well as sensitization capacities of pectate lyase pollen allergens in cohorts from distinct geographic regions.MethodsThe clinically relevant pectate lyase pollen allergens Amb a 1, Art v 6, Cup a 1, Jun a 1, and Cry j 1 were purified from aqueous pollen extracts, and patients´ sensitization pattern of cohorts from Austria, Canada, Italy, and Japan were determined by IgE ELISA and cross-inhibition experiments. Moreover, we performed microarray experiments and established a mouse model of sensitization.ResultsIn ELISA and ELISA inhibition experiments specific sensitization pattern were discovered for each geographic region, which reflected the natural allergen exposure of the patients. We found significant cross-reactivity within Asteraceae and Cupressaceae pectate lyase pollen allergens, which was however limited between the orders. Animal experiments showed that immunization with Asteraceae allergens mainly induced antibodies reactive within the order, the same was observed for the Cupressaceae allergens. Cross-reactivity between orders was minimal. Moreover, Amb a 1, Art v 6, and Cry j 1 showed in general higher immunogenicity.ConclusionWe could cluster pectate lyase allergens in four categories, Amb a 1, Art v 6, Cup a 1/Jun a 1, and Cry j 1, respectively, at which each category has the potential to sensitize predisposed individuals. The sensitization pattern of different cohorts correlated with pollen exposure, which should be considered for future allergy diagnosis and therapy.     

An aeropalynological survey in the city of Van,a high altitudinal region,East Anatolia-Turkey

Pollen concentrations in the atmosphere of Van city has been monitored for two consecutive years (2010–2011). This was the first detailed aeropalynological study for the elevated East Anatolia Region of Turkey. The sampling was performed by Hirst-type volumetric sampler, and pollen grains of 35 taxa were identified. The main pollen producers of the pollen flora were recorded as: Poaceae (20.94 %), Cupressaceae (10.53 %), Fraxinus (8.56 %), Chenopodiaceae/Amaranthaceae (7.77 %), Populus (7.75 %), Quercus (6.70 %), Platanus (6.68 %), Morus (5.57 %), Plantago (3.03 %). The pollen spectrum reflected the floristic diversity of the region, and the highest pollen concentration was recorded in April. There were a great percentage of allergenic taxa found in the city atmosphere, otherwise many of them scored under threshold values for risk of pollinosis. Statistical analyses were performed for correlating daily pollen concentrations of dominated pollen types concurrent with the data of meteorological parameters in MPS periods and number of significant correlations found. In addition, comparing 2-year data in terms of pollen concentrations and meteorological factors in MPS durations, many variables were found explanatory and concordant with the data. MPS starting dates of many plant taxa were found nearly a month later compared with western sites and lower altitudes of the country as well as Mediterranean countries; this case is mostly thought the ecological factors of the study area which directly affects the plant growth about the timing.     

Efficient and sensitive identification and quantification of airborne pollen using next‐generation DNA sequencing

Pollen monitoring is an important and widely used tool in allergy research and creation of awareness in pollen‐allergic patients. Current pollen monitoring methods are microscope‐based, labour intensive and cannot identify pollen to the genus level in some relevant allergenic plant groups. Therefore, a more efficient, cost‐effective and sensitive method is needed. Here, we present a method for identification and quantification of airborne pollen using DNA sequencing. Pollen is collected from ambient air using standard techniques. DNA is extracted from the collected pollen, and a fragment of the chloroplast gene trnL is amplified using PCR. The PCR product is subsequently sequenced on a next‐generation sequencing platform (Ion Torrent). Amplicon molecules are sequenced individually, allowing identification of different sequences from a mixed sample. We show that this method provides an accurate qualitative and quantitative view of the species composition of samples of airborne pollen grains. We also show that it correctly identifies the individual grass genera present in a mixed sample of grass pollen, which cannot be achieved using microscopic pollen identification. We conclude that our method is more efficient and sensitive than current pollen monitoring techniques and therefore has the potential to increase the throughput of pollen monitoring.     

Antiproliferative,Antifungal, and Antibacterial Activities of Endophytic Alternaria Species from Cupressaceae

Recent research has shown the bioprospecting of endophytic fungi from Cupressaceae. Here, we further uncover that the healthy cypress plants such as Cupressus arizonica, Cupressus sempervirens var. cereiformis, and Thuja orientalis host highly bioactive endophytic Alternaria fungal species. Indeed, endophytic Alternaria alternata, Alternaria pellucida, and Alternaria tangelonis were recovered from healthy Cupressaceous trees. Biodiversity and bioactivity of recovered endophytic Alternaria species were a matter of biogeography and host identity. We further extracted such Alternaria’s metabolites and highlighted their significant antiproliferative, growth inhibitory, and antibacterial activities against the model target fungus Pyricularia oryzae and the model pathogenic bacteria Bacillus sp., Erwinia amylovora, and Pseudomonas syringae. In vitro assays also indicated that endophytic Alternaria species significantly inhibited the growth of cypress fungal phytopathogens Diplodia seriata, Phaeobotryon cupressi, and Spencermartinsia viticola. In conclusion, since the recovered Alternaria species were originally reported as pathogenic and allergenic fungi, our findings suggest a possible ecological niche for them inside the foliar tissues of Cupressaceous trees. Moreover, in this study, the significant bioactivities of endophytic Alternaria species in association with Cupressaceae plant family are reported.     

Free orbicules of Cupressaceae detected in daily aerobiological samples by optical and confocal microscopy

Members of Cupressaceae and Taxaceae are known to release large amounts of highly allergenic pollen grains into the atmosphere, which are responsible for the onset of pollinosis in many countries throughout the world. In addition to pollen grains, their pollen sacs produce orbicules, which are submicron particles reported to carry allergens and which are potentially able to reach much further down the respiratory tract than pollen grains. Previous research has postulated the presence of orbicules in the atmosphere; however, direct observations have not yet been reported. The aim of this research was to provide the first direct evidence that Cupressaceae orbicules are released into the atmosphere by detecting them in daily aerobiological samples. We observed pollen sacs, pollen grains, and orbicules of nine species of Cupressaceae using scanning electron microscope (SEM). We then used a light and confocal microscope, to examine daily aerobiological samples. Under SEM, we measured the orbicule size (0.494–0.777 µm) and detected unknown nanometric particles (130–200 nm). Under the light microscope, aerobiological samples showed clusters of stained dots surrounding the pollen grains of Cupressaceae. Under the confocal microscope, the same clusters were resolved into submicron particles with the same autofluorescence as the pollen grains. These features enabled us to identify them as orbicules. We believe that our findings help to explain the onset of pollinosis and allergic asthma related to Cupressaceae pollen grains in many countries, even before pollen grains are actually detected or after they are no longer observed in aerobiological monitoring samples.     

First aerobiological study in Mediterranean part of Croatia (Dalmatia): pollen spectrum and seasonal dynamics in the air of Split

The aim of this study was to investigate aerobiological dynamics of pollen in the Mediterranean part of Croatia in the air of the city of Split. Pollen monitoring during the period from 2005 to 2013 was performed using a Hirst volumetric trap. Among the identified pollen of 50 taxa, 21 were allergenic. The average annual pollen index was 33,513. Three pollination seasons were established: early winter season dominated by tree pollen, spring–summer season dominated by herbaceous plants and summer–autumn season with lower amounts of Parietaria and Cupressaceae pollen. According to the abundance, the main taxa were: Cupressaceae, Parietaria/Urtica, Pinus, Quercus, Olea, Carpinus/Ostrya, Poaceae, Platanus and Ambrosia. The annual pollen index together with the daily maximum concentrations showed an upward trend for selected taxa during the study period. The highest monthly pollen index and the highest biodiversity were recorded in April and the lowest during the late autumn and winter months. The pollen calendar created for the city of Split confirmed Mediterranean features of the pollen spectrum. The longest pollen seasons were recorded for Cupressaceae, Parietaria/Urtica and Poaceae pollen types. The correlations between pollen concentrations and meteorological parameters were analyzed. The correlations between pollen concentrations and temperature were positive, while the humidity and the precipitation mostly showed negative influence.     

A change from grass pollen induced allergy to tree pollen induced allergy: 30 years of pollen observation in Switzerland

Pollen monitoring with a Hirst like pollen trap for 30 years in Basel, Switzerland allows understanding of the behaviour of the different plants producing allergenic pollens. It becomes evident that in this time period the different tree pollen like Hazel and Birch increased, whereas in the control, the observed amount of a herb pollen like Artemisa did not change. Moreover, it was observed that in the different trees flowering took place earlier namely at the beginning of the year. This was especially impressive for Hazel, where flowering was shifted ofmore than one month. These aerobiological observations were supported and confirmed by some epidemiological studies, showing that tree pollen allergy has become more important in the last years compared to grass pollen allergy and herb pollen allergy. The reason for the increase of tree pollen counts and the shift in the flowering time depends on the observed increase of the annual temperature, an increased input of nutrients like CO₂or NO_x but probably also a real increase of certain trees, like birch for example, in parks and avenues. This revised version was published online in June 2006 with corrections to the Cover Date.     

Spatial and temporal variations of pollen concentrations in Islamabad (Pakistan): effect of meteorological parameters and impact on human health

This study focuses on the identification and quantification of airborne pollen grains from allergenic plant species and their relationship with meteorological factors, i.e. maximum and minimum daily temperature, relative humidity, rainfall and wind speed in the city of Islamabad, Pakistan. An aerobiological data set (2010–2012), collected using rotorod samplers in five different sectors of the city, was supplied by the Pakistan Meteorological Department. Pollen of eight allergenic species was identified amongst which Broussonetia papyrifera exceeded the highest pollen level and, therefore, likely played a key role in aggravating the symptoms of pollen allergy in the city. The mean weekly pollen counts were next correlated with the weekly number of allergic patients visiting hospitals during 2010–2011. Clinical data were acquired from the Pakistan Institute of Medical Sciences. The highest number of allergic patients visiting hospital was usually observed during weeks with high pollen level. These results suggest a close relationship between the pollen concentration in the air and the allergy symptoms. Spearman’s rank correlation analysis was performed to establish the relationships between meteorological parameters and daily average pollen counts. A pollen calendar for the Islamabad city was also prepared to provide a guide for the timing and duration of season for all encountered pollen types.     

Effect of environmental pollution on the proteins, allergenic bands, ontogeny and structure of Avicennia marina (Forsk.) Vierh (Avicenniaceae) pollen grains

In Bushehr province of Iran, Avicennia marina trees have grown in Bordekhoon (Mond Protected Area) and Assaluyeh (Marine National Park of Nayband). Contrary to Bordekhoon, Assaluyeh is a petrochemical region with environmental pollution. This study was aimed to studying protein profiles, allergenic bands, ontogeny, structure and elemental composition of tectum of A. marina pollens in Assaluyeh and Bordekhoon. Pollens were collected from two regions and extracted in PBS, and protein profiles of pollens were determined by sulfate–polyacrylamide gel electrophoresis (SDS-PAGE). As an experimental model, 20 female 6–8-week-old Balb/C mice were divided into two groups. The mice of first and second groups were sensitized by Bordekhoon and Assaluyeh pollen extracts, respectively, and mice serum samples were used for immunoblotting. Pollen characteristics were studied using light and scanning electron microscopes. SDS-PAGE showed some differences between pollen protein profiles of two regions. Immunoblotting assay detected that pollens have two allergenic bands and the protein band at 100 KD is the common allergenic protein in two regions. Light microscopy revealed that the development of anther wall was basic type and some abnormalities were observed in microspores and pollens of Assaluyeh. Scanning electron microscopy studies showed that the apertures in considerable numbers of Assaluyeh pollens were closed. The comparison of elemental composition of pollen tectum between two regions showed that pollens of Assaluyeh have accumulated Cu on their tectum. Results obtained indicated that environmental pollution can affect protein profile, allergenic bands, structure and elemental composition of tectum of A. marina pollens.