Immunochemical characterization of rapid and slowly released allergens from the pollen of Parthenium hysterophorus

Allergens from the pollen of Parthenium hysterophorus (American feverfew), responsible for high incidence of allergic rhinitis, were found by immunoprint analysis to be localized on the surface of the pollen grains. The allergens were released very rapidly when extracted in vitro. The allergenic activity of the rapid (10 s) and slowly (20 h) released pollen proteins was comparable by in vivo skin test and ELISA inhibition assay. The isoelectric focusing patterns of the rapid and slowly released proteins, were also identical. SDS-PAGE and Western blot analysis revealed that all the major pollen allergens with molecular weight 14, 28, 31, 37 and 45 kDa were eluted within 10 s of extraction. Periodate-Schiff staining showed that the 28, 31 and 45 kDa components of the pollen extract are glycoproteins. The pollen allergens released after different periods of extraction lost 75% of IgE binding activity when subjected to in situ sodium m-periodate oxidation under controlled conditions, while 80% of the allergenic activity was still retained after extensive proteolysis. Our results support the clinical observation of a rapid onset of symptoms of allergic rhinitis in patients sensitive to Parthenium pollen, mediated predominantly by glycoproteins.

DNA from pollen: principles and potential

This paper describes our recent extraction of ancient DNA (aDNA) from Holocene pollen and discusses the potential of the technique for elucidating timescales of evolutionary change. We show that plastid DNA is recoverable and usable from pollen grains of Scots pine Pinus sylvestris from 10 ka and 100 years ago. Comparison of the ancient sequences with modern sequences, obtained from an extant population, establish a first genetic link between modern and fossil samples of Scots pine, providing a genetic continuity through time. One common haplotype is present in each of the three periods investigated, suggesting that it persisted near the lake throughout the postglacial. The retrieval of aDNA from pollen has major implications for palaeoecology by allowing (i) investigation of population level dynamics in time and space, and (ii) tracing ancestry of populations and developing phylogenetic trees that include extinct as well as extant taxa. The method should work over the last glacial oscillation, thus giving access to ancestry of populations over a crucial period of time for the understanding of the relationship between speciation and climate change.

Regulation of the pollen-specific actin-depolymerizing factor LIADF1

Pollen tube growth is dependent on a dynamic actin cytoskeleton, suggesting that actin-regulating proteins are involved. We have examined the regulation of the lily pollen-specific actin-depolymerizing factor (ADF) LIADF1. Its actin binding and depolymerizing activity is pH sensitive, inhibited by certain phosphoinositides, but not controlled by phosphorylation. Compared with its F-actin binding properties, its low activity in depolymerization assays has been used to explain why pollen ADF decorates F-actin in pollen grains. This low activity is incompatible with a role in increasing actin dynamics necessary to promote pollen tube growth. We have identified a plant homolog of actin-interacting protein, AIP1, which enhances the depolymerization of F-actin in the presence of LIADF1 by similar to60%. Both pollen ADF and pollen AIP1 bind F-actin in pollen grains but are mainly cytoplasmic in pollen tubes. Our results suggest that together these proteins remodel actin filaments as pollen grains enter and exit dormancy.

Regional Importance of Alnus Pollen as an Aeroallergen: A Comparative Study of Alnus Pollen Counts from Worcester (UK) and Poznań (Poland)

Daily average Alnus pollen counts (1996-2005) from Worcester (UK) and Poznań (Poland) were examined with the aim of assessing the regional importance of Alnus pollen as an aeroallergen. The average number of Alnus pollen grains recorded annually at Poznań was more than 2.5 times that of Worcester. Furthermore, daily average Alnus pollen counts exceeded the thresholds of 100, 500 and 1,000 grains/m3 more times at Poznań than Worcester. Skin prick test results (1996-2005) and allergen-specific IgE(asIgE) measurements using the CAP (Pharmacia) system (2002-2005), were supplied by the Allergic Diseases Diagnostic Centre in Poznań. The annual number of positive skin prick tests to Alnus pollen allergens was significantly related (p<0.05) to seasonal variations in the magnitude of the Alnus pollen catch recorded at Poznań (r=0.70). The symptoms of patients with positive skin prick tests to Alnus pollen allergens were: 51% pollinosis, 43% atopic dermatitis, 4% asthma, 1% chronic urticaria and 1% eczema. On a scale of 0-6, 20.5% of patients examined for serum asIgE in relation to Alnus pollen allergens had asIgE measurements in classes 5 and 6. Alnus pollen is generally considered to be mildly allergenic. However, the amount of Alnus pollen released into the atmosphere in places such as Poznań may increase its impact on the population and make it one of the more important aeroallergens present.

Examining Ambrosia Pollen Episodes at Poznań (Poland) Using Back-trajectory Analysis

The pollen grains of Ambrosia spp. are considered to be important aeroallergens in parts of southern and central Europe. Back-trajectories have been analysed with the aim of finding the likely sources of Ambrosia pollen grains that arrived at Poznań (Poland). Temporal variations in Ambrosia pollen at Poznań from 1995–2005 were examined in order to identify Ambrosia pollen episodes suitable for further investigation using back-trajectory analysis. The trajectories were calculated using the transport model within the Lagrangian air pollution model, ACDEP (Atmospheric Chemistry and Deposition). Analysis identified two separate populations in Ambrosia pollen episodes, those that peaked in the early morning between 4 a.m. and 8 a.m., and those that peaked in the afternoon between 2 p.m. and 6 p.m.. Six Ambrosia pollen episodes between 2001 and 2005 were examined using backtrajectory analysis. The results showed that Ambrosia pollen episodes that peaked in the early morning usually arrived at Poznań from a southerly direction after passing over southern Poland, the Czech Republic, Slovakia and Hungary, whereas air masses that brought Ambrosia pollen to Poznań during the afternoon arrived from a more easterly direction and predominantly stayed within the borders of Poland. Back-trajectory analysis has shown that there is a possibility that long-range transport brings Ambrosia pollen to Poznań from southern Poland, the Czech Republic, Slovakia and Hungary. There is also a likelihood that Ambrosia is present in Poland, as shown by the arrival of pollen during the afternoon that originated primarily from within the country.

Comparison of Two Pollen Counting Methods of Slides from a Hirst Type Volumetric Trap.

Two of the most frequently used methods of pollen counting on slides from Hirst type traps are evaluated in this paper: the transverse traverse method and the longitudinal traverse method. The study was carried out during June–July 1996 and 1997 on slides from a trap at Worcester, UK. Three pollen types were selected for this purpose: Poaceae, Urticaceae and Quercus. The statistical results show that the daily concentrations followed similar trends (p < 0.01, R-values between 0.78–0.96) with both methods during the two years, although the counts were slightly higher using the longitudinal traverses method. Significant differences were observed, however, when the distribution of the concentrations during 24 hour sampling periods was considered. For more detailed analysis, the daily counts obtained with both methods were correlated with the total number of pollen grains for the taxon over the whole slide, in two different situations: high and low concentrations of pollen in the atmosphere. In the case of high concentrations, the counts for all three taxa with both methods are significantly correlated with the total pollen count. In the samples with low concentrations, the Poaceae and Urticaceae counts with both methods are significantly correlated with the total counts, but none of Quercus counts are. Consideration of the results indicates that both methods give a reasonable approximation to the count derived from the slide as a whole. More studies need be done to explore the comparability of counting methods in order to work towards a Universal Methodology in Aeropalynology.

Pollen Production in the Poaceae Family.

Total pollen production per inflorescence was studied in the most important species of the Poaceae family in the city of Córdoba in order to further our knowledge of the partial contribution of each species of this family to the total amount of pollen released into the atmosphere. The contribution of grasses in a given area was estimated by counting the number of inflorescences in an area of one square meter. Four different representative areas of the city were selected. The number of pollen grains per anther and flowers per inflorescence was also estimated in order to obtain total pollen production per inflorescence. Pollen production per inflorescence ranged from 14,500 to more than 22,000,000 pollen grains, the amount being clearly higher in the perennial species. Pollen production per square meter was higher in the mountains near the city and lower in areas of abandoned crops. Only a few species are responsible for the majority of pollen produced. A phenological study is necessary in order to determine the temporal distribution of this pollen production and subsequent shedding.

The Pannonian Plain as a Source of Ambrosia Pollen in the Balkans

This study aims to find likely sources of Ambrosia pollen recorded during 2007 at five pollen-monitoring sites in central Europe, Novi Sad, Ruma, Negotin and Nis (Serbia) and Skopje (Macedonia). Ambrosia plants start flowering early in the morning and so Ambrosia pollen grains recorded during the day are likely to be from a local source. Conversely, Ambrosia pollen grains recorded at night or very early in the morning may have arrived via long-range transport. Ambrosia pollen counts were analysed in an attempt to find possible sources of the pollen and to identify Ambrosia pollen episodes suitable for further investigation using back-trajectory analysis. Diurnal variations and the magnitude of Ambrosia pollen counts during the 2007 Ambrosia pollen season showed that Novi Sad and Ruma (Pannonian Plain) and to a lesser degree Negotin (Balkans) were located near to sources of Ambrosia pollen. Mean bi-hourly Ambrosia pollen concentrations peaked during the middle of the day and concentrations at these sites were notably higher than at Nis and Skopje. Three episodes were selected for further analysis using back-trajectory analysis. Back-trajectories showed that air masses brought Ambrosia pollen from the north to Nis and, on one occasion, to Skopje (Balkans) during the night and early morning after passing to the east of Novi Sad and Ruma during the previous day. The results of this study identified the Southern part of the Pannonian Plain around Novi Sad and Ruma as being a potential source region for Ambrosia pollen recorded at Nis and Skopje in the Balkans.

Investigating Ambrosia Pollen Episodes in Poland Using Back-Trajectory Analysis

Background: The pollen grains of Ambrosia spp. are considered to be important aeroallergens. Previous studies have shown that the long-range transport of Ambrosia pollen to Poland is intermittent and mainly related to the passage of air masses over the Carpathian and Sudetes mountains from sources to the south, e.g. the Czech Republic, Slovakia and Hungary. In this study, Ambrosia pollen counts and back-trajectories from specific episodes in 1999 and 2002 have been analysed with the aim of identifying possible new sources of Ambrosia pollen arriving at three sites in Poland. Method: The combination of Ambrosia pollen measurements (daily average and bi-hourly concentrations) and air mass trajectory calculations were used to investigate two Ambrosia pollen episodes recorded at Rzeszow, Krakow and Poznań on the 4th and 5th September 1999 and 3rd September 2002. Ambrosia pollen counts were recorded by volumetric spore traps of the Hirst design. Trajectories were calculated using the transport model within the Lagrangian air pollution model, ACDEP (Atmospheric Chemistry and Deposition). Results: The collective results of pollen measurements and back-trajectory analysis indicate plumes of Ambrosia pollen travelling up through Poland from the southeast during the investigated episodes. In 1999, the plume was first recorded at Rzeszow in Southeastern Poland during the morning of the 4th September. Its route can be followed as it passed Krakow during the afternoon of the 4th, and later on the 4th and 5th September at Poznań. Similarly, back-trajectories calculated during the morning and afternoon from Krakow and Rzeszow on the 3rd September 2002 indicates that the air masses arrived at these sites from the East or Southeast. Conclusion: This study shows the progress of Ambrosia plumes into Poland from the southeast. Ambrosia pollen release occurs mainly during the day and so a midday peak in Ambrosia pollen concentrations may indicate a local source. However, if the plume of Ambrosia pollen tracked along its northwesterly path over Poland during investigated episodes did not originate from inside Poland, then it is likely that it came from the Ukraine. This identifies a possible new source of ragweed pollen for Poland. Trajectory analysis can only show the path along which an air mass travels, not the specific source area. Further investigation could therefore include source based transport models such as 3D Eulerian atmospheric transport models.

Release of Bet v 1 from Birch Pollen 1 from 5 European 2 Countries. Results from the HIALINE Study

Exposure to allergens is pivotal in determining sensitization and allergic symptoms in individuals. Pollen grain counts in ambient air have traditionally been assessed to estimate airborne allergen exposure. However, the exact allergen content of ambient air is unknown. We therefore monitored atmospheric concentrations of birch pollen grain and the matched major birch pollen allergen Bet v 1 simultaneously across Europe within the EU-funded project HIALINE (Health Impacts of Airborne Allergen Information Network). Pollen count was assessed with Hirst type pollen traps at 10 l/min at sites in France, United Kingdom, Germany, Italy and Finland. Allergen concentrations in ambient air were sampled at 800l/min with a Chemvol high-volume cascade impactor equipped with stages PM>10μm, 10 μm>PM>2.5μm, and in Germany also 2.5 μm>PM>0.12μm. The major birch pollen allergen Bet v 1 was determined with an allergen specific ELISA. Bet v 1 isoform patterns were analyzed by 2D-SDS-PAGE blots and mass spectrometric identification. Basophil activation was tested in an FcεR1-humanized rat basophil cell line passively sensitized with serum of a birch pollen lmptomatic patient. Compared to 10 previous years, 2009 was a representative birch pollen season for all stations. About 90% of the allergen was found in the PM>10μm fraction at all stations. Bet v 1 isoforms pattern did not varied substantially neither during ripening of pollen nor between different geographical locations. The average European allergen release from birch pollen was 3.2 pg Bet v 1/pollen and did not vary much between the European countries. However, in all countries a >10-fold difference in daily allergen release per pollen was measured which could be explained by long range transport of pollen with a deviating allergen release. Basophil activation by ambient air extracts correlated better with airborne allergen than with pollen concentration. Although Bet v 1 is a mixture of different isoforms, its fingerprint is constant across Europe. Bet v 1 was also exclusively linked to pollen. Pollen from different days varied >10-fold in allergen release. Thus exposure to allergen is inaccurately monitored by only monitoring birch pollen grains. Indeed, a humanized basophil activation test correlated much better with allergen concentrations in ambient air than with pollen count. Monitoring the allergens themselves together with pollen in ambient air might be an improvement in allergen exposure assessment.

A Mechanism For Long Distance Transport of Ambrosia Pollen From the Pannonian Plain

The pollen grains of ragweed are important aeroallergens that have the potential to be transported longdistances through the air. The arrival of ragweed pollen in Nordic countries from the Pannonian Plain canoccur when certain conditions are met, which this study aims to describe for the first time. Atmosphericragweed pollen concentrations were collected at 16 pollen-monitoring sites. Other factors included inthe analysis were the overall synoptic weather situation, surface wind speeds, wind direction and tem-peratures as well as examining regional scale orography and satellite observations. Hot and dry weatherin source areas on the Pannonian Plain aid the release of ragweed pollen during the flowering seasonand result in the deep Planetary Boundary Layers needed to lift the pollen over the Carpathian Moun-tains to the north. Suitable synoptic conditions are also required for the pollen bearing air masses tomove northward. These same conditions produce the jet-effect Kosava and orographic foehn winds thataid the release and dispersal of ragweed pollen and contribute towards its movement into Poland andbeyond.

Modelling Atmospheric Concentrations of Ragweed Pollen From Local and Distant Sources

Pollen grains from the genus ragweed (Ambrosia spp.) are important aeroallergens. In Europe, the largest sources of atmospheric ragweed pollen are the Rhône Valley (France), parts of Northern Italy, the Pannonian Plain and Ukraine. Episodes of Long Distance Transport (LDT) of ragweed pollen from these centres can cover large parts of Europe and are predominantly studied using receptor based models (Smith et al., (2013) and references therein). The clinical impact of allergenic ragweed pollen arriving from distant sources remains unclear (Cecchi et al. 2010). Although a recent study has found the major allergens of ragweed in air samples collected in Poznań, Poland, during episodes of long-distance transport from the Pannonian Plain (Grewling et al. 2013). The source orientated models SILAM, DEHM, COSMO-Art, METRAS and ENVIRO-HIRLAM currently report having the capability of modelling atmospheric concentrations of pollen in Europe. The performance of such source-orientated models is strongly dependent on the quality of the emissions data, which is a focus of current research (e.g. Thibaudon et al. (2014)). The output from these models are important for warning allergy sufferers in areas polluted by ragweed, but could also be used to warn the public of ragweed pollen being transported into areas where the plant is not abundant. Areas outside of the main areas of ragweed infection that contain considerable local populations must, however, also include local scale models. These models can be used to predict local concentrations, even when LDT is not present. This concept of combined LDT and local scale calculations has been shown to be work for air pollutants and is considered usable for urban scale calculations of aeroallergens once urban scale maps of aeroallergen sources have been produced.

Risk of Exposure to Airborne Ambrosia Pollen from Local and Distant Sources in Europe – an Example from Denmark

Background. Ambrosia artemisiifolia L. is a noxious invasive alien species in Europe. It is an important aeroallergen and millions of people are exposed to its pollen. Objective. The main aim of this study is to show that atmospheric concentrations of Ambrosia pollen recorded in Denmark can be derived from local or more distant sources. Methods. This was achieved by using a combination of pollen measurements, air mass trajectory calculations using the HYPLIT model and mapping all known Ambrosia locations in Denmark and relating them to land cover types. Results. The annual pollen index recorded in Copenhagen during a 15-year period varied from a few pollen grains to more than 100. Since 2005, small quantities of Ambrosia pollen has been observed in the air every year. We have demonstrated, through a combination of Lagrangian back-trajectory calculations and atmospheric pollen measurements, that pollen arrived in Denmark via long-distance transport from centres of Ambrosia infection, such as the Pannonian Plain and Ukraine. Combining observations with results from a local scale dispersion model show that it is possible that Ambrosia pollen could be derived from local sources identified within Denmark. Conclusions. The high allergenic capacity of Ambrosia pollen means that only small amounts of pollen are relevant for allergy sufferers, and just a few plants will be sufficient to produce enough pollen to affect pollen allergy sufferers within a short distance from the source. It is necessary to adopt control measures to restrict Ambrosia numbers. Recommendations for the removal of all Ambrosia plants can effectively reduce the amount of local pollen, as long as the population of Ambrosia plants is small.

Mesoscale Atmospheric Transport of Ragweed Pollen Allergens from Infected to Uninfected Areas

Allergenic ragweed (Ambrosia spp.) pollen grains, after being released from anthers, can be dispersed by air masses far from their source. However, the action of air temperature,humidity and solar radiation on pollen grains in the atmosphere could impact on the ability of long distance transported (LDT) pollen to maintain allergenic potency. Here, we report that the major allergen of Ambrosia artemisiifolia pollen (Amb a 1) collected in ambient air during episodes of LDT still have immunoreactive properties. The amount of Amb a 1 found in LDT ragweed pollen grains was not constant and varied between episodes. In addition to allergens in pollen sized particles, we detected reactive Amb a 1 in subpollen sized respirable particles. These findings suggest that ragweed pollen grains have the potential to cause allergic reactions, not only in the heavily infested areas but, due to LDT episodes, also in the regions unaffected by ragweed populations.

The Long Distance Transport of Airborne Ambrosia Pollen to the UK and the Netherlands from Central and South Europe

Background: The invasive alien species Ambrosia artemisiifolia (common or short ragweed) is increasing its range in Europe. In the UK and the Netherlands airborne concentrations of Ambrosia pollen are usually low. However, more than 30 Ambrosia pollen grains per cubic metre of air (above the level capable to trigger allergic symptoms) were recorded in Leicester (UK) and Leiden (NL) on 4 and 5 September 2014. Objective: The aims of this study were to determine whether the highly allergenic Ambrosia pollen recorded during the episode could be the result of long distance transport, to identify the potential sources of these pollen grains and describe the conditions that facilitated this possible long distance transport. Methods: Airborne Ambrosia pollen data were collected at 10 sites in Europe. Back trajectory and atmospheric dispersion calculations were performed using HYSPLIT_4. Results: Back trajectories calculated at Leicester and Leiden show that higher altitude air masses (1500m) originated from source areas on the Pannonian Plain and Ukraine. During the episode, air masses veered to the west and passed over the Rhône Valley. Dispersion calculations showed that the atmospheric conditions were suitable for Ambrosia pollen released from the Pannonian Plain and the Rhône Valley to reach the higher levels and enter the air stream moving to Northwest Europe where they were deposited at ground level and recorded by monitoring sites. Conclusions: The study indicates that the Ambrosia pollen grains recorded during the episode in Leicester and Leiden were probably not produced by local sources, but transported long distances from potential source regions in East Europe, i.e. the Pannonian Plain and Ukraine, as well as the Rhône Valley in France.