Localisation of allergens in ryegrass pollen and in airborne micronic particles

In Melbourne, Australia, grass pollen allergens, especially from ryegrass, are a major cause of allergic hayfever and asthma. This review outlines recent developments in our understanding of how grass pollen allergens find their way into the atmosphere and how they are transported in particulate form. Much of this work has relied on antibody technology in immunological and immunocytochemical investigations. The localisation of allergens in situ has proved difficult due to their water-soluble character. Recently, allergens have been localised in developing ryegrass pollen by dryfixation, rapid-freeze and freeze-substitution techniques. This involved anthers being substituted in a mixture of aldehydes, organic solvents, and 2,2-dimethoxypropane. Incubation in dimethylsulfoxide prior to embedding in LR Gold resin provided good infiltration with freeze-substituted material. Immunogold-labelled sections show that the major allergens, Lol p 1 and Lol p 5, are synthesised in the pollen cytoplasm from the early bicellular stage, soon after the first starch granules are formed. From the early tricellular stage, Lol p 5 moves into the starch granules where it remains until maturity. Lol p 1 is localised in the cytoplasm of mature pollen grains. The incidence of airborne grass pollen, as measured in pollen traps, correlates with hayfever symptoms. Forecasting models which rely on rainfall and temperature data have been produced for the grass pollen (daily and seasonal) counts in Melbourne. Research over the past six years has shed light on the causes of grass-pollen-induced asthma. Micronic particles in the atmosphere may be starch granules originating from pollen grains osmotically ruptured by rainwater. Ultrastructural and immunological characterisation of micronic particles collected from outdoor air filters confirm the presence of airborne starch granules. These are loaded with grass pollen allergens, occur in the atmosphere especially after rainfall, and correlate significantly with instances of allergic asthma. Diesel particles might also play a role in the transmission of grass pollen allergens and thus become an extra asthma trigger. A variation in the mode of release of micronic particles occurs in other species, such as birch, where such particles are derived from burst birch pollen tubes. These particles are positive for Bet v 1 and are starch granules which are released into the atmosphere after light rain as a result of pollen germination on, e.g., leaves. After subsequent rupture of pollen tubes their contents are released when conditions become drier.

The temporal sequence of allergic sensitization and onset of infantile eczema

Background: Eczema is commonly associated with sensitization in infants, but the causative role of sensitization in the development of eczema has been questioned.

Objective: To determine if allergic sensitization increases the risk of developing eczema, or alternatively, if eczema increases the risk of developing allergic sensitization.

Methods: We used data from the Melbourne Atopy Cohort Study, a prospective birth cohort of 552 infants with a family history of atopic disease. The main outcomes were risk of developing eczema from 6 months to 7 years of age in asymptomatic infants; and risk of developing sensitization, as measured by skin prick tests to milk, egg white, peanut, house dust mite, rye grass pollen and cat extracts, in previously unsensitized infants.

Results: Sensitization to food extracts at 6 months was associated with an increased risk of developing eczema [hazard ratio (HR) 1.63, 95% confidence interval 1.13–2.35] up to 7 years of age, after excluding infants with eczema in the first 6 months. However, eczema in the first 6 months was also associated with increased risk of new sensitization at both 1 year (HR 2.34, 1.38–3.98) and 2 years (HR 3.47, 1.65–7.32).

Conclusion: In some infants, sensitization precedes and predicts the development of eczema, while in others eczema precedes and predicts the development of sensitization. This indicates that there are multiple pathways to atopic eczema.

Thunderstorm asthma

Thunderstorms have often been linked to epidemics of asthma, especially during the grass flowering season; however, the precise mechanisms explaining this phenomenon are unknown. Evidence of high respirable allergen loadings in the air associated with specific meteorologic events combined with an analysis of pollen physiology suggests that rupture of airborne pollen can occur. Strong downdrafts and dry, cold outflows distinguish thunderstorm rain from frontal rain. The weather system of a mature thunderstorm likely entrains grass pollen into the cloud base, where pollen rupture would be enhanced, then transports the respirablesized fragments of pollen debris to ground level where outflows distribute them ahead of the rain. The conditions occurring at the onset of a thunderstorm might expose susceptible people to a rapid increase in concentrations of pollen allergens in the air that can readily deposit in the lower airways and initiate asthmatic reactions.

Source of Bet v 1 loaded inhalable particles from birch revealed

Allergenic proteins present in pollen grains, when inhaled, interact with the airways to cause an attack of asthma in susceptible humans. In one system, grass pollen grains rupture osmotically in rainfall, releasing allergen-containing inhalable particles into the atmosphere. In contrast, birch tree pollen grains do not rupture under these conditions, yet the major allergen, Bet v 1, has been detected in the atmosphere in inhalable particles of unknown origin. It is possible that Bet v 1 may diffuse from intact settled pollen grains and the allergenic material may again become airborne, interacting with settled fine particles from other sources prior to resuspension. This study investigates the mechanism for the release of birch pollen allergen-containing inhalable particles from pollen grains. We propose the hypothesis that (1) airborne birch pollen grains settle on nearby leaf surfaces; (2) then, following light rainfall, the grains germinate and, (3) later, pollen tubes burst, releasing inhalable particles carrying Bet v 1 into the atmospheric aerosol.   We used microscopic analyses of pollen behaviour following anther opening, a Burkard volumetric trap for pollen counts and a high volume air sampler with a two-stage cascade impactor for quantitative immunochemical analyses of Bet v 1. On dry days of high birch pollen count (48 grains/m3, 1.5 ng/m3 of Bet v 1), we found that the surfaces of birch leaves became coated with pollen. This ”pollen rain” is a source of secondary emission of allergens into the atmosphere. We observed that following light rainfall (<1 mm per day), about 80% of the birch pollen grains germinated, producing pollen tubes, especially in the sticky surface secretions of leaf glands. These pollen tubes may grow up to 300 μm in length prior to rupturing, each releasing about 400 starch granules coated with allergen molecules that may, after drying, be dispersed into the aerosol. On these days following light rainfall, the highest atmospheric levels of Bet v 1 (1.18 ng/m3) are associated with inhalable particles. Following heavy rainfall, both pollen and inhalable particles are washed from the atmosphere. Immunoprinting studies show that Bet v 1 is associated with starch granules rather than the smaller orbicules. Bet v 1 is present in the atmosphere in large particles, i.e. in particular pollen grains and in inhalable particles, i.e. in particular starch granules.

Development of plastids in pollen and tapetum of rye-grass, Lolium perenne L

Proplastids of both tapetal cells and microsporocytes were present early in anther development. Tapetal proplastids differentiated—probably into elaioplasts—at late microspore stage. The tapetal cytoplasm was completely resorbed by early tricellular pollen stage. Microspore proplastids differentiated into amyloplasts at early bicellular stage, and were present in both vegetative and generative cells. In the generative cell, the amyloplasts were ephemeral and apparently degenerated within autophagic vacuoles. Plastids were absent from sperm cells. Vegetative cell amyloplasts increased in number apparently by fission such that one amyloplast produced one amyloplast and one proplastid per division. Mature pollen grains were estimated to contain between 550 and 820 amyloplasts with only one starch granule per plastid.

Effects of electric fields on pollen rupture

RATIONALE: To determine whether the potential for previous termpollennext term fragmentation is increased during thunderstorms by exploring the previous termeffectsnext term of previous termelectricnext termprevious termfieldsnext term, with magnitude as found in the outdoor environment.

METHODS: Fresh previous termpollennext term grains were collected from bermudagrass flowers. A light microscope was modified with the addition of an previous termelectricnext termprevious termfieldnext term generated from a DC source (0-20 V) that was applied to the stage. Water was added to test for previous termpollennext termprevious termrupturenext term and to assess previous termpollennext term viability.

RESULTS: Bermuda grass previous termpollennext term did not previous termrupturenext term within 1 h of contact with water. Only after exposure to an previous termelectricnext termprevious termfieldnext term did Bermudagrass previous termpollennext term show a considerable amount of rupturing immediately upon immersion in water. The higher the voltage the previous termpollennext term is exposed to before coming into contact with water, the higher the percentage of previous termrupturenext term of the previous termpollennext term. previous termElectricnext termprevious termfieldsnext term, generated in the laboratory and of magnitude found during thunderstorms, affected the previous termpollennext term after as little as a 5 s exposure. The highest percentage of previous termrupturenext term occurred after exposures of at least 10 s: 80% previous termrupturenext term occurred after 10 s exposure at 10kVolts/m. This previous termeffectnext term is sustained for at least 15 min.

CONCLUSIONS: Thunderstorm regularly generate previous termelectricnext termprevious termfieldsnext term up to 5 kV/m in strength, and can reach 10kV/m, and cover several km in distance. The magnitude of the previous termelectricnext termprevious termfieldsnext term that affects the previous termpollennext term grains in the laboratory is low enough to be commonly found in the outdoor environment during thunderstorms. These previous termelectricnext termprevious termfields prime previous termpollen grains for more rapid release of allergenic particles.

Exotic grass fuel in south eastern Australia

One potentially serious impact of environmental weed invasion is an alteration to the fuel characteristics and subsequent fire regimes of an area. Considerable global evidence indicates that environmental weed invasion may produce 'new' fuel types, but there is little documented evidence of the impact of invasive exotic species on fuel characteristics and fire regimes in south-eastern Australia. This study on the impact of invasive exotic grasses in southeastern Australia has revealed major changes to fuel characteristics, particularly fuel loads.