Ragweed is in the Air: Ambrosia L. (Asteraceae) and Pollen Allergens in a Changing World.

Globally Ambrosia species (Asteraceae), commonly called ragweed, are recognized to be one of the most problematic groups of invasive weeds and one of the main allergenic genus. Climate and land-use change and air pollution are expected to promote ragweed spread, increase airborne ragweed pollen concentrations (the source of allergens), extend the pollen season, and promote longdistance transport of pollen or sub-pollen particles containing allergens. The allergenicity of pollen itself is going to increase. Likely, all these factors will have meaningful effects in the exacerbation of the sensitization to ragweed pollen and the severity of allergy symptoms. Globally the major health concern regards A. artemisiifolia, because of its very wide global distribution and highly invasive behavior. Together with A. artemisiifolia, also A. trifida and A. psilostachya are species of health concern distributed across different continents, widespread and invasive in several regions. The present review summarizes the characteristics of these species and gives an overview of factors contributing to their allergenicity.

Integrated biological and chemical characterisation of a pair of leonardesque canal lock gates.

The Museo Nazionale della Scienza e della Tecnologia "Leonardo da Vinci" in Milan is exposing two pairs of canal lock gates, used to control the water flow in Milan canal system, whose design appears in the Leonardo's Codex Atlanticus. The wood present in the gates has been deeply characterised by mean of a multidisciplinary investigation involving i) DNA barcoding of wood fragments; ii) microbial community characterisation, and iii) chemical analyses. DNA barcoding revealed that two fragments of the gates belonged to wood species widely used in the middle age: Fagus sylvatica and Picea abies. The chemical characterisations were based on the use of ionic liquid as dissolving medium in order to analyse the entire cell wall material by means of Gel Permeation Chromatography (GPC) and 2D-NMR-HSQC techniques. This multidisciplinary analytical approach was able to highlight the complex nature of the degradation occurred during the gate operation (XVI-XVIII centuries): an intricate interplay between microbial populations (i.e. Shewanella), inorganic factors (i.e. iron from nails), physical factors and the lignocellulosic material.

Allergenicity at component level of sub-pollen particles from different sources obtained by osmolar shock: A molecular approach to thunderstorm-related asthma outbreaks.

BACKGROUND: The so-called "thunderstorm asthma" (TA) is an uncommon but dramatic outbreak of asthma attacks occurring during a thunderstorm in the pollen and moulds season. Mechanisms which make the pollen able to enter the deeper airways and provoke severe asthma symptoms are still unclear. OBJECTIVE: To test the hypothesis that sub-pollen particles (SPPs) originated from the rupture by an osmotic shock of pollen associated with TA contain allergens. METHODS: After hydration, SPPs released from pollen grains of grass, pellitory, olive, cypress, ragweed and birch were isolated and determined by microscopy. Allergens were determined by in vitro ELISA inhibition tests indirectly using the sera from 10 polyreactive patients. An inhibition <50% was considered as negative, 50%-75% moderate and > 75% complete. RESULTS: The inhibition experiments showed that the SPPs from birch and cypress were unable to inhibit serum IgE reactivity to Bet v 1 and Cup a 1, respectively. Ragweed SPPs inhibited ragweed pollen extract and Amb a 1 by 75.8 ± 0.11% and 81.2 ± 0.15%, respectively. Olive and pellitory SPPs retained almost the whole IgE-binding capability in all cases tested. Grass SPPs inhibited 32 ± 0.06% of Lolium perenne Lol p 1 and 65% of Phleum pratense extracts, but results were highly variable for individual allergens (97.5%-0.03% for Phl p 2, 45.3 ± 0.12% for Phl p 5, 24.7 ± 0.22% for Phl p 6, and 38.3 ± 0.2% for Phl p 1). CONCLUSIONS: Inhibition experiments confirm the hypothesis that SSPs obtained after the osmotic shock of pollen involved in TA, namely grass, pellitory and olive tree pollen, contain allergens and therefore they can induce severe asthma attacks during thunderstorms.

Ambrosia artemisiifolia L. temperature-responsive traits influencing the prevalence and severity of pollinosis: a study in controlled conditions.

BACKGROUND: Ambrosia artemisiifolia L. is one of the most important sources of allergenic pollen in many regions of the world. Its health impact increased over the last decades and is expected to further increase in consequence of climate change. However little information is available on the specific role played by temperature on allergy rising. The aim of this work was to evaluate the effect of temperature on A. artemisiifolia growth, flowering and pollen allergenicity, the major plant functional traits influencing the prevalence and severity of pollinosis. RESULTS: Plants were grown in controlled conditions at three thermal regimes: "Low" (LT: 18-14 °C light-dark), "Intermediate" (IT: 24-20 °C light-dark) and "High" (HT: 30-26 °C light-dark). During plant development, plant vegetative and reproductive morpho-functional traits were measured and, at the end of plant life-cycle, mature pollen was collected and analyzed for its allergenic properties by slot blot, 1D- and 2D-western blot (by using a pool of sera from ragweed-allergic patients) and liquid chromatography-tandem mass spectrometry. A. artemisiifolia showed a great development plasticity leading to a broad temperature tolerance. Shoot architecture, growth rate, number of male inflorescence and pollen allergenicity were temperature-responsive traits. Pollen allergenicity increased in parallel with temperature and differences were related to allergen synthesis and Amb a 1-IgE-binding. Flavonoids whose concentration in pollen decreased with the increase of temperature, were recognized as the cause of the negligible Amb a 1-IgE binding in LT pollen. CONCLUSIONS: Results show that temperature governs plant development and pollen allergenicity influencing the temporal and spatial magnitude of subject exposure to allergens.

Nitration of pollen aeroallergens by nitrate ion in conditions simulating the liquid water phase of atmospheric particles.

Pollen aeroallergens are present in atmospheric particulate matter (PM) where they can be found in coarse biological particles such as pollen grains (aerodynamic diameter dae>10µm), as well as fragments in the finest respirable particles (PM2.5; dae<2.5µm). Nitration of tyrosine residues in pollen allergenic proteins can occur in polluted air, and inhalation and deposition of these nitrated proteins in the human respiratory tract may lead to adverse health effects by enhancing the allergic response in population. Previous studies investigated protein nitration by atmospheric gaseous pollutants such as nitrogen dioxide and ozone. In this work we report, for the first time, a study on protein nitration by nitrate ion in aqueous solution, at nitrate concentrations and pH conditions simulating those occurring in the atmospheric aerosol liquid water phase. Experiments have been carried out on the Bovine serum albumin (BSA) protein and the recombinant Phleum pratense allergen (Phl p 2) both in the dark and under UV-A irradiation (range 4-90Wm-2) to take into account thermal and/or photochemical nitration processes. For the latter protein, modifications in the allergic response after treatment with nitrate solutions have been evaluated by immunoblot analyses using sera from grass-allergic patients. Experimental results in bulk solutions showed that protein nitration in the dark occurs only in dilute nitrate solutions and under very acidic conditions (pH<3 for BSA; pH<2.2 for Phl p 2), while nitration is always observed (at pH0.5-5) under UV-A irradiation, both in dilute and concentrated nitrate solutions, being significantly enhanced at the lowest pH values. In some cases, protein nitration resulted in an increase of the allergic response.

Is ragweed pollen allergenicity governed by environmental conditions during plant growth and flowering?

Pollen allergenicity is one of the main factors influencing the prevalence and/or severity of allergic diseases. However, how genotype and environment contribute to ragweed pollen allergenicity has still to be established. To throw some light on the factors governing allergenicity, in this work 180 ragweed plants from three Regions (Canada, France, Italy) were grown in both controlled (constant) and standard environmental conditions (seasonal changes in temperature, relative humidity and light). Pollen from single plants was characterized for its allergenic potency and for the underlying regulation mechanisms by studying the qualitative and quantitative variations of the main isoforms of the major ragweed allergen Amb a 1. Results showed a statistically higher variability in allergenicity of pollen from standard conditions than from controlled conditions growing plants. This variability was due to differences among single plants, regardless of their origin, and was not ascribed to differences in the expression and IgE reactivity of individual Amb a 1 isoforms but rather to quantitative differences involving all the studied isoforms. It suggests that the allergenic potency of ragweed pollen and thus the severity of ragweed pollinosis mainly depends on environmental conditions during plant growth and flowering, which regulate the total Amb a 1 content.

Concomitant sensitization to ragweed and mugwort pollen: who is who in clinical allergy?

BACKGROUND: In many areas of Europe, double sensitization to ragweed and mugwort is common, and because of the overlapping flowering periods of the 2 plants, it is not possible to diagnose the primary sensitizing allergen source and hence to determine the proper immunotherapy. OBJECTIVES: To elucidate whether double-sensitized patients are cosensitized or cross-sensitized and, in the latter case, to define the primary sensitizer. METHODS: Serum samples from 34 patients with late summer respiratory allergy underwent skin prick testing with whole ragweed, and mugwort extracts were analyzed for their reactivity to recombinant Art v 1 and Amb a 1 by ImmunoCAP and then to Amb a 1, Art v 6, and Art v 1 isoforms by a proteomic approach. In double reactors, the primary sensitizing sources were detected by inhibition experiments. RESULTS: Serum samples from patients monosensitized to ragweed contained IgE to epitopes specific of all Amb a 1 isoforms. In contrast, serum samples from double reactors found to be primarily sensitized to mugwort reacted to Art v 1 and Art v 6 and cross-reacted to a few Amb a 1 isoforms. Finally, serum samples from double reactors found to be primarily sensitized to ragweed contained IgE reacting to all Amb a 1 isoforms, part of which cross-reacted to Art v 6. We did not find cosensitized patients. CONCLUSION: This study found that Art v 6 plays an important role in mugwort allergy and that the cross-reactivity between Art v 6 and Amb a 1 is frequent, bidirectional, and clinically relevant in the area of Milan.

Sporophytic and gametophytic functions of the cell cycle-associated Mob1 gene in Arabidopsis thaliana L.

Mob1 genes are primarily involved in the cell cycle progression and mitosis exit in yeasts and animals. The function of a Mob1-like gene (At5g45550) from Arabidopsis thaliana was investigated using RNAi and immunological staining. AtMob1-like RNAi silenced lines showed a reduced radial expansion of the inflorescence stem and a reduced elongation zone of the primary root. Morphological features of plant organs were accompanied by a reduction in cell size. The fertility of AtMob1-like RNAi silenced lines was very low as seed production was strongly reduced. About 2% of the progeny of AtMob1-like RNAi silenced plants were tetraploid. The female and male sporogenesis was affected differentially. The ovules developed irregularly and one third of the megaspores and embryo sacs degenerated prematurely. Up to 20% of the ovules produced binucleated megaspores that failed to develop further, being their degeneration likely accompanied with a delayed programmed cell death. The anthers produced about 30% of aborted pollen grains, showing also a strong variation in their size. Together, the results show that Arabidopsis MOB1-like is required to regulate cell expansion and cell division, presumably by affecting the mitotic as well as the meiotic cell cycle.

Genetic and DNA-methylation changes induced by potassium dichromate in Brassica napus L.

This study evaluated genetic and DNA methylation alteration induced by potassium dichromate in Brassica napus L. plants. Amplified fragment length polymorphism (AFLP) and selective amplification of polymorphic loci (SAMPL) tests revealed dose-related increases in sequence alterations in plantlets exposed to 10-200 mg/l potassium dichromate. Individual plantlets exposed to chromium under similar conditions showed different AFLP and SAMPL DNA profiles. These observations suggest random DNA mutation in response to potassium dichromate and argue against preferential sites for mutation. DNA methylation changes in response to chromium treatment were also evaluated. Two complementary approaches were applied: (i) immunolabelling, using a monoclonal antibody against 5-methylcytosine; and (ii) methylation-sensitive amplified polymorphism (MSAP). Immunolabelling showed cytosine-hypermethylation in the Brassica napus L. genome when plants were treated with potassium dichromate. MSAP analysis showed extensive methylation changes in CCGG-sequences, with the net result being genome-wide hypermethylation. These results showed a clear DNA alteration in plants as a response to chromium exposure and the effect was dose-dependent. DNA polymorphism detected by different markers supports the effectiveness of the use of these tools for the investigation of environmental toxicology and for evaluating the concentration of pollutants by DNA analysis in plants.