Relationship between IgE Levels Specific for Ragweed Pollen Extract, Amb a 1 and Cross-Reactive Allergen Molecules.

Ragweed (Ambrosia artemisiifolia) pollen is a major endemic allergen source responsible for severe allergic manifestations in IgE-sensitized allergic patients. It contains the major allergen Amb a 1 and cross-reactive allergen molecules, such as the cytoskeletal protein profilin, Amb a 8 and calcium-binding allergens Amb a 9 and Amb a 10. To assess the importance of Amb a 1, profilin and calcium-binding allergen, the IgE reactivity profiles of clinically well-characterized 150 ragweed pollen-allergic patients were analysed regarding specific IgE levels for Amb a 1 and cross-reactive allergen molecules by quantitative ImmunoCAP measurements, IgE ELISA and by basophil activation experiments. By quantifying allergen-specific IgE levels we found that Amb a 1-specific IgE levels accounted for more than 50% of ragweed pollen-specific IgE in the majority of ragweed pollen-allergic patients. However, approximately 20% of patients were sensitized to profilin and the calcium-binding allergens, Amb a 9 and Amb a 10, respectively. As shown by IgE inhibition experiments, Amb a 8 showed extensive cross-reactivity with profilins from birch (Bet v 2), timothy grass (Phl p 12) and mugwort pollen (Art v 4) and was identified as a highly allergenic molecule by basophil activation testing. Our study indicates that molecular diagnosis performed by the quantification of specific IgE to Amb a 1, Amb a 8, Amb a 9 and Amb a 10 is useful to diagnose genuine sensitization to ragweed pollen and to identify patients who are sensitized to highly cross-reactive allergen molecules present in pollen from unrelated plants, in order to enable precision medicine-based approaches for the treatment and prevention of pollen allergy in areas with complex pollen sensitization.

Ragweed Major Allergen Amb a 11 Recombinant Production and Clinical Implications.

Ragweed pollen is highly allergenic and elicits type I hypersensitivity reactions in the exposed populations. Amb a 11 is a recently discovered component of this pollen, and its biological role in allergy is still being researched. In our study, ragweed allergy patients were recruited prospectively over a three-year period; a comprehensive questionnaire was administered, and sera were collected and stored. The production of recombinant Amb a 11 was achieved in parallel with patients' recruitment. The gene coding for mature protein was inserted in E. coli and in Sf9 Spodoptera frugiperda cells. The recombinant allergens (designated eAmb a 11 and iAmb a 11) were tested for His-tag presence in Western blot. IgE reactivity was evaluated in 150 patients' sera for both recombinant allergen forms in ELISA, with 5 positive sera being tested further by hRBL (humanized rat basophilic leukemia) hexosaminidase release assay. Both allergen forms were proven to be IgE-reactive His-tagged proteins, with an extensive overlap of positive sera (92 toward the former recombinant allergen, 100 toward the latter) and an overall Amb a 11 sensitization prevalence estimated at 68.67%. The hRBL mediator release assay revealed a significant, slightly weaker effect of recombinant allergens when compared with nAmb a 1. Sensitization to this major allergen appears to be associated with more severe asthma symptoms (OR = 4.71, 95% CI = 1.81-12.21). In conclusion, recombinant Amb a 11 is a bona fide allergen, which is IgE-reactive and an inducer of hRBL degranulation. It is an important IgE-reactive component from ragweed pollen, with high IgE sensitization prevalence in the sample population and allergenicity of the recombinant allergen comparable to Amb a 1.

A new cysteine protease allergen from Ambrosia trifida pollen: proforms and mature forms.

Giant ragweed (Ambrosia trifida) pollen is closely associated with respiratory allergy in late summer and autumn, and the prevalence of giant ragweed pollen allergy progressively increases. Compared with short ragweed (Ambrosia artemisiifolia), allergenic components from giant ragweed pollen are poorly investigated. To promote component-resolved diagnosis and treatment for giant ragweed pollen allergy, it becomes necessary to identify and characterize unknown allergens from giant ragweed pollen. In the present study, we identified and characterized a new cysteine-protease (CP) allergen from giant ragweed pollen, named as Amb t CP. The cloned Amb t CP gene encoded 387 amino acids. Recombinant Amb t CP (rAmb t CP) and natural Amb t CP (nAmb t CP) were purified by high-affinity Ni2+ resin and immunoaffinity chromatography respectively. During refolding, purified rAmb t CP could autocatalytically converted to its mature forms displaying a higher enzymatic activity. Moreover, the autocatalytic conversion of proforms to mature forms of nAmb t CP could cause their amount to change in giant ragweed pollen extracts. Then, the allergenicity of Amb t CP was characterized: 23 (33.8%) of 68 Chinese patients with ragweed pollen allergy showed positive IgE binding to nAmb t CP by enzyme-linked immunosorbent assay (ELISA); the result of subsequent ELISA showed that IgE-binding activity of proforms and mature forms of rAmb t CP was different, with positive rate of 39.1% (9/23) and 47.8% (11/23) respectively; Amb t CP showed IgE cross-reactivity with the CP components from short ragweed, Artemisia annua and Artemisia sieversiana pollen. Our findings will help to promote component-resolved diagnosis and treatment for giant ragweed pollen allergy, standardize allergen products and individualize allergen-specific immunotherapy.

Glyphosate resistance in Ambrosia trifida: Part 2. Rapid response physiology and non-target-site resistance.

BACKGROUND: The glyphosate-resistant rapid response (GR RR) resistance mechanism in Ambrosia trifida is not due to target-site resistance (TSR) mechanisms. This study explores the physiology of the rapid response and the possibility of reduced translocation and vacuolar sequestration as non-target-site resistance (NTSR) mechanisms. RESULTS: GR RR leaf discs accumulated hydrogen peroxide within minutes of glyphosate exposure, but only in mature leaf tissue. The rapid response required energy either as light or exogenous sucrose. The combination of phenylalanine and tyrosine inhibited the rapid response in a dose-dependent manner. Reduced glyphosate translocation was observed in GR RR, but only when associated with tissue death caused by the rapid response. Nuclear magnetic resonance studies indicated that glyphosate enters the cytoplasm and reaches chloroplasts, and it is not moved into the vacuole of GR RR, GR non-rapid response or glyphosate-susceptible A. trifida. CONCLUSION: The GR RR mechanism of resistance is not associated with vacuole sequestration of glyphosate, and the observed reduced translocation is likely a consequence of rapid tissue death. Rapid cell death was inhibited by exogenous application of aromatic amino acids phenylalanine and tyrosine. The mechanism by which these amino acids inhibit rapid cell death in the GR RR phenotype remains unknown, and it could involve glyphosate phytotoxicity or other agents generating reactive oxygen species. Implications of these findings are discussed. The GR RR mechanism is distinct from the currently described glyphosate TSR or NTSR mechanisms in other species. © 2017 Society of Chemical Industry.

Characterization and standardization of allergen extracts.

This paper summarizes the development of the extraction and characterization of allergens responsible for the induction of immunoglobulin (lg) E-induced allergies from the beginning of the 20th century, including the nomenclature of allergens. The majority of papers characterizing allergens and allergen extracts state that the lack of standardization of allergen extracts is the reason for the paper, and so it has been for more than 100 years. A natural part of that process might be the isolation of an allergen molecule and this starts the speculation of 'what makes that allergen an allergen?' To achieve the perfect standardization is a desirable end that is still awaited. So far none of these problems have been finally solved. I started in allergy shortly after the discovery of IgE in 1967. Since that time the history as I remember it is based on the literature, my interpretation of it, and of course may be a little biased due to personal prejudice! The history of the last 10-15 years has still not matured and it might be a little early to draw conclusions. However, at the end of this chapter I do dare to make a few conclusions after having followed the development in this field for 40 years. As this is history it is not meant to be either comprehensive or technically and scientifically precise in all aspects, but rather draws on some thoughts as to what in my mind have been important developments until now. Specific techniques are only mentioned by name and not intended to be discussed in depth. This activity has, however, pushed me to reflect on my hopes and speculations at the time of my introduction to the field of allergen chemistry. To my surprise I realize that far more than I ever expected at that time has been fulfilled. It has been extremely exciting to be a part of that development.

Changes in sensitization rate to weed allergens in children with increased weeds pollen counts in Seoul metropolitan area.

The prevalence of allergic diseases in children has increased for several decades. We evaluated the correlation between pollen count of weeds and their sensitization rate in Seoul, 1997-2009. Airborne particles carrying allergens were collected daily from 3 stations around Seoul. Skin prick tests to pollen were performed on children with allergic diseases. Ragweed pollen gradually increased between 1999 and 2005, decreased after 2005 and plateaued until 2009 (peak counts, 67 in 2003, 145 in 2005 and 83 grains/m(3)/day in 2007). Japanese hop pollen increased between 2002 and 2009 (peak counts, 212 in 2006 and 492 grains/m(3)/day in 2009). Sensitization rates to weed pollen, especially ragweed and Japanese hop in children with allergic diseases, increased annually (ragweed, 2.2% in 2000 and 2.8% in 2002; Japanese hop, 1.4% in 2000 and 1.9% in 2002). The age for sensitization to pollen gradually became younger since 2000 (4 to 6 yr of age, 3.5% in 1997 and 6.2% in 2009; 7 to 9 yr of age, 4.2% in 1997 and 6.4% in 2009). In conclusion, sensitization rates for weed pollens increase in Korean children given increasing pollen counts of ragweed and Japanese hop.

Ozone affects pollen viability and NAD(P)H oxidase release from Ambrosia artemisiifolia pollen.

Air pollution is frequently proposed as a cause of the increased incidence of allergy in industrialised countries. We investigated the impact of ozone (O(3)) on reactive oxygen species (ROS) and allergen content of ragweed pollen (Ambrosia artemisiifolia). Pollen was exposed to acute O(3) fumigation, with analysis of pollen viability, ROS and nitric oxide (NO) content, activity of nicotinamide adenine dinucleotide phosphate (NAD[P]H) oxidase, and expression of major allergens. There was decreased pollen viability after O(3) fumigation, which indicates damage to the pollen membrane system, although the ROS and NO contents were not changed or were only slightly induced, respectively. Ozone exposure induced a significant enhancement of the ROS-generating enzyme NAD(P)H oxidase. The expression of the allergen Amb a 1 was not affected by O(3), determined from the mRNA levels of the major allergens. We conclude that O(3) can increase ragweed pollen allergenicity through stimulation of ROS-generating NAD(P)H oxidase.

Transitory effects of elevated atmospheric CO2 on fine root dynamics in an arid ecosystem do not increase long-term soil carbon input from fine root litter.

Experimental increases in atmospheric CO2 often increase root production over time, potentially increasing soil carbon (C) sequestration. Effects of elevated atmospheric CO2 on fine root dynamics in a Mojave desert ecosystem were examined for the last 4.5 yr of a long-term (10-yr) free air CO2 enrichment (FACE) study at the Nevada desert FACE facility (NDFF). Sets of minirhizotron tubes were installed at the beginning of the NDFF experiment to characterize rooting dynamics of the dominant shrub Larrea tridentata, the codominant shrub Ambrosia dumosa and the plant community as a whole. Although significant treatment effects occurred sporadically for some fine root measurements, differences were transitory and often in opposite directions during other time-periods. Nonetheless, earlier root growth under elevated CO2 helped sustain increased assimilation and shoot growth. Overall CO2 treatment effects on fine root standing crop, production, loss, turnover, persistence and depth distribution were not significant for all sampling locations. These results were similar to those that occurred near the beginning of the NDFF experiment but unlike those in other ecosystems. Thus, increased C input into soils is unlikely to occur from fine root litter under elevated atmospheric CO2 in this arid ecosystem.

Forecasting ragweed pollen characteristics with nonparametric regression methods over the most polluted areas in Europe.

Nonparametric time-varying regression methods were developed to forecast daily ragweed pollen concentration, and the probability of the exceedance of a given concentration threshold 1 day ahead. Five-day and 10-day predictions of the start and end of the pollen season were also addressed with a nonparametric regression technique combining regression analysis with the method of temperature sum. Our methods were applied to three of the most polluted regions in Europe, namely Lyon (Rhône Valley, France), Legnano (Po River Plain, Italy) and Szeged (Great Plain, Hungary). For a 1-day prediction of both the daily pollen concentration and daily threshold exceedance, the order of these cities from the smallest to largest prediction errors was Legnano, Lyon, Szeged and Legnano, Szeged, Lyon, respectively. The most important predictor for each location was the pollen concentration of previous days. The second main predictor was precipitation for Lyon, and temperature for Legnano and Szeged. Wind speed should be considered for daily concentration at Legnano, and for daily pollen threshold exceedances at Lyon and Szeged. Prediction capabilities compared to the annual cycles for the start and end of the pollen season decreased from west to east. The order of the cities from the lowest to largest errors for the end of the pollen season was Lyon, Legnano, Szeged for both the 5- and 10-day predictions, while for the start of the pollen season the order was Legnano, Lyon, Szeged for 5-day predictions, and Legnano, Szeged, Lyon for 10-day predictions.

Biosynthesis of polyacetylenes in Ambrosia maritima hairy roots.

Thiophene A and thiophene A diol are the major polyacetylenes isolated from the hairy roots of Ambrosia maritima (Asteraceae) cultured under continuous light irradiation. The biosynthesis of thiophene A was studied using [1-(13)C]-, [2-(13)C]-, [1,2-(13)C]-acetates. The biosynthesis of thiarubrine A, produced by hairy roots cultured in the dark, was studied using [1,2-(13)C]-acetates and [18-(13)C]-linoleic acid. Our results suggest a catabolic pathway for polyacetylenes biosynthesis from linoleic acid in hairy roots of Ambrosia maritima.

Interaction of the onset of spring and elevated atmospheric CO2 on ragweed (Ambrosia artemisiifolia L.) pollen production.

Increasing atmospheric carbon dioxide is responsible for climate changes that are having widespread effects on biological systems. One of the clearest changes is earlier onset of spring and lengthening of the growing season. We designed the present study to examine the interactive effects of timing of dormancy release of seeds with low and high atmospheric CO2 on biomass, reproduction, and phenology in ragweed plants (Ambrosia artemisiifolia L.), which produce highly allergenic pollen. We released ragweed seeds from dormancy at three 15-day intervals and grew plants in climate-controlled glass-houses at either ambient or 700-ppm CO2 concentrations, placing open-top bags over influorescences to capture pollen. Measurements of plant height and weight; inflorescence number, weight, and length; and days to anthesis and anthesis date were made on each plant, and whole-plant pollen productivity was estimated from an allometric-based model. Timing and CO2 interacted to influence pollen production. At ambient CO2 levels, the earlier cohort acquired a greater biomass, a higher average weight per inflorescence, and a larger number of influorescences; flowered earlier; and had 54.8% greater pollen production than did the latest cohort. At high CO2 levels, plants showed greater biomass and reproductive effort compared with those in ambient CO2 but only for later cohorts. In the early cohort, pollen production was similar under ambient and high CO2, but in the middle and late cohorts, high CO2 increased pollen production by 32% and 55%, respectively, compared with ambient CO2 levels. Overall, ragweed pollen production can be expected to increase significantly under predicted future climate conditions.

[Absorption and accumulation of heavy metals by plants around a smelter].

The study on the absorption and accumulation of heavy metals lead, zinc, copper and cadmium by 8 plant species around a smelter showed that the metals accumulation by plants differed with plant species, their parts, and kinds of metals. Abutilon theophrasti had a higher capability of absorbing and accumulating Pb, Conyza canadensis, Ambrosia trifida, Polygonumn lapathifolium, A. theophrasti, Solanum nigrum, Chenopodium acuminatum and Helianthus tuberosus had a higher capability of absorbing and accumulating Zn, C. acuminatunz and A. theophrasti had a higher capability of absorbing and accumulating Cu, and S. nigrum, C. acuminatum, A. theophrasti, P. lapathifolium and C. canadensis had a higher capability of absorbing and accumulating Cd. These plants had TF values higher than 1, and were suitable for phytoextraction to remedy polluted soil. As for the plants with TF values lower than 1, they were suitable as the phytostabilizers of heavy metals-contaminated lands.

[Gas exchange features of Ambrosia artemisiifolia leaves and fruits and their correlations with soil heavy metals].

Ambrosia artemisiifolia can survive well in the habitats of heavy human disturbance and partial soil pollution. Weather its photosynthetic features benefit their survival is worthwhile to concern. With a refuse dump in Changchun City (43 degrees 50'N, 125 degrees 23'E) as study site, this paper analyzed the contents of soil Cu, Pb, Zn, Mn, Cr, Co, Ni, Cd, As, Sb and Hg at ten plots, and measured in situ the gas exchange in A. artemisiifolia leaves and young fruits. The results showed that the study site was slightly contaminated by Ni, but the contents of other soil heavy metals were approached to or substantially lower than their threshold values. The net photosynthetic rate of leaves ranged from 1.88 to 9.41 micromol x m(-2) x s(-1), while that of young fruits could be up to 2. 81 micromol x m(-2) s(-1). Averagely, the respiration rate, stomatal conductance, photosynthetic rate, and water utilization efficiency of leaves were 1.81 micromol x m(-2) x s(-1), 75.7 mmol x m(-2) x s(-1), 6.05 micromol x m(-2) x s(-1), and 4.72 micromol CO2 x mmol(-1) H2O, being 5.26, 0.64, 1.31 and 1.69 times as much as those of young fruits, respectively, indicating that the respiratory and photosynthetic capacities and water use efficiency of A. artemisiifolia young fruits were equivalent to or higher than those of its leaves. Many test heavy metals, such as Cu, Pb, Zn, Cd, As, Sb and Hg, had no significant effects on the gas exchange features of leaves and fruits, but there were significant correlations of Ni and Cr with the stomatal conductance and water use efficiency of leaves and young fruits, Cr with the gross photosynthesis of leaves, and As with the stomatal conductance of young fruits, suggesting that a majority of test soil heavy metals had no direct effects on the gas exchange in A. artemisiifolia leaves and fruits, but soil Ni, Cr and As with the contents approached to or substantially lower than the threshold values could affect the gas exchange features of A. artemisiifolia.