Atmospheric exposure to the major Artemisia pollen allergen (Art v 1): Seasonality, impact of weather, and clinical implications.

Artemisia pollen grains are important aeroallergens worldwide. The amount of allergenic proteins produced by pollen, or pollen allergenicity, is regulated by both genes and the environment. As a result, even closely related plant taxa may release pollen with distinctly different allergen contents. Here, we determined the variability in atmospheric exposure to the major Artemisia pollen allergen, Art v 1, during the pollination seasons of two common species, i.e., A. vulgaris (early flowering species) and A. campestris (late flowering species), in Poznan, Poland (2013-2015). Artemisia pollen grains were collected using Hirst-type volumetric trap, while Art v 1 was collected by a two-stage cascade impactor (PM10 and PM>10 air fractions) and quantified by immunoenzymatic analysis. The results showed that daily Art v 1 levels correlated significantly with mean daily concentrations of Artemisia pollen (from r = 0.426 to r = 0.949, depending on air fraction and peak of the season). Significant differences were observed between 1) the median pollen allergenicity in different seasons (from 2.5 to 4.7 pg Art v 1/pollen) and 2) the median pollen allergenicity in different peak periods of the season (from 1.8 to 6.7 pg Art v 1/pollen). During the late peak (flowering of A. campestris), the median pollen allergenicity was significantly higher (on average by 63%, p < 0.05) than that during A. vulgaris flowering. The highest mean seasonal pollen allergenicity was observed during the wettest season, while the lowest was observed during the driest season (from July-August). In summary, our study showed distinct differences in Artemisia pollen allergenicity, that were not only related to daily and seasonal variability, which may exceed 800% and 80%, respectively but also noticeable when two common Artemisia species were compared. Therefore, we argue that variability in pollen allergenicity (both seasonal and species-specific) should be considered in future studies assessing pollen exposure.

Cross-sensitization to Artemisia and Ambrosia pollen allergens in an area located outside of the current distribution range of Ambrosia.

INTRODUCTION: The role of long-distance transported (LDT) Ambrosia pollen in inducing new sensitization and affecting sensitization rates in Artemisia-sensitized patients is unclear. AIM: The aim of this study was to estimate the degree of cross-sensitization to Ambrosia/Artemisia allergens in citizens of Poznan (Western Poland). This area is covered by extensive Artemisia populations but does not currently have local Ambrosia populations. MATERIAL AND METHODS: Sera of 119 patients were tested by fluoroenzyme immunoassay (CAP-FEIA system) against pollen allergen extracts of Artemisia vulgaris and Ambrosia artemisiifolia, an allergenic component of A. vulgaris (nArt v 1), and an allergenic component of A. artemisiifolia (nAmb a 1). Skin prick tests (SPTs, n = 86) were performed with pollen allergen extracts of A. vulgaris and A. artemisiifolia. Artemisia and Ambrosia pollen in ambient air was collected (1996-2013) by a Hirst type volumetric trap sited at roof level (33 m). RESULTS: The SPT showed that the prevalence of sensitization to Ambrosia and Artemisia pollen exceeded 3.5%, and 10.5%, respectively. The measurements of IgE in blood serum (CAP-FEIA) revealed that among Ambrosia-sensitized patients 90.1% (20/22 patients) were concomitantly sensitized to Artemisia. 59.1% (13/22) of these patients reacted to nArt v 1, suggesting primary sensitization to Artemisia pollen. Only 2 (9.1%) patients were mono-sensitized to Ambrosia pollen extract, but surprisingly not to nAmb a 1. CONCLUSIONS: The LDT Ambrosia pollen had a negligible effect on the rate of sensitization to Ambrosia allergens in Poznan and did not increase the prevalence of sensitization to Artemisia pollen in this region. However, the majority of patients showing hypersensitization to Artemisia pollen might also present symptoms during elevated episodes of LDT of Ambrosia pollen.

Flowering phenology and potential pollen emission of three Artemisia species in relation to airborne pollen data in Poznan (Western Poland).

Artemisia pollen is an important allergen in Europe. In Poznan (Western Poland), three Artemisia species, A. vulgaris, A. campestris and A. absinthium, are widely distributed. However, the contributions of these species to the total airborne pollen are unknown. The aim of the study was to determine the flowering phenology and pollen production of the three abovementioned species and to construct a model of potential Artemisia pollen emission in the study area. Phenological observations were conducted in 2012 at six sites in Poznan using a BBCH phenological scale. Pollen production was estimated by counting the pollen grains per flower and recalculating the totals per inflorescence, plant and population in the study area. Airborne pollen concentrations were obtained using a Hirst-type volumetric trap located in the study area. Artemisia vulgaris began to flower the earliest, followed by A. absinthium and then A. campestris. The flowering of A. vulgaris corresponded to the first peak in the airborne pollen level, and the flowering of A. campestris coincided with the second pollen peak. The highest amounts of pollen per single plant were produced by A. vulgaris and A. absinthium. A. campestris produced considerably less pollen, however, due to its common occurrence, it contributed markedly (30 %) to the summation of total of recorded pollen. A. vulgaris is the most important pollen source in Poznan, but the roles of two other Artemisia species cannot be ignored. In particular, A. campestris should be considered as an important pollen contributor and likely might be one of the main causes of allergic reactions during late summer.

Isoenergetic microarrays to study the structure and interactions of DsrA and OxyS RNAs in two- and three-component complexes.

Information on the secondary structure and interactions of RNA is important to understand the biological function of RNA as well as in applying RNA as a tool for therapeutic purposes. Recently, the isoenergetic microarray mapping method was developed to improve the prediction of RNA secondary structure. Herein, for the first time, isoenergetic microarrays were used to study the binding of RNA to protein or other RNAs as well as the interactions of two different RNAs and protein in a three-component complex. The RNAs used as models were the regulatory DsrA and OxyS RNAs from Escherichia coli, the fragments of their target mRNAs (fhlA and rpoS), and their complexes with Hfq protein. The collected results showed the advantages and some limitations of microarray mapping.

LNA-modified primers drastically improve hybridization to target RNA and reverse transcription.

Knowledge about the structure of RNA is crucial to understanding its biological activities. Very often, the presence of unusually thermodynamically stable structural fragments in RNAs, such as hairpins, makes it impossible to apply primer extension to visualize the results of chemical mapping experiments. However, replacement of DNA primers with LNA-modified primers overcomes this limitation. This approach was tested successfully on regulatory OxyS RNA and DsrA RNA from Escherichia coli.

The thermodynamics of 3'-terminal pyrene and guanosine for the design of isoenergetic 2'-O-methyl-RNA-LNA chimeric oligonucleotide probes of RNA structure.

To facilitate design of short isoenergetic hybridization probes for RNA, we report the influence of adding 5'- or 3'-terminal 2'-O-methylguanosine (GM), LNA-guanosine (GL), or 3'-terminal pyrene pseudo-nucleotide (PPN) on the thermodynamic stability of 2'-O-methyl-RNA/RNA (2'-O-Me-RNA/RNA) duplexes with sequences 5'CMGMGMCMAM/3'AAXGCCGUXAA, where X is A, C, G, or U. A 3'-terminal GM or GL added to the 2'-O-Me-RNA strand to form a G-A, G-G or G-U mismatch enhances thermodynamic stability (DeltaDeltaG degrees 37) of the 2'-O-Me-RNA/RNA duplexes on average by 0.7 and 1.5 kcal/mol, respectively. A 3'-terminal GM or GL in a GM-C or GL-C pair stabilizes the 2'-O-Me-RNA/RNA duplex by 2.6 and 3.4 kcal/mol, respectively. A 5'-terminal GM or GL in a G-A or G-G mismatch provided less stabilization in comparison with a 3'-terminal G-A or G-G mismatch, but more stabilization in a G-C or G-U pair. In contrast to guanosine derivatives, pyrene residue (P) as PPN at the 3'-terminal position enhances thermodynamic stability of the 2'-O-Me-RNA/RNA duplexes on average by 2.3 +/- 0.1 kcal/mol, relatively independent of the type of ribonucleotide placed in the opposite strand. The thermodynamic data can be applied to design 2'-O-Me-RNA/RNA duplexes with enhanced thermodynamic stability that is also sequence independent. This is useful for design of hybridization probes to interrogate RNA structure and/or expression by microarray and other methods.