Epicutaneous immunization with ovalbumin and CpG induces TH1/TH17 cytokines, which regulate IgE and IgG2a production

Background: Subcutaneous allergen-specific immunotherapy is a standard route for the immunotherapy of allergic diseases. It modulates the course of allergy and can generate long-term remission. However, subcutaneous allergen-specific immunotherapy can also induce anaphylaxis in some patients, and therefore additional routes of administration should be investigated to improve the safety and tolerability of immunotherapy. Objective: We sought to determine whether epicutaneous treatment with antigen in the presence of a Toll-like receptor 9 agonist can suppress TH2-mediated responses in an antigenspecific manner. Methods: Epicutaneous immunization was performed by applying a skin patch soaked with ovalbumin (OVA) plus CpG, and its suppressor activity was determined by using the mouse model of atopic dermatitis. Finally, adoptive cell transfers were implemented to characterize the regulatory cells that are induced by epicutaneous immunization. Results: Epicutaneous immunization with OVA and CpG reduces the production of OVA-specific IgE and increases the synthesis of OVA-specific IgG2a antibodies in an antigen-specific manner. Moreover, eosinophil peroxidase activity in the skin and production of IL-4, IL-5, IL-10, and IL-13 are suppressed. The observed reduction of IgE synthesis is transferable with T-cell receptor (TCR) ab1CD41CD252 cells, whereas IgG2a production is dependent on both TCRab1 and TCRgd1 T cells. Further experiments show that the described phenomenon is myeloid differentiation primary response 88, IFN-g, and IL-17A dependent. Finally, the results suggest that epicutaneous immunization with OVA and CpG decreases the synthesis of OVA-specific IgE and skin eosinophil peroxidase activity in mice with ongoing skin allergy. Conclusion: Epicutaneous application of protein antigen in the presence of adjuvant could be an attractive needle-free and self-administered immunotherapy for allergic diseases

Application of photochemical parameters and several indices based on phenotypical traits to assess intraspecific variation of oat (Avena sativa L.) tolerance to drought

Functionality of the photosynthetic system under water stress is of major importance in drought tolerance. Oat (Avena sativa L.) doubled haploid (DH) lines obtained by pollination of F_{1} oat crosses with maize were used to assess the differences in plant genotypic response to soil drought. The investigations were based on the measurements of gas exchange and chlorophyll a fluorescence kinetics. Drought was applied to 17-day-old seedlings by withholding water for 14 days and subsequent plant recovery. Non-stressed optimally watered plants served as controls. Yield components were determined when plants reached full maturity. It was shown differences among the oat lines with respect to drought stress susceptibility (SI) and stress tolerance index mean productivity and drought susceptibility index. Sensitivity to drought of individual DH lines was significantly different, as demonstrated by the correlation between drought susceptibility index and yield components, such as dry weight (GW) or grain number (GN) of the harvested plants. GW and GN were lower in drought-sensitive genotypes exposed to drought stress compared to those resistant to drought. The principal component analysis allow to separate three groups of lines differing in their sensitivity to drought stress and indicated that tolerance to drought in oat has a common genetic background

The effect of light intensity on the production of oat (Avena sativa L.) doubled haploids through oat × maize crosses

Oat haploid embryos were obtained by wide crossing with maize. The effect of light intensity during the growing period of donor plants (450 and 800 µmol m−2 s−1) and in vitro cultures (20, 40, 70 and 110 µmol m−2 s−1) was examined for the induction and development of oat DH lines. Oat florets (26008) from 32 genotypes were pollinated with maize and treated with 2,4-dichlorophenoxyacetic acid. All the tested genotypes formed more haploid embryos when donor plants were grown in a greenhouse (9.4%) compared to a growth chamber (6.1%). The light intensity of 110 µmol m−2 s−1 during in vitro culture resulted in the highest percentage of embryo germination (38.9%), conversion into plants (36.4%) and DH line production (9.2%) when compared with lower light intensities (20, 40 and 70 µmol m−2 s−1). The results show that the growth conditions of the donor plant and light intensity during in vitro culture can affect the development of haploid embryos. This fact may have an impact on oat breeding programs using oat × maize crosses

Obtaining of winter rye (Secale cereale L. ssp. cereale) haploid embryos through hybridization with maize (Zea Mays L.)

The aim of this study was to determine the effect of selected factors on rye (Secale cereale L.) haploid embryo production by the wide crossing method. The study was performed on fifteen winter rye genotypes. This is the first time for rye when besides the genotype, on the enlargement of ovaries and haploid embryo production, such factors as: type of auxin analogues 2,4-dichlorophenoxyacetic acid (2,4-D), 3,6-dichloro-2-methoxybenzoic acid (dicamba) and 4-amino-3,5,6-trichloropyridine-2-carboxylic acid (picloram), and the time between florets emasculation and pollination were investigated. All factors had a significant impact on rye ovary enlargement, however the haploid embryo formation depended only on rye genotype, not on kind of auxin and days between emasculation to pollination. In total, twenty one haploid embryos were formed by six genotypes of fifteen tested. On average, 13.86% (after 2,4-D treatment) to 20.05% (after dicamba treatment) enlarged ovaries per emasculated florets were obtained. Most of the ovaries enlarged when florets were pollinated 4 and 6 days after emasculation. Most of the haploid embryos formed when florets were pollinated 6 days after emasculation. The obtained haploid embryos did not germinate

Chlorophyll a fluorescence parameters in the evaluation of oat DH lines yield components

Chlorophyll a fluorescence can provide insight into the ability of plants to tolerate environmental conditions that can damage photosynthetic apparatus and decrease yield. The aim of the study was to determine the relationship between chlorophyll a fluorescence parameters and yield components of oat DH lines. All DH lines significantly differed in chlorophyll a fluorescence parameters and yield components. The overall performance index of PSII photochemistry (PI), showed the highest variation between DH lines, whereas the lowest had the ratio of variable to maximum fluorescence (Fv/Fm). The highest differences were observed in the number of grains per plant (21.3 to 600). Thousand-grain weight varied from 17.82 g to 41.01 g and the biomass from 8.01 g to 29.31 g. The highest negative correlations were found between Fv/Fm, Area (pool size of electron acceptors from PSII), PI and grain number per plant and biomass. Positive correlations were observed between light energy absorption (ABS/CS), grain number per plant and biomass, as well as the amount of excitation energy trapped in PSII reaction centers (TRo/CS) and biomass. Principal component analysis of chlorophyll a fluorescence parameters, together with yield components, discriminated two oat DH lines groups according to their photosynthetic efficiency and yield

Complex characterization of oat (Avena sativa L.) lines obtained by wide crossing with maize (Zea mays L.)

Background. The oat×maize addition (OMA) lines are used for mapping of the maize genome, the studies of centromere-specific histone (CENH3), gene expression, meiotic chromosome behavior and also for introducing maize C4 photosynthetic system to oat. The aim of our study was the identification and molecular-cytogenetic characterization of oat × maize hybrids. Methods. Oat DH lines and oat × maize hybrids were obtained using the wide crossing of Avena sativa L. with Zea mays L. The plants identified as having a Grande- 1 retrotransposon fragment, which produced seeds, were used for genomic in situ hybridization (GISH). Results. A total of 138 oat lines obtained by crossing of 2,314 oat plants from 80 genotypes with maize cv. Waza were tested for the presence of maize chromosomes. The presence of maize chromatin was indicated in 66 lines by amplification of the PCR product (500 bp) generated using primers specific for the maize retrotransposon Grande-1. Genomic in situ hybridization (GISH) detected whole maize chromosomes in eight lines (40%). All of the analyzed plants possessed full complement of oat chromosomes. The number of maize chromosomes differed between the OMA lines. Four OMA lines possessed two maize chromosomes similar in size, three OMA-one maize chromosome, and one OMA-four maize chromosomes. In most of the lines, the detected chromosomes were labeled uniformly. The presence of six 45S rDNA loci was detected in oat chromosomes, but none of the added maize chromosomes in any of the lines carried 45S rDNA locus. Twenty of the analyzed lines did not possess whole maize chromosomes, but the introgression of maize chromatin in the oat chromosomes. Five of 66 hybrids were shorter in height, grassy type without panicles. Twenty-seven OMA lines were fertile and produced seeds ranging in number from 1-102 (in total 613). Sixty-three fertile DH lines, out of 72 which did not have an addition of maize chromosomes or chromatin, produced seeds in the range of 1-343 (in total 3,758). Obtained DH and OMA lines were fertile and produced seeds. Discussion. In wide hybridization of oat with maize, the complete or incomplete chromosomes elimination of maize occur. Hybrids of oat and maize had a complete set of oat chromosomes without maize chromosomes, and a complete set of oat chromosomes with one to four retained maize chromosomes

Evaluation of DNA variants associated with androgenetic alopecia and their potential to predict male pattern baldness

Androgenetic alopecia, known in men as male pattern baldness (MPB), is a very conspicuous condition that is particularly frequent among European men and thus contributes markedly to variation in physical appearance traits amongst Europeans. Recent studies have revealed multiple genes and polymorphisms to be associated with susceptibility to MPB. In this study, 50 candidate SNPs for androgenetic alopecia were analyzed in order to verify their potential to predict MPB. Significant associations were confirmed for 29 SNPs from chromosomes X, 1, 5, 7, 18 and 20. A simple 5-SNP prediction model and an extended 20-SNP model were developed based on a discovery panel of 305 males from various European populations fitting one of two distinct phenotype categories. The first category consisted of men below 50 years of age with significant baldness and the second; men aged 50 years or older lacking baldness. The simple model comprised the five best predictors: rs5919324 near AR, rs1998076 in the 20p11 region, rs929626 in EBF1, rs12565727 in TARDBP and rs756853 in HDAC9. The extended prediction model added 15 SNPs from five genomic regions that improved overall prevalence-adjusted predictive accuracy measured by area under the receiver characteristic operating curve (AUC). Both models were evaluated for predictive accuracy using a test set of 300 males reflecting the general European population. Applying a 65% probability threshold, high prediction sensitivity of 87.1% but low specificity of 42.4% was obtained in men aged <50 years. In men aged ≥50, prediction sensitivity was slightly lower at 67.7% while specificity reached 90%. Overall, the AUC=0.761 calculated for men at or above 50 years of age indicates these SNPs offer considerable potential for the application of genetic tests to predict MPB patterns, adding a highly informative predictive system to the emerging field of forensic analysis of externally visible characteristics

UV-B radiation modifies the acclimation processes to drought or cadmium in wheat

Under natural conditions plants are often subjected to multiple stress factors. The main aim of the present work was to reveal how UV-B radiation affects acclimation to other abiotic stressors. Wheat seedlings grown under normal light conditions or normal light supplemented with UV-B radiation were exposed to drought or Cd stress and were screened for changes in the contents of salicylic acid and its putative precursor ortho-hydroxy-cinnamic acid, and in the activity of the key synthesis enzyme, phenylalanine ammonia lyase. Certain other protective mechanisms, such as antioxidant enzyme activities and polyamines, were also investigated. PEG treatment under UV-B radiation did not cause wilting, but resulted in more pronounced salicylic acid accumulation, which may provide protection against drought stress in wheat plants. In contrast, the high level of salicylic acid accumulation in Cd-treated plants was not further enhanced by UV-B stress, but resulted in pronounced oxidative stress and the activation of antioxidant systems and polyamine synthesis. Changes in the levels of phenolic compounds are accompanied by increased phenylalanine ammonia lyase activity in the roots, but not in the leaves. The similar pattern observed for stress-induced changes in salicylic acid and ortho-hydroxy-cinnamic acid contents suggested that salicylic acid may play a decisive role via ortho-hydroxy-cinnamic acid. The results indicated that UV-B radiation might have either a positive or negative impact under the same conditions in wheat, depending on the type of secondary abiotic stress factor. The protective or damaging effects observed may be related to changes in the levels of phenolic compounds