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We evaluated whether protected European butterflies can potentially be at risk if transgenic maize is extensively grown in Central Europe. We explored potential consequences of both insect resistant (IR) and herbicide resistant (HR) transgenic maize. IR maize can produce pollen that is toxic to lepidopteran larvae, and this puts butterfly species at possible risk if the presence of young larvae coincides with maize flowering, during which large quantities of maize pollen can be deposited on vegetation. By considering the timing of maize flowering in Europe and the phenology of the protected Lepidoptera species, we found that 31 species had at least one generation where 50% of the larval stage overlapped with maize flowering, and 69 species for which first instar larvae were present during maize pollen shedding. HR maize allows high concentration herbicide treatments on fields without seasonal limitation, which can drastically reduce weed densities. In cases where such weed species are host plants for protected butterflies, reduced host plant/food availability can result, causing population decreases. By using published information, we first identified the important weed species in major maize‐growing European countries. Subsequently, we checked whether the host plants of protected Lepidoptera included species that are common maize weeds. We identified 140 protected species having food plants that are common weeds in one or more of the major European maize‐growing countries. If HR maize is grown in Europe, there is a potential hazard that their food plants will seriously decline, causing a subsequent decline of these protected species. [ABSTRACT FROM AUTHOR]

Studies focused on the seasonal distribution of pollen and spores in semiarid cities are scarce. At these sites, climate change potentiates the emission and transport of fine (PM 10 ) to ultrafine particles (PM 2.5 ), easily attached to pollen surfaces, causing allergen's release. This study examines the potential correlation of seasonal variations of pollen, fungal spores, PM 10 , and meteorological parameters with allergic reactions of 150 people living in a Sonoran desert city. We collected PM 10 , airborne pollen, and spores during a year. We also studied topsoil and road dust samples as potential PM-emission sources. We obtained dust-mineralogy, chemistry, and particle size attached to pollen by X-ray diffraction and scanning electron microscope. Results show that seasonal high PM-loading in the urban atmosphere coincides with aeroallergens promoting micro- to nanoparticles' attachment to pollen's surface. A collapsed membrane was observed in several samples after individual grains show the following maximum wall coverage: Poaceae 28%, Asteraceae 40%, Chenopodiaceae-Amaranthacea 29%, Fabaceae 18%. Most of the particles covering pollen's surface have a geogenic origin mixed with metals linked to traffic (bromide, chlorine, and antimony). Mineralogical, granulometric analysis, and main wind-direction show that two local soil-types are the main contributors to PM. A high frequency of positive sensitization to pollen with high particle loading was detected. These results suggest that climate-driven dust emissions may alter pollen and spore surfaces' physicochemical characteristics with the further consequences in their allergenic potential.

In the present work, a novel sample preparation method, micro salting-out assisted matrix solid-phase dispersion (μ-SOA-MSPD), was developed for the determination of bisphenol A (BPA) and bisphenol B (BPB) contaminants in bee pollen. The proposed method was designed to combine two classical sample preparation methodologies, matrix solid-phase dispersion (MSPD) and homogenous liquid-liquid extraction (HLLE), to simplify and speed-up the preparation process. Parameters of μ-SOA-MSPD were systematically investigated, and results indicated the significant effect of salt and ACN-H 2 O extractant on the signal response of analytes. In addition, excellent clean-up ability in removing matrix components was observed when primary secondary amine (PSA) sorbent was introduced into the blending operation. The developed method was fully validated, and the limits of detection for BPA and BPB were 20 μg/kg and 30 μg/kg, respectively. Average recoveries and precisions were ranged from 83.03% to 94.64% and 1.76% to 5.45%, respectively. This is the first report on the analysis of bisphenol contaminants in bee pollen sample, and also on the combination of MSPD and HLLE. The present method might provide a new strategy for simple and fast sample preparation of solid and semi-solid samples.

Stressful conditions during development can have sub-lethal consequences on organisms aside from mortality. Using previously reported in-hive residues from commercial colonies, we examined how multi-pesticide exposure can influence honey bee (Apis mellifera) queen health. We reared queens in beeswax cups with or without a pesticide treatment within colonies exposed to treated or untreated pollen supplement. Following rearing, queens were open-mated and then placed into standard hive equipment in an "artificial swarm" to measure subsequent colony growth. Our treated wax had a pesticide Hazard Quotient comparable to the average in beeswax from commercial colonies, and it had no measurable effects on queen phenotype. Conversely, colonies exposed to pesticide-treated pollen had a reduced capacity for viable queen production, and among surviving queens from these colonies we observed lower sperm viability. We found no difference in queen mating number across treatments. Moreover, we measured lower brood viability in colonies later established by queens reared in treated-pollen colonies. Interestingly, royal jelly from colonies exposed to treated pollen contained negligible pesticide residues, suggesting the indirect social consequences of colony-level pesticide exposure on queen quality. These findings highlight how conditions during developmental can impact queens long into adulthood, and that colony-level pesticide exposure may do so indirectly.

Bee-pollen is a functional food sold for human and animal consumption but also is a favorable microhabitat for many spore-forming bacteria. Among them, Bacillus cereus can produce several toxins and other virulence factors, causing an emetic or diarrheal syndrome after ingestion. The study involved 36 bee-pollen samples obtained from different sampling points throughout the production process (collecting, freezing, drying, and cleaning) in Argentina. Fifty isolates of B. cereus yielded 24 different fingerprint patterns with BOX and ERIC primers. Only three fingerprint patterns were maintained throughout the production process. In contrast, others were lost or incorporated during the different steps, suggesting that cross-contamination occurred as shown by differences in fingerprint patterns after freezing, drying, and cleaning steps compared to the initial collection step. Genes encoding for cereulide (ces), cytotoxin K (cytK), sphingomyelinase (sph), the components of hemolysin BL (hblA, hblB, hblC, hblD) and non-hemolytic complex (nheAB) were studied. All the isolates displayed one or more enterotoxin genes. The most frequent virulence genes detected belong to the HBL complex, being the most abundant hblA (98%), followed by hblD (64%), hblB (54%), and hblC (32%), respectively. Ten strains (20%), present at all sampling points, carried all the subunits of the HBL complex. The non-hemolytic enterotoxic complex (nheAB) was found in 48 strains (96%), while seven strains (14%) present at all sampling points showed the amplification product for sphingomyelinase (sph). One cereulide-producer was isolated at the cleaning step; this strain contained all the components for the hemolytic enterotoxin complex HBL, the NHE complex, and cytotoxin K related to the foodborne diarrhoeal syndrome. In total, 11 different virulence patterns were observed, and also a correlation between rep-fingerprint and virulence patterns. The results suggest that bee-pollen can be contaminated at any point in the production process with potential enterotoxic B. cereus strains, emphasizing the importance of hygienic processing., Competing Interests: Declaration of competing interest The authors declare that no conflict of interest exists., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Objective: Particulate matter (PM), such as air pollutants and pollens, are known to cause skin ageing through skin inflammation. It is important to develop formulations which protect the skin from PM. We previously developed a conventional water-in-oil emulsion with a synthetic surfactant, distearyldimonium chloride, modified bentonite (C-W/O), which protects skin from allergens. In the present study, we developed a novel water-in-oil emulsion with a natural surfactant, lecithin, modified bentonite (N-W/O)., Methods: The microarray analysis was performed using total RNA extracted from a reconstructed human epidermis (RHE) stimulated with urban aerosols or cedar pollen for 6 h in order to develop an epidermal inflammation model by PM for the evaluation of topical formulations. We then compared the efficacy of N-W/O and C-W/O to prevent epidermal degradation. Tissues and culture media were collected 24 h after the urban aerosol or cedar pollen stimulation for a histological assay, and the quantification of MMP1 and IL-8 secretion., Results: The expression levels of proinflammatory cytokines and chemokines, such as IL1A and CXCL8, and matrix metalloproteinases, including MMP1, MMP3 and MMP9, were significantly up-regulated by the PM stimulation. As a result of ranking based on the pathway enrichment analysis, oxidative stress-related pathways, such as MAPK-mediated signalling, HIF-1 signalling, IL-1 signalling and ROS-induced cellular signalling, were ranked high in the urban dust- and cedar pollen-treated groups. A thickened stratum corneum, thinned vital layer and cleaved E-cadherin were observed by haematoxylin and eosin staining and immunohistochemical staining of E-cadherin in the PM treated groups. The secretion of MMP1 and IL-8 into the media was significantly increased by the PM stimulation. N-W/O prevented the degradation of epidermal integrity and secretion of inflammatory proteins more effectively than C-W/O., Conclusion: The present results showed that N-W/O made using natural surfactant is useful at protecting skin from PM, such as urban aerosols and cedar pollen., (© 2020 The Authors. International Journal of Cosmetic Science published by John Wiley & Sons Ltd on behalf of Society of Cosmetic Scientists and the Société Française de Cosmétologie.)

Many bees are effective pollen collectors; however, pollen grains collected by bees for larval food are lost for plant sexual reproduction. Recognition of these conflicting interests between bees and flowers is essential for understanding of reproduction for both bees and flowers [1-3]. Plant defense compounds in pollen may function to reduce pollen waste by deterring ineffective pollinators [4-6], but this hypothesis remains unexamined. Here, we provide evidence that secondary metabolites in pollen function as chemical defense by deterring some bees from gathering pollen. In two Dipsacus species, a defense compound, dipsacus saponin [7], occurs in pollen but not in nectar. We observed that bumblebees disliked grooming bitter-tasting pollen with a high saponin content. Manipulation of saponin concentrations in nectar and measurements of corbicular pollen showed that the bumblebee species differed in their tolerance to saponin. Those species susceptible to saponin groomed little Dipsacus pollen into their pollen loads, and their ungroomed pollen was observed to be effectively delivered to stigmas. By rewarding bees with edible nectar, but not pollen, plants solve the conflict of pollen partitioning between sexual and reward functions. Ungroomed toxic pollen on the bee body promotes pollen transfer efficiency, facilitating pollination., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Pollens are well-known triggers of respiratory allergies and asthma. The pollen burden in today's ambient air is constantly increasing due to rising climate change and air pollution. How pollens interact with the respiratory mucosa remains largely unknown due to a lack of representative model systems. We here demonstrate how pollen proteases of Kentucky bluegrass, white birch and hazel selectively destroy integrity and anchorage of columnar respiratory epithelial cells, but not of basal cells, in both ex vivo respiratory mucosal explants and in vitro primary equine respiratory epithelial cells (EREC). In turn, this pollen protease-induced damage to respiratory epithelial cell anchorage resulted in increased infection by the host-specific and ancestral alphaherpesvirus equine herpesvirus type 1 (EHV1). Pollen proteases of all three plant species were characterized by zymography and those of white birch were fully identified for the first time as serine proteases of the subtilase family and meiotic prophase aminopeptidase 1 using mass spectrometry-based proteomics. Together, our findings demonstrate that pollen proteases selectively and irreversibly damage integrity and anchorage of columnar respiratory epithelial cells. In turn, alphaherpesviruses benefit from this partial loss-of-barrier function, resulting in increased infection of the respiratory epithelium.

The ladybird Propylea japonica , adults of the green lacewing Chrysoperla nipponensis and the honey bee Apis mellifera are common pollen feeders in many crop systems. They could therefore be directly exposed to Cry proteins in Bacillus thuringiensis ( Bt )-transgenic crop fields by ingestion of pollen. They, or closely related species, are therefore often selected as surrogate test species in non-target risk assessment of Bt plants. In the current study, we evaluated the potential effects of the ingestion of Bt maize pollen containing the Cry1Ab/Cry1Ac fusion protein on various life-table parameters of the three pollen-feeding non-target species in laboratory feeding assays. The results showed that pupation rate and male adult fresh weight of P. japonica were significantly increased when fed pollen from Bt maize compared to control maize pollen, but other test life-table parameters were not affected. For the other two species, none of the tested life-table parameters (survival, pre-oviposition period, fecundity and adult fresh weight for C. nipponensis; survival and mean acinus diameter of hypopharyngeal glands for A. mellifera ) differed between non- Bt and Bt maize pollen treatments. ELISA measurements confirmed the stability and uptake of the Cry protein by all three species during the feeding bioassays. In addition, a sensitive insect bioassay confirmed the bioactivity of the Cry1Ab/Cry1Ac protein in the Bt maize pollen used. Overall, the results suggested that the three pollen feeders are not sensitive to the Cry1Ab/Cry1Ac protein, and planting of the Bt maize variety will pose a negligible risk to P. japonica , adult C. nipponensis and adult A. mellifera .

Honeybees may be exposed to insecticidal proteins from transgenic plants via pollen during their foraging activity. Assessing effects of such exposures on honeybees is an essential part of the risk assessment process for transgenic Bacillus thuringiensis (Bt) cabbage. Feeding trials were conducted in a laboratory setting to test for possible effects of Cry1Ba3 cabbage pollen on Italian-derived honeybees Apis mellifera L. Newly emerged A. mellifera were fed transgenic pollen, activated Cry1Ba3 toxin, pure sugar syrup (60% w/v sucrose solution), and non-transgenic cabbage pollen, respectively. Then the effects on survival, pollen consumption, weight, detoxification enzyme activity and midgut enzyme activity of A. mellifera were monitored. The results showed that there were no significant differences in survival, pollen consumption, weight, detoxification enzyme activity among all treatments. No significant differences in the activities of total proteolytic enzyme, active alkaline trypsin-like enzyme and weak alkaline trypsin-like enzyme were observed among all treatments. These results indicate that the side-effects of the Cry1Ba3 cabbage pollen on A. mellifera L. are unlikely.

Background: Previous studies have revealed the interactive effects of airborne pollen and particulate matter on the daily consultations for pollinosis, but it is uncertain which compositions are responsible. This study aimed to investigate the interactive effects of specific PM 2.5 compositions and airborne pollen on the daily number of clinic visits for pollinosis in Fukuoka., Methods: We obtained daily data on pollen concentrations, PM 2.5 compositions, PM 2.5 mass, gaseous pollutants (SO 2 , NO 2 , CO, and O 3 ), and weather variables monitored in Fukuoka between February and April, 2002-2012. In total, 73,995 clinic visits for pollinosis were made at 10 clinics in Fukuoka Prefecture during the study period. A time-stratified case-crossover design was applied to examine the interactive effects. The concentrations of PM 2.5 and its compositions were stratified into low (<15th percentile), moderate (15th-85th percentile), and high (>85th percentile) levels, and the association between airborne pollen and daily clinic visits for pollinosis was analyzed within each level., Results: We found a significant interaction between specific PM 2.5 compositions and airborne pollen. Specifically, the odds ratio of daily clinic visits for pollinosis per interquartile increase in pollen concentration (39.8 grains/cm 2 ) at the average cumulative lag of 0 and 2 days during high levels of non-sea-salt Ca 2+ was 1.446 (95% CI: 1.323-1.581), compared to 1.075 (95% CI: 1.067-1.083) when only moderate levels were observed. This result remained significant when other air pollutants were incorporated into the model and was fairly persistent even when different percentile cut-off points were used. A similar interaction was found when we stratified the data according to non-sea-salt SO 4 2- levels. This finding differed from estimates made according to PM 2.5 and NO 3 - levels, which predicted that the effects of pollen were strongest in the lower levels., Conclusions: Associations between airborne pollen and daily clinic visits for pollinosis could be enhanced by high levels of specific PM 2.5 compositions, especially non-sea-salt Ca 2+ ., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Recent efforts to evaluate the contribution of neonicotinoid insecticides to worldwide pollinator declines have focused on honey bees and the chronic levels of exposure experienced when foraging on crops grown from neonicotinoid-treated seeds. However, few studies address non-crop plants as a potential route of pollinator exposure to neonicotinoid and other insecticides. Here we show that pollen collected by honey bee foragers in maize- and soybean-dominated landscapes is contaminated throughout the growing season with multiple agricultural pesticides, including the neonicotinoids used as seed treatments. Notably, however, the highest levels of contamination in pollen are pyrethroid insecticides targeting mosquitoes and other nuisance pests. Furthermore, pollen from crop plants represents only a tiny fraction of the total diversity of pollen resources used by honey bees in these landscapes, with the principle sources of pollen originating from non-cultivated plants. These findings provide fundamental information about the foraging habits of honey bees in these landscapes.

Maize plants expressing dsRNA for the management of Diabrotica virgifera virgifera are likely to be commercially available by the end of this decade. Honey bees, Apis mellifera, can potentially be exposed to pollen from transformed maize expressing dsRNA. Consequently, evaluation of the biological impacts of RNAi in honey bees is a fundamental component for ecological risk assessment. The insecticidal activity of a known lethal dsRNA target for D. v. virgifera, the vATPase subunit A, was evaluated in larval and adult honey bees. Activity of both D. v. virgifera (Dvv)- and A. mellifera (Am)-specific dsRNA was tested by dietary exposure to dsRNA. Larval development, survival, adult eclosion, adult life span and relative gene expression were evaluated. The results of these tests indicated that Dvv vATPase-A dsRNA has limited effects on larval and adult honey bee survival. Importantly, no effects were observed upon exposure of Am vATPase-A dsRNA suggesting that the lack of response involves factors other than sequence specificity. The results from this study provide guidance for future RNAi risk analyses and for the development of a risk assessment framework that incorporates similar hazard assessments., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Thirty-two honeybee (Apis mellifera) colonies were studied in order to detect and measure potential in vivo effects of neonicotinoid pesticides used in cornfields (Zea mays spp) on honeybee health. Honeybee colonies were randomly split on four different agricultural cornfield areas located near Quebec City, Canada. Two locations contained cornfields treated with a seed-coated systemic neonicotinoid insecticide while the two others were organic cornfields used as control treatments. Hives were extensively monitored for their performance and health traits over a period of two years. Honeybee viruses (brood queen cell virus BQCV, deformed wing virus DWV, and Israeli acute paralysis virus IAPV) and the brain specific expression of a biomarker of host physiological stress, the Acetylcholinesterase gene AChE, were investigated using RT-qPCR. Liquid chromatography-mass spectrometry (LC-MS) was performed to detect pesticide residues in adult bees, honey, pollen, and corn flowers collected from the studied hives in each location. In addition, general hive conditions were assessed by monitoring colony weight and brood development. Neonicotinoids were only identified in corn flowers at low concentrations. However, honeybee colonies located in neonicotinoid treated cornfields expressed significantly higher pathogen infection than those located in untreated cornfields. AChE levels showed elevated levels among honeybees that collected corn pollen from treated fields. Positive correlations were recorded between pathogens and the treated locations. Our data suggests that neonicotinoids indirectly weaken honeybee health by inducing physiological stress and increasing pathogen loads.

Japanese cedar (JC) pollinosis is caused by Japanese cedar pollen (JCP) and most common seasonal allergic disease in Japan. Subcutaneous immunotherapy (SCIT) with allergen extract of JCP (JCP-allergen extract) is well established for JC pollinosis treatment with improvement of symptoms. However, major drawbacks for SCIT are repeated painful injections, frequent hospital visits and anaphylactic risk. Currently, sublingual immunotherapy (SLIT) has received much attention as an advanced alternative application with lower incidence of systemic reactions because the liquid or tablet form of allergen is placed under the tongue. The aim of this study was safety evaluation of standardized JCP-allergen extract currently developed for SLIT in JC pollinosis. JCP-allergen extract showed no potential genotoxicity. No systemic effects were observed in rats administered JCP-allergen extract orally for 26 weeks followed by 4-week recovery period. Mild local reactions such as hyperplasia and increased globule leukocytes resulting from vehicle (glycerin)-induced irritation were observed in stomach. No-observed-adverse-effect level was greater than 10,000 JAU/kg/day for systemic toxicity, equivalent to 300-fold the human dose. No local irritation was found in rabbits oral mucosae by 7-day sublingual administration. These results demonstrate the safe profile of standardized JCP-allergen extract, suggesting it is suitable for SLIT in JC pollinosis., (Copyright © 2015 Elsevier Inc. All rights reserved.)

In recent literature, little has been said regarding the morphological changes that occur in lung cells after treatment with particles and nanoparticles. Using an in vitro model of type-II lung epithelium (A549), we studied the effects of submicron particles (PM1.0), Parietaria officinalis (ALL), and PM1.0 + ALL together. To date several biochemical effects have been described, instead few data exist in literature regarding morphological events following these treatments, in particular we focused on the morphological changes and distribution of mitochondria, tonifilaments and rough endoplasmic reticulum, using a transmission electron microscopic (TEM) approach. After exposure to PM1.0 particles (PM1.0), Parietaria officinalis as allergen, and PM1.0 with P. officinalis, changes in the cytoplasmic area were observed, such as damage to mitochondria and morphological alterations of the tonifilaments and rough endoplasmic reticulum. The data obtained strongly support the hypothesis that cells in contact with submicron particles (PM1.0), or P. officinalis, undergo alteration of their metabolism., (Copyright © 2014 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.)

Pollen is the most common aeroallergen to cause seasonal conjunctivitis. The result of allergen exposure is a strong Th2-mediated response along with conjunctival mast cell degranulation and eosinophilic infiltration. Oleanolic acid (OA) is natural a triterpene that displays strong anti-inflammatory and immunomodulatory properties being an active anti-allergic molecule on hypersensitivity reaction models. However, its effect on inflammatory ocular disorders including conjunctivitis, has not yet been addressed. Hence, using a Ragweed pollen (RWP)-specific allergic conjunctivitis (EAC) mouse model we study here whether OA could modify responses associated to allergic processes. We found that OA treatment restricted mast cell degranulation and infiltration of eosinophils in conjunctival tissue and decreased allergen-specific Igs levels in EAC mice. Th2-type cytokines, secreted phospholipase A2 type-IIA (sPLA2-IIA), and chemokines levels were also significantly diminished in the conjunctiva and serum of OA-treated EAC mice. Moreover, OA treatment also suppressed RWP-specific T-cell proliferation. In vitro studies, on relevant cells of the allergic process, revealed that OA reduced the proliferative and migratory response, as well as the synthesis of proinflammatory mediators on EoL-1 eosinophils and RBL-2H3 mast cells exposed to allergic and/or crucial inflammatory stimuli such as RWP, sPLA2-IIA or eotaxin. Taken together, these findings demonstrate the beneficial activity of OA in ocular allergic processes and may provide a new intervention strategy and potential therapy for allergic diseases.

In the recent literature there has been an increased interest in the effects of particulate matter on the respiratory tract. The objective of this study was to use an in vitro model of type II lung epithelium (A549) to evaluate the cell ability to take up sub-micron PM(1.0) particles (PM(1.0)), Parietaria officinalis (ALL), and PM(1.0) + ALL together. Morphological analysis performed by Transmission Electron Microscope (TEM) showed that PM and ALL interacted with the cell surface, then penetrating into the cytoplasm. Each single treatment was able to point out a specific change in the morphology. The cells treated appear healthy and not apoptotic. The main effect was the increase of: multilamellar bodies, lysosomal enzymes, microvilli, and presence of vesicle/vacuoles containing particles. These observations demonstrate morphological and functional alterations related to the PM(1.0) and P. officinalis and confirm the induction of the inflammatory response in lung cells exposed to the inhalable particles., (Copyright © 2012. Published by Elsevier Ltd.)

The cultivation of Lepidoptera-resistant Bt-maize may affect nontarget butterflies. We assessed the risk posed by event MON89034 × MON88017 (expressing Cry1A.105 and Cry2Ab2 against corn borers) to nontarget Lepidoptera. Using the small tortoiseshell Aglais urticae, a butterfly species common in central Europe, as a test organism we (i) assessed the toxicity of Bt-maize pollen on butterfly larvae; (ii) measured pollen deposition on leaves of the host plant Urtica dioica; (iii) mapped the occurrence and distribution of host plants and larvae in two arable landscapes in Germany during maize anthesis; and (iv) described the temporal occurrence of a 1-year population of A. urticae. (i) Larvae-fed 200 Bt-maize pollen grains/cm(2) had a reduced feeding activity. Significant differences in developmental time existed at pollen densities of 300 Bt-maize pollen grains/cm(2) and in survival at 400 grains/cm(2). (ii) The highest pollen amount found was 212 grains/cm(2) at the field margin. Mean densities were much lower. (iii) In one region, over 50% of A. urticae nests were located within 5 m of a maize field, while in the other, all nests were found in more than 25 m distance to a maize field. (iv) The percentage of larvae developing during maize anthesis was 19% in the study area. The amount of pollen from maize MON89034 × MON88017 found on host plants is unlikely to adversely affect a significant proportion of larvae of A. urticae. This paper concludes that the risk of event MON89034 × MON88017 to populations of this species is negligible., (© 2012 Blackwell Publishing Ltd.)

Honeybees may be exposed to insecticidal proteins from transgenic plants via pollen. An assessment of the impact of such exposures on the honeybee is an essential part of the risk assessment process for transgenic Bacillus thuringiensis corn. A field trial was conducted to evaluate the effect of transgenic Bt cry1Ah corn on the honeybee Apis mellifera ligustica. Colonies of honeybees were moved to Bt or non-Bt corn fields during anthesis and then sampled to record their survival, development and behavior. No differences in immature stages, worker survival, bee body weight, hypopharyngeal gland weight, colony performance, foraging activity or olfactory learning abilities were detected between colonies that were placed in non-Bt corn fields and those placed in Bt corn fields. We conclude that cry1Ah corn carries no risk for the survival, development, colony performance or behavior of the honeybee A. mellifera ligustica., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Asthma is currently defined as a chronic inflammatory disease of the airway. Several evidence indicate that vehicle emissions in cities is correlated with the allergic respiratory diseases. In the present study, we evaluated in the A549 cells the production and release of IL-4, IL-5 and IL-13 after treatment with sub-micron PM(1.0) particles (PM(1.0)), Parietaria officinalis (ALL), and PM(1.0) + ALL together. Our data demonstrated that PM(1.0) + ALL together exhibited the greatest capacity to induce A549 cells to enhance the expression of IL-4 and IL-5 compared with the only PM(1.0) or ALL treatment. Interestingly, IL-13 that is necessary for allergen-induced airway hyper responsiveness, is increased in cells treated with PM(1.0) + ALL together, but is higher expressed when the cells are treated only with the allergen. Our data support the hypothesis that the urban environment damage the acinar lung units and activates cells of the immune system., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Transgenic Cry1Ac+CpTI cotton (CCRI41) is a promising cotton cultivar throughout China but side effects and especially sublethal effects of this transgenic cultivar on beneficial insects remain poorly studied. More specifically potential sublethal effects on behavioural traits of the honey bee Apis mellifera L. have not been formally assessed despite the importance of honey bees for pollination. The goal of our study was to assess potential effects of CCRI41 cotton pollen on visual and olfactory learning by honey bees. After a 7-day oral chronic exposure to honey mixed with either CCRI41 pollen, imidacloprid-treated conventional pollen (used as positive sublethal control) or conventional pollen (control), learning performance was evaluated by the classical proboscis extension reflex (PER) procedure as well as a T-tube maze test. The latter assay was designed as a new device to assess potential side effects of pesticides on visual associative learning of honey bees. These two procedures were complementary because the former focused on olfactory learning while the latter was involved in visual learning based on visual orientation ability. Oral exposure to CCRI41 pollen did not affect learning capacities of honey bees in both the T-tube maze and PER tests. However, exposure to imidacloprid resulted in reduced visual learning capacities in T-tube maze evaluation and decreased olfactory learning performances measured with PER. The implications of these results are discussed in terms of risks of transgenic CCRI41 cotton crops for honey bees.

Transgenic Cry1Ac + CpTI cotton (CCRI41) is increasingly planted throughout China. However, negative effects of this cultivar on the honey bee Apis mellifera L., the most important pollinator for cultivated ecosystem, remained poorly investigated. The objective of our study was to evaluate the potential side effects of transgenic Cry1Ac + CpTI pollen from cotton on young adult honey bees A. mellifera L. Two points emphasized the significance of our study: (1) A higher expression level of insecticidal protein Cry1Ac in pollen tissues was detected (when compared with previous reports). In particular, Cry1Ac protein was detected at 300 ± 4.52 ng g(-1) [part per billion (ppb)] in pollen collected in July, (2) Effects on chronic mortality and feeding behaviour in honey bees were evaluated using a no-choice dietary feeding protocol with treated pollen, which guarantee the highest exposure level to bees potentially occurring in natural conditions (worst case scenario). Tests were also conducted using imidacloprid-treated pollen at a concentration of 48 ppb as positive control for sublethal effect on feeding behaviour. Our results suggested that Cry1Ac + CpTI pollen carried no lethal risk for honey bees. However, during a 7-day oral exposure to the various treatments (transgenic, imidacloprid-treated and control), honey bee feeding behaviour was disturbed and bees consumed significantly less CCRI41 cotton pollen than in the control group in which bees were exposed to conventional cotton pollen. It may indicate an antifeedant effect of CCRI41 pollen on honey bees and thus bees may be at risk because of large areas are planted with transgenic Bt cotton in China. This is the first report suggesting a potential sublethal effect of CCRI41 cotton pollen on honey bees. The implications of the results are discussed in terms of risk assessment for bees as well as for directions of future work involving risk assessment of CCRI41 cotton.

Transgenic insect-resistant cotton has been planted in China in a large scale and may have adverse impacts on honeybees. Pollens from the transgenic Cry1Ac+CpTI cotton Zhong-41 and the parental cotton Zhong-23 were collected from the field and their impacts on adult worker bees were assessed. Experimental results showed that Zhong-41 pollen had no acute oral toxic effect on worker bees. No significant differences were observed in the superoxide dismutase activity or in the longevity of worker bees fed with diets containing the two cotton pollens. The main reasons for the outcome may be the low expression level of the transgenic proteins Cry1Ac and CpTI in the pollen of Zhong-41 as well as the substantial equivalence in the amounts of gross protein and soluble saccharides for the two cotton pollens. The implications of these results are discussed and further work to be carried out is put forward.

Long range transport of airborne pollen has been seldom studied in South America. Backward trajectories of Celtis and Nothofagus pollen grains trapped over a meteorological station outside Mar del Plata City were calculated in one-hour steps using the regional hybrid model developed by the NOAA (HYSPLIT 4.5) and the data of the NCEP filed in the NOAA server. Results showed that the observed trajectories agree with the location of vegetation sources of the collected tree species. In the case of Celtis, the transport was associated to anticyclones located east of the city, generating winds with a N-NE component, which produce pollen cloud advection from the Celtis forests located some tens of kilometers to the N and NE of the city. The sources of Nothofagus pollen correspond to a narrow strip on the Andes slopes between 39 degrees and 55 degrees S, at least 1100 km to the SW of Mar del Plata. The transport was associated to eastward displacement of the troughs corresponding to the Westerlies circulation and the presence of an anticyclone system that brings back Nothofagus pollen towards Mar del Plata area.

Background: Grape farmers are exposed to a variety of agents capable of inducing occupational skin disease. We conducted a study to measure the prevalence of skin symptoms and work-related skin symptoms among grape farmers in the Malevisi region of Crete and to provide data on associated risk factors., Methods: One hundred twenty grape farmers and 100 controls participated in the study. The protocol consisted of a questionnaire, skin prick tests for 16 common allergens, and measurement of specific IgE antibodies against 8 allergens., Results: Self-reported itchy rash (OR, 2.31; 95%CI, 1.10-4.84, P<0.05) within the last 12 months, and work-related itchy rash (OR, 4.08; 95%CI, 1.01-20.33, P<0.05) were significantly higher in grape farmers than in controls, after adjusting for age and sex. Sensitization to pollens (OR, 4.20; 95% CI, 1.41-12.82, P<0.01) and allergic rhinitis (OR, 3.06; 95% CI, 1.21-8.28, P<0.05) were found to be significantly associated with self-reported itchy rash in the grape farmers group., Conclusions: Grape farmers reported skin symptoms more frequently than non-exposed controls, and IgE-mediated sensitization to pollens was found to be significantly associated with the reported symptoms. Further studies are needed to evaluate the impact of specific occupational agents on skin diseases among grape farmers., (Copyright (c) 2005 Wiley-Liss, Inc.)

The aim of the study was to compare the concentration of 3 allergenic taxa pollen (ash, birch and oak) in the cities of Lublin (eastern Poland) and Szczecin (western Poland) during the years 2004 and 2005 and examine correlations between weather factors and pollen concentration. The meteorological parameters analysed were maximum and mean air temperature, relative humidity, amount of precipitation and wind speed. Measurements were performed by the volumetric method (Lanzoni 2000 pollen sampler). Pollen seasons were defined as the period in which 95 % of the total catch occurred. The pollen seasons of the studied taxa started earlier in Szczecin than in Lublin. In both cities much higher Betula pollen concentration and slightly higher Fraxinus pollen concentration were recorded in 2004 compared to 2005. But for Quercus, higher concentrations were recorded in Lublin and in Szczecin in 2005. Annual totals and maximum daily pollen grain concentrations of the tree taxa in question were higher in Lublin than in Szczecin. In 2005, maximum Betula and Fraxinus pollen concentrations occurred in each of the cities at the same time: in Lublin on 18 April, in Szczecin on 16 April. Among the meteorological parameters analysed, maximum temperature and relative humidity belong to the most important factors which affect ash, birch and oak pollen concentrations in the air.

An aeropalynological study was carried out in the atmosphere of Estepona, a very popular tourist resort situated in the "Costa del Sol", (southern Spain) based on the data obtained during a three year air-monitoring programme (March 1995 to March 1998) using a volumetric pollen trap. The 34 taxa that reached a 10-day mean air pollen concentration equal to or greater than 1 grain of pollen/m(3) of air are reflected in the calendar. The first 10 taxa, in order of abundance, were: Cupressaceae, Olea europaea, Quercus, Poaceae, Urticaceae, Plantago, Pinus, Chenopodiaceae-Amaranthaceae, Ericaceae and Castanea, the first 3 of which accounted for approximately 56 % of the annual total pollen count. The greatest diversity of pollen type occurred during spring, while the highest pollen concentrations were reached from February-June, when approximately more than 80 % of the annual total pollen was registered. The lowest concentrations were obtaining during January, August and September. The annual quantity of pollen collected, the intensity and the dates on which the maximum peaks were recorded differed for the 3 years studied, which can be explained by reference to various meteorological parameters, especially rainfall and temperature. The pollen calendar spectrum is typically Mediterranean and similar to those of nearby localities, in which many pollen types are represented and the long tails indicating long flowering periods.

The problem of allergy to decorative plants is still poorly known. Reports on occupational allergy to flowers are scarce and usually concern gardeners, greenhouse workers and florists. The handling, smelling and caring of flowers may cause rhinoconjunctivitis, asthma, urticaria and also contact dermatitis. Plants of the Compositae family that includes many different species have been frequently described. Chrysanthemum flowers, the strongest sensitizer among ornamental Compositae plants, especially contribute to the development of contact dermatitis. Allergy to Liliaceae, mostly to tulips, hyacinths, lilies and crocuses has quite often been reported as well as sensitization to rose pollen in rose planters. Occupational sensitization to flowers of other families, among which spathe flowers, primulas, weeping fig or Stephanotis floribunda should be mentioned, is less frequent. Exposure to ornamental flowers is common in the general population. Persons occupationally involved in cultivation of flowers and who demonstrate allergic symptoms are often forced to change their jobs. Candidates to these occupations with diagnosed atopy should be informed about the risk of developing allergy to flowers, which could make them unable to perform the job.

The aims of this study were to describe and compare the characteristics of grass and ragweed airborne pollen in rural and urban areas in the southern Panonnian Valley. Airborne pollen data were collected by using Hirst type volumetric samplers simultaneously in rural and urban localities. If rural and urban environment are considered, both grass and ragweed daily pollen concentrations showed a significant degree of association. Observed parameters (pollen index, maximum daily concentration, number of days during which the pollen is recorded in the air and start day of main pollen season), showed year-to-year variations for both grass and ragweed aeropollen. Average values of these parameters were higher in the rural environment, but the difference was statistically significant only for grass pollen index. Such a low difference indicates the possibility for conducting dose response clinical trials based on data obtained from one sampling station. The least year-to-year variations as well as the least difference between rural and urban environment, have been observed in the case of start date of the MPS. Such a situation suggests the possibility for using data obtained in one type of environment for the development of long-term forecast models for an entire region.

Background: Allergic eye diseases are complex inflammatory conditions of the conjunctiva that are becoming increasingly prevalent and present an increasing economic burden because of direct and indirect health expenditures., Objective: We sought to identify factors that may synergize with antigen-induced allergic inflammation and lead to allergic conjunctivitis. We used a murine model of allergic conjunctivitis to test the effect of oxidative stress generated by pollen oxidases using nicotinamide adenine dinucleotide (reduced) or nicotinamide adenine dinucleotide phosphate (reduced) (NAD[P]H) as an electron donor present in pollen grains., Methods: Reactive oxygen species (ROS) generation by hydrated Ambrosia artemisiifolia pollen (short ragweed pollen; RWP) grains was determined by using 2'-7'-dihydro-dichlorofluorescein diacetate, nitroblue tetrazolium reduction, and Amplex Red assay. The RWP-induced changes in intracellular ROS levels were examined in A549 cells, human primary bronchial epithelial cells, and murine conjunctiva., Results: Ragweed pollen grains contain NAD(P)H oxidase activity, which is diphenyleneiodonium-sensitive and quinacrine-sensitive and sodium azide-resistant. These NAD(P)H oxidases generate a superoxide anion that can be converted to H2O2 by pollen grain-associated superoxide dismutase. These diffusible oxygen radicals from pollen grains increase intracellular ROS levels in cultured epithelial cells and murine conjunctiva. Similar phenomena were observed in sensitized and naive mice, indicating that the RWP-induced oxidative stress in conjunctival epithelium is independent of adaptive immunity. Inactivation of NAD(P)H oxidase activity in RWP decreases the immediate-type hypersensitivity and inflammatory cell infiltration into the conjunctiva., Conclusion: Our data suggest that ROS generated by NAD(P)H oxidases in pollen grains intensify immediate allergic reactions and recruitment of inflammatory cells in murine conjunctiva.

Purpose: Social status influences asthma morbidity but the mechanisms are not well understood. To determine if sociodemographics influence the susceptibility to ambient aeroallergens, we determined the association between daily hospitalizations for asthma and daily concentrations of ambient pollens and molds in 10 large Canadian cities., Methods: Daily time-series analyses were performed and results were adjusted for day of the week, temperature, barometric pressure, relative humidity, ozone, carbon monoxide, sulfur dioxide, and nitrogen dioxide. Results were then stratified by age, gender, and neighborhood family education and income., Results: There appeared to be age and gender interactions in the relation between aeroallergens and asthma. An increase in basidiomycetes equivalent to its mean value, about 300/m3, increased asthma admissions for younger males (under 13 years of age) by 9.3% (95% CI, 4.8%, 13.8%) vs. 4.2% (95% CI, - 0.1%, 8.5%) for older males. The reverse was true among females with increased effect in the older age group: 2.3% (95% CI, 1.2%, 5.8%) in those under 13 years vs. 7.1% (95% CI, 4.1%, 10.1%) for older females. Associations were seen between aeroallergens and asthma hospitalization in the lowest but not the highest education group., Conclusions: Our results suggest that younger males and those within less educated families may be more vulnerable to aeroallergens as reflected by hospitalization for asthma.

In recent decades an increased prevalence of allergic conditions has been observed in developed countries. Although lifestyles, exposure to infection, and diet are all likely important factors, many studies have also shown a strong link between industrialization and allergy. The aim of this study was to investigate which extract fractions from traffic particulate matter (TPM, collected in a tunnel in Prague) have the greatest impact on different inflammatory and immunological parameters, such as cytokine production, levels of immunoglobulin E (IgE) antibodies, and bronchial hyperresponsiveness (BHR) in mice, when the extracts are used together with birch pollen for immunization. BP2 mice were immunized with birch pollen and different fractions of TPM (fractions 1-8). They were provoked intranasally with a mixture of pollen and TPM or pollen alone before they were challenged with methacholine. The BHR was evaluated in a whole-body plethysmograph. Th2 cytokines and fibronectin concentrations were measured, and differential cell counts were performed in the bronchioalveolar lavage (BAL) fluid. Sera were collected for determination of antibody titers. The highest titers of IgE and the highest BHR were found in the positive control mice (immunized and provoked with a mixture of pollen and TPM), followed by mice immunized with pollen and fraction 2 (which contains organic acids). Fraction 2 also induced the highest number of eosinophils and increased levels of interleukin 5 (IL-5) in the BAL fluid. The highest levels of IL-5, in BAL fluid and sera, were obtained in mice immunized with fraction 6 (moderately polar compounds), a somewhat surprising result since those mice did not produce any IgE, did not have any eosinophils in their BAL, or showed almost no BHR. Our data demonstrate that fractions 2 (organic acids) and 7 (highly polar compounds) seem to contain potential adjuvants stimulating the IL-5 production, the IgE synthesis, the eosinophil recruitment, and the bronchial hyperreactivity. Further characterization at the molecular level is now necessary to be able to identify the exact nature of those potential adjuvants. This will be of help in the future to improve the quality of the urban air aerosol.

Environmental agents, when applied in combination or sequentially, can induce a wide variety of adverse health effects in humans. To determine the effects of sequential allergen challenge and acid exposure on human bronchial epithelial cell function, we subjected normal, nonallergic control and ragweed-allergic individuals to bronchoscopic segmental ragweed challenge in vivo. We harvested bronchial epithelial cells by brush biopsy both before challenge and 24 hr after challenge and exposed cells to an acid stress in vitro (pH 5 for 3 hr), followed by a 1-hr recovery period at normal pH. In normal, nonallergic subjects, segmental allergen challenge produced no effects on ciliary activity; pH 5 exposure produced reduced ciliary activity (a decrease in the percent of the initially active area), with significant recovery after cells were returned to a normal pH. Ciliary activity from allergic subjects was also inhibited by pH 5 exposure; however, activity was not recovered when cells were placed in medium of normal pH. Ciliary activity in allergics who developed a stress response postantigen challenge, as determined by an induction of the 27 kDa stress (heat shock) protein, displayed no ciliary dysfunction when exposed to a pH 5 stress. In this case, a stress sufficient to provoke a heat shock (stress) protein (HSP) response (but not one that produced more severe lung injury and did not provoke an HSP response) protected cells from a subsequent acid stress. Because of our observations and recent findings reported in the literature, we suggest that in order to define the wide variety of health effects of environmental agents, control as well as at-risk populations should be studied and the ability to define potentially beneficial as well as detrimental effects should be built into the experimental design. Inclusion of different and novel end points also should be considered.

Objective: Extracts of environmental allergens, moulds and plant pollens are known to consume haemolytic complement (huC) in human serum in vitro through the antibody-independent engagement of the first component C1 of the classical pathway. The present work was undertaken to establish the nature and characteristics of the complement activating agents in allergenic extracts and to probe their relationship with the IgE-binding allergens., Materials and Methods: A large series of different > 10 kDa allergenic products was investigated for their capacity to consume haemolytic complement in the sera of allergic patients with specific anti-allergen IgE antibodies, as well as in normal control sera. UV-spectroscopy was used for categorizing the non-protein components in the extracts., Results: The experiments confirmed that huC is consumed in an antibody-independent fashion and in a qualitatively similar, but quantitatively distinct manner. The ratio of the complement activating potencies among the different allergenic preparations thereby remained constant and independent of the serum source, while an overt relationship with specific IgE-antibodies could not be established. UV-spectroscopy of the allergenic preparations revealed the presence of chemical decomposition products in nearly every extract and roughly in proportion to the complement activating potencies., Conclusion: The data support the idea that huC-activation by traditional allergenic extracts is mainly due to by-stander degradation products of the melanoidin (Maillard) or tannin type, which may or may not occur in physical association with the IgE-binding protein allergens.

A quantitative skin-test assay of allergenic potency was developed and tested on 28 allergic patients. The best-fit linear regression line was calculated from the sum of erythema or wheal diameters obtained from four intradermal threefold serial dilutions near the endpoint. Each line must have a correlation coefficient greater than 0.85 and the slopes of reference and test extract lines must not be significantly different. The relative potency can be calculated from the horizontal distance between the reference and test lines. The between-assay reproducibility was determined by comparing references against themselves. The 95% confidence limits were 54% to 186% when erythema was used and 27% to 367% when wheal was used. The narrower limits for the assay using erythema maybe due to the 5.5 times steeper slope of erythema lines than wheal lines. Potency results on five commercial extracts were highly correlated with potency results by RAST, AgE assay, or rye I assay. Extracts with isolectric focusing patterns dissimilar from the references were of significantly lower potency by in vivo and in vitro assays. The measured potency of two low-potency short ragweed extracts varied significantly with patient sensitivity to heat-stable and heat-labile ragweed allergens, indicating that patient selection may significantly affect assay results. Dose-response lines using either histamine or short ragweed by puncture (P) and intradermal (ID) techniques in the same patient showed that P/ID dose for equal erythema response was 909 for histamine and 31,250 for ragweed. This quantitative skin-test assay is highly re[rpdicob;e, yields potency data comparable to those of in vitro tests, can be applied to studies of extract stability, and is suitable as a primary bioassay of allergenic activity.

Observation of 100 patients with atopic dermatitis due to hypersensitivity to pollen over a period of 12 years emphasized certain important diagnostic and therapeutic features. The incidence was higher in females than in males and higher in middle and old age than in the earlier years. Pollen dermatitis may be the sole or major manifestation of allergy; 43 patients gave no history of other allergic symptoms. It may involve any or all areas of the body. The site or the distribution of lesions or the nature of the lesions gave no clue as to the diagnosis of pollen sensitivity. The character of the eruption varied widely from patient to patient and in given patients from week to week at times. Atopic dermatitis due to pollen sensitivity may be purely seasonal, perennial with seasonal exacerbations or perennial without seasonal variation. Reactions to skin testing with pollens suspected as allergens may be positive, equivocal or negative. In 58 patients there were positive correlative skin reactions to pollens. The diagnosis of atopic dermatitis due to pollen sensitivity, and the composition of the desensitizing antigen or antigens, must be based primarily on the clinical history and the area of residence. Most patients could tolerate only very weak dilutions at the beginning of desensitization therapy. Strong dilutions caused exacerbation of the dermatitis. Good or excellent results were obtained with perennial pollen desensitization therapy administered over long periods. In 13 patients good results took four to eight years of desensitization therapy. Fifty required less than two years. Tolerance of the patient for a given dose of antigen should determine the maximum dilution used in therapy.