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Epidemiology of allergic diseases in children.

The one-year-prevalence rate of bronchial asthma in children varies from 1-3%, when investigated in general practice, to 5-7% in population studies. The prevalence rate is highest in young boys. Eighty percent of the asthmatic children are allergic, house-dust-mite allergy being the most common allergy. The one-year-prevalence rate of rhinitis is 5-10% in general practice, and 10-12% in population studies. Again, the prevalence rate is highest in young boys. About 90% of children with rhinitis symptoms are allergic, with pollen allergy as the most common allergy. Risk factors for developing allergic diseases are many. The predisposition is probably the most prevailing risk factor. Period of birth, sex, race, diet, the presence of other allergic diseases, tobacco smoking, pollution, and allergens in the environment, all these factors alone or in combination almost double the risk. There is no doubt that both asthma and hay-fever prevalences have steadily increased within the last 50 years. Also, admissions to hospitals for childhood asthma have continued to increase, while the mortality of asthma in children has not risen statistically. This increase is in contrast to the effective medication available for both asthma and allergic rhinitis, and to the number of preventive factors known to us today. The time has come to try to change it at all costs. The outcome of allergic rhinitis and asthma shows that only 10% are cured, 50% ameliorate, 30% remain unchanged, and 10% deteriorate. Factors determining the outcome are age, immunotherapy, sex, mother's age at childbirth, infections, other allergic diseases, and signs and symptoms of food allergy.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Enhancement of development of larval Anopheles arabiensis by proximity to flowering maize (Zea mays) in turbid water and when crowded.

To determine whether proximity to flowering maize enhances the development of larval anopheline mosquitoes breeding in turbid water and when crowded, we evaluated the development of larval Anopheles arabiensis under various conditions of turbidity, larval density, and proximity to pollen-shedding maize in simulated breeding puddles in a malaria-endemic site. In naturally formed puddles, water turbidity, as well as larval density, increased as the rainy season progressed. In sites remote from flowering maize, more pupae developed and the resulting adults were larger in relatively clear water than in turbid water, and larval crowding inhibited development. In close proximity to flowering maize, however, larval development was little affected by water turbidity and larval crowding. Larvae of this member of the African An. gambiae complex of mosquitoes develop readily in turbid water and when crowded, provided that their breeding sites are located where maize pollen is abundant.

Animals↗

Immunoglobulin G4-antibodies to rBet v 1 and risk of sensitization and atopic disease in the child.

BACKGROUND: In 1993 extremely high levels of birch-pollen were recorded in Stockholm, Sweden. This provided an opportunity to evaluate the effects of aeroallergen exposure (exp.) on the early immune response. OBJECTIVE: To assess the influence of exp. to birch-pollen during pregnancy and infancy on the allergen-specific IgE- and IgG4-antibody (ab) response and the development of atopic disease in children. METHODS: A total of 970 children with atopic heredity and born in Stockholm 1992, 1993 or 1994 were investigated at age 4.5-5 years. They were divided into five groups; high-dose exp. at 1 year of age, high-dose exp. at 0-3 months, low-dose exp. at 0-3 months, high-dose exp. during pregnancy and low-dose exp. during pregnancy. The children were examined and skin prick tested with inhalant and food allergens. IgE abs (against birch-pollen and recombinant Bet v 1(rBet v1)) and IgG4 abs (against rBet v 1) were analysed in serum. All children were assembled in one group to assess the effects of different ab responses (IgE/IgG4) on the development of atopic disease. RESULTS: Children exposed to high doses of birch-pollen during the first 3 months of life more often had detectable levels of IgG4 abs to rBet v 1 than the children in the other groups (P < 0.001), independent of sensitization to birch. Overall, the risk of allergic rhinoconjunctivitis was increased among children sensitized to birch-pollen and appeared more pronounced in children without detectable levels of IgG4 ab to rBet v 1 (Odds ratio 9.4; 95% Confidence interval: 5.5-16.1). IgE sensitization to birch-pollen seemed to have a stronger influence on the development of atopic disease than the IgG4-ab response. CONCLUSION: Exposure to high doses of inhalant allergens during the early postnatal period is associated with detectable levels of allergen-specific IgG4 ab even at 5 years of age. An immune modulating effect by IgG4 on symptoms of allergic rhinoconjunctivitis is suggested in children sensitized to birch.

Air Pollutants↗

Standardisation of glutaraldehyde-modified tyrosine-adsorbed tree pollen vaccines containing the Th1-inducing adjuvant, monophosphoryl lipid A (MPL).

BACKGROUND: a new range of allergy vaccines has been developed by the introduction of a relatively new Th1-inducing adjuvant known as 3-deacylated monophosphoryl lipid A (MPL). MPL adjuvant is of natural origin, derived from the lipopolysaccharide of Salmonella minnesota R595. This adjuvant is incorporated in a glutaraldehyde-modified pollen extract adsorbed to L-tyrosine (Pollinex Quattro). A major potential benefit provided by MPL adjuvant is the promotion of a Th1 response which enhances the efficacy of allergy vaccination and can consequently allow a reduction in the number of injections required for treatment. The standardisation of Pollinex Quattro tree pollen allergy vaccine is described and we include details of some innovative analytical procedures. METHODS AND RESULTS: an essential feature of the analytical strategy is the assay of the MPL adjuvant using a recently developed HPLC technique. The adjuvant has a complex chemical structure and the analysis is illustrated in detail. We give a full picture of the vaccine standardisation by describing biochemical and immunological characterisation of the allergen extract, together with some brief manufacturing details. CONCLUSIONS: a high overall level of standardisation is illustrated by a number of different tests applied to all stages of vaccine manufacture. Tree pollen allergen potency is measured following the pollen extraction, chemical modification and formulation as a tyrosine adsorbate. Good batch-to-batch reproducibility is shown. The HPLC assay for MPL adjuvant showed high quality resolution which did not vary when measuring raw material or when incorporated in the vaccine and the technically complex assay is shown to be reliable.

Adjuvants, Immunologic↗

Food hypersensitivity among adult patients: epidemiological and clinical aspects.

Food hypersensitivity (FH) is lesser frequent among adult patients than in childhood. Foods implicated in hypersensitivity reactions vary with sociocultural and diet habits from a geographic place to other. We studied 142 adult patients sensitized to foods, among 7698 patients visited at our Outpatient Clinic. Hundred and twenty patients referred clinical symptoms after consumption of one or more foods consistently. From the latest, 107 patients (89.2%) were atopics (92 of them sensitizes to dust mites) and 54 (45%) referred atopic familiar background. Most frequent recorded symptoms were: urticaria/angioedema 84 cases (70%), oral syndrome 65 (54%), asthma 48 (37%) and anaphylaxis 33 patients (27.5%). Shellfish sensitization occurred in 50 patients, fresh fruits in 33 and nuts in 29 cases. Shrimp (48 patients), squid (33), kiwi (14), papaya (14), avocado (13) and banana (12 cases) were the most frequent causes of FH. Significant statistical association between foods and inhalants was observed for fresh fruits and latex (p < 0.001), fresh fruits and pollens (p < 0.01), and shellfish and Blatta germanica (p < 0.001). Prevalence of FH among patients at our Area is around 1.6%. Tropical fruits, as other kind of fruits, seem to share common IgE-epitopes to pollens. High prevalence of shellfish and cockroach hypersensitivity could be more easily developed by previous domestic mites sensitization.

Adult↗

Developmental regulation of cell interactions in the Arabidopsis fiddlehead-1 mutant: a role for the epidermal cell wall and cuticle.

Although the plant epidermis serves primarily a protective role, during plant development some epidermal cells specialize, becoming competent to interact not only with pollen but also with other epidermal cells. In the former case, these interactions mediate recognition, germination, and pollen growth responses and, in the latter case, result in interorgan fusions which, most commonly, alter floral architecture in ways that are thought to promote reproductive success. In either case, all of the initial signaling events must take place across the cell wall and cuticle. In Arabidopsis, mutation of the FIDDLEHEAD gene alters the shoot epidermis such that all epidermal cells become competent to participate in both types of interactions. In fdh-1 mutants, epidermal cells manifest not only a contact-mediated fusion response but also interact with pollen. Since carpel epidermal derivatives manifest both of these properties, we postulated that fdh-1 epidermal cells were ectopically expressing a carpel-like program. In this report we demonstrate that manifestation of the fdh-1 phenotype does not require the product of the AGAMOUS gene, indicating that the phenotype is either independent of the carpel development program or that fdh-1 mutations activate a carpel-specific developmental program downstream of the AG gene. Furthermore, we demonstrate that plants bearing mutations in the fdh-1 gene show significant changes in cell wall and cuticular permeability. Biochemical analyses of the lipid composition of the crude cell wall fraction reveal that fdh-1 cell walls differ from wild-type and manifest significant changes in high-molecular-weight lipid peaks. These results suggest that cell wall and cuticular permeability may be important determinants in developmental signaling between interacting cells and implicate lipids as important factors in modulating the selectivity of the permeability barrier presented by the epidermal cell wall and cuticle.

Arabidopsis↗

Developmental staging of maize microspores reveals a transition in developing microspore proteins.

A method for the preparation of developmentally staged microspores and young pollen from maize (Zea mays) has been devised. The preparations are of sufficient purity and quantity for biochemical analysis, including the analysis of steady-state protein and RNA populations associated with each stage. A major transition in protein populations occurs during the developmental period that encompasses microspore mitosis, the asymmetric nuclear division producing the vegetative and generative nuclei. Several differences between early and late stage proteins can be detected by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Two-dimensional gel electrophoresis of proteins reveals that over half of the steady-state proteins differ between the younger and older stages, either quantitative or qualitative. One protein that increases in relative abundance about fourfold is actin. In vitro translation of RNA isolated from staged microspores demonstrates changes in microspore gene expression during the same developmental period.

Journal Article↗

Analysis of pollen and nectar of Arbutus unedo as a food source for Bombus terrestris (Hymenoptera: Apidae).

The mineral, total amino acid, and sterol compositions of pollen collected by Apis mellifera L. were compared with the pollen of a plant consumed by Bombus terrestris (L.): Arbutus unedo L. This plant provides the predominant food resource for the main autumn generation of B. terrestris in southern France. Honey bees also forage on this plant, although only for nectar. The mineral composition of 30 pollen samples collected by honey bees is close to the presently known requirements of A. mellifera, except for Cu and Mn, which are substantially lower. The total amino acid mean composition of a set of 54 pollen samples fits the basic requirements of honey bees except for valine, isoleucine, and methionine, which are present in lower concentrations in all the samples. For pollen of A. unedo, the amino acid balance is not very different from that of the survey. The main sterolic component in pollen of A. unedo, beta-sitosterol, is known to have antifeedant effects on A. mellifera. Honey bees cannot dealkylate C29 sterols like beta-sitosterol or delta5-avenasterol to obtain C27 cholesterol and ecdysteroids. Because these phytosterols as well as cholesterol are nearly absent from pollen of A. unedo, the metabolic capabilities of Apis seem unadapted to this plant. On the contrary, pollen of A. unedo is freely consumed by B. terrestris, which develops huge autumn populations solely on this food. These data indicate that the sterolic metabolisms of B. terrestris and A. mellifera differ, allowing separation in foraging activity.

Amino Acids↗

Expression of ACC oxidase genes differs among sex genotypes and sex phases in cucumber.

Ethylene has been implicated as a sex-determining hormone in cucumber: its exogenous application increases femaleness, and gynoecious genotypes were reported to produce more ethylene. In this study, three full-length ACC oxidase cDNAs were isolated from cucumber floral buds. RFLP analysis of a population that segregates for the F(femaleness) locus indicated that CS-ACO2 is linked to F at a distance of 8.7 cM. Expression of two of the genes, CS-ACO2 and CS-ACO3, was monitored in flowers, shoot tips and leaves of different sex genotypes. In situ mRNA hybridization indicated different patternsof tissue- and stage-specific expression of CS-ACO2 and CS-ACO3 in developing flowers. CS-ACO3 expression in mid-stage female flowers was localized to the nectaries, pistil and in the arrested staminoids, whereas CS-ACO2 transcript levels accumulated later and were found in placental tissue, ovary and staminoids. In male flowers, petals and nectaries expressed both genes, whereas ACO2 expression was strong in pollen of mature flowers. In young buds, strong expression was observed along developing vascular bundles. Four sex genotypes were compared for CS-ACO2 and CS-ACO3 expression in the shoot apex and young leaf. FF genotypes had higher transcript levels in leaves but lower levels in the shoot apex and in young buds, as compared to ff genotypes; the shoot-tip pattern is, therefore, inversely correlated with femaleness, and the possibility of a feedback inhibition mechanism underlying such correlation is discussed. The two CS-ACO genes studied displayed a differential response to ethrel treatment in different organs and sex genotypes, further demonstrating the complexity of the mechanisms controlling ethylene production during cucumber floral development.

Amino Acid Oxidoreductases↗

Study of the Th1/Th2 balance, including IL-10 production, in cultures of peripheral blood mononuclear cells from birch-pollen-allergic patients.

BACKGROUND: Excessive production of interleukin (IL)-4, IL-5, IL-10, and IL-13 is thought to be important in the development of allergy and asthma. The objective of this investigation was to study Th1/Th2-like cytokine profiles in vitro in seven patients allergic to birch pollen and six nonallergic controls during the birch-pollen season. METHODS: Peripheral blood mononuclear cells (PBMC) were isolated and cultured with birch-pollen extract (BPE) or tetanus toxoid (TT) for 7 days, harvested, and restimulated with the mitogens phytohemagglutinin (PHA) and phorbol 12-myristate 13-acetate (PMA) for 24 h. Cytokine production was determined by ELISA, and logarithmic cytokine ratios were compared between the two groups and between the antigens. RESULTS: In the allergic group, the cultures prestimulated with BPE had a more Th2-like cytokine response than the TT-prestimulated cultures; i.e., lower IFN-gamma and higher IL-10 production (P<0.05), as well as higher IL-5/IFN-gamma and IL-13/ IFN-gamma ratios (P<0.05). There were also significantly higher IL-4/IFN-gamma (P<0.005) and IL-5/IFN-gamma (P<0.05) ratios in BPE-stimulated cultures in the allergic group than in the control group. The IL-4 and IL-13 production in vitro correlated with the specific serum IgE levels. CONCLUSIONS: BPE stimulation induces a Th2-like cytokine response by PBMC isolated during the pollen season from birch-pollen-allergic patients, indicating a Th2-type immune response to birch pollen in vivo.

Adult↗

Hypomethylation promotes autonomous endosperm development and rescues postfertilization lethality in fie mutants.

In most flowering plants, fertilization is necessary for development of the central cell into endosperm, but in the fie-1 mutant of Arabidopsis, the central cell can proliferate autonomously. However, autonomous fie-1 endosperms do not develop completely: They have fewer nuclei than sexually produced endosperms, cellularization does not take place, and no clear distinction is seen between the different endosperm compartments. Here, we show that autonomous endosperm develop much further in hypomethylated than normally methylated fie-1 mutants, undergoing cellularization and regional specification to resemble endosperm in sexually produced wild-type seeds. Therefore, the combination of maternal hypomethylation and loss of FIE function enables formation of differentiated endosperm without fertilization. A maternal fie-1 mutation is also lethal to sexual seeds, even if the pollen donor is wild type. We report that sexual mutant fie-1 endosperms fail to cellularize and overproliferate, consistent with the hypothesis that embryo abortion may be due, at least in part, to a defect in endosperm development. Finally, we show that pollen from hypomethylated plants rescues fie-1 mutant seeds provided that it also donates a wild-type paternal FIE allele. These results are discussed in light of models for parent-of-origin effects on seed development.

Arabidopsis↗

A flourishing time for flower development. Workshop on Flower Development, sponsored by Fundación Juan March, Madrid, Spain, March 11-13, 1991.

Molecular genetics has recently erupted in the field of flower development, an area of research traditionally cultivated by plant physiologists. The isolation and molecular characterization of seven homeotic genes (four in Antirrhinum majus and three in Arabidopsis thaliana) that control both floral organogenesis and the transition from inflorescences to floral meristems is leading to major breakthroughs in the understanding of the mechanisms governing flower development. This has already had a great impact among plant physiologists, who are incorporating mutant analysis into studies of floral induction and flower development. We are still missing data about the nature of the pollen product of the S-locus in self-incompatibility systems, although current experimental approaches might provide this information in the near future. Gene technology appears to have a high potential in hybrid seed production through the construction of male sterile plants as well as of plants able to restore fertility. The study of genes regulating pigment formation in flowers continues to provide interesting data on gene expression in plants, in which phenomena such as co-suppression and methylation seem to play an important role. Altogether, one can predict that very exciting times are coming in the field of flower development.

Gene Expression Regulation↗

Heterogeneity of grass pollen allergens (Dactylis glomerata) recognized by IgE antibodies in human patients sera by a new nitrocellulose immunoprint technique.

A new nitrocellulose immunoprint technique has been developed to detect specific antigens or/and allergens present among a heterogeneous solution such as a water-soluble crude extract of a grass pollen (Dactylis glomerata). The antigens are separated by isoelectric focusing (IEF) in an agarose gel and characterized by their isoelectric point (pI). These antigens are transferred and immobilized on a nitrocellulose sheet. They are recognized by the binding of specific antibodies contained in an unfractionated serum to be studied. Finally, the binding of these antibodies is visualized by species- or/and class-specific antibodies themselves labeled by an enzyme or by radioactivity. So one can detect the allergens recognized by the specific serum IgE antibodies and also the other antigens recognized by specific IgG, IgA or IgM antibodies.

Allergens↗

Discontinuous IgE-binding epitopes contain multiple continuous epitope regions: results of an epitope mapping on recombinant Hol l 5, a major allergen from velvet grass pollen.

The knowledge of IgE-binding epitopes on allergen molecules is important for better understanding allergen-antibody interactions and, thus, for developing new strategies for immunotherapy. Our purpose was to more precisely define the number and structure of IgE-binding epitopes of a paradigmatic major grass pollen allergen. We performed an IgE-binding epitope mapping of rHol l 5, a group V pollen allergen of velvet grass (Holcus lanatus), with overlapping fragments (length between 15 and 186 amino acids), which were expressed in E. coli as MBP fusion proteins. Using sera of 65 grass pollen allergic patients, the fragments were analysed by immunoblotting for IgE reactivity. Specificity of antibody binding was confirmed by competitive blot inhibition assays. At least four different continuous IgE-binding epitopes were identified on small fragments (about 30 amino acids), and at least five different discontinuous IgE-binding epitopes on larger fragments, which were destroyed by further fragmentation. The fragments were differentially recognized by individual patients' sera. By investigating IgE-binding to one of the small fragments in more detail, we found further epitope regions on this fragment. It was noteworthy that IgE reactivity to small fragments was weak compared to large fragments or to the complete molecule. Competitive blot inhibition experiments showed that binding of IgE antibodies to the small fragments was specific but with lower avidity than to the complete rHol l 5. rHol l 5 harbours multiple discontinuous as well as continuous IgE-binding epitopes spread over the whole molecule, which were individually recognized by IgE antibodies from different patients. Low avidity of IgE antibodies to small fragments suggests that the continuous epitope regions do not represent the complete epitope and are most probably parts of discontinuous epitopes.

Allergens↗

A human model of allergic conjunctivitis.

Using the method of refractometry to measure protein concentration in tears, a simple model was developed to evaluate the allergic response in humans. Timothy grass pollen was instilled into the cul de sac of a human subject, and the protein content of the subject's tears was sampled every 15 minutes for three hours. Once the time course of this experiment was predictable, various eye medications were instilled into the subject's eye 30 minutes after allergic challenge. It was noted that 1.0% and 0.1% prednisolone, each combined with 0.12% phenylephrine hydrochloride, as well as 1% epinephryl borate eye drops, produced an almost immediate return to normal of tear protein levels, whereas 1.0% medrysone, 0.1% fluorometholone, 1.0% prednisolone, and 0.12% prednisolone had less pronounced effects on reduction of elevated tear protein concentration.

Administration, Topical↗

Automated histamine analysis for in vitro allergy testing. I. A method utilizing allergen-induced histamine release from whole blood.

A sensitive, automated, histamine assay system has been developed and applied for in vitro allergy testing. Nine common pollen and environmental allergens were used at three log dilutions for in vivo studies utilizing small volumes of blood (15-20 ml). The clinical evaluation was correlated with the results of the histamine release. two different procedures were utilized. The first is the commonly used histamine release from washed leukocytes. There was excellent correlation between the clinical evaluation and the results of histamine release from washed leukocytes in 17 different individuals. The second and simpler method utilized whole heparinized blood which might better reflect the immunologic reaction which occurs in vivo. Aliquots of blood and allergen were incubated for 1 hr at 37 degrees C and each supernatant was then analyzed for histamine release. There was excellent correlation between the two tests in 29 patients tested by both the whole blood and washed leukocyte methods. There was also good correlation between the clinical evaluation of the patients amd the intro tests. The precision, accuracy, and sensitivity of the automated histamine assay make feasible its routine application in the clinical study of allergic patients.

Allergens↗

Global expression profiling applied to plant development.

Plant development is controlled by both endogenous genetic programs and responses to exogenous signals. Microarray experiments are being used to identify the genes involved in these developmental processes. Most of the analyses conducted to date have been conducted on whole organs. Although these studies have provided valuable information, they are limited by the composite nature of plant organs that consist of multiple cell types. Technical advances that have made it possible to study global patterns of gene expression in individual cell types promise to increase greatly the information revealed by microarray experiments.

Flowers↗