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PubMed2006

Sensitization to different pollens and allergic disease in 4-year-old Swedish children.

BACKGROUND: Although the relationship between sensitization to different inhalant allergens in adolescents and adults has been intensively studied, information concerning sensitization in children is scarce in particular to pollens. OBJECTIVES: In 4-year-old children to elucidate the pollen immunoglobulin (IgE) antibody profile (birch only, timothy only and combinations of three pollens (birch, timothy or mugwort) and to relate the results to other inhalant and food allergens, as well as the presence of allergic diseases. METHODS: A total of 2551 4-year-old children belonging to a prospective birth cohort, which has been followed longitudinally (BAMSE), were investigated with respect to IgE antibodies to pollen and other inhalant and food allergens, and expression of allergic disease, based on questionnaire data. RESULTS: Eleven percent (n=285) of the children were sensitized to pollen. Birch was the dominating cause of pollen sensitization (birch sensitization only, n=133); followed by timothy grass pollen (n=56) and a combination of two (n=64) or three (n=30) pollens. A remarkably high proportion of the children sensitized only to birch was also sensitized to other inhalant allergens. This was not seen for children sensitized only to timothy. The highest frequencies of IgE reactivity to food were found in the group of children sensitized to the combination of birch, timothy and mugwort pollen. Children sensitized to timothy only, exhibited symptoms of allergic disease significantly less frequently compared with children sensitized to birch only. Sensitization to birch pollen was found to be closely associated with rhinitis and eczema compared with asthma. The highest frequency of asthma and/or rhinitis and/or eczema was reported in children sensitized to at least two pollens. CONCLUSION: Our results demonstrate that birch is the dominating source of pollen sensitization at the age of four in Sweden. This might associate with the pattern of sensitization to other inhalant and food allergens as well as influence on the expression of allergic disease in this particular age group.

Air Pollutants↗
PubMed1981

Selective suppression of antibody production with the aid of radiolabelled birch pollen allergen.

In accordance with the clonal selection theory we intended to prevent the development of artificially induced birch pollen allergy in rabbits with the aid of the radiolabelled pollen allergen (75-1000 microCi125 I-pollen/animal) intravenously administered prior to pollen sensitization. The birch pollen allergen, is accordance with Burnet's working hypothesis, reacts only with a genetically determining B cell subpopulation. The fixation of the radiolabelled birch pollen allergen to the receptors of the competent B cell clone causes the lesion of the latter. Compared with the control group, this group of rabbits showed an extensive suppression of anaphylactic reagin-like PCA-antibodies, and haemagglutinating antibodies in the blood as well as in nasal secretion. In addition, we tried to influence the already ongoing synthesis of the antibodies with the aid of subsequent intravenously administered radiolabelled birch pollen allergen (750-100 microCi 125 I-pollen/animal). An intensive suppression of the synthesis of antibodies could also be proved in this case. The simultaneous immunization of the control rabbits with birch pollen and egg albumin resulted in the production of antibodies against both antigens, as expected. The hot-labelled birch pollen antigen intravenously injected before or after immunization with egg albumin and birch pollen led selectively to suppression of anti-birch-pollen PCA antibodies. The synthesis of anti-egg albumin PCA antibodies was unaffected.

Allergens↗
PubMed1998

Characterization of cogon grass (Imperata cylindrica) pollen extract and preliminary analysis of grass group 1, 4 and 5 homologues using monoclonal antibodies to Phleum pratense.

BACKGROUND: Previous studies have established the role of Imperata cylindrica (Ic) pollen in type I allergic disorders. However, no systematic information is available on the allergen composition of Ic pollen extract. OBJECTIVES: To characterize the IgE-binding proteins of Ic pollen extract and to detect the presence of grass group 1, 4 and 5 allergen homologues, if any. METHODS: Pollen extract of Ic was analyzed by in vivo and in vitro procedures such as intradermal tests (ID), enzyme-linked immunosorbent assay (ELISA), ELISA-inhibition, thin-layer isoelectric focusing (TLIEF), sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. Dot blot assay was carried out to check the presence of well-known group 1, 4, and 5 allergen homologues in Ic pollen extract. RESULTS: Out of 303 respiratory allergies patients skin-tested, 27 showed sensitivity to Ic pollen extract. Specific IgE levels were elevated in all 15 serum samples tested. The extract prepared for this study was found to be highly potent since it required only 400 ng of homologous proteins for 50% inhibition of binding in ELISA inhibition assays. TLIEF of Ic pollen extract showed 44 silver-stained bands (pI 3.5-7.0) while SDS-PAGE resolved it into 24 Coomassie-Brilliant-Blue-stained bands (MW 100-10 kD). Immunoblotting with individual patient sera recognized 7 major IgE-binding bands (MW 85, 62, 57, 43, 40, 28 and 16 kD) in Ic pollen extract. A panel of monoclonal antibodies, specific to group 1, 4 and 5 allergens from Phleum pratense pollen extract identified group 5 and group 4 homologues in Ic pollen extract. CONCLUSION: Ic pollen extract was characterized for the protein profile by TLIEF and SDS-PAGE. IgE reactivity was determined by ELISA and immunoblot. Monoclonal antibodies to group 5 and group 4 allergens reacted weakly showing that this pollen contains group 5 and group 4 homologous allergens.

Adolescent↗
PubMed2005

Group 13 allergens as environmental and immunological markers for grass pollen allergy: studies by immunogold field emission scanning and transmission electron microscopy.

BACKGROUND: Polygalacturonases were recently identified as important grass pollen allergens and designated group 13 allergens. The objective of the present study was to investigate the presence of group 13 grass pollen allergens in different grass species, their release and ultrastructural location in dry and hydrated grass pollen. METHODS: Nitrocellulose-blotted allergen extracts from 12 wild and cultivated grass genera were probed with a rabbit antiserum raised against purified recombinant timothy grass pollen allergen, Phl p 13. The release kinetics of Phl p 13 from timothy grass pollen hydrated for 0.5 min to 3 h were analyzed by immunoblotting. Phl p 13 was localized in dry and hydrated grass pollen grains by immunogold field emission scanning and transmission electron microscopy. RESULTS: Group 13 allergens were detected in all 12 wild and cultivated grass genera representing the major subfamilies of the Poaceae. Ultrastructurally, the allergen was located in the wall and in the cytoplasm of timothy grass pollen grains. In the cytoplasm, Phl p 13 was associated with polysaccharide particles and as yet undescribed stacks of microtubule-like structures. After hydration in rain water, pollen grains expel cytoplasmic particles of respirable size containing Phl p 13, which becomes detectable in aqueous supernatants already after 0.5 min. CONCLUSIONS: Group 13 allergens represent one set of marker allergens which specifically occur in pollen of the major grass subfamilies and are rapidly released in association with respirable particles after pollen hydration. They may be considered as environmental markers for grass pollen exposure and group 13-specific IgE antibodies as immunological markers for genuine grass pollen sensitization.

Allergens↗
PubMed2006

The major allergen of olive pollen Ole e 1 is a diagnostic marker for sensitization to Oleaceae.

BACKGROUND: Trees of the family Oleaceae are important allergen sources, with a strongly varying geographic distribution. For example, olive pollen is an important allergen source in Mediterranean countries, whereas ash pollen dominates in Northern and Central Europe and North America. The aim of this study was to compare the profiles of olive and ash pollen allergens and to study the degree of cross-reactivity using populations of allergic patients selectively exposed to olive or ash pollen. METHODS: Olive and ash pollen extracts were analyzed by IgE immunoblotting using sera from Spanish patients highly exposed to olive pollen and Austrian patients without olive but ash pollen exposure. IgE cross-reactivity was studied by qualitative immunoblot inhibition assays and semiquantitative ELISA inhibitions using olive, ash, birch, mugwort, timothy grass pollen extracts and the major olive pollen allergen, Ole e 1. RESULTS: Spanish and Austrian patients exhibited an almost identical IgE-binding profile to olive and ash pollen allergens, with major reactivity directed against Ole e 1, and its homologous ash counterpart, Fra e 1. IgE inhibition experiments demonstrated extensive cross-reactivity between olive and ash pollen allergens. However, whereas cross-reactions between profilins and calcium-binding allergens also occurred between unrelated plant species, cross-reactivity to Ole e 1 was confined to plants belonging to the Oleaceae. CONCLUSIONS: Ole e 1 is a marker allergen for the diagnosis of olive and ash pollen allergy.

Allergens↗
PubMed2005

Transgene dispersal through pollen.

Techniques used for the transfer of novel genes into host plant genomes have created new possibilities for crop improvement. The implementation of transgenic crop species into agriculture has introduced the possibility of transgene escape into the environment via pollen dispersal. Although the movement of pollen is a critical step in transgene escape, there is currently no system to monitor transgenic pollen movement under field conditions. The development of an effective in vivo monitoring system suitable for use under field conditions is needed for research and commercial purposes so potential risks can be quantified and evaluated. This chapter describes the development of a model system using green fluorescent protein (GFP) expression in pollen as a marker to monitor pollen distribution patterns. A pollen specific promoter was used to express the GFP gene in tobacco (Nicotiana tabacum L.). GFP was visualized in pollen and growing pollen tubes using fluorescent microscopy. Furthermore, the goal of this research was to compare the dynamics of pollen movement with that of gene flow by using another method of whole plant expression of GFP to estimate out-crossing frequencies by progeny analysis. Pollen movement and gene flow were quantified under field conditions. Pollen traps were collected and screened for presence of GFP-tagged pollen using fluorescence microscopy. Progeny from wild type plants were screened with a hand held ultraviolet light for detection of the GFP phenotype.

Gene Transfer Techniques↗
PubMed2001

[Long-term study of airborne allergic pollen count, C. Japonica and cupressaceae in Japan].

We have investigated the distribution of airborne pollen at the different eleven points in Japan from 1987 to 1998 using gravity sampler. To clarify the characteristics causative pollen for Japanese cedar pollinosis, we examined annual change of pollen counts, dispersing period and geographical difference of C. japonica and Cupressaceae pollen. C. japonica pollen occupied much more in Central Japan and Cupressaceae in the west of Japan than the other area. In Hamamatsu City, both of pollen counts were most of all and we found a tendency that the more pollen counts the longer dispersing period. As they reported that at the starting day of pollen in this method some patients had already suffered from allergic symptoms, we considered pollen grains were dispersing in spite of being continuously captured. In these twelve years we found that patients with Japanese pollinosis are exposured by causative agents during about 100 days every spring. But we could not observe the trend of increasing pollen counts and earlier starting day because of global warming. Further more we found that the pollen counts of C. japonica in autumn was increasing since 1994. As one of the factors of increasing patients with pollinosis, we thought that total exposure period of causative pollen every year were longer than that of 1980s.

Air Pollutants↗
PubMed2001

Comparative analysis of pollen counts of Corylus, Alnus and Betula in Szczecin, Warsaw and Lublin (2000-2001).

The aim of the study was to compare the airborne concentrations of allergenic pollen produced by three early flowering tree taxa (Corylus, Alnus, Betula) in the cities of Warsaw (central Poland), Lublin (eastern Poland) and Szczecin (western Poland) during the years 2000-2001. Measurements were performed by the volumetric method. Pollen seasons were defined as the periods in which 95% of the total catch occurred. The highest concentration and annual pollen count of Corylus was measured in Lublin in both seasons, while the highest annual pollen counts of Alnus and Betula were noted in Warsaw, where the annual pollen count of Betula in 2001 was four times higher than in 2000 and equalled 5,376 grains in m3 per 24 h. Significant differences in the pollen count of the examined taxa were observed between two seasons: the pollen count of Corylus was higher in 2000 than in 2001, while for Alnus and Betula the opposite was the case. The longest pollen seasons were observed at low annual pollen counts for the pollen of Corylus. Results of the study reveal significant differences between the seasons and the cities. The differences concern the dates of the appearance of pollen grains in the air, the duration of the presence of sporomorphs and the maximum concentrations in particular seasons. The pollen counts of alder, birch and hazel trees are determined by the weather, diversity of local flora and specific rhythm of pollination of particular taxa.

Air Pollution↗
PubMed1990

Cross-reactivity among the pollen proteins of birch and apple trees.

In the spring of 1986, the pollen were collected from apple trees in full blossom, and were investigated for their allergenicity. The patients selected for study were subjects with a combined inhalant allergy to birch pollen and an oral allergy to apple fruit. The apple pollen extract yielded about the same percentage of nondialysable substance as obtained from birch pollen. In contrast to the latter, UV-spectroscopy revealed no flavonoids adsorbed to the apple pollen proteins. Patients with a combined allergy to birch pollen and apple fruit showed positive skin reactions to both birch and apple pollen extract. Inhibition of IgE-binding in RAST to birch pollen was observed by apple pollen extract at a 1000-fold lower potency than the homologous birch allergens. Immunoblotting demonstrated IgG-antibodies in birch-allergic sera cross-reactive with apple pollen components. It is concluded that minor allergenic determinants cross-reactive with birch pollen epitopes occur not only in the fruit, but also in the pollen of the apple tree.

Cross Reactions↗
PubMed1996

[Effects of global climate change on Japanese cedar pollen concentration in air--estimated results obtained from Yamagata City and its surrounding area].

The effects of global warming on Japanese cedar pollen concentration in air were examined. There was a significant increase in total pollen count in years where small or average values of pollen dispersion were observed and where summertime temperatures were postulated to have risen from 2 to 5 degrees C. There were no remarkable differences in total pollen count in years where large values of pollen dispersion were observed. On the other hand, a drastic decrease in total pollen count was expected when summertime temperatures were postulated to have dropped from 2 to 5 degrees C. A major factor controlled the value of total pollen count was examined by use of a simulation method of airborne Cryptomeria japonica pollen. The values of total pollen count mainly controlled by the amount of male flower at basins and in Japanese cedar forested areas, and the manner in which plains isolated from pollen sources are considered in part to be subjected to meteorological conditions. The difference of initiation time of the pollen season has no great influence on the values of total pollen count.

Air Pollution↗
PubMed1997

Foraging currencies for non-energetic resources: pollen collection by bumblebees

Animals sometimes forage for resources whose benefits are not energy-based. In such cases, the benefits and costs of foraging cannot be directly compared, making it difficult to evaluate the currency of fitness maximized by the animal's behaviour. For bees, pollen represents such a resource, because they collect pollen as the sole protein source for developing larvae, rather than as a source of energy. This study compared three currencies that could be maximized during pollen collection by bumblebees (Bombus spp.) foraging on different lupin species (Lupinus spp.): pollen collected per inflorescence, pollen collection rate (gross benefits/time) and pollen collection efficiency (gross benefits/costs). Bees visiting lupin inflorescences begin foraging low on an inflorescence and then move upward through a gradient of increasing pollen availability. To maximize each currency, we predicted that a bee would begin foraging at a different position along a lupin inflorescence, depending on the relative influences of the time and energy costs associated with handling flowers and flight. Based on comparisons of observed and predicted starting positions for seven situations, maximization of gross pollen-collection efficiency predicted observed behaviour better than either pollen collected per inflorescence or gross rate of pollen collection. Such maximization of pollen-collection efficiency would enhance a bee's lifetime input of pollen to her colony, as has been demonstrated for nectar-collecting bees. Hence, the economics of foraging for non-energetic resources do not differ qualitatively from foraging for energy.Copyright 1997 The Association for the Study of Animal Behaviour1997The Association for the Study of Animal Behaviour

Journal Article↗
PubMed2000

Perception of the pollen need by foragers in a honeybee colony.

Honeybees, Apis mellifera, adjust their pollen foraging activity according to the need for pollen within the colony, determined by the amount of stored pollen and young brood present in the hive. To clarify how pollen foragers detect the supply of pollen, we followed individual honeybees while they were returning with pollen. Pollen foragers deposited their loads on the frame where most of the unsealed brood was, independent of the position of this frame within the hive. They also inspected more cells on that frame and spent most of their time there, indicating that pollen foragers may individually evaluate the pollen requirements of the colony. In 18 normal-sized colonies we also tested whether olfactory cues provided by a frame of hungry young brood or an additional pollen frame covered by cages affect foraging activity. These experiments showed that olfactory stimulation within the colony is insufficient to increase or decrease the foraging effort, but suggest that foragers must have direct contact with the brood and pollen area to regulate their foraging activity according to the conditions in the colony. The different mechanisms by which foragers may gather the information about pollen supply are discussed. Copyright 2000 The Association for the Study of Animal Behaviour.

Journal Article↗
PubMed2001

Pollen digestion by flower-feeding Scarabaeidae: protea beetles (Cetoniini) and monkey beetles (Hopliini).

Pollen protoplasm is very nutritious, but the hard and highly resistant outer wall (exine) of the pollen grain presents an obstacle that pollen-feeders must overcome to benefit from the valuable protoplasm. Pollen digestion in three pollen-eating scarab beetles, the green protea beetle Trichostetha fascicularis and two monkey beetle species, Peritrichia cinerea and Pachynema flavolineata, was investigated. Adult beetles were collected for observations of feeding mechanisms, scanning electron microscopy of mouthparts, and histological examination of gut contents. Serial sections of different regions along the gut were stained for different nutrients and examined to determine the appearance of the pollen grains and the removal of carbohydrates, proteins and lipids from the pollen. During feeding, beetles used dense brushes of setae on the tips of their maxillae to mop up nectar and sweep pollen into their mouths. Grains were ingested intact. Nutrients were effectively removed from the grains as they passed along the gut and most pollen grains in the hindgut were empty, with broken exines. All the beetles excreted the undigested exine. Comparison with other studies on pollen digestion by a variety of insect and vertebrate pollen-feeders suggests that digestive enzyme penetration of ingested pollen grains and exudation of the products of digestion is the most likely method used by these beetles.

Journal Article↗
PubMed1999

Analysis of mRNA stabilities during pollen development and in BY2 cells

RNA stability is important in post-transcriptional gene expression. However, its role during plant male gametophyte development has not been studied in detail, probably because of technical difficulties and because earlier work suggested that, in general, mRNAs expressed during late pollen development would be stable. We determined mRNA stabilities of 12 pollen-expressed genes. We measured pollen mRNA decay during in planta development and in vitro maturation after transcriptional inhibition with ActinomycinD (ActD). Because five of these genes were also expressed in somatic cells we also measured RNA decay in BY2 cells after transcriptional inhibition with ActD. Ten of the mRNAs were highly stable during pollen development (t1/2 > 30 h), including all seven pollen-specific mRNAs. However, two mRNAs showed a logarithmic decrease as pollen development proceeded, both in planta and in vitro. In fact, for one of these two, GUT8-2b, the mRNA level decreased significantly within 3 h. Our results show that many pollen-expressed mRNAs are indeed long-lived but that at least some mRNAs undergo decay during pollen maturation. Thus, there is no overall cytoplasmic condition in pollen that stabilizes mRNAs and specific mRNA turnover mechanisms must exist. We detected reciprocal mRNA stabilities with two genes that are expressed in both BY2 cells and pollen. GUT15 mRNA, known to be unstable (t1/2 < 3 h) in BY2 cells, was highly stable in pollen. In contrast, GRP2 mRNA decayed in pollen but was highly stable in BY2 cells. Therefore, we conclude that mRNA stability can be tissue-specific.

Journal Article↗
PubMed1991

Action of the Style Product of the Self-Incompatibility Gene of Nicotiana alata (S-RNase) on in Vitro-Grown Pollen Tubes.

The products of the S-locus expressed in female tissues of Nicotiana alata are ribonucleases (S-RNases). The arrest of growth of incompatible pollen tubes in styles may result from entry of the S-RNase into the pollen tube and degradation of pollen tube RNA. We investigated the action of isolated S-RNases on pollen tubes grown in vitro and found that S-RNase is taken up by the pollen without substantial alteration. The S-RNases inhibit incorporation of exogenously added radioactive amino acids into protein by the germinated pollen. The S-RNases also inhibit in vitro translation of pollen tube RNA in a wheat germ cell-free extract. We found no evidence for a specific mRNA substrate for the S-RNases, which implies that if RNase activity is involved in the control of self-incompatibility, allelic specificity is more likely to depend on the selective uptake of S-RNases into pollen tubes or their selective activation or inactivation by pollen factors, rather than cleavage of a specific substrate. Heat treating S2-RNase largely destroys its RNase activity but increases its inhibitory effect on in vitro pollen tube growth. This effect is not due to an increased uptake of S2-RNase by the pollen but is associated with a greatly enhanced accumulation of S2-RNase on the outer surface of the pollen grains.

Journal Article↗
PubMed2000

Absence of conspecific pollen advantage in the dynamics of an Ipomopsis (Polemoniaceae) hybrid zone.

The frequency of hybrid formation in angiosperms depends on how often heterospecific pollen is transferred to the stigma and on the success of that heterospecific pollen at fertilizing ovules. Even if heterospecific pollen is capable of effecting fertilization it may perform poorly when conspecific pollen is also available on the stigma. We applied pollen mixtures to stigmas to determine how pollen interactions affect siring success and the frequency of hybrid formation between two species of Ipomopsis (Polemoniaceae) in Colorado. Plants of both parental species and natural hybrids were pollinated with I. aggregata and I. tenuituba pollen in ratios of 100:0, 80:20, 50:50, 20:80, and 0:100 by mass. Plants were homozygous for different alleles at an isozyme marker, allowing us to distinguish the type of pollen parent for 2166 viable seeds from 273 fruits. In contrast to studies of many other hybridizing taxa, there was no evidence of an advantage to conspecific pollen, nor did composition of the stigmatic pollen load affect seed set. Instead, the frequency of seeds sired by a given species was proportional to its representation in the pollen load. In this hybrid zone, both the frequency of first-generation hybrid formation and the relative male fitness of the two parental species should be predictable from the rates of pollen transfer to stigmas.

Journal Article↗
PubMed1996

A study on the nutritive value of pollen from the Chinese Masson Pine (Pinus massoniana) and its effect on fecal characteristics in rats.

A digestion experiment with growing rats was conducted to study the effect of native and broken pollen of Chinese Masson Pine (Pinus massoniana) on fecal composition and digestibilities of dry matter, crude protein and crude ash. For that issue each 10 of 30 animals were fed a semisynthetic diet containing either no pollen or and addition of 6% native or broken pollen for 17 days at restricted amounts. In a second experiment each 6 of 12 growing rats received a semisynthetic diet containing either no or 5% broken pollen. At day 4, 11 and 18 samples of the feces were collected and analyzed for mesophilic aerobic bacteria. The pollen contained large amounts of cell wall constituents (26-30% lignin and 10-15% cellulose, 2-13% hemicellulose) with higher values found in native pollen. The contents of crude protein, crude fat, total lipids, available carbohydrates (starch and sugars), crude ash, gross energy and metabolizable energy were 13%, 2-10%, 8-10%, 17-18%, 3.1-3.5%, 21-22 kJ/g and 5.7-9.0 kJ/g. The addition of native pollen to the diet increased the total amount of feces by 71% as compared to the control level. The fecal contents of dry matter and of crude protein and crude ash in fecal dry matter changed by -5, +4 and -5 percentage units. The apparent digestibilites of dry matter and crude protein decreased by 3 and 5 percentage units, respectively, while the apparent digestibility of crude ash remained unchanged. Broken pollen acted in the same direction, however the effects were about 20% less pronounced as compared to native pollen. The pollen feeding reduced the fecal germ contents of Proteus mirabilis and Escherichia coli, while the amounts of alpha-hemolysing streptococci was increased. In total, the observed effects of an addition of native broken or pollen to the diet seemed to be based mainly on the increased intake of cell wall constituents.

Animals↗
PubMed1995

Pollen viability and transgene expression following storage in honey.

Transgenic plants of tobacco and Arabidopsis that produce genetically marked pollen, expressing the reporter gene uidA (gusA), were generated to determine whether pollen proteins can be expressed and stable in honey, a potential route by which foreign proteins might enter the wider environment. Hydrated tobacco pollen was found to lose viability rapidly in honey, while pollen in the natural dehydrated form remained viable for at least several days and in some cases several weeks, as determined by FDA staining activity and germinability. Dehydrated pollen was found to be capable of transient foreign gene expression, following microprojectile bombardment, after incubation in honey for at least 120 h. PCR amplification of transgene sequences in pollen of transgenic plants revealed that pollen DNA can remain relatively intact after 7 weeks in honey. GUS enzyme activity analysis and SDS-PAGE of pollen proteins revealed that foreign and native pollen proteins are stable in pollen incubated in honey for at least 6 weeks. We conclude that pollen may represent an ecologically important vector for transgenic protein products.

Arabidopsis↗