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At least 271 records · Page 15Linked to original sources

Localization of calcium-dependent ATPase in germinating pollen grain and pollen tube in Vicia faba.

It was demonstrated by a cytochemical method that the calcium-dependent ATPase was present in the germinating pollen grain. Immediately after hydration the enzyme was observed in the pollen plasmalemma. During germination, the chief sites of the enzyme activity were the plasmalemma around the aperture site and the cisternae of endoplasmic reticulum. In the pollen grain with a grown tube, the enzyme was localized in the plasmalemma of the tube tip, in the endoplasmic reticulum below the tip, and in the tonoplast of the vacuoles formed in the pollen grain after tube germination.

Calcium-Transporting ATPases↗

[Pollen information of airborne Japanese cedar pollen using a simulation method].

We have developed a simulation method of airborne Cryptomeria japonica pollen distribution on a map displayed visually on a TV screen. Each patient can be available the information where the place he or she lives. The pollen season in 1995, we served the information about airborne pollen distribution on a map and C. japonica flowering areas on a map to a local resident through TV broadcasting. To verify the simulation method, comparison was made between the result from actual pollen counting and from simulation. It was clarified that both results were comparatively agreed on daily basis. Problem about compatibility among personal computers were solved to rewrite the program of displaying the image using Visual Basic for MS-Windows and create image files. The files can be read continuously by animation software. We think the information can be offered to local resident, local clinicians and patients waiting at the clinics by use of computer networks.

Computer Communication Networks↗

Endogenous Gibberellins of Pine Pollen: II. Changes during Germination of Pinus attenuata, P. coulteri, and P. ponderosa Pollen.

The endogenous gibberellins (GAs) of pollen of Pinus attenuata, P. coulteri, and P. ponderosa were bioassayed at hour 0, 3, 15, 24, 48 and 72 of germination. Dormant pollen showed relatively high GA activity throughout the elution spectrum (i.e. ranging from relatively nonpolar to highly polar). The maximum GA activity was obtained at hour 15 in more polar regions and especially in the zone corresponding to GA(3) (for P. attenuata estimated as 250 micrograms of GA(3)/kilogram pollen). It is probable that the "nonpolar" GAs present in high quantities in dormant pollen and in early stages of germination were converted to "more polar" GAs as germination progressed. The amount of all GAs decreased after hour 15 of germination and by hour 72 no GAs could be detected. Among the species tested P. attenuata showed the highest over-all GA activity.

Journal Article↗

cDNA cloning and expression of timothy grass (Phleum pratense) pollen profilin in Escherichia coli: comparison with birch pollen profilin.

Profilin, an actin-binding protein, was previously described as a ubiquitous allergen which is responsible for cross-reactivities in about 20% of pollen and food allergic patients. A complete cDNA clone coding for timothy grass (Phelum pratense) pollen profilin was isolated using allergic patients IgE. The deduced amino acid sequence of timothy grass profilin shares a sequence identity of 79% with birch profilin and other plant profilins and a lower average sequence identity of 35% with other eukaryotic profilins. The high degree of homology among different plant profilins at the DNA and protein level explains the extensive cross-reactivities observed in profilin allergic patients. Recombinant timothy grass pollen profilin was expressed in Escherichia coli as a beta-galactosidase fusion protein and shown to bind IgE from profilin allergic patients similar to recombinant birch profilin. Slight differences regarding the IgE-binding capacity of birch and timothy grass profilin indicate that not all IgE-epitopes of the two profilins are conserved. It is speculated that profilin allergic patients were initially sensitized against a certain profilin and then cross-react with the homologous proteins.

Allergens↗

A gene showing sequence similarity to pectin esterase is specifically expressed in developing pollen of Brassica napus. Sequences in its 5' flanking region are conserved in other pollen-specific promoters.

Differential screening of a Brassica napus genomic library led to the isolation of the clone named Bp 19 containing a gene which is highly expressed during microspore development. The accumulation of Bp19 mRNA starts in uninucleate microspores, increases during development reaching a peak in the late stages but declines considerably in mature pollen. The nucleotide sequence of the entire coding region and of extended portions of the 5' and 3' flanking regions was determined. Several homologous cDNA clones were also isolated and sequenced. The Bp 19 gene contains a single intron of 137 bp and gives origin to a mRNA of ca. 1.9 kb which codes for a polypeptide of 584 amino acids. Bp 19 protein has an estimated molecular weight of 63 kilodaltons and has a highly hydrophobic amino terminal region which shows features of a signal peptide. The carboxy half of the Bp 19 protein, starting at amino acid 269, has striking sequence similarity to the pectin esterases of tomato and of the plant pathogen Erwinia chrysanthemi. Four short domains are extremely well conserved in all the three proteins and therefore could represent catalytic sites responsible for enzyme activity. Comparison of the 5' flanking region of the Bp 19 gene with the sequence of other pollen-specific promoters revealed the presence of several conserved regions. These short promoter sequences could correspond to regulatory elements responsible for pollen-specific gene expression.

Amino Acid Sequence↗

Comparison of in-house allergen extracts of Phleum pratense (timothy grass) pollen with the international standard and investigation of IgE specificities of a grass-pollen serum pool from West Germany and of the one recommended by the World Health Organization.

We compared our in-house reference extract (RE) and a production extract (PE) with the international reference preparation (IRP) of International Union of Immunological Societies of timothy-grass pollen, using various biochemical and immunochemical methods. Furthermore, we compared the IgE composition of our in-house grass-pollen serum pool (West Germany) with the serum pool recommended by the World Health Organization, using the RE for crossed radioimmunoelectrophoresis (CRIE). Our extracts (RE and PE) were nearly comparable with the IRP. Only by CRIE one more allergen could be detected in RE and PE than in IRP and in another extract purchased from an American company. This finding may depend on the region where the source materials were harvested and which source materials were used for the preparation of the extracts. Furthermore, by high-performance liquid chromatography in the low-molecular-weight range, two distinct peaks could be detected in the nondiafiltered IRP, which were detected in the diafiltered RE and PE as traces only. The IgE composition of our in-house grass-pollen serum pool was comparable with the serum pool recommended by the World Health Organization, as detected by CRIE using RE.

Allergens↗

Homology of the major birch-pollen allergen, Bet v I, with the major pollen allergens of alder, hazel, and hornbeam at the nucleic acid level as determined by cross-hybridization.

To investigate the relationship of the major allergens of birch (Bet v I), alder (Aln g I), hazel (Cor a I), and hornbeam (Car b I) at the nucleic acid level, a cDNA clone coding for the complete Bet v I protein was used for Northern and Southern blot experiments. RNAs were isolated from pollen of birch (Betula verrucosa), alder (Alnus glutinosa), hazel (Corylus avellana), and hornbeam (Carpinus betulus). Hybridization was performed at different stringencies. At high stringency, comparable binding of the complete Bet v I cDNA probe to pollen RNAs from birch, alder, and hazel could be observed, indicating high homology of the mRNAs coding for these allergens. With the 3' and 5' half fragments of the Bet v I cDNA, both probes bound to transcripts of all four tree pollens, but most strongly to birch RNA. In Southern blots, distinct binding patterns of genomic DNA digests of birch, alder, hazel, and hornbeam were observed. Most bands were observed with birch DNA digests and less with alder, whereas in genomic DNA digests of hornbeam and hazel, only one band was observed. The result of these cross-hybridization experiments indicate a high homology at the nucleic acid level of the four major allergens of trees belonging to the order Fagales. The sequence similarity presented here further corroborates earlier observations of immunologic cross-reactivity at the protein level. Therefore, in the case of the major allergens of the Betulaceae, an extract with only one major allergen, preferentially Bet v I, instead of all four major allergens, should be sufficient for diagnostic and therapeutic purposes.

Allergens↗

Allergens of Arizona cypress (Cupressus arizonica) pollen: characterization of the pollen extract and identification of the allergenic components.

Species of the Cupressaceae family are an important cause of respiratory allergies in countries with a Mediterranean climate. An allergenic extract from Cupressus arizonica pollen was prepared with two extraction steps followed by ammonium sulfate precipitation, giving a protein yield of about 3%. Cupressus arizonica pollen extract was also characterized by means of sodium dodecylsulfate-polyacrylamide gel electrophoresis, followed by IgE and IgG immunoblotting and lectin blotting. IgE reactivity was restricted to six components, whereas IgG binding showed a more complex pattern. A 43 kd component, predominant both in its intensity and frequency of recognition by human IgE antibodies, was identified as the major allergen of C. arizonica. Four of the six IgE-binding components, including the major allergen, seem to be glycoproteins, as confirmed by the lectin blotting analysis. The extract produced inhouse was used to set up an immunoenzymatic test to evaluate the specific IgE binding in a panel of sera from 33 immunotherapy-free subjects who were monosensitized to cypress pollen. The percent of positivity obtained was much higher than that reported in the literature for commercial immunoassays.

Adult↗

Zymolyase removes callose from germinating pollen and pollen tube walls.

The impermeable callosic wall that surrounds germinating pollen grains and pollen tubes is removed by brief treatment with Zymolyase, an enzyme preparation that hydrolyzes linear glucose polymers with beta-1,3 linkages. Zymolyase treatment does not interfere with pollen tube growth or cylosis in the vegetative cell.

Cell Wall↗

Molecular cloning of a cDNA encoding a pollen extracellular protein as a potential source of a pollen allergen in Brassica rapa.

A polyclonal antiserum was raised against the extracellular pollen proteins of Brassica rapa and used for screening the expression cDNA libraries made from immature anthers. We obtained five groups of cDNA clones, including cDNAs similar to PCP1, thioredoxin, and lipid transfer protein (LTP). Recombinant protein of the cDNA clone showing sequence similarity to LTP was demonstrated to bind IgE of a patient allergic to Brassica pollen. The cDNA clone reported here, therefore, represents a novel pollen allergen of Brassica rapa.

Allergens↗

Studies on the physiology of pollen and pollen tube growth. 1. pinus roxburghii.

Pollen grains and pollen tubes of Pinus roxburghii were subjected to histochemical technique with a view to determining the distributional pattern of reserve substances (ascorbic acid, proteins starch, polysaccharides, nucleic acids) and some enzymes (acid phosphatase, glucose-6-phosphatase, peroxidase, esterase, cytochrome oxidase, succinate dehydrogenase). An attempt is made to correlate the activity of different enzymes in relation to pollen tube growth.

Enzymes↗

Allergic rhinitis to grass pollen: measurement of inflammatory mediators of mast cell and eosinophils in native nasal fluid lavage and in serum out of and during pollen season.

BACKGROUND: In allergic rhinitis, mast cells, activated by cross-linking of allergen to mast cell-bound specific IgE, release both vasoactive mediators related to the early nasal symptoms and chemotactic mediators that attract inflammatory cells, such as eosinophils, related to the late-phase response. OBJECTIVE: We have analyzed, during and out of pollen season, in blood and nasal fluid from patients allergic to grass pollen, histamine and tryptase to monitor the early phase markers and eosinophil and eosinophil cationic protein (ECP) to monitor the late phase. METHODS: Twenty patients were enrolled in the study. As a control, we studied 10 nonatopic subjects. Mediators and eosinophils were assessed in blood and nasal fluid. Histamine was tested only in nasal fluid. RESULTS: During pollen season, tryptase but not histamine increased in nasal fluids from patients (2.96 vs 0.22 U/ml, p = 0.001) and correlated with symptom scores (r(s) = 0.63, p = 0.003). Tryptase was not detected in serum. Eosinophils increased in nasal cytology (17.0% vs 2.0%, p = 0.001) and in the blood (265 vs 12.7 x 10(6) L, p = 0.001) from patients, but they did not correlate with symptom scores. ECP increased only in the nasal lavage (1633 vs 1.30 ng/ml, p = 0.001) and correlated with symptom scores (r(s) = 0.53, p = 0.016). CONCLUSIONS: Both ECP and tryptase increase in nasal secretion in natural disease. Therefore the measurement of tryptase and ECP levels in nasal fluid might be a useful clinical test for monitoring disease activity and the effects of therapeutic agents.

Adult↗

Functional dissection of the promoter of the pollen-specific gene NTP303 reveals a novel pollen-specific, and conserved cis-regulatory element.

Regulatory elements within the promoter of the pollen-specific NTP303 gene from tobacco were analysed by transient and stable expression analyses. Analysis of precisely targeted mutations showed that the NTP303 promoter is not regulated by any of the previously described pollen-specific cis-regulatory elements. However, two adjacent regions from -103 to -86 bp and from -86 to -59 bp were shown to contain sequences which positively regulated the NTP303 promoter. Both of these regions were capable of driving pollen-specific expression from a heterologous promoter, independent of orientation and in an additive manner. The boundaries of the minimal, functional NTP303 promoter were determined to lie within the region -86 to -51 bp. The sequence AAATGA localized from -94 to -89 bp was identified as a novel cis-acting element, of which the TGA triplet was shown to comprise an active part. This element was shown to be completely conserved in the similarly regulated promoter of the Bp 10 gene from Brassica napus encoding a homologue of the NTP303 gene.

Base Sequence↗

Cloning of PCP1, a member of a family of pollen coat protein (PCP) genes from Brassica oleracea encoding novel cysteine-rich proteins involved in pollen-stigma interactions.

The pollen coatings of both Brassica oleracea and Brassica napus contain a small family of basic 6-8 kDa proteins which are released on to the stigmatic surface on pollination. Following partial amino-acid sequencing of one of these pollen coat proteins (PCPs), PCR primers were constructed to isolate the PCP sequence from anther mRNA using RT-PCR. A cDNA was obtained which, in Northern hybridization experiments, revealed a characteristic pattern of expression during late stages of anther development. Interestingly, in situ hybridization revealed expression of this sequence to be confined to the cytoplasm of the trinucleate pollen grains: no signal was detected in the tapetum. Southern hybridization experiments have shown the gene (PCP1) to be a member of a large family of between 30 and 40 PCP genes in the genome of Brassica oleracea. Surprisingly, RFLP experiments showed reduced copy number (one to two copies) in some of the F2 segregants, perhaps resulting from the clustering of PCP sequences. PCP1 contains a single intron and encodes a small, basic peptide 83 amino acids in length featuring a hydrophobic signal peptide sequence separated from the more hydrophilic, cysteine-rich mature protein. The central part and C-terminal region of the peptide contain a characteristic and invariant pattern of eight cysteines which show clear homology with a number of other anther-specific genes; the remainder of the sequence shows little similarity to other sequences on the data bases. The product of PCP1 is a member of a large family of similar proteins, some of which have been demonstrated to bind specifically to S-locus glycoproteins, but does not appear to be genetically linked to the S-locus.

Amino Acid Sequence↗

Two-generation analysis of pollen flow across a landscape. II. Relation between phi(ft), pollen dispersal and interfemale distance.

We study the behavior of Phi(ft), a recently introduced estimator of instantaneous pollen flow, which is basically the intraclass correlation of inferred pollen cloud genetic frequencies among a sample of females drawn from a single population. Using standard theories of identity by descent and spatial processes, we show that Phi(ft) depends on the average distance of pollen dispersal (delta) and on the average distance between sampled mothers (x(1)). Provided that mothers are sampled far enough apart (x(1) > 5delta), Phi(ft) becomes independent of x(1) and is then inversely proportional to the square of delta. Provided that this condition is fulfilled, delta is directly estimable from Phi(ft). Even when x(1) < 5delta, estimation can easily be achieved via numerical evaluation. We show that the relation between Phi(ft) and delta is only modestly affected by the shape of the distribution function, a result of importance, since this shape is generally unknown. We also study the impact of adult density within the population on Phi(ft), showing that to achieve the correct inference of delta from Phi(ft) it must be taken into account, but that it has no effect on the distance at which mothers must be sampled.

Alleles↗

Some carbohydrates found in pollen and pollen substitutes are toxic to honey bees.

Carbohydrates in some pollen substitutes (galactose, lactose, raffinose, stachyose, glucuronic acid, galacturonic acid, polygalacturonic acid, and pectin) were toxic to caged adult Apis mellifera L. These toxins can be diluted to safe levels by sucrose. Collected nectar apparently dilutes the toxic sugars in pollen thus permitting assimilation of essential nutrients from pollen.

Animals↗

Comparision of a glutaraldehyde-modified pollen--tyrosine adsorbate with an alum-precipitated pollen vaccine in the treatment of hay fever.

Nineteen patients with hay fever symptoms received nine weekly injections of pyridine-extracted alum-precipitated grass pollen vaccine (Alavac P). The results were compared with agroup of twenty patients who received a three injection series of a glutaraldehyde-modified pollen-tyrosine vaccine (Pollinex). In both groups 65% had a subjective improvement in their symptoms and no significant difference in symptom scores was obtained throughout the pollen season. In the Pollinex group a lower incidence of systemic side effects was recorded.

Adolescent↗

Immunotherapy of tree pollen allergy with a modified alginate conjugated birch pollen extract compared to an aluminium adsorbed extract.

The safety and efficacy of two birch pollen extracts, one chemically conjugated to alginate (Anjuvac) the other adsorbed to aluminium hydroxide (Alutard), were investigated in an open multicentre comparative study of 63 birch pollen allergic patients. Both extracts decreased the nasal symptoms during the birch pollen season. The changes in specific IgE and IgG were much the same in both treatment groups. The adverse reactions recorded were mild in both groups, but more frequent in the Anjuvac group, probably because of a more aggressive dose schedule though there were twice as many asthmatics in the Anjuvac group. The two investigated allergen extracts were useful alternatives for immunotherapy.

Adolescent↗