Minor pollinosis fromCryptomeria japonica in Varese,northern Italy. Aerobiological monitoring and main clinical features

In the last 2 years, we have conducted an aerobiological monitoring program ofCryptomeria japonica, a plant belonging to the family of Taxodiaceae that sometimes causes pollinosis in the period from February to April. Throughout 1994, we checked the incidence of its sensitization and the clinical effects in 85 subjects with correlated seasonal symptoms, who gave a positive skin prick test (SPT) for Betulaceae and/or Corylaceae. Twenty-five patients (29.4%; 19 M; 6 F; mean age, 38.8 years) all with oculorhinitis, were SPT positive to an allergenic extract ofCryptomeria. RAST confirmed this positivity in 44% of the cases. No patients showed monosensitization forCryptomeria to either SPT or RAST. Two subjects gave a positive result on specific nasal provocation. RAST inhibition showed no cross-reaction betweenCryptomeria and birch pollen. During the pollen season each patient made a list, scoring symptoms and specifying any drugs used, so we could correlate these elements with aerobiological observations. The pollen concentration probably exceeded the allergizing threshold forCryptomeria on 8 days during 3 months of recording. The intensity and duration of symptoms seemed to be mainly influenced by sensitization to Betulaceae and Corylaceae. It is thus possible that a combination of ‘minor’ pollinosis may produce seasonal symptoms in allergic patients.     

Minor pollinosis fromCryptomeria japonica in Varese,northern Italy. Aerobiological monitoring and main clinical features

In the last 2 years we have conducted an aerobiological monitoring program ofCryptomeria japonica, a plant belonging to the family of Taxodiaceae that sometimes causes pollinosis in the period from February to April. Throughout 1994, we checked the incidence of its sensitization and the clinical effects in 85 subjects with correlated seasonal symptoms, who gave a positive skin prick test (SPT) for Betulaceae and/or Corylaceae. Twenty-five patients (29.4%; 19 M; 6 F; mean age, 38.8 years) all with oculorhinitis, were SPT positive to an allergenic extract ofCryptomeria. RAST confirmed this positivity in 44% of the cases. No patients showed monosensitization forCryptomeria to either SPT or RAST. Two subjects gave a positive result on specific nasal provocation. RAST inhibition showed no cross-reaction betweenCryptomeria and birch pollen. During the pollen season each patient made a list, scoring symptoms and specifying any drugs used, so we could correlate these elements with aerobiological observations. The pollen concentration probably exceeded the allergizing threshold forCryptomeria on 8 days during 3 months of recording. The intensity and duration of symptoms seemed to be mainly influenced by sensitization to Betulaceae and Corylaceae. It is thus possible that a combination of minor pollinosis may produce seasonal symptoms in allergic patients.     

Allergenic pollen and pollinosis in Warsaw

Summary Our study of fifty two hay fever patients included twenty six solely allergic to grass pollen and twenty six exhibiting allergy to various pollen species, such as hazel, birch, oak, poplar, andArtemisia. Their total and specific IgE response was evalutated by the immunoenzymatic method, while clinical reactivity was assessed by recording nasal and bronchial symptom scores between mid-March and mid-July. Simultaneously pollen counts were made. Polysensitized patients showed significantly higher levels of both total and specific IgE, which testifies to the enhanced quantitative and qualitative IgE. Multisensitized patients reacted earlier than patients sensitized to grass pollen only, which confirms that non-grass plants flowering only in the spring cause the priming effect on the nasal and bronchial mucosa. The early symptoms may be attributable to tree pollen sensitivity or may refletct higher grass pollen IgE levels in the polysensitized group. Characteristically, nasal symptoms preceded bronchial symptoms of several weeks.On comparing nasal washing from the polysensitized patients to washing from patients with grass pollen, we found much cytological material with the predominance of eosinophils.     

The gene coding for the major birch pollen allergen Betv1, is highly homologous to a pea disease resistance response gene.

Pollen of the white birch (Betula verrucosa) is one of the main causes of Type I allergic reactions (allergic rhinoconjunctivitis, allergic bronchial asthma) in Middle and Northern Europe, North America and the USSR. Type I allergies are a major threat to public health in these countries, since 10-15% of the population suffer from these diseases. BetvI, an allergenic protein with an Mr of 17 kd is a constituent of the pollen of white birch and is responsible for IgE binding in more than 95% of birch pollen allergic patients. Here, we report the complete nucleotide sequence and deduced amino acid sequence of a cDNA clone coding for the major pollen allergen (BetvI) of white birch. It is similar to the N-terminal peptide sequences of the allergens of hazel, alder and hornbeam (close relatives) but it has no significant sequence homology to any other known allergens. However, it shows 55% sequence identity with a pea disease resistance response gene, indicating that BetvI may be involved in pathogen resistance of pollen.     

Clinical significance of allergic sensitization toOlea europaea L. pollen in Naples area,Italy

Summary The aim of this study is the evaluation of clinical significance of the allergic sensitization toOlea europaea L. pollen allergens in the Naples area, because in the last 10 years an increasing frequency of IgE mediated sensitization to these allergens has been observed. 4124 patients of both sexes (2989 adults and 1153 children) consecutively examined in our Allergological Center from January 1, 1988 to December 30, 1989 have been considered in this study. The patients were selected on the basis of a history of rhinitis (R) and/or asthma (A) and were evaluated by clinical examination, SPT, RAST and specific or aspecific BPT. Among 2267 adults and 879 children sensitized to almost one aeroallergen we found respectively 306 (13.49%) and 75 (8.53%) subjects with an allergic sensitization toOlea europaea L. pollen. The frequency of clinical manifestation was respectively R+A (58.16%), R (39.54%), A (2.28%) in adults; R+A (61.33%), R (26.66%), A (12%) in children. A monosensitization toOlea europaea L. pollen was found in 1.33% of children and in 2.28% of adults examined. The association with other allergens especially with mites and/or other pollen was very common.     

Comparison between Urticaceae (Parietaria) pollen count and hay fever symptoms: assessment of a «threshold-value»

Summary A comparison betweenParietaria pollen count and allergic symptoms of rhino-conjunctivitis in the early season was used in utilized to determine a «threshold-value» for this pollen. Clinical data were obtained from diary-cards of 34 allergic patients and pollen data from a volumetric sampling, carried out by means of a Hirst-Burkard pollen-trap. A significant correlation (r=0.98) was found between pollen count and symptom scores. Mild symptoms were registered with concentrations above 10–15 pollens/m³. Severe symptoms occurred when pollen count exceeded 80/m³/24 h., and over 90% of patients recorded symptoms. The importance of the late reactions and of the total allergenic airborne content are emphasized.     

Biochemical and immunochemical characterization of hop-hornbeam (Ostrya Carpinifolia Scop.) pollen

In the last few years Ostrya carpinifolia pollen is consideredas an important cause of respiratoryallergy in Mediterranean areas. The concentration ofthe pollen was measured over a period of fifteen yearsfrom 1981 to 1996 in an area around Genoa; the resultsof this study have clearly indicated an increasingtrend that correlate with persons sensitization.In this study we sought to define the immunochemical andbiochemical properties of hop-hornbean pollen. Soluble proteins extracted from Ostryacarpinifolia pollen and from the taxonomicallyrelated species Corylus Avellana, were analyzedby polyacrylamide gel electrophoresis (SDS-PAGE), byhorizontal isoelectrofocusing (IEF) and by twodimensions electrophoresis (2D-PAGE). Allergenicproteins were identified with sera of sensibilizedpatients and cross-reactivity was evaluated byimmunoblotting techniques. The electrophoreticanalysis showed a partial identity between theproteins from Ostrya and Corylus extracts. The immunoblotting assay, developed withhuman IgE from subjects allergic to hop-hornbeampollen, displayed the major IgE reactivity for acomponent with a molecular weight of 17 kDa expressedin both Ostrya and Corylus extracts. This reactivity is consistent with the presence ofBet v 1 that is described as the major pollen allergenin the Betulaceae and Corylaceae families. Sera fromsubjects allergic to Ostrya were then preadsorbed with recombinant Bet v 1 immobilized in the Pharmacia CAP System; a significant reduction ofthe IgE binding activity was observed after thetreatment. We therefore suggest that Bet v 1 couldbe one of the allergenic proteins present in theOstrya pollen possibly being responsible forcross-reactivity with other members of taxonomicallyrelated families.     

Betulaceae and Corylaceae in Trieste (NE-Italy):Aerobiological and clinical data

Airborne pollen produced by Betulaceae and Corylaceaeis present in the Trieste area for a long period fromJanuary to June, but only in April it does representa considerable proportion (436 p/m³) of the totalpollen count (1193 p/m³). In the years considered (1995–1997), there was a gradual increase in thepollen count of Corylaceae and Betulaceae: frommaximum levels of 580 p/m³ in 1995 to 1218 p/m³ in 1997, and the taxon making the mostsignificant contribution to the pollen concentrationcurve was Ostrya carpinifolia. Sensitization to Betulaceae and Corylaceae wasanalyzed in 2213 subjects visiting our clinic betweenJanuary 1st 1995 and December 31th 1997, withallergic symptoms believed to be IgE mediated. Of thegroup, 1292 (58.4%) were atopic by skin prick testand 328 of them (25.4%) were sensitized to Betulaceaeand Corylaceae. Of the 328 subjects, 72.6% werecosensitized to Gramineae, 56.1% to Oleaceae, 42.1%to Compositae, and just over half to house-dust mites(52.1%). Only ten cases were mono-sensitized (3%).Of the subjects sentitized to Betulaceae andCorylaceae, 163 complained of rhinitis (72%) and 110of asthma (33.6%), often in association withrhinitis; 177 (54%) had only seasonal symptoms.Sensitization to Betulaceae and Corylaceae is high,but its role in inducing allergic respiratorysymptoms is difficult to evaluate because almost allpatients were sensitized to other pollen types. Inconclusion, the role played by this family of trees indetermining allergic respiratory symptoms could becomeincreasingly important in this geographical area inthe future, if pollen levels continue to rise at theirpresent rates. For the moment, sensitivity toBetulaceae-Corylaceae among the population is presentalmost exclusively in subjects sensitized also toother pollen types.     

The first approach towards recognising allergy-provoking flora in Belluno and its relationship with allergic phenomena

Summary The town of Belluno has a high number of wild and cultivated plants whose pollen has various degrees of importance from an allergological point of view.In this preliminary study we have taken into consideration the distribution and the flowering period of these species, correlating them with the data on the quantity and type of pollen collected by a sampler placed on the roof of the City Hospital, and also with the results of the allergometric tests (Prick Tests) carried out in the Broncho-Pneumological Department during the first 9 months of 1990.The wild specis of spring-flowering trees which can be found in the woods surrounding the city (Fagaceae, Betulaceae, Corylaceae) and along the gravel banks of the Piave river (Salicaceae) as well as the Graminaceae which are widespread both in the urban area and in the fields outside the city and some cultivated species (Robinia, Tilia, Acer) which flower a little later, are the species which produce most pollen and which are held responsible for allergies. However, only the pollen of the Graminaceae gave rise to cases of mono-sensitivity (16%) (apart from one case of Betulaceae and Corylaceae and one case ofArtemisia as well asDermatophagoides) (9.7%). For the other species, the positive reactions were associated with the positive reactions to the pollen of other species belonging to different families and/or to other factors.     

Reproductive biology ofFrangula alnus (Rhamnaceae) in southern Spain

Various aspects of the reproductive biology of the perennial hermaphroditeFrangula alnus Mill. were studied in two populations located in the province of Cádiz, southern Spain. Flowering extends from the second week of May to early July. The small, whitegreenish, entomophilous flowers are incompletely protandrous and last 8–10 days, but pollen transfer takes place only on days 1–3. A very weak, pleasant odour along with nectar and pollen attract over 45 insect species, of which 21 are probable pollinators. These are mainlyDiptera but also includeHymenoptera and the unusual (as flower visitors)Mecoptera. Reproduction is exclusively sexual and strictly xenogamous, with pollen transfer depending solely on insect vectors. Although flower morphology and individual flower phenology do not fully prevent self-pollination, and geitonogamy can easily take place, the level of autogamy was negligible. Therefore, some self-incompatibility mechanism is operative in this species. Only 2.8% of open-pollinated flowers set fruit. At the flower level, fruit initiation was apparently limited by availability of cross-pollen, which in turn seemed influenced by the structure of the population. The seed/ovule ratio in ripe fruits was ca. 50%. Predispersal (maternal) reproductive success (percent ovules becoming filled seeds) was 1.42%. For mature individuals this corresponded to ca. 430 to 1560 potential offspring per year.     

TheCaMV 35S promoter is highly active on floral organs and pollen of transgenic strawberry plants

We have evaluated the expression of the reporter -glucuronidase (GUS) gene driven by the cauliflower mosaic virus 35S (CaMV 35S) promoter in flowers and pollen from 14 independent transgenic strawberry lines. Of the 14 lines evaluated, 13 (92.8%) showed GUS activity—as estimated by the histochemical GUS assay—in some floral organs, with expression being most common in the flower stem, sepals, petals, ovary and stigma. Ten of these thirteen transgenic lines (77%) showed GUS activity in pollen, although the percentages of positive pollen per flower varied greatly among the different lines. A study of the GUS expression during pollen maturation showed that the (CaMV 35S) promoter showed low expression in pollen from flower buds before anthesis but was activated in mature pollen following anther dehiscence. The percentages of pollen grains that showed GUS activity ranged from 2.1% to 46.3%. These percentages were similar or even higher when mature pollen was stored dry at room temperature for 2 weeks. After 5 weeks of storage, the percentages of GUS-positive pollen decreased in two of the six lines analysed but remained at similar values in the other four lines. GUS activity was also measured in protein extracts of mature pollen by means of the fluorometric GUS assay, with the values obtained ranging from 3.8 mol MU mg protein^–1 h^–1 to 0.26 mol MU mg protein^–1 h^–1. Contrary to the generally held view that the CaMV 35S promoter is virtually silent in pollen, we conclude that it is highly expressed in transgenic strawberry pollen.Abbreviations CaMV 35S Cauliflower mosaic virus promoter - GUS -Glucuronidase (EC 3.2.1.31) - MU 4-Methyl umbelliferone - nos Nopaline synthase promoter - nptII Neomycin phosphotransferase - X-Gluc 5-Bromo-4-chloro-3-indolyl--d-glucuronic acid     

Airborne birch pollen in Neuchâtel (Switzerland): onset, peak and daily patterns

An accurate forecast of the starting point of thebirch pollen season in Neuchâtel can be made byadding the positive daily average air temperature fromFebruary 1st onward until the figure 270 is reached.At this point, the birch trees are ready to bloom.After that, the daily average temperature has toexceed 10 °C to allow pollen release.Today, the birch pollen season starts some 19 daysearlier in the year than in the 1980's, a consequenceof a recent climate change.The daily patterns of airborne birch pollen isirregular. Moreover, pollen concentrations frequentlyexceed the threshold of the appearance of allergicsymptoms, except during rainfall. Therefore, the onlybehavioral recommendation that can be given to peopleallergic to birch pollen is to shorten as much aspossible the contact with outdoor air during the mainbirch pollen season.     

Seasonal variation in food allergy to apple

The aim of the study was to investigate the possibility of a seasonal variation in reactivity to apples in 27 birch pollen allergic patients. Before and during the birch pollen season 1998, the patients were subjected to double-blind, placebo-controlled food challenges (DBPCFCs) with grated fresh Golden Delicious apple followed by an open food challenge with whole fresh apple. The clinical reactions elicited during the challenges were evaluated both by the patients and the investigators. Moreover, the skin reactivity and the in vitro reactivity to apple were evaluated by skin prick test (SPT), leukocyte histamine release (HR), measurement of specific IgE, and immunoblotting experiments. The sensitivity of the DBPCFC, when compared with the result of the open challenge, was 0.74 (14/19) before the season and 0.80 (16/20) during the season. None of the patients reacted to the blinded challenge without a subsequent reaction to the open challenge. One placebo reaction was registered both before and in season, but not in the same patient. The patient scores of the first positive challenges, and the maximal scores of each combined blinded and open challenge session, were significantly increased during the pollen season (P<0.05). The scores of the open challenge were significantly higher than the scores of the DBPCFC both before the season and during the in-season challenges (P<0.05). Specific IgE against Golden Delicious increased during season (P<0.05), while neither SPT, HR, nor immunoblotting experiments could confirm an increase in reactivity. In conclusion, the results of the oral challenge tests indicated an increase in clinical reactivity to apples during the birch pollen season in birch pollen allergic individuals.     

Applicability of the co-inoculation technique using Agrobacterium tumefaciens shooty-tumour strain 82.139 in silver birch

In the co-inoculation technique, genetic transformation is performed using a mixture of Agrobacterium strains – shoot regeneration is induced by the wild-type strain 82.139, while the transferable genes are provided in a binary plasmid by another, non-oncogenic Agrobacterium strain. The aim of the present work was to study the applicability of co-inoculation under both in vitro and greenhouse conditions for in planta transformation in silver birch (Betula pendula Roth). In addition to the original method, several modifications of the technique including an genetically engineered 82.139 strain harbouring the binary pGUSINT were tested. The co-inoculations resulted in a gall formation frequency of 52–94% with greenhouse seedlings, and 4–63% with tissue-cultured plantlets, the shoot induction percentage varying by 0–13 in the greenhouse and 42–75 in vitro. PCR analysis verified that the majority of the regenerated material was non-transgenic, with a few individuals showing integration of the oncogenic T-DNA. According to the histochemical tests, however, some of the numerous differentiating buds and small shoots on gall tissues were transgenic, and contained the GUS reporter gene. The results show that it would have been necessary to apply selection pressure during differentiation in order to recover shoots transformed with the desired genes from the binary plasmid. The morphology and growth of all the regenerated plantlets was normal, suggesting that the oncogenic T-DNA was not expressed even though it was present. In conclusion, it was possible to obtain transgenic silver birch plantlets using the A. tumefaciens strain 82.139, but the co-inoculation method is not directly applicable as in planta transformation protocol.     

Annual, daily and intradiurnal variation of Celtis pollen in the city of La Plata, Argentina

The behaviour of Celtis airborne pollenwas studied for a period of three years(1998–2000) in the city of La Plata. The pollengrains were captured with a Lanzoni trap andthe maximum pollination period was observed tooccur during the end of winter and spring inthe three years. The annual values of pollenconcentration varied, and a significantdecrease was observed during 2000. The greatestairborne pollen record was in October with anaverage of 82.3% in relation to its totalconcentration during 1998–2000. Based on theintradiurnal behaviour analysis, it wasobserved that the maximum pollination peakoccurs at 2H when the temperaturereaches its maximum values. Considering thethree sampling years, the meteorologicalvariables that most influenced the processes of pollen emission,dispersion and transportation in the atmospherewere: maximum and minimum temperature andrainfall.     

Notes on the pollination mechanisms ofMoraea inclinata andM. brevistyla (Iridaceae)

Individual flowers ofMoraea inclinata are nectariferous and last about six hours. They appear to be pollinated largely by bees in the familyHalictidae (Lasioglossum spp.,Nomia spp.,Zonalictus) and to a lesser extent by bees in the familyAnthophoridae (Amegilla). The mechanism of bee-pollination inM. inclinata is the Iris type; i.e., each flower consists of three pollination units (an outer tepal, a partly exserted anther, and the opposed style branch which terminates in a pair of petal-like crests). Bees rarely visit more than one pollination unit per flower. Transferral of pollen to the bee is passive and nototribic although all bees collected on the flowers were female and 55% of the bees carried pollen loads with 2–5 pollen taxa in their scopae.Moraea brevistyla flowers are nectariferous but lack scent and last two days. They are visited infrequently by bees and only one femaleLasioglossum spec. carried the pollen ofM. brevistyla. Unlike flowers ofM. inclinata those ofM. brevistyla deposit pollen only on the head and thorax. Bee-mediated autogamy in both species is avoided due to the erratic foraging patterns of the bees and the flexibility of each stigma lobe as the bee backs out of the flower. Approximately 2–4 flowers in the inflorescences of both species (6–8 flowers/infloresence) develop into capsules.     

Ragweed pollen: The aeroallergen is spreading in Italy

Presently in Europe, ragweed pollen as an aeroallergen is not as important as Poaceae,Parietaria or Betulaceae, even if in some countries the plant is beginning to influence the local composition of the airborne pollen spectra. In northern Italy, the presence of ragweed airborne pollen has only been reported since the beginning of the 1980’s and it is increasingly spreading from year to year. Given this situation, the allergologists have begun to regard the potential risk of sensitisation to ragweed pollen with much attention. Up to now, such pollen has not been included in the routine allergological tests. In 1995 in some sites of northern Italy (Turin, Milan, Trieste), the concentration values of ragweed pollen were remarkable (∼ 20–30 p/m³) and on the increase with respect to the previous years. This investigation aims at focusing the atmospheric concentration trend on this new aeroallergen (Ambrosia sp.) in Italy from 1991 throughout 1995.     

A mathematical model for mugwort (Artemisia vulgaris L.) pollen forecasts

Pollen forecasts are a fundamental prerequisite to obtain prophylactic measures for allergic individuals. Mugwort belongs to the most relevant allergenic pollen types after grasses and birch. An approach to modeling of mugwort pollen concentrations has not been attempted previously in Germany. A process-oriented mathematical model for the relative local daily average mugwort airborne pollen concentration was developed on the basis of pollen and weather data measured during a 6-year period. The model depends on the daily minimum and maximum temperature, amount of precipitation and atmospheric pressure, which have to and can be supplied by measurement and prediction. The comparison of modeling results and pollen counting for an additional year confirms the fitness of the model. A computer program was written, which rests upon the model and supplies daily predictions of mugwort pollen flight during the period of the weather forecast. The latter should allow a pollen forecasting period of about 5 days, with an accuracy of about 32–63% explained variance, which in view of the low mugwort pollen counts (nine grains/m³ maximum in the validation year) represents a high relative measurement error. The mathematical model may serve to improve and rationalize of present pollen forecasts.     

Airborne pine (Pinus spp.) pollen in the atmosphere of Perugia (Central Italy): Behaviour of pollination in the two last decades

The phenology of many species, which grow intemperate climate, is principally regulated bythe temperature and the plants respond withvariations in the beginning, in the durationand in the intensity of the various phenophasestowards every climate change. We have analysedthe data of Pinus pollination in Perugia,Central Italy, during last 2 decades(1982–2001), in a period during which theannual mean temperature significantly increasedby about 0.8 °C.The pine pollination started, on average,between the end of March and mid-April andended in the last days of June, with a meanduration of 65 days. The start dates showed asignificant negative correlation with theaverage air temperature in March andsignificant trends towards an earlier beginningof pollination by 18 days (–0.9 day/year) and ashorter duration of the pollen season by 10days (–0.6 day/year) were found over thestudied period. Moreover, the trend of thedaily pollen counts showed, on average, analmost normal distribution, but the analysis ofeach yearly trend revealed significantdifferences correlated with the meantemperature during the pollen season. Theseobserved trends in pine pollination suggest theuse of aerobiological monitoring of thisairborne pollen as indicator of temperaturechange in Central Italy over a relatively longperiod.     

Pollen morphology ofSonchus and related genera,and a general discussion

The pollen morphology of the taxa belonging to the generaAetheorhiza Cass.,Launaea Cass.,Reichardia Roth andSonchus L. in the Iberian Peninsula has been studied with light and electron microscopy. The pollen is 3(-4)-zonocolporate and echinolophate (without polar lacunae, but in general with prelacunae), with equatorial ridges and 15–20 lacunae: 3–4 poral, 6–8 abporal and 6–8 paraporal. Small to medium size, P × E = 19–36 × 23–42 µm; sometimes two different sizes have been found. Exine 3–9 µm thick and ornamentation microreticulate and echinate. The results clearly show the relationships between genera. Moreno-Socías, E., Mejías, J. A., Díez, M. J., 1994: Morfología polínica deLactuceae (Asteraceae) en la Península Ibérica, I.Lactuca y géneros relacionados. — Acta Bot. Malacitana.19: 103–113.