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Background Pollen exposure has acute adverse effects on sensitized individuals. Information on the prevalence of respiratory diseases in areas with different pollen concentrations is scanty. Aim We performed an ecologic analysis to assess whether the prevalence of allergic rhinitis and asthma in young adults varied across areas with different pollen concentrations in Italy. Methods A questionnaire on respiratory diseases was delivered to random samples of 20–44 year-old subjects from six centers in 2005–2010. Data on the daily air concentrations of 7 major allergologic pollens (Poaceae, Urticaceae, Oleaceae, Cupressaceae, Coryloideae, Betula and Ambrosia) were collected for 2007–2008. Center-specific pollen exposure indicators were calculated, including the average number of days per year with pollens above the low or high concentration thresholds defined by the Italian Association of Aerobiology. Associations between pollen exposure and disease prevalence, adjusted for potential confounders, were estimated using logistic regression models with center as a random-intercept. Results Overall, 8834 subjects (56.8%) filled in the questionnaire. Allergic rhinitis was significantly less frequent in the centers with longer periods with high concentrations of at least one (OR per 10 days = 0.989, 95%CI: 0.979–0.999) or at least two pollens (OR = 0.974, 95%CI: 0.951–0.998); associations with the number of days with at least one (OR = 0.988, 95%CI: 0.972–1.004) or at least two (OR = 0.985, 95%CI: 0.970–1.001) pollens above the low thresholds were borderline significant. Asthma prevalence was not associated with pollen concentrations. Conclusions Our study does not support that the prevalence of allergic rhinitis and asthma is greater in centers with higher pollen concentrations. It is not clear whether the observed ecologic associations hold at the individual level.

Background Ragweed allergy represents a challenge for allergists due to its increase in some geographic areas, mainly northern Italy. Objectives To compare two allergy centers in northern Italy by analyzing both the pollen count and the patient characteristics (sensitization and allergy prevalence) over a 10-year period. Methods Pollen counts for 10 years (2004 –2013) were evaluated and compared in the two centers. Characteristics of patients who were sensitized and allergic to ragweed were analyzed. Results There was a significantly higher pollen count in Busto Arsizio than in Novi Ligure (p < 0.001). The patients who were seen in the Busto Arsizio center showed more frequent sensitization (p < 0.01) and allergy (p < 0.05) to ragweed than patients who were seen in the Novi Ligure center. The pollen count was significantly (p < 0.01) associated with sensitization and allergy difference. Allergic asthma was more frequent (p < 0.05) in Busto Arsizio. Conclusions The pattern of sensitization and allergy prevalence seemed to be associated with ragweed pollen pressure, such as quantity. The knowledge of the pollen count is relevant in clinical practice, mainly for the management of patients with allergy.

Summary Introduction Madelung's disease, also known as Multiple Symmetric Lipomatosis (MSL) or the Launois-Bensaude syndrome, is characterized by subcutaneous lipid deposits, which, unlike lipomas, are nonencapsulated. The disease is diagnosed mainly in Mediterranean countries, and 90% of all cases involve male alcoholics. These masses are usually found on the neck, chest, abdomen, and arms. The symptoms are the result of acute sensorimotor axonal neuropathy caused by compression. Treatment (surgical and/or medical) does not produce significant or lasting results. Case report We describe the case of a 57-year old male alcoholic (daily ethanol intake: approximately 2 L of wine), who has been treated and followed by our staff for 6 years. Discussion and conclusions As this case illustrates, management of MSL is often associated with serious difficulties related to the patient's alcoholism. The effects of adipose tissue masses on the sensitive and motor nervous system are complicated by the effects of alcoholic neuropathy, requiring complex long-term monitoring. Treatment is often compromised by poor compliance related to patient's drinking problems. Madelung's disease is currently regarded as a rare disease, although we suspect that the incidence in Italy (and in the rest of the world) has probably been underestimated for various reasons.

Sensitization to Cupressaceae pollen has become one of the most important causes of pollinosis in Western countries during winter and early spring. However, the characterization of the extracts, the allergens involved and the cross-reactivity with other pollen sources still remain poorly studied; in the case of Cupressus arizonica only two allergens have been described so far. A new allergen from C. arizonica pollen, Cup a 4, was cloned and expressed in Escherichia coli as an N-terminally His-tag recombinant protein that was characterized biochemically, immunologically and by circular dichroism spectroscopy. The new allergen has high sequence identity with Prickly Juniper allergen Jun o 4 and contains four EF-hand domains. The recombinant protein has structural similarities with other calcium binding allergens such as Ole e 3, Ole e 8 and Phl p 7. Cup a 4 is expressed in mature pollen grains and shares antigenic properties with the recombinant form. Sera from 9.6% C. arizonica allergic patients contain specific IgE antibodies against recombinant Cup a 4.

Seasonal Allergic rhinitis (SAR) is characterized by runny nose, congestion, sneezing and sinus pressure. A clinical study was performed to demonstrate the efficacy of Lertal®, an innovative food supplement containing Quercetin, Perilla frutescens and Vitamin D3 formu-lated in a double layer "fast-slow" release tablet form, in the relief of symptoms of seasonal allergic rhinitis and in the reduction of consumption of anti-allergic drugs.23 subjects enrolled in the open clinical study had at least one year history of allergic rhinitis and positive skin prick test or RAST to Parietaria officinalis pollen. At baseline, the subjects had symptoms of nasal and/or ocular seasonal allergic rhinitis. The activity of the food supplement was evaluated using the Total Symptoms Score at first (baseline) and second (final) visit, after one month of supplementation. The consumption of anti-allergic drugs was also evaluated.All subjects enrolled completed the study. The comparison of the scores obtained in the two visits (baseline and final) showed a highly significant reduction of the overall symptoms: approximately 70% for symptom scores and 73% in use of anti-allergic drugs. Sneezing, rhinorrhea, nasal obstruction, ocular itching, lacrimation and congestion of the conjunctiva, all showed a highly significant reduction. No noteworthy side effect was recorded and all patients finished the study with good compliance.The results showed a clear efficacy of the food supplement Lertal® in reducing nasal and/or eye symptoms. This activity was objectively confirmed by the reduction in the consumption of anti-allergic drugs used to relieve symptoms. (www.actabiomedica.it).

Background:Holoptelea integrifolia and Parietaria judaica belong to the family Urticaceae, but are geographically distantly located. H. integrifolia is an important pollen allergen of India and sensitizes almost 10% of the atopic population in Delhi. P. judaica, on the other hand, is a very dominant pollen allergen of the Mediterranean region, sensitizing almost 80% of the allergic population. Since both these important pollen allergens belong to the family Urticaceae, the objective of the present study was to assess cross-reactivity between these two pollen allergens from different geographical regions. Methods: Cross-reactivity between these two pollen allergens was assessed on the basis of skin prick tests and ELISA, ELISA inhibition and immunoblot inhibition studies. Results: Out of 44 atopic Indian patients skin prick tested with H. integrifolia extract, 34% were found to be sensitized. All the patients sensitized to H. integrifolia also showed varying degrees of skin positivity to P. judaica pollen extract. ELISA and ELISA inhibition studies suggested strong cross-reactivity between H. integrifolia and P. judaica pollen. Immunoblot inhibition studies revealed that 14-, 16-, 28-, 38-, 42- and 46-kDa proteins are the cross-reactive proteins in H. integrifolia and P. judaica. However, Par j 1, the major allergen of P. judaica, is absent in H. integrifolia pollen. Conclusion:H. integrifolia and P. judaica pollens share cross-reactive as well as unique epitopes. The major allergen of P. judaica, Par j 1, seems to be absent in H. integrifolia pollen allergen.

Background: Recombinant DNA technology does provide pure, well-defined and reproducible products to be used for clinical purposes, by cloning and expressing the cDNA of allergens present in a specific extract. Ole e 5 is a pollen allergen of Olea europaea with an IgE-binding frequency of about 35%, which has been identified as a superoxide dismutase (SOD). The aim of this study was to clone the cDNA of Ole e 5, to express Ole e 5 in Escherichia coli and to characterize its immunoreactivity. Methods: cDNA of Ole e 5 was amplified by nested 3′-RACE PCR and cloned in pGEX vector 6P expression vector. After sequencing of some clones and homology analysis, the rOle e 5 was produced in an E. coli strain as a fusion protein with GST and purified. Then, the protein immunoreactivity was evaluated by patients’ IgE binding (ELISA, ELISA inhibition, and immunoblotting) and by rabbit anti-rOle e 5 binding (immunoblotting and immunoblotting inhibition). Results: The sequence analysis of Ole e 5 cDNA confirmed that Ole e 5 is a Cu/Zn SOD, with an identity from 90 to 80% with SOD from other species. rOle e 5 was recognized by IgE from 39% of olive pollen-allergic patients tested; moreover, this binding was inhibited by the olive pollen extract. An anti-rOle e 5 antiserum raised in rabbit strongly reacted with a natural component of about 16-kDa molecular weight present in the olive pollen extract; moreover, this binding was inhibited by the recombinant protein. Conclusions: Ole e 5 is the first Cu/Zn SOD identified as an allergen in a pollen source. Due to the widespread presence of this enzyme, rOle e 5 allergen, cloned and expressed in a complete form in E. coli, could represent a good tool to investigate the allergen cross-reactivity between O. europaea pollen and other allergenic sources, such as plant foods and other pollens.

Summary Background Cypress pollinosis is an important cause of respiratory allergies. Recently, the Cupressus arizonica major allergen, Cup a1, has been cloned and expressed. The native counterpart of this allergen has been purified and characterized by our group. It has been suggested that sugar moieties play a role in the in vitro IgE binding on Cupressus arizonica pollen extract. Objective To characterize the immunoreactivity of the recombinant major allergen in comparison with its native counterpart. To evaluate the role of carbohydrate moieties in the IgE-mediated in vitro histamine release from basophils by using the native glycosylated Cup a1 as compared with the recombinant one. Methods Recombinant Cup a1 was expressed in E. coli. IgE reactivity of Cupressaceae-allergic patients on the native as well as the recombinant molecule was investigated by immunoblotting, ELISA experiments and histamine release test from passively sensitized basophils. Results Fourteen out of 17 Cup a1-positive sera had IgE antibodies reactive with the native molecule only and lost their reactivity after periodate deglycosylation of the allergen. Moreover, only native molecule was capable of inducing histamine release by this group of sera. Both the recombinant and the native molecules were recognized by three out of the 17 sera and were equally capable of triggering degranulation. Conclusion A large number of sera reactive with the major allergen recognize carbohydrate epitopes only. IgE from these sera are able to induce histamine release from basophils and they might play a functional role in the clinical symptoms of allergy.

Background: Airborne insect antigens represent important aeroallergens which have been widely investigated. Although it has been demonstrated that house dust contains significant silverfish (Lepisma saccharina) levels, none of the extracts obtained so far has been extensively characterized. Thus, we have prepared and characterized a silverfish extract and investigated its IgE-reactive components by testing the reactivity of sera from patients allergic to inhalant insect allergens. Methods: The extract from silverfish insect bodies was prepared by homogenizing frozen silverfish in Tris-HCl buffer. The soluble material (Sup) was filtered and the insoluble material (Ppt) was resuspended in 100 mM Tris pH 10.6. The two fractions were characterized by biochemical and immunochemical methods. IgE reactivity was investigated on both fractions before and after periodate treatment. Results: Protein content and total carbohydrates was 2 and 3% w/w for Sup and 1 and 0.3% w/w for Ppt. The SDS-PAGE profile of the two fractions showed a different pattern in the MW range of 5–175 kD. Sup and Ppt, probed with allergic sera, showed a complex pattern of IgE reactivity. When periodate-treated fractions were tested, IgE reactivity was either completely abrogated, reduced or not affected, depending on the allergic serum employed. Conclusions: The results obtained indicate that the classic aqueous-extraction procedures that have been used up to now for other insects might not be completely satisfactory, since several allergenic components are not soluble at the normally used pH. We developed a dedicated extraction procedure allowing the detection of a certain degree of reactivity in sera negative to allergens extracted following classic procedures.

In this study, we used the affected sibling-pairs approach to investigate the linkage of HLA (human leukocyte antigen)-DRB* with phenotypes related to allergy to Parietaria, the most common pollinosis in Mediterranean countries. The study population consisted of 51 nuclear families (235 subjects). Linkage was detected with Parietaria skin test positivity (p(0.01), presence of IgG and IgE antibodies specific for the major allergen Par o 1 (p0.020 and p0.025, respectively), and absence of Par o 1-specific IgE (p0.020). High levels of Par o 1-specific IgG were associated with DRB1*1101 and/or DRB1*1104 (p0.0001 and p0.0119, respectively) in parents and probands. High levels of Par o 1-specific IgE were associated with DRB*1104 in parents (p0.017) and with DRB1*1101 in probands (p0.0146). When siblings were categorized according to high/low total IgE levels (or =125 IU/ml and125 IU/ml, respectively), high IgE antibody response was associated with DRB1*1104 in siblings with low total IgE (p0.034) and with DRB1*1101 in siblings with high total IgE (p0.05). These results demonstrate that HLA-DRB1*, or genes in linkage disequilibrium, contributes to susceptibility to Parietaria allergy and that total IgE levels can discriminate population subsets where different alleles (at the HLA region or at loci in linkage disequilibrium) contribute to control allergen-specific IgE synthesis.

The authors examined 734 sensitised patients living in four localities in Liguria (Genoa, Savona, Pietra Ligure and Sanremo). The commonest source of sensitisation (62.7%) was Urticaceae (Parietaria), followed by Poaceae (52.5%) andOlea europaea L. (24.0%). A survey of airborne pollens revealed a greater presence of Urticaceae and Poaceae in Genoa and of Oleaceae in Pietra Ligure and Sanremo.

Trends of the atmospheric dispersion of the pollens of Urticaceae, Poaceae, Asteraceae,Plantago, Chenopodiaceae-Amaranthaceae, Polygonaceae, Oleaceae, Betulaceae-Corylaceae,Quercus, Castanea, and Cupressaceae-Taxaceae were studied from 1981 to 1989 in two localities (Genoa and Sanremo) about 120 km apart on the coast of Liguria in Italy. Statistical analysis revealed, in particular, and overall increase in pollen rain in Genoa during the period considered. The following observations were carried out on the pollens of greatest allergological interest: Urticaceae, a slightly increasing trend in Genoa; Poaceae, a moderately increasing trend in Sanremo; Oleaceae, almost parallel, uniform trendsin both centres. The values of this study lies in its ability to point out any increase which may occur over the years in the dispersal of certain allergenic pollens.

We have taken advantage of the temporary exposure of Marseilles population to castor bean seed proteins to follow 26 allergic patients more than 20 years after sensitization. Skin tests, specific immunoglobulin E (IgE) antibody assays, and specific immunoblots were performed. Skin test reactivity to Ricinus Communis and specific IgE concentrations decreased progressively and almost completely disappeared after 20 years. Specific IgE concentration displayed a fairly exponential decrease, with a half-life of 4.7 years. Thus, in the absence of any antigenic stimulation, directly by castor bean, or indirectly by cross-reactivity to other Euphorbiaceae, especially latex, IgE sensitization is bound to disappear.

Only a few studies analyzed the pharmacoeconomics of allergen immunotherapy compared with drug treatment in subjects with allergic rhinitis and asthma. This study was aimed at evaluating whether allergen immunotherapy has an economic advantage on standard antiallergic drugs in patients with pollen-induced rhinitis and asthma. Thirty patients with rhinitis and asthma caused by Parietaria pollen were included in the study, 20 (11 men and 9 women; mean age, 35.45 +/- 10.45 years) were treated with subcutaneous immunotherapy by a Parietaria judaica extract (Alustal, Stallergénes, Antony, France) by a conventional build-up schedule in 12 weeks and a maintenance treatment every 4 weeks for 3 years, and 10 (6 men and 4 women; mean age, 31.90 +/- 10.97 years) were treated with antiallergic drugs. Each patient was evaluated before starting the treatment and annually for 6 years in the pollen period of Parietaria by means of nose, eyes, and lung symptom scores, along with drug consumption registered in diary cards. In other specifically designated cards general practitioner's or specialist's visits, the number of desensitizing injections and the number of boxes of antiallergic drugs were registered. A significant difference in favor of immunotherapy plus drug treatment versus drug treatment alone was observed, reaching a cost reduction of approximately 15% the second year and 48% the third year, with a highly statistical significance that was maintained up to the sixth year, i.e., 3 years after stopping immunotherapy, when an 80% reduction was found. The net saving for each patient at the final evaluation corresponded to euros 623 (dollars 830)/year. These findings confirm some previous observations in studies from Germany and the United States that subcutaneous immunotherapy has significant economic advantages over antiallergic drug treatment in the long term.

Summary Background The presence of specific IgE antibodies to invertebrates is common among patients with rhinitis and asthma. Tropomyosin has been described as an invertebrate cross-reactive allergen. We have recently characterized an allergenic extract from silverfish (Lepisma saccharina). Since this insect could be a new source of tropomyosin in the indoor environment, we have thought important to clone and characterize the tropomyosin from it. Methods Recombinant tropomyosin was cloned and characterized by means of immunoblotting with tropomyosin-specific monoclonal antibodies, rabbit polyclonal antibodies and IgE from allergic patients. Its allergenic activity was investigated in histamine release assays. Immunoblotting and ELISA inhibition were carried out to identify the natural tropomyosin in the silverfish extract and to study the cross-reactivity among other arthropod tropomyosins. Results Tropomyosin-specific antibodies recognized in immunoblotting the natural tropomyosin in the insoluble fraction of silverfish extract. The silverfish tropomyosin (Lep s 1) was cloned and fully expressed. It shared high homology with other arthropod tropomyosins. rLep s 1 was recognized by tropomyosin-specific monoclonal and polyclonal antibodies and by IgE of allergic patients. It was able to inhibit the IgE binding to the insoluble fraction of silverfish extract, and to induce histamine release by an arthropod-allergic serum. Inhibition experiments revealed IgE cross-reactivity between rLep s 1 and other arthropod tropomyosins. Conclusion rLep s 1 is the first allergen cloned and characterized from silverfish extract. It enabled us to identify the natural counterpart in the insoluble fraction of silverfish extract, suggesting that the tropomyosin is not readily extractable with a classic aqueous extraction procedure. rLep s 1 displayed biological activity, suggesting that it could be regarded as a useful tool to study the role of silverfish tropomyosin in the sensitization to invertebrate allergic sources.

Recombinant DNA technology does provide pure, well-defined and reproducible products to be used for clinical purposes, by cloning and expressing the cDNA of allergens present in a specific extract. Ole e 5 is a pollen allergen of Olea europaea with an IgE-binding frequency of about 35%, which has been identified as a superoxide dismutase (SOD). The aim of this study was to clone the cDNA of Ole e 5, to express Ole e 5 in Escherichia coli and to characterize its immunoreactivity.cDNA of Ole e 5 was amplified by nested 3'-RACE PCR and cloned in pGEX vector 6P expression vector. After sequencing of some clones and homology analysis, the rOle e 5 was produced in an E. coli strain as a fusion protein with GST and purified. Then, the protein immunoreactivity was evaluated by patients' IgE binding (ELISA, ELISA inhibition, and immunoblotting) and by rabbit anti-rOle e 5 binding (immunoblotting and immunoblotting inhibition).The sequence analysis of Ole e 5 cDNA confirmed that Ole e 5 is a Cu/Zn SOD, with an identity from 90 to 80% with SOD from other species. rOle e 5 was recognized by IgE from 39% of olive pollen-allergic patients tested; moreover, this binding was inhibited by the olive pollen extract. An anti-rOle e 5 antiserum raised in rabbit strongly reacted with a natural component of about 16-kDa molecular weight present in the olive pollen extract; moreover, this binding was inhibited by the recombinant protein.Ole e 5 is the first Cu/Zn SOD identified as an allergen in a pollen source. Due to the widespread presence of this enzyme, rOle e 5 allergen, cloned and expressed in a complete form in E. coli, could represent a good tool to investigate the allergen cross-reactivity between O. europaea pollen and other allergenic sources, such as plant foods and other pollens.

Allergen cross-reactions among three strongly sensitizing Euphorbiaceae species, i.e., the rubber tree (Hevea brasiliensis), castor bean (Ricinus communis), and the Mediterranean weed Mercurialis annua were studied in Finnish patients (n = 25) allergic to natural rubber latex (NRL), but with no known exposure to castor bean or M. annua, and French patients allergic to castor bean (n = 26) or to M. annua (n = 9), but not to NRL. In immunoglobulin E (IgE)-immunoblotting, 28% of NRL-allergic patient sera recognized castor bean seed and 48% reacted to castor bean pollen proteins. Likewise, 35% of the NRL-allergic patient sera bound to M. annua pollen allergens. Nineteen percent of castor bean-allergic patients showed IgE to NRL and 8% to M. annua proteins. Sera from patients allergic to M. annua reacted in 44% to NRL, in 56% to castor bean seed, and in 78% to castor bean pollen proteins. In immunoblotting, castor bean seed extract inhibited the binding of NRL-reactive IgE to 20 kDa, 30 kDa of NRL, and 55 kDa of proteins; NRL extract, in turn, inhibited the binding of castor bean-reactive IgE to 14, 21-22, 29, and 32-34 kDa of castor bean proteins. In ELISA inhibition, NRL extract inhibited 33% of the binding of M. annua--reactive IgE of pooled sera to M. annua pollen. In conclusion, allergen cross-reactivity in vitro was observed among three botanically related Euphorbiaceae members, H. brasiliensis, R. communis, and M. annua, but the molecular specificity of the observed cross-reactions as well as their clinical significance remains to be elucidated. Allergen cross-reactivity should be taken into account in diagnostic work.

Background: A rapid method for the purification of the major 43-kDa all_ergen of Cupressus arizonica pollen, Cup a 1, was developed. Methods: The salient feature was a wash of the pollen in acidic buffer, followed by an extraction of the proteins and their purification by chromatography. Immunoblotting, ELISA, and lectin binding were tested on both the crude extract and the purified Cup a 1. Biochemical analyses were performed to assess the Cup a 1 isoelectric point, its partial amino-acid sequence, and its glycan composition. Results: Immunochemical analysis of Cup a 1 confirmed that the all_ergenic reactivity is maintained after the purification process. Partial amino-acid sequencing indicated a high degree of homology between Cup a 1 and all_ergenic proteins from the Cupressaceae and Taxodiaceae families displaying a similar molecular mass. The purified protein shows one band with an isoelectric point of 5.2. Nineteen out of 33 sera (57%) from patients all_ergic to cypress demonstrated significant reactivity to purified Cup a 1. MALDI-TOF mass spectrometry indicated the presence of three N-linked oligosaccharide structures: GnGnXF3 (i.e., a horseradish peroxidase-type oligosaccharide substituted with two nonreducing N-acetylglucosamine residues), GGnXF3/GnGXF3 (i.e., GnGnXF with one nonreducing galactose residue), and (GF)GnXF3/Gn(GF)XF3 (with a Lewisa epitope on one arm) in the molar ratio 67:8:23. Conclusions: The rapid purification process of Cup a 1 all_owed some fine studies on its properties and structure, as well as the evaluation of its IgE reactivity in native conditions. The similarities of amino-acid sequences and some complex glycan stuctures could explain the high degree of cross-reactivity among the Cupressaceae and Taxodiaceae families.

Sono stati confrontati i rilievi aeropalinologici, ottenuti con campionamenti volumetrici, delle citta di Genova e Sanremo (anno 1983), entrambe affacciate sul mare della Riviera ligure e distanti tra loro di circa 115 chilometri in linea d'aria. Le entita prese in esame sono state esclusivamenteUrticaceae, Poaceae edOleaceae per la loro preminente importanza allergologica in Liguria. Dal raffronto dei dati pollinici si puo osservare, oltreche la massiccia presenza in ambedue le zone delle tre entita, una particolarmente piu spiccata diffusione aerea ed un piu precoce inizio della fase di massima pollinazione dei taxa considerati in Genova rispetto a Sanremo.