Genetic variation of a global germplasm collection of chickpea (Cicer arietinum L.) including Italian accessions at risk of genetic erosion.

Chickpea (Cicer arietinum L.) is one of the most important legumes worldwide. We addressed this study to the genetic characterization of a germplasm collection from main chickpea growing countries. Several Italian traditional landraces at risk of genetic erosion were included in the analysis. Twenty-two simple sequence repeat (SSR) markers, widely used to explore genetic variation in plants, were selected and yielded 218 different alleles. Structure analysis and hierarchical clustering indicated that a model with three distinct subpopulations best fits the data. The composition of two subpopulations, named K1 and K2, broadly reflects the commercial classification of chickpea in the two types desi and kabuli, respectively. The third subpopulation (K3) is composed by both desi and kabuli genotypes. Italian accessions group both in K2 and K3. Interestingly, this study highlights genetic distance between desi genotypes cultivated in Asia and Ethiopia, which respectively represent the chickpea primary and the secondary centres of diversity. Moreover, European desi are closer to the Ethiopian gene pool. Overall, this study will be of importance for chickpea conservation genetics and breeding, which is limited by the poor characterization of germplasm collection.

Genetic structure and natural variation associated with host of origin in Penicillium expansum strains causing blue mould.

Blue mould, caused by Penicillium expansum, is one of the most economically damaging postharvest diseases of pome fruits, although it may affect a wider host range, including sweet cherries and table grapes. Several reports on the role of mycotoxins in plant pathogenesis have been published, but few focussed on the influence of mycotoxins on the variation in host preference amongst producing fungi. In the present study the influence of the host on P. expansum pathogenicity/virulence was investigated, focussing mainly on the relationship with patulin production. Three P. expansum strain groups, originating from apples, sweet cherries, and table grapes (7 strains per host) were grown on their hosts of isolation and on artificial media derived from them. Strains within each P. expansum group proved to be more aggressive and produced more patulin than the other two groups under evaluation when grown on the host from which they originated. Table grape strains were the most aggressive (81% disease incidence) and strongest patulin producers (up to 554µg/g). The difference in aggressiveness amongst strains was appreciable only in the presence of a living host, suggesting that the complex pathogen-host interaction significantly influenced the ability of P. expansum to cause the disease. Incidence/severity of the disease and patulin production proved to be positively correlated, supporting the role of patulin as virulence/pathogenicity factor. The existence of genetic variation amongst isolates was confirmed by the High Resolution Melting method that was set up herein, which permitted discrimination of P. expansum from other species (P. chrysogenum and P. crustosum) and, within the same species, amongst the host of origin. Host effect on toxin production appeared to be exerted at a transcriptional level.

Oral hyposensitization to nickel induces clinical improvement and a decrease in TH1 and TH2 cytokines in patients with systemic nickel allergy syndrome.

Some patients with nickel (Ni) allergic contact dermatitis suffer from systemic (intestinal or cutaneous) symptoms after ingestion of Ni-rich foods and experience symptoms reduction with low-Ni diet, a condition termed Systemic Ni Allergy Syndrome (SNAS). We aimed at evaluating whether oral administration of low nickel doses improved clinical conditions and modulated immunological aspects of SNAS, without significant side effects. Thirty-six SNAS patients were enrolled. Treatment started after 1-month of low-Ni diet and consisted in an incremental oral NiOH dose phase (0.3ng to 1.5 microg/week) followed by a 12-months maintenance phase (1.5 microg/week). Randomly, twenty-four patients added Ni therapy to low-Ni diet and 12 remained with diet alone. All patients were allowed rescue medications (antihistamines and topical steroids). After 4 months, Ni-rich foods were gradually reintroduced. In vitro allergen-driven IL13, IL5 and IFN-gamma release by peripheral blood mononuclear cells was evaluated before and after treatment. Twenty-three patients receiving NiOH and the 12 control patients completed the study. Evaluation of SNAS clinical severity (by VAS and drug consumption) showed a significant difference in favor of NiOH-treated patients compared to controls. Twenty of 23 patients in the NiOH group and none in the control group tolerated Ni-rich food reintroduction. Release of all studied cytokines in culture supernatants was significantly lower after NiOH treatment. In conclusion NiOH is effective in reducing symptoms and drug consumption in SNAS and is able to modulate inflammatory parameters.

Applicability of extracellular electrical impedance tomography in monitoring respiratory tract inflammation.

BACKGROUND: The presence of persistent mild inflammation is widely considered to provide the immunopathogenic basis for bronchial, nasal, or sinusal inflammation between critical phases and in asymptomatic periods. Exhaled nitric oxide (NO) is currently the most reliable marker of rhinobronchial inflammation, but its routine assessment is difficult as the test is available only in highly specialized centers. OBJECTIVE: The aim of this study was to evaluate the agreement between a new diagnostic method (extracellular electrical impedance tomography) and immunological and clinical symptom scores, anterior rhinomanometry, peak expiratory flow rate (PEFR), serum eosinophil cationic protein (ECP) level, and blood eosinophil count in the clinical monitoring of respiratory tract inflammation before and after treatment of asthma or rhinitis. PATIENTS AND METHODS: Eighty-seven patients were studied; 73 had mild persistent asthma (PEFR > or = 20% below predicted) and 14 had rhinitis. At baseline (TO), the patients underwent a medical examination to record symptom scores, PEFR, anterior rhinomanometry, an extracellular electrolytic conductivity test (bioimpedance tomography), serum ECP level and blood eosinophil count. Appropriate treatment was prescribed, following the guidelines of the Global Initiative for Asthma and the Allergic Rhinitis and Its Impact on Asthma. After 21 days of therapy (T1), the patients were re-evaluated for the same parameters. RESULTS AND CONCLUSIONS: This study demonstrates the good agreement (Cohen's kappa = 0.689) between the symptom scores of patients with rhinitis and the findings of extracellular tomography and very good agreement (kappa = 0.846) between symptom scores of asthma patients and extracellular tomography. These findings validate the use of this new technique for the real-time monitoring and adjustment of treatment in these clinical settings.

Practice parameters for sublingual immunotherapy.

The efficacy and safety of sublingual immunotherapy (SLIT) are currently supported by clinical trials, meta-analysis and post-marketing surveys. Practice parameters for clinical use of SLIT are proposed here by a panel of Italian specialists, with reference to evidence based criteria. Indications to SLIT include allergic rhinoconjunctivitis, asthma, and isolated conjunctivitis (strength of recommendation: grade A). As to severity of the disease, SLIT is indicated in moderate/severe intermittent rhinitis, persistent rhinitis and mild to moderate asthma (grade D). SLIT may be safely prescribed also in children aged three to five years (grade B), and its use in subjects aged more than 60 years is not prevented when the indications and contraindication are ascertained (grade D). The choice of the allergen to be employed for SLIT should be made in accordance with the combination of clinical history and results of skin prick tests (grade D). Polysensitisation, i.e. the occurrence of multiple positive response does not exclude SLIT, which may be done with the clinically most important allergens (grade D). As to practical administration, co-seasonal, pre co-seasonal, and continuous schedules are available, being the latter recommended for perennial allergens or for pollens with particularly prolonged pollination, such as Parietaria (grade D). For pollens with relatively short pollination, such as grasses and trees (cypress, birch, alder, hazelnut, olive) the pre co-seasonal and perennial schedules are preferred (grade C). The build-up phases suggested by manufacturers can be safely used (grade A), but they can be modified according to the patient's tolerance (grade C). A duration of SLIT of 3-5 years is recommended to ensure a long-lasting clinical effect after the treatment has been terminated (grade C).

The efficacy of sublingual immunotherapy for respiratory allergy is not affected by different dosage regimens in the induction phase.

BACKGROUND: Sublingual administration of allergens is a safe and effective alternative to subcutaneous immunotherapy in patients with respiratory allergies. A drawback to this therapeutic approach is the relatively long and complex management of the induction phase. AIM OF THE STUDY: To determine whether different induction regimens affect the outcome of sublingual immunotherapy. METHODS AND RESULTS: Adult and pediatric patients with allergic rhinoconjunctivitis and/or asthma were included in the study. Ten subjects served as controls and received symptomatic treatments. Forty-three subjects were allocated to sublingual immunotherapy, with three different induction protocols (8-, 15- and 20-day, respectively). Symptom and medication scores, skin test results and (in asthmatic patients) FEV1 values were monitored for two years. Adverse effects were recorded. All induction regimens produced a significant improvement in symptom and medication usage (p < 0.0001); skin test scores decreased (p < 0.0001) and FEV1 improved (p < 0.05). In contrast, symptom and skin test scores did not significantly change in controls. No relevant adverse effects were observed with any of the induction regimens. CONCLUSIONS: For patients with respiratory allergies, sublingual immunotherapy with an 8-day induction protocol is safe and effective. Our results encourage the usage of shorter induction regimens, which produce better compliance with this therapy.

A model for management of sublingual immunotherapy.

UNLABELLED: High dose sublingual immunotherapy (SLIT) is generally well tolerated, since most side effects are local reactions involving the oral or gastrointestinal mucosa. However, such reactions, if not adequately managed, may lead the patient to stop SLIT. OBJECTIVE: We introduce a method to manage side effects of SLIT, which was successfully applied on a group of children undergoing SLIT. METHODS: SLIT was performed with Staloral 300 (Stallergénes, Antony, France) by the schedule in 11 days indicated by the manufacturer with a tap dose of 8 pressures of the 300 IR concentration. The treatment was managed by flow charts suggesting adjustment of dose, regimen, or kind of assumption depending on the kind of reactions (oral, gastrointestinal, respiratory). RESULTS: An overall number of 200 children (120 males, 80 females, mean age 9 yrs, range 3-14 yrs) were treated. Of them, 155 (77.5%) had no reaction, while 45 (22.5%) had side effects, consisting of 20 oral itching or burning, 15 gastrointestinal symptoms and 10 rhinitis. No systemic reaction was observed. All side effects were successfully managed by the proposed method, thus reaching the maintenance dose in all patients but one, reporting repeated intense oral itching, who stopped SLIT. CONCLUSIONS: High dose SLIT is well tolerated in treated patients. In most patients with side effects, which are mainly oral itching or gastrointestinal disturbances due to contact with the allergen extract, a proper management allows to reach the maintenance dose with no further reactions.

Long-lasting effect of sublingual immunotherapy in children with asthma due to house dust mite: a 10-year prospective study.

BACKGROUND: Subcutaneous immunotherapy for respiratory allergy has shown a long-lasting efficacy after its discontinuation, whereas this evidence is still lacking for sublingual immunotherapy, despite the fact that it is widely used. OBJECTIVE: We aimed to evaluate whether a long-lasting effect of SLIT occurs, in a prospective parallel group controlled study. METHODS: Sixty children (mean age 8.5 years) suffering from allergic asthma/rhinitis due to mites were subdivided into two matched groups: 35 underwent a 4- to 5-year course of SLIT with standardized extract and 25 received only drug therapy. The patients were evaluated at three time points (baseline, end of SLIT and 4 to 5 years after SLIT discontinuation) regarding presence of asthma, use of anti-asthma drugs, skin prick tests and specific IgE. RESULTS: We found that in the SLIT group there was a significant difference vs. baseline for the presence of asthma (P

Post-marketing surveillance study on the safety of sublingual immunotherapy in pediatric patients.

BACKGROUND: Immunotherapy (IT) is the only causal treatment for allergic subjects recognized to be effective and to offer long-lasting efficacy. The noninjective routes, aimed at improving the safety of the treatment, have been validated as effective in adults, but documentation of their safety in children is still poor. The aim of the present survey study was to assess the safety of sublingual immunotherapy in pediatric patients, by evaluating a large population. METHODS: A total of 268 children (aged 2-15 years), receiving sublingual IT for respiratory allergy, were followed-up over a period ranging from 3 months to 7 years (mean 34 months). The side-effects possibly due to the treatment were recorded on a proper diary card; self-assessment of the clinical outcome was also evaluated. RESULTS: About 96000 doses of extract were globally administered. Local side-effects were of no clinical relevance. Eight side-effects were reported (3% of patients; 0.083 per 1000 doses). Seven systemic side-effects (abdominal pain, conjunctival itching, and rhinitis) were mild and required no treatment. One case of urticaria was well controlled with oral antihistamines. No life-threatening event occurred. The clinical outcome was judged excellent or good by 80% of the patients. CONCLUSIONS: The sublingual IT herein investigated appeared to be well tolerated and safe in pediatric patients. The risk/benefit ratio was therefore favorable.

Grass pollen specific sublingual/swallow immunotherapy in children: open-controlled comparison among different treatment protocols.

BACKGROUND: sublingual-swallow immunotherapy has been recognized as safe and effective in adults. Different treatment schedules have been proposed and used, but the current experience in children is insufficient. We carried out a study to evaluate the clinical efficacy and the safety in children allergic to grass pollen, following different schedules. METHODS: an open study was performed with 3 parallel treated groups and a control group, each including 12 children suffering from rhinoconjunctivitis due to grass pollen. The patients underwent either a preseasonal treatment or a pre-coseasonal treatment during the grass pollen season. The analysis of the clinical outcome was based on the number of days with symptoms and/or drug consumption in the pollen peak month. RESULTS: significantly less days with symptoms and/or drug intake in all of the groups submitted to SLIT in comparison to the control group were found (p < 0.0001). Among the three schedules used, the best clinical results were obtained with the two pre-coseasonal treatments (p < 0.0001), with a preference for the schedule with the lowest maintenance dose according to the drug intake (p = 0.0267). Two patients belonging to the coseasonal schedule with the highest maintenance dose interrupted the treatment because of systemic mild side effects. CONCLUSIONS: our data suggests that in children with rhinoconjunctivitis to grass pollen, SLIT is effective and well tolerated and that the pre-coseasonal schedule should be preferred to the preseasonal treatment. According to our data, the best maintenance dose during the pollen season for children, both for tolerance and clinical outcome, corresponds to 0.4 microg of the grass major allergen Group 5 administered three times a week.

Nasal secretion specific IgE in rhinitis patients.

Forty-five subjects with rhinitis of suspected allergic origin underwent immune and allergy testing including skin-tests and immuno-enzymatic assay of serum and nasal secretion specific IgE (RAST). Nasal secretion was collected using a cotton tampon inserted in both middle meati. A statistical analysis comparing the results of skin-tests and serum RAST was performed: the results obtained with these two techniques were very homogeneous (86.7% accuracy). An overall concordance (82.2%) was also observed by comparing skin-tests and nasal secretion RAST. Finally, the results of serum and nasal secretion RAST were compared: in this case totally superimposed data were reported (overall accuracy: 93.3%). In all cases the comparison between RAST classes in serum and nasal secretion was carried out: the difference between the semiquantitative evaluation of reaginic concentration classes using these two methods is not statistically significant (chi-square test). This technique of nasal secretion collection is simple and with no risk involved.

Serum IgE and atopy in drug-sensitive patients.

One of the most interesting aspects of the pathogenesis of the allergic and pseudo-allergic reactions to drugs is the one concerning the possible roles of genetic factors, in general, and the relationship with atopy, in particular. We decided to study the correlation between the serum level of total IgE, the personal and familial history of atopy, and the allergologic tests (skin test, PRIST, RAST, etc) for one or more drugs taken by 465 patients with personal history of allergic reactions. From the data, we observed that the average amount of total IgE lies within the norm, 125.4 KU/L (+/- 209.2), without any significant difference between the allergic (132 KU/L) and the pseudo-allergic (121.8 KU/L) patients. The IgE levels tend to be higher in that part of the tested population which presented a circumstantial atopic status (11.8%) and they are not influenced by an allergic or pseudo-allergic reaction to drugs. According to these results, the relationship between drug hypersensitivity and atopy needs further investigation.

Pancuronium allergy: a case report.

A case of anaphylactic shock to pancuronium bromide is described in which a type I immunological reaction occurred. A 33-yr-old male anaesthetist, with no family or personal history of allergy, collapsed soon after induction of anaesthesia. The drugs used included fentanyl, droperidol, thiopentone and pancuronium. Diffuse erythema occurred, with probable cardiac arrest. The patient was treated with cardiac massage, sympathomimetics and cortisone, and made a complete recovery. Examination (skin tests, passive transfer according to Prausnitz and Küstner, in vitro lymphocyte transformation test, total IgE assay) revealed that the allergic reaction was to pancuronium bromide.

Status of patch and other skin tests in diagnosis of systemic penicillin allergy.

It is generally accepted that cell-mediated immunity is responsible for contact dermatitis, but controversy still exists about the involvement of Type IV immune-reactions in other clinical manifestations following inhalant, oral and parenteral administration of penicillin and semi-synthetic penicillin or cephalosporins. The aim of the present study was to contribute to a better understanding of the pathogenetic mechanism responsible for these reactions. Detailed history taking, clinical and immuno-allergological investigation on 161 patients with a history of an allergic reaction to penicillin and/or semisynthetic penicillins and/or cephalosporins was carried out with the following tests: skin tests, patch tests, RAST, PRIST and lymphocyte transformation test. The statistical evaluation of data was performed by X2 test. Comparative studies were focused on groups of patients who presented a positive reaction to patch test (37) skin test (scratch and intradermal) (42), simultaneous skin and patch test (5) and negative skin test (87). No significant difference was observed among the groups studied, although in positive and skin test, a higher frequency is observed to be associated with anaphylactic shock and eczematous type of reaction. It is not possible however not only, to establish whether this is due to a combined immune-reaction (Type I and IV), orto a causal coexistence of the two mechanisms but even to determine either of allergic or pseudoallergic phenomenon without any accurate immunoallergological test. It is suggested that patch tests be carried out as routine investigations on patients with a systematic reaction to penicillins and/or cephalosporins, regardless of the route of administration.

ASA disease: the clinical relationship of nasal polyposis to ASA intolerance.

We studied the complex relationship between nasal polyposis and ASA (acetylsalicylic acid) intolerance in 154 patients with nasal polyps. The clinical histories of all patients were reviewed, and diagnostic tests for immune or allergic causes and the responsiveness of patients to challenges with ASA-substitutive drugs were analyzed. A third of our patients tested were found to have positive personal histories of atopy and 35% showed ASA intolerance. Although 40% had bronchial asthma, only 16.8% of all patients had positive tests for allergy. We were unable to find a specific mechanism to explain the relationship between nasal polyposis and ASA intolerance and further investigations are still required.

Oral specific hyposensitization in the management of patients allergic to food.

Results are presented of a protocol of hyposensitizing treatment per os in 19 cases of food allergy (allergy to milk in 8, to egg in 8, to fish in 2, and to orange in one). Treatment, performed in the out-patients service, was successful in 14 out of the 15 patients who followed it correctly (i.e. 93.3% of cases). Treatment lasted between 3 and 12 months.