Accelerated Aging Effects on Color Stability of Potentially Color Adjusting Resin-based Composites.

The aim of this study was to compare the effects of accelerated aging on the overall color stability of potentially color adjusting commercial resin-based composite resins. Thirty specimens (10 mm diameter and 2.5 mm thick; n=6) were fabricated using five different materials: Estelite Omega, GC Kalore, Venus Pearl, Harmonize, and Omnichroma. Color measurements were taken for each sample using a spectrophotometer before and after submitting samples through the artificial aging process (Q-sun Xenon Test Chamber, 102 min light at 63°C black panel temperature; 18 min light and water spray per ASTM G155) for a total of 300 hours (12.5 days). The total color difference (ΔE*ab) was calculated using SpectraMagic NX software and analyzed using one-way analysis of variance and Tukey test. The results for color change (ΔE*ab) were statistically significant. Omnichroma and Venus Pearl presented superior color stability and the lowest overall color change, whereas GC Kalore and Harmonize presented significant color change that would be considered clinically unacceptable (ΔE*ab > 3.3).

Which allergen extract for grass pollen immunotherapy? An in vitro study.

BACKGROUND: Grass pollen is a major cause of allergy throughout the world. The only treatment targeting the causes and not only the symptoms of allergy is specific immunotherapy (IT). A number of controlled trials demonstrated the efficacy of IT in grass pollen allergic subjects, most using extracts of multiple grasses but some using extracts of a single grass. The optimal grass extract for IT has not yet been established. METHODS: This study is aimed at investigating the IgE-binding pattern in sera from IT-naïve patients from central Italy with allergic rhinitis and/or asthma caused by grass pollen. A 5-grass extract was used (containing Dactylis glomerata, Poa pratensis, Lolium perenne, Antoxanthum odoratum and Phleum pratense) and compared to Phleum pratense alone, which is the most frequently used single grass extract, by the RAST-inhibition technique. RESULTS: The 5-grass extract showed, by RAST-inhibition, a significantly higher binding compared to the Phleum pratense extract for Antoxanthum odoratum and Poa pratensis, while the two extracts for immunotherapy showed similar binding affinity for Phleum pratense and the non-Pooideae grass, Cynodon dactylon. CONCLUSIONS: The use of a mixed-grass pollen extract seems to be the optimal choice when applying specific IT in grass pollen-allergic subjects from the Mediterranean area.

[Minimally invasive techniques for treatment of metastatic cancer]. / Minimal-invasive Techniken zur Behandlung metastasierter Karzinome.

Many percutaneous image-guided ablative techniques have been used in the treatment of cancer-related bone metastases. These techniques are rapidly becoming a focus in the palliative and curative treatment of patients with both benign and malignant bone cancer.In this article we will briefly review the principles of radiofrequency ablation and our experience with this technique, including its use in combination with surgery; we will also discuss other minimally invasive techniques such as cryoablation and osteoplasty in the treatment of bone metastases.

Lack of inflammatory cells in the oral mucosa of subjects undergoing sublingual immunotherapy.

The cells involved in allergic inflammation, such as mast cells, basophils, and eosinophils, have been thoroughly studied in the nose, the lungs and the skin, demonstrating an evident increase in response to the introduction of the specific allergen, while little is known in the mucosal system and particularly in the oral mucosa. We investigated such tissue by using the model of sublingual immunotherapy (SLIT), by which high doses of the specific allergen enter the mouth. Oral biopsies were carried out on seven subjects allergic to grass pollen and treated with SLIT by a grass extract. In biopsies carried out before SLIT there was a very low number of mast cells and eosinophils both in the epithelium and subepithelium layers, and insignificant changes were detected after SLIT. These findings show the lack of allergic inflammation in the oral mucosa upon contact with the specific allergen and confirm the role of the mouth as a tolerogenic site, which is conceivable considering the different attitude of the mouth, where the antigens transit to undergo digestion, in respect to the airways or the skin, where the antigen absorption is potentially dangerous.

Allergy and the gastrointestinal system.

The gastrointestinal system plays a central role in immune system homeostasis. It is the main route of contact with the external environment and is overloaded every day with external stimuli, sometimes dangerous as pathogens (bacteria, protozoa, fungi, viruses) or toxic substances, in other cases very useful as food or commensal flora. The crucial position of the gastrointestinal system is testified by the huge amount of immune cells that reside within it. Indeed, gut-associated lymphoid tissue (GALT) is the prominent part of mucosal-associated lymphoid tissue (MALT) and represents almost 70% of the entire immune system; moreover, about 80% of plasma cells [mainly immunoglobulin A (IgA)-bearing cells] reside in GALT. GALT interacts strictly with gastrointestinal functions in a dynamic manner; for instance, by increasing intestinal permeability in replay to particular stimulations, or orientating the immune response towards luminal content, allowing either tolerance or elimination/degradation of luminal antigens, or sometimes provoking damage to the intestinal mucosa, such as in coeliac disease or food allergy. The immune mechanisms implicated in these actions are very complex and belong to both innate and adaptive immunity; innate immunity supplies an immediate non-specific response that is indispensable before specific adaptive immunity, which needs 7-10 days to be efficacious, takes place. The results of their interactions depend upon different contexts in which contact with external agents occurs and may change according to different genetic settings of the hosts.

Comparison between continuous or intermittent schedules of sublingual immunotherapy for house dust mites: effects on compliance, patients satisfaction, quality of life and safety.

Sublingual immunotherapy (SLIT) is indicated in the treatment of allergic rhinitis and asthma. However, an issue scantly investigated is the patients satisfaction and the consequent compliance. This study is aimed at evaluating the possible differences of SLIT administered continuously or intermittently on several parameters: clinical efficacy, Quality of Life (QoL), satisfaction, compliance and safety. Forty allergic patients were treated for 12 months. The treatment was carried out by sublingual administration of an allergen extract of a 50% mixture of Dermatophagoides pteronyssinus and Dermatophagoides farinae at 10 and 300 IR/ml concentrations. Patients were randomly treated continuously or intermittently (i.e. 2 month treatment alternate to 2 month suspension). Both schedules were significantly effective in reducing allergic symptoms and improving QoL. Compliance and satisfaction were good in both groups. Local and systemic reactions were few, self-resolving, and mild in both schedules. Intergroup analysis did not reveal any difference between the two groups regarding these parameters. In conclusion, this preliminary study provides the evidence that also intermittent SLIT is as effective and safe as traditional continuous treatment. In addition, compliance and satisfaction are super-imposable in the two groups.

Sublingual immunotherapy: from safety to mechanism of action.

Allergen specific immunotherapy is an important option for the treatment of respiratory allergy and its clinical efficacy has been clearly demonstrated by several studies. However, the injective route of administration and the possibility of severe side effects has limited its use in children and led to the introduction of new forms of administration. Sublingual immunotherapy (SLIT) has proven to be an effective and safe treatment for respiratory allergy. However, its mechanism of action is still debated. Pharmacokinetic studies showed that, differently from nasal mucosa, allergen extracts administered by SLIT are not immediately adsorbed but are long retained before being drained to local lymph nodes. This difference may be responsible of the absence of severe side effects and instead of short-lasting local symptoms. Studies by biopsies of the oral mucosa should greatly help in defining the presence and the role of cells involved in the mechanisms of oral tolerance.

Lower airway inflammation before and after house dust mite nasal challenge: an age and allergen exposure-related phenomenon.

BACKGROUND: Upper and lower airways allergic disease is currently considered unitarily. Allergic inflammation in one site can extend to other sites of the respiratory tract. OBJECTIVE: To evaluate bronchial inflammation before and after allergen-specific nasal challenge (ASNC) in rhinitic and asthmatic children, considering the different levels of allergen exposure, i.e. summer (low) and winter (high). METHODS: Fourteen children with rhinitis and 15 with rhinitis and asthma, all monosensitized to mites and 10 healthy controls were studied. Nasal IgE were measured before ASNC in summer and in winter season. Nasal clinical score, eosinophil cationic protein (ECP), nasal tryptase, bronchial clinical score, FEV(1), PEF, sputum ECP, sputum tryptase and exhaled nitric oxide (eNO) were evaluated before and after ASNC in summer and winter season. RESULTS: Nasal scores significantly increased after ASNC in rhinitic and asthmatic children in both seasons. Nasal IgE were significantly higher in summer compared to winter. Bronchial symptoms, FEV(1) and PEF showed no mean differences in rhinitic and asthmatic children after ASNC, with an increase of bronchial symptoms and a decrease of FEV(1) and PEF occurring in 3/15 asthmatic children. In both groups nasal tryptase and ECP after ASNC significantly increased in summer and winter, while sputum tryptase was undetectable before or after ASNC in both groups. Sputum ECP and eNO at baseline were significantly higher in patients than in controls (summer P=0.002, winter P=0.001). Sputum ECP significantly increased after ASNC in 3/15 asthmatics in summer and in 11/15 in winter, as well as in 3/14 rhinitics in summer and in 4/14 in winter. eNO significantly increased after ASNC in 3/15 asthmatics in summer and in 10/15 in winter, and in 1/14 rhinitics in summer and in 4/14 in winter. A significant median increase of sputum ECP (P=0.0007) and eNO (P=0.0012) after ASNC in asthmatic and of eNO (P=0.013) in rhinitic children was also found in winter. CONCLUSIONS: Basal sputum ECP and eNO values, significantly higher before ASNC in rhinitic patients compared to control subjects, confirm the inflammatory link of upper and lower airways. The more frequent detection of inflammatory changes induced by ASNC in winter suggests that allergen exposure favours the transfer of nasal inflammation to lower airways.

Quadriceps-sparing versus mini-subvastus approach in total knee arthroplasty.

In a prospective randomized double-blind study we compared the postoperative recovery and early results of two groups of 30 patients having total knee arthroplasty with minimally invasive techniques using either a mini-subvastus or a modified "quadriceps-sparing" approach. All knees were implanted with the same posterior-stabilized prosthesis (LPS-Flex, Zimmer, Warsaw, IN) by the same surgeon with the same dedicated set of downsized instruments. Epidural anesthesia with the same postoperative analgesia and rehabilitation protocol was used in all patients. Evaluation was performed preoperatively, postoperatively in the first week, and at 1 and 3 months. In five cases in the "quadriceps- sparing" group, the incision was extended a few cm to facilitate exposure. Tourniquet time, estimated blood loss, and postoperative pain were similar in the two groups. Active straight leg raising was achieved half a day earlier, on average, in the mini-subvastus group (1.9 vs 1.4 days). Average maximum active flexion was similar in the two groups at each interval, and reached 117 degrees and 119 degrees at 3 months for the mini-subvastus and "quadriceps-sparing" group, respectively. We believe there was no difference between the mini-subvastus and "quadriceps-sparing" approach in relation to short term recovery or early results.

Dose dependence of efficacy but not of safety in sublingual immunotherapy.

Sublingual immunotherapy (SLIT) currently represents, as indicated by meta-analysis of its efficacy and safety, a valid option to the generally used traditional subcutaneous immunotherapy (SCIT) for treating respiratory allergy. Regarding efficacy, recent studies demonstrated that, similar to what has already been observed in SCIT as well as in experimental and clinical studies about the magnitudo of allergen exposure, the effectiveness on both clinical symptoms and immunologic changes depends on the amount of allergen administered during treatment. In addition, in vitro studies addressed with the role of dendritic cells, currently considered to be of pivotal importance in orienting toward tolerance the immune response to allergens, showed that the internalisation of allergen molecules, which is followed by tolerogenic presentation to T cells, depends on the amount of allergen. However, such dose dependence is not apparent concerning the safety. In fact, the comparison of studies respectively conducted with high and low allergen doses did not show differences in the rate of systemic reactions, which in any case never had the presentation of anaphylaxis, and instead a significant difference in the rate of local reactions, following the oral and gastrointestinal contact with the allergen extract, in favour of high dose studies.

Sublingual reactivity to rBET V1 and rPHL P1 in patients with oral allergy syndrome.

Oral Allergy Syndrome (OAS) in patients with pollen-induced rhinoconjunctivitis is caused by specific IgE recognizing cross-reacting epitopes of fruits and plants, which were clearly shown in vitro, but failed to be demonstrated in vivo by cross-challenges in the target organs. Considering the hypothesis of degradation of such epitopes in natural extracts, challenges with recombinant pollen allergens were done to evaluate the reactivity of the oral mucosa in OAS patients. Seventeen patients with OAS and rhinitis from birch (10) and grass pollen (7) and 10 non-atopic controls were studied by skin prick tests (SPT), allergen specific nasal challenges (ASNC) and allergen specific sublingual challenges (ASSC) with birch and timothy extracts and with rBet v1 and rPhl p1 at increasing concentrations from 1 to 1000 mcg/ml. None of the healthy subjects in the control group had any positive test for birch and timothy extracts or for recombinant allergens. In the OAS group the following results were observed: SPTs with recombinant allergens were positive in all patients, mostly at 10 mcg/ml concentration; ASNC with rBet v1 were positive in all patients, mostly at 100 mcg/ml; ASSC with natural pollen extracts were positive in only 2 of 17 patients, but in 15 of 17 with rBet v1 and rPhl p1, mostly at 500 mcg/ml and 1000 mcg/ml. ASSC with rBet v1 and rPhl p1 were positive with a mean concentration of 677 and 533 mcg/ml, respectively. The results of sublingual challenges with rBet v1 and rPhl p1 showed the in vivo cross-reactivity between pollens and foods in patients with OAS, but high concentrations of the recombinant allergens were needed to reproduce oral symptoms, thus explaining the failure of challenges performed with natural extracts, which have concentrations of major allergens lower than 50 mcg/ml. This indicates that sublingual mucosa is much less reactive to allergens than other surfaces, such as skin and nasal mucosa, probably because of its anatomic and immunologic peculiarity.

Mini-subvastus versus medial parapatellar approach in total knee arthroplasty.

UNLABELLED: We compared short-term clinical results of the mini-subvastus approach with the standard parapatellar approach for total knee arthroplasty. Can one gain adequate access through the mini-subvastus approach without exceeding operating time, incurring additional complications, and maintaining correct implant geometry? In a prospective, observer-blinded study, 120 consecutive patients having total knee arthroplasty were operated on by one surgeon using either the mini-subvastus approach without patella eversion or the standard parapatellar approach with patella eversion. Patients were matched according to age, gender, body mass index, knee flexion, deformity, and pre-existing high tibial osteotomy. The mini-subvastus approach was technically more demanding. Reduced access and visibility prolonged the tourniquet time by an average of 15 minutes and led to two intraoperative complications. Patients in the mini-subvastus group lost on average 100 mL less blood and had better pain scores on day one [visual analogue scale (VAS): mean 2.4 versus 3.89]. They reached 90 degrees knee flexion earlier (2.8 versus 4.5 days), and an active straight-leg raise earlier (3.2 versus 4.1 days). Their average flexion at 30, 60 and 90 days was slightly better (100 degrees , 110 degrees , and 112 degrees versus 94 degrees , 106 degrees , and 109 degrees ). All patients including those with complications had good results with good component geometry and leg alignment. The mini-subvastus approach offers early but short-lived benefits for patients at the expense of a longer operation and a higher risk of complications. LEVEL OF EVIDENCE: Therapeutic study, Level II-1 (prospective comparative study). See the Guidelines for Authors for a complete description of levels of evidence.

Three-year follow-up of clinical and inflammation parameters in children monosensitized to mites undergoing sub-lingual immunotherapy.

Parallel follow-up of clinical and inflammatory markers during sub-lingual immunotherapy (SLIT) is highly beneficial. Twenty-four children (age 4-16) monosensitized to house dust mite were randomized to receive either active or placebo SLIT for 1 yr in a double-blind placebo controlled design (Marcucci et al., Allergy 2003: 58: 657-62). Thereafter, for 2 yr they all received active treatment. Symptom scores for rhinitis, asthma, and drug usage were daily recorded. Eosinophil cationic protein (ECP) and tryptase in sputum and nasal secretions, serum and nasal mite-specific immunoglobulin E (IgE) were recorded before treatment and at 10-12 months intervals. Nasal ECP and nasal tryptase after specific nasal provocation tests were significantly reduced as compared to baseline values (p = 0.0043 and 0.0195, respectively) in the third year of active treatment. None of the other inflammatory parameters was increased. In placebo treated patients all these parameters tended to decrease only after switching to active treatment. Clinical scores did not improve in treated vs. placebo patients in the double-blind placebo-controlled phase of the study. In both cohorts a clinical benefit was observed as intra-group score reduction as compared to baseline. A significant difference was reached in patients treated for 2 yr for rhinitis and asthma (p = 0.0009 and 0.0019, respectively) but not for drug usage and in patients treated for 3 yr for rhinitis, asthma, and drug usage (p = 0.0105, 0.0048, and 0.02, respectively). SLIT in children monosensitized to mites reverted the spontaneous increase in nasal IgE and in local parameters of allergic inflammation. These outcomes were followed by a consolidated clinical improvement in the second and third year of treatment.

Dose dependence of immunological response to sublingual immunotherapy.

BACKGROUND: Sublingual-swallow immunotherapy (SLIT) is an accepted treatment for allergic rhinitis but its optimal dosage is scantly investigated. We studied the dose dependence of clinical efficacy and immunological response to SLIT by administering two different dosages of the same allergen in rhinitic children monosensitized to grass pollen. METHODS: Seventy-one patients with comparable age and symptoms were randomized to receive SLIT by the same grass pollen extract from Stallergenes (Antony, France), 40 of them with the 100 IR and 31 with the 300 IR extract. All patients recorded diary cards for symptoms, medications and side-effects of the treatment, and had measurements of specific IgE and IgG4 in serum by the CAP System FEIA (Pharmacia, Uppsala, Sweden) and in nasal secretion by an in situ incubation method with the same reagents of CAP System FEIA. RESULTS: Symptom/medication scores during the pollen season were significantly higher in patients treated with the lower dosage compared with those treated with the 300 IR dosage. Side-effects occurred with a comparable rate (25.8%vs 27.5%) in the two groups. Serum-specific IgE and IgG4 had no significant changes after 3 months of SLIT in both groups, while a significant seasonal increase of nasal IgE (P = 0.015) and IgG4 (P = 0.019) was found only in patients treated with the lower dosage. CONCLUSIONS: A rise of specific IgG4 and a blunting of seasonal increase of specific IgE in serum was repeatedly reported during subcutaneous immunotherapy (SCIT) with pollen extracts. Our findings show such blunting of specific nasal IgE along with a low symptom/medication score in patients treated with SLIT with the higher dosage, but not a concomitant rise of specific nasal IgG4. This suggests a local immunological effect of SLIT, different from systemic mechanisms of SCIT.

The safety of sublingual-swallow immunotherapy: an analysis of published studies.

BACKGROUND: As the main target of sublingual immunotherapy (SLIT) is to reduce at most the occurrence of adverse events (AE), safety represents a critical issue. This aspect deserves particular mention when a higher dose of allergen extract than traditional subcutaneous immunotherapy (SCIT) is required to be effective: that may be up to 500 times that employed for SCIT. OBJECTIVE: All published controlled studies concerning SLIT-swallow were analysed to evaluate AE rates. METHODS: Studies were subdivided in two groups: (i) studies using low allergen dose (LAD), i.e. ranging from 1 to 50 times the dose commonly administered with SCIT, and (ii) studies with high allergen dose (HAD), i.e. ranging from 50 to 500 times the dose administered with SCIT. RESULTS: Twenty-five studies were altogether analysed: 13 studies belonged to the low-dose group, 12 belonged to the high-dose group. We considered all patients with at least one AE. Local reactions were significantly more frequent in the LAD group than in the HAD group (P<0.0001), while there was no difference in the rate of systemic reactions. Severe systemic reactions were never reported. CONCLUSION: This study represents the first analysis of the safety of SLIT concerning the allergen dose employed in the treatment. There is evidence that AE occurrence is substantially not dose-dependent. This fact highlights two main clinical aspects: the elevated tolerability of SLIT in general and the safety of HAD regimen.

Physical exercise does not favour adverse reactions to allergen immunotherapy by the sublingual route.

It is generally recommended in consensus documents on allergen immunotherapy to avoid any kind of physical exercise in the 24 hours following the administration of the allergen extract but such recommendation is not supported by scientific evidence. We evaluated the risk of developing adverse reactions in a group of patients submitted to sublingual immunotherapy by performing a controlled exercise test. Eleven patients were included in the study, 8 treated with grass pollen and 3 with Parietaria pollen extract by Staloral300 (Stallergénes, Antony, France), with the build-up phase in 11 days suggested by the manufacturer and a top dose of 300 IR. At the first maintenance dose, in all patients were measured heart rate, blood pressure, FEV1, and tryptase in blood. Then the maintenance dose was assumed and the physical exercise performed by cycloergometer maintaining a heart rate around 100 b/min and measuring again tryptase, blood pressure and FEV1. The results showed that no patient had adverse reaction following physical exercise, neither showed significant changes in FEV1, blood pressure, or tryptase level, which was 5.57 +/- 4.54 mcg/l before and 5.81 +/- 4.34 mcg/l after exercise. This suggests that there is no reason to advise against physical exercise, and particularly sporting activity, in patients treated with sublingual immunotherapy thus avoiding to interfere with the subject's lifestyle and consequently with his quality of life.

Evaluation of food-pollen cross-reactivity by nose-mouth cross-challenge in pollinosis with oral allergy syndrome.

BACKGROUND: Oral allergy syndrome (OAS) is often associated with pollen-induced rhinitis, and there are preferential associations between causative substances. If OAS and rhinitis are both immunoglobulin (Ig)E-mediated and there are cross-reacting proteins, it is expected that similar reactions can be elicited in the nose and mouth. In order to test this hypothesis we performed a series of 'cross-challenges' with foods and pollens in both the nose and the mouth. METHODS: Nine patients with ascertained OAS due to vegetables and rhinitis due to pollens were studied. On the first day a nasal challenge with pollen extracts and an oral challenge with fresh food was carried out. After a week, washout nasal challenge with food and an oral challenge with pollens were performed. Immediate symptoms, mucosal tryptase and soluble eosinophil cationic protein (ECP) were assessed after each challenge. RESULTS: The administration of pollen into the nose and food into the mouth elicited symptoms as expected, but the cross-challenge had no clinical effect. In parallel, tryptase and ECP increased after nasal challenge with pollens, whereas foods did not elicit a measurable response. CONCLUSION: The cross-reactivity between foods and pollens, when evaluated at the shock organ, was not clinically evident. This data can be explained with a low concentration of cross-reagent epitopes in pollen extracts and food homogenized because of degradation. The different behaviour upon challenge suggests that different immunological mechanisms may act in the nose and mouth.

Measurement of nasal IgE in an epidemiological study: assessment of its diagnostic value in respiratory allergy.

BACKGROUND: Specific questionnaire and skin prick test (SPT) are the most used methods in epidemiological studies on respiratory allergy. SPT, however, can be positive in many subjects without evidence of any allergic disease. Nasal IgE determination has been suggested by some authors as a valuable diagnostic method, which may overcome this lack of specificity. OBJECTIVE: The aim of this study was to evaluate sensitivity and specificity of nasal specific IgE for the seven most common inhalant allergens in order to verify its reliability as a screening test. METHODS: 126 children, involved in an epidemiological study on prevalence of respiratory allergic disease, were evaluated. All children were assessed with a specific questionnaire, SPT and nasal specific IgE. Nasal specific IgE were determined with a previously described method modified for screening purposes, in order to test seven allergens at the same time. When discordant results were obtained between questionnaire, SPT and nasal IgE, an allergen specific nasal challenge (ASNC) was performed and nasal tryptase was also determined before and after challenge. RESULTS: The questionnaire was positive for respiratory allergy in 28/126 children. SPT was positive in 21 of the 28 children, but also in 5/10 children with atopic dermatitis (AD), and in 12/88 children without allergic symptoms. Nasal IgE were positive in 22/28 and also in 2/10 with AD. Nasal challenge and tryptase confirmed the negativity of nasal IgE in 12/17 children with positive SPT but totally negative for allergic respiratory disease. Moreover nasal IgE was found to be positive to dermatophagoides in one of seven children with negative SPT despite a clinical history suggestive for mite respiratory allergy. In this patient and in 2 of the 5 children with AD the positive nasal IgE to mites was confirmed by a positive ASNC and tryptase. CONCLUSIONS: Nasal IgE have shown a specificity significantly higher than SPT (98% vs. 83%) and a good sensibility. This screening test may also be useful to detect the beginning of upper airways sensitization in patients with AD.

Do tryptase, ECP and specific IgE measurement by nasal incubation increase the specific nasal provocation test sensitivity?

The specific Nasal Provocation Test (sNPT) is a third level diagnostic tool. Fitted to reproduce natural exposure condition to pick the responsible allergen for nasal symptoms out, it is applied when prick test and RAST responses are doubtful. SNPT results have been evaluated measuring nasal resistance (anterior rhinomanometry) and nasal symptoms (clinical score), reaching 50% of sensitivity. This study focused on the determination of allergic response markers, triggered by nasal challenge: tryptase levels in the nose, specific IgE and ECP (Eosinophil Cationic Protein). The aim was to increase sNPT sensitivity. Twenty patients suffering from allergic rhinitis and 16 age-matched-nonallergic subjects were enrolled in the study. Tryptase, specific IgE and ECP were determined in nasal mucosa applying a new method, based on in situ incubation, before and after sNPT. The latter was performed following a standardized method. Tryptase levels increased in 13 patients (65%), were unchanged in four patients (20%), and slightly decreased in three patients (15%). The increase recorded was significant in mite allergic patients (p=0.005), but not significant (p> 0.05) in pollen allergic patients. ECP values increased in 13 patients (65%), were unchanged in two patients (10%), and highly decreased in five patients (25%). ECP increase was not significant (p> 0.05). Specific IgE levels increased in seven patients (35%), were unchanged in 11 patients (55%) and decreased in two patients (10%). The IgE increase was significant in pollen-allergic patients (p<0.05), while it was not significant in mite-allergic patients (p>0.05). Tryptase, ECP, and specific IgE were not detected in the control group. The data obtained showed a positive sNPT response in 12 patients (60%). Comparing our results with those derived from classical-parameter employment, we gathered an improvement of 10%. On the basis of the usual parameters, in fact, we recorded 50% positivity, while the use of mediators provided an additional 10% improvement in sNPT sensitivity: taking together the usual parameters and nasal allergic mediators values, we reached an sNPT over-all sensitivity of 85%.