Aneuploidy identification in pre-B acute lymphoblastic leukemia patients at diagnosis by Multiplex Ligation-dependent Probe Amplification (MLPA).

Three-quarters of the patients with acute lymphoblastic leukemia (ALL), show numerical or structural chromosomal alterations, which are important factors in leukemogenesis. The use of Multiplex Ligation-dependent Probes Amplification (MLPA) has been mainly limited for searching copy number alterations of genes, suggesting that MLPA could detect numerical alterations in cancer. However, the use of MLPA in pediatrics to analyze subtelomeric sequences for aneuploidy detection has not been considered in previous studies. The aim of this study was to identify aneuploidy for the first time using MLPA and correlate the results with karyotype and DNA-index (DI), from preB ALL patients. Forty-two bone marrow samples were analyzed by cytogenetics and flow cytometry to determine the DI. The chromosomal gains and/or losses were detected by the SALSA MLPA P036 Subtelomere Mix 1 probemix®. The chromosomal number matched in 36 out of 42 samples between MLPA and karyotype (R2=0.7829, p=3.7×10-10), 18/42 between MLPA and DI (R2=0.1556, p=0.023), and 20/42 between karyotype and DI (R2=0.1509, p=0.015). MLPA results correlated with karyotype and DI. The use of MLPA led us to identify a gained marker chromosome. Our results indicate that MLPA could be a useful and fast alternative tool for aneuploidy identification in pediatric leukemia.

Neuropatía óptica secundaria a mucocele esfeno-etmoidal: caso clínico / Optic neuropathy secondary to a sphenoid-ethmoidal mucocele: case report

CASO CLÍNICO: Describimos el caso de un paciente varón de 51 años de edad, con antecedente de trauma contuso en la región frontal y desde hace 1 año con disminución de la agudeza visual, proptosis y paresia del músculo recto lateral del ojo derecho. Se le diagnosticó de un mucocele etmoidal derecho y esfenoidal bilateral, drenándose por abordaje externo y endoscópico, con mejoría de los síntomas y sin recurrencias a los 10 meses. DISCUSIÓN: Los mucoceles esfenoidales representan el 1% de todos los mucoceles y pueden presentarse con diferentes manifestaciones clínicas. La escisión quirúrgica es el tratamiento de elección y una intervención temprana está indicada para prevenir complicaciones

CASE REPORT: The case is presented of a 51-year old male patient with a history of blunt trauma in the frontal region and a one-year history of vision loss, proptosis and paresis of the lateral rectus muscle of the right eye. A right ethmoid and bilateral sphenoid mucocele was diagnosed. Drainage was performed using an external and endoscopic approach with improvement in symptoms without recurrence at 10 months. DISCUSSION: Sphenoid mucoceles represent 1% of all mucoceles and may present with different clinical manifestation. Surgical excision is the treatment of choice, and early intervention is indicated to prevent complications

Optic neuropathy secondary to a sphenoid-ethmoidal mucocele: Case report.

CASE REPORT: The case is presented of a 51-year old male patient with a history of blunt trauma in the frontal region and a one-year history of vision loss, proptosis and paresis of the lateral rectus muscle of the right eye. A right ethmoid and bilateral sphenoid mucocele was diagnosed. Drainage was performed using an external and endoscopic approach with improvement in symptoms without recurrence at 10 months. DISCUSSION: Sphenoid mucoceles represent 1% of all mucoceles and may present with different clinical manifestations. Surgical excision is the treatment of choice, and early intervention is indicated to prevent complications.

Pregnancy associated plasma protein-A2: a novel biomarker for Down syndrome.

INTRODUCTION: In an effort to improve prenatal screening for Trisomy 21, we evaluated pregnancy associated plasma protein-A2 (PAPP-A2) as a potential novel second trimester biomarker for Trisomy 21. METHODS: Trisomy 21 and normal control mid-trimester placental samples were subjected to quantitative rt PCR analysis of seven genes we had previously found to be differentially expressed in Trisomy 21 placentae. The localization and differential expression of PAPP-A2 in second trimester placentae from normal and Trisomy 21 pregnancies was determined by immunohistochemistry. PAPP-A2 maternal serum protein levels in ten Trisomy 21 and ten diploid pregnancies were compared by Western blotting. Maternal serum PAPP-A2 levels were measured in 30 Down syndrome cases and 142 normal controls, using ELISA. Regression analysis was used to determine the correlation of PAPP-A2 with other existing markers of Trisomy 21. RESULTS: PAPP-A2 (aka PLAC 3) mRNA and protein expression were both increased in Down syndrome placentae as compared to diploid placentae. PAPP-A2 was also increased in maternal serum from Down syndrome pregnancies as compared to diploid pregnancies. PAPP-A2 expression correlated weakly with established markers. DISCUSSION: This work takes advantage of our previously performed systematic approach to the discovery of novel maternal serum biomarkers for Trisomy 21, using cDNA microarray analysis. Beginning with the validation of the microarray results, we have tracked PAPP-A2 overexpression in Down syndrome from placental mRNA to maternal serum protein. CONCLUSION: PAPP-A2 could serve as an additional maternal serum marker in prenatal screening for Trisomy 21.

Lipid transfer protein syndrome: clinical pattern, cofactor effect and profile of molecular sensitization to plant-foods and pollens.

BACKGROUND: Multiple plant-food sensitizations with a complex pattern of clinical manifestations are a common feature of lipid transfer protein (LTP)-allergic patients. Component-resolved diagnosis permits the diagnosis of the allergen sensitization profile. OBJECTIVE: We sought to clinically characterize and describe the plant-food and pollen molecular sensitization profile in patients with LTP syndrome. METHODS: Forty-five subjects were recruited, after being diagnosed with multiple plant-food allergies sensitized to LTP, but not to any other plant-food allergen, according to the molecular allergen panel tested (Pru p 3 (LTP), Pru p 1 (Bet v 1-like), Pru p 4 (profilin) and those included in a commercial microarray of 103 allergenic components). IgE-mediated food-allergy symptoms and pollinosis were collected. Patients were skin prick tested with a plant-food and pollens panel, and specific IgE to Tri a 14 was evaluated. RESULTS: A heterogeneous group of plant-foods was involved in local and systemic symptoms: oral allergy syndrome (75.6%), urticaria (66.7%), gastrointestinal disorders (55.6%) and anaphylaxis (75.6%), 32.4% of which were cofactor dependent (Non-Steroidal Anti-inflammatory Drugs, exercise). All tested subjects were positive to peach and Pru p 3, Tri a 14 and to some of the LTPs included in the microarray. Pollinosis was diagnosed in 75.6% of subjects, with a broad spectrum of pollen and pollen-allergen sensitization. Plane tree and mugwort were the statistically significant pollens associated with Pru p 3. CONCLUSIONS AND CLINICAL RELEVANCE: Several plant-foods, taxonomically unrelated, independent of peach involvement, are implicated in LTP syndrome. Local symptoms should be evaluated as a risk marker for anaphylaxis because they are frequently associated with cofactor-dependent anaphylaxis. The association of these symptoms with pollinosis, especially plane tree pollinosis, could be part of this syndrome in our area.

Enhanced diagnosis of pollen allergy using specific immunoglobulin E determination to detect major allergens and panallergens.

BACKGROUND: Pollen is one of the main causes of allergic sensitization. It is not easy to make an etiological diagnosis of pollen-allergic patients because of the wide variety of sensitizing pollens, association with food allergy, and increasing incidence of polysensitization, which may result from the presence of allergens that are common to different species, as is the case of panallergens. OBJECTIVE: To compare the results of skin prick tests (SPT) using whole pollen extract with specific immunoglobulin (Ig) E determination for several allergens (purified panallergens included) in the diagnosis of polysensitized pollen-allergic patients. METHODS: The study sample comprised 179 pollen-sensitized patients who underwent SPT with pollen extract and allergen-specific IgE determination against different allergens. RESULTS: The level of concordance between the traditional diagnostic test (SPT) and IgE determination was low, especially in patients sensitized to the panallergens profilin and polcalcin. In the case of SPT, the results demonstrated that patients who are sensitized to either of these panallergens present a significantly higher number of positive results than patients who are not. However, IgE determination revealed that while patients sensitized to polcalcins are sensitized to allergens from a higher number of pollens than the rest of the sample, this is not the case in patients sensitized to profilins. On the other hand, sensitization to profilin or lipid transfer proteins was clearly associated with food allergy. CONCLUSIONS: Sensitization to panallergens could be a confounding factor in the diagnosis of polysensitized pollen-allergic patients as well as a marker for food allergy. However, more studies are required to further investigate the role of these molecules.

Wheat allergens associated with Baker's asthma.

Baker's asthma is a frequent occupational allergic disease caused mainly by inhalation of cereal flour, particularly wheat flour. This review deals with the current diagnosis and immunomodulatory treatments, as well as the role of wheat allergens as molecular tools to enhance management and knowledge of this disease. The review also discusses the current status of several salt-soluble proteins (albumins and globulins)--cereal alpha-amylase/trypsin inhibitors, peroxidase, thioredoxin, nonspecific lipid transfer protein, serine proteinase inhibitor, and thaumatin-like protein-as well as salt-insoluble storage proteins (prolamins, namely, gliadins and glutenins) as allergens associated with baker's asthma. Finally, current limitations to using these proteins as molecular tools for diagnosis and immunotherapy are highlighted.

Developments in the field of allergy in 2009 through the eyes of Clinical and Experimental Allergy.

In 2009 the journal published in the region of 200 papers including reviews, editorials, opinion pieces and original papers that ran the full gamut of allergic disease. It is instructive to take stock of this output to determine patterns of interest and where the cutting edge lies. We have surveyed the field of allergic disease as seen through the pages of Clinical and Experimental Allergy (CEA) highlighting trends, emphasizing notable observations and placing discoveries in the context of other key papers published during the year. The review is divided into similar sections as the journal. In the field of Asthma and Rhinitis CEA has contributed significantly to the debate about asthma phenotypes and expressed opinions about the cause of intrinsic asthma. It has also added its halfpennyworth to the hunt for meaningful biomarkers. In Mechanisms the considerable interest in T cell subsets including Th17 and T regulatory cells continues apace and the discipline of Epidemiology continues to invoke a steady stream of papers on risk factors for asthma with investigators still trying to explain the post-second world war epidemic of allergic disease. Experimental Models continue to make important contributions to our understanding of pathogenesis of allergic disease and in the Clinical Allergy section various angles on immunotherapy are explored. New allergens continue to be described in the allergens section to make those allergen chips even more complicated. A rich and vibrant year helpfully summarized by some of our associate editors.

Characterization of peach thaumatin-like proteins and their identification as major peach allergens.

BACKGROUND: Peach is the most important fruit related to food allergy in the Mediterranean area. Pru p 3, its lipid transfer protein, has been described as the principal allergen responsible for cross-reactivities with other foods and pollen and the severity of clinical symptoms. However, the involvement of other allergenic families cannot be ruled out. Thaumatin-like proteins (TLPs) have been described as food allergen in several fruits, such as apple, cherry, kiwi and banana, and pollen. OBJECTIVE: To identify members of the TLP family in peach fruit and to characterize putative allergens. METHODS: Through two-dimensional (2D) electrophoresis of peach extract and immunodetections with a pool of peach-allergic patients, IgE-binding spots were identified and the corresponding proteins purified and characterized as allergens by in vitro and in vivo assays. Three isoforms, belonging to the TLP family, were purified by different chromatographic systems and characterized by N-terminal amino acid sequences, molecular weight determination (MALDI) and enzymatic activity analysis (beta-1,3-gluconase test and inhibition growth of fungi). In the same way, their IgE-binding capacity and allergenic activity were tested by ELISA assays, basophil activation tests and skin prick tests (SPT). RESULTS: Two peach-TLPs, Pru p 2.0101 and Pru p 2.0201, were identified as IgE-binding spots by 2D electrophoresis. Another peach-TLP, Pru p 2.0301, was cloned and produced as recombinant protein in a yeast system. The three isoforms were purified and characterized as TLPs by immunoblotting with anti-chestnut TLP antibodies and anti-plant N-asparagine complex glycan (anti-cross-reactive carbohydrate determinant). All of them showed beta-1,3-glucanase activity and inhibition of fungal growth. The three TLPs were recognized by around 50% of the sera from 31 patients analysed in ELISA experiments. All three gave a positive response to an SPT and/or in basophil activation experiments. CONCLUSION: Three isoforms, belonging to the TLP family, were identified in peach as principal allergens. Their prevalence, observed in in vitro, ex vivo and in vivo analyses, suggests that they are important allergens and should therefore be included in the routine diagnosis of peach allergy, at least in the Mediterranean area.

Effect of dietary crude protein modification on ammonia and nitrous oxide concentration on a tie-stall dairy barn floor.

Dietary crude protein (CP) reduction is considered a useful strategy to minimize cow N excretion and NH(3) and N(2)O emissions. The aim of the current work was to relate dietary CP modification to whole-animal N balance and subsequent NH(3) and N(2)O concentrations on a tie-stall barn floor. The effect of temperature on NH(3) and N(2)O concentration was also studied. Three Holstein mid to late lactating cows were confined in separate tie-stalls and randomly assigned to 3 diets with varying CP content [low CP (LCP): 14.1%; moderate CP (MCP): 15.9%; high CP (HCP): 16.9%]. Increasing N intake (from 438.6 to 522.8 g of N/d) improved milk yield (from 22.1 to 24.2 kg/d). However, N use efficiency tended to decrease with increasing dietary CP, as shown by milk N use efficiency (from 23.9 to 22.6%), milk urea N (from 15.4 to 18.7 mg/dL), and excreted N per milk yield unit (from 14.7 to 16.4 g of N/kg of milk). Because of higher N excretion, NH(3) concentration on the dairy barn floor increased (LCP: 7.1mg of NH(3)/m(3); MCP: 10.4 mg of NH(3)/m(3); HCP: 10.8 mg of NH(3)/m(3)). In contrast, N(2)O concentration did not respond to dietary manipulation (mean 1.1mg of N(2)O/m(3)). Temperature, which ranged between 12.6 and 18.0 degrees C, did not affect NH(3) and N(2)O concentrations at the stall level. However, when fecal and urinary samples were incubated at 4, 19, and 29 degrees C in the laboratory, ammonia concentration increased for all diets, especially for the MCP and HCP diets, as the temperature increased. In contrast, N2O concentration was negatively related to increasing temperature. In conclusion, data from the current trial demonstrate that lowering dietary CP minimizes NH(3) concentration on dairy stall floors although temperature controls the rate of NH(3) volatilization. On the other hand, N(2)O concentration is not affected by dietary treatments on tie-stall floors.

Supervivencia y resultado funcional a largo plazo de prótesis de rodilla no cementadas / Uncemented knee arthroplasty: supervivorship and long-term functional outcome

Objetivo: Analizar la supervivencia a largo plazo y el resultado funcional de un modelo de prótesis de rodilla no cementada. Material y método: Realizamos un estudio prospectivo observacional de pacientes operados entre los años 1989 y 1996 con el modelo Low Contact Stress Mobile-Bearing Total Knee Replacement(R) (Depuy, Warsaw, IN, EE. UU.). Las variables estudiadas fueron edad, sexo, tiempo de seguimiento, puntuación del Knee Society Score (KSS), supervivencia del implante y motivos de la revisión. Un total de 96 artroplastias de 127 completaron el seguimiento. La media de edad fue de 79 años. La media de seguimiento fue de 14,5 años. Once prótesis se reintervinieron (6 por aflojamiento aséptico de implantes y 5 por problemas de componentes móviles). Se obtuvieron curvas de supervivencia al considerado el tiempo de seguimiento, el tiempo desde la cirugía primaria hasta la reintervención y el episodio «recambio por aflojamiento de componente metálico» y «necesidad de reintervención». Resultados: Las curvas de supervivencia arrojaron una supervivencia del 96,45% a los 9 años y del 92,78% a los 12 años, y se consideró la «necesidad de reintervención» como punto final. Al considerar la necesidad de recambio de componentes metálicos, la supervivencia a los 14 años y medio fue del 93,75%. La puntuación media del KSS funcional fue de 89,2. Conclusión: En nuestra experiencia y con el modelo protésico utilizado, las prótesis totales de rodilla no cementadas proporcionan un resultado clínico y funcional excelente, valorado por el KSS, y una supervivencia del 93,75% a los 14 años y medio al considerar la supervivencia del implante metálico (AU)

Purpose: To analyze the long term survivorship and functional results of a model of uncemented knee prosthesis. Materials and methods: We carried out a prospective observational study of patients implanted between 1989 and 1996 with the Low Contact Stress Mobile-Bearing Total Knee Replacement (Depuy, Warsaw, IN, EEUU). Variables studied included: age, gender, follow-up, Knee Society Score, implant survivorship and reasons for revision. A total of 96 arthroplasties out of 127 reached the end of follow-up Mean age was 79 years. Mean follow-up was 14 years. Eleven prostheses were revised (6 following aseptic loosening and 5 because of problems with the mobile bearings). Survivorship curves were obtained considering length of follow-up, time elapsed between the primary and the revision surgery and the “revision following loosening of a metal component” and “need for revision” events. Results: When "need for revision" was considered as the endpoint, survivorship curves revealed a survivorship rate of 96.45% at 9 years and 92.78% at 12 years. When the endpoint was the "need to revise metal components" 14.5-year survivorship was 93.75%. Mean Knee Society Score was 89.2 points. Conclusion: The use of the uncemented LCS knee has resulted in excellent clinical and functional results as measured by the Knee Society Score, as well as 93.75% survivorship at 14 years and a half, considering the survivorship of the metal implant (AU)

Sublingual immunotherapy in peach allergy: monitoring molecular sensitizations and reactivity to apple fruit and Platanus pollen.

BACKGROUND: Peach allergy is prevalent, persistent, and potentially severe and as such is a target for immunotherapy. Our aims were to evaluate the profile of sensitization to Rosaceae allergens and the effects of sublingual peach immunotherapy on immunoglobulin (Ig) E levels to these allergens, to monitor for neosensitizations, and to check if this treatment modified other Rosaceae fruit and pollen-related sensitizations. METHODS: A double-blind placebo-controlled trial was conducted on 56 peach-allergic patients who received, sublingually, a standardized peach extract quantified in mass units of Pru p 3, or placebo for 6 months. IgE to recombinant (r) Mal d 1, rMal d 4, rPru p 3, and natural (n) Art v 3 and skin prick test (SPT) reactivity to Platanus pollen and apple extracts evaluated before treatment (T0), after 1 month (T1) and after and 6 months (T6) were recorded. RESULTS: In total, 18.5% of patients recognized rMal d 1, 83.3%, rPru p 3, 24.1%, rMal d 4, and 25.9% nArt v 3. IgE to Pru p 3 rose from T0 to T1 in both the active group (P = .003) and the placebo group (P = .022), and remained elevated at T6 in the active group (P = .001). IgE to other purified allergens did not change significantly and no relevant neosensitizations were detected. SPT reactions to peach decreased from T0 to T6 in the active group (P < 0.05). Reactivity to peach (T1 and T6) and apple (T6) was lower in the active group than in the control group. CONCLUSIONS: The main allergen was Pru p 3. Changes in rPru p 3 IgE levels and in peach and apple extract SPT were induced by sublingual immunotherapy.

Component-resolved diagnosis of pollen allergy based on skin testing with profilin, polcalcin and lipid transfer protein pan-allergens.

BACKGROUND: Allergy diagnosis needs to be improved in patients suffering from pollen polysensitization due to the existence of possible confounding factors in this type of patients. OBJECTIVE: To evaluate new diagnostic strategies by comparing skin responses to pan-allergens and conventional allergenic extracts with specific IgE (sIgE) to purified allergen molecules. METHODS: One thousand three hundred and twenty-nine pollen-allergic patients were diagnosed by a combination of an in vitro method with a panel of 13 purified allergens, including major allergens and pan-allergens, using a high-capacity screening technology (ADVIA-Centaur) and skin prick test (SPT) to pan-allergens and conventional extracts. RESULTS: There was a high concordance (kappa index) between in vitro (sIgE to major allergens) and in vivo (SPT to conventional extracts) methods in patients who were not sensitized to pan-allergens, but SPT with conventional extracts failed to diagnose patients with sensitization to pan-allergens. In patients who were simultaneously sensitized to polcalcins and profilins, there was a duplication both in the number of sensitizations to major allergens and in the years of disease evolution. There was a statistical association between sensitization to profilins and/or lipid transfer proteins and food allergy (P<0.0001). CONCLUSION: The novel diagnostic strategy has proven to be a valuable tool in daily clinical practice. Introduction of routine SPT to pan-allergens is a simple and feasible way of improving diagnostic efficacy. Patients sensitized to pan-allergens should be tested by an adequate panel of allergenic molecules in order to identify the allergens that are responsible for the allergic disease.

Recombinant lipid transfer protein Tri a 14: a novel heat and proteolytic resistant tool for the diagnosis of baker's asthma.

BACKGROUND: Baker's asthma is an important occupational allergic disease. Wheat lipid transfer protein (LTP) Tri a 14 is a major allergen associated with wheat allergy. No panel of wheat recombinant allergens for component-resolved diagnosis of baker's asthma is currently available. OBJECTIVE: To evaluate the potential role of recombinant Tri a 14 as a novel tool for the diagnosis of baker's asthma, and to test the heat and proteolytic resistance of the wheat LTP allergen. METHODS: A cDNA encoding Tri a 14 was isolated and sequenced, the recombinant allergen produced in Pichia pastoris and purified by chromatographic methods. Physicochemical and immunological comparison of the natural and recombinant forms of Tri a 14 was carried out by N-terminal amino acid sequencing, matrix-assisted laser desorption/ionization mass spectrometry, circular dichroism (CD) analysis, IgE immunodetection, and specific IgE determination and ELISA-inhibition assays using a pool or individual sera from 26 patients with baker's asthma. Thermal denaturation and simulated gastrointestinal digestion of both Tri a 14 forms were checked by spectroscopic and electrophoretic methods, respectively, and biological activity by basophil activation test (BAT). RESULTS: Natural and recombinant Tri a 14 were similarly folded, as indicated by their nearly identical CD spectra and heat denaturation profiles. A high interclass correlation coefficient (0.882) was found between specific IgE levels to both Tri a 14 proteins in individual sera from baker's asthma patients, but a slightly lower IgE-binding potency of rTri a 14 was detected by ELISA-inhibition assays. Natural and recombinant Tri a 14 elicited positive BAT in two and one out of three patients, respectively. Heat denaturation profiles and simulated gastrointestinal digestion assays indicated that Tri a 14 displayed a high heat and digestive proteolytic resistance, comparable to those of peach Pru p 3, the model food allergen of the LTP family. CONCLUSIONS: Recombinant Tri a 14 is a potential tool for baker's asthma diagnosis, based on its physicochemical and immunological similarity with its natural counterpart. Wheat Tri a 14 shows a high thermal stability and resistance to gastrointestinal digestion.

Component-resolved in vitro diagnosis in peach-allergic patients.

BACKGROUND: The in vitro diagnosis of pollen-related food allergy presents low specificity and reproducibility with many conventional extracts. This can be improved using natural purified allergens, recombinant purified allergens, or both. OBJECTIVE: We compared specific immunoglobulin (Ig) E determination (slgE), the basophil activation test (BAT), the histamine release test (HRT), and the cellular allergen stimulation test (CAST) using natural and recombinant allergens in the diagnosis of peach allergy. METHODS: Thirty-two peach allergic patients were studied. Skin prick tests were performed with commercial peach and extract with Mal d 1, nPru p 3, and profilin (nPho d 2). slgE, BAT, CAST, and HRT were determined using rPru p 3, rMal d 3, rBet v 1, rMal d 1, and rMal d 4. RESULTS: Agreement between the techniques was good with all the allergens, except HRT with rMal d 1 and rMal d 4. With rPru p 3, slgE, CAST, BAT, and HRT showed sensitivity values of 88%, 81%, 72%, and 69% and specificity values of 100%, 93%, 97%, and 83%, respectively. In patients with systemic symptoms or contact urticaria, the values were 100%, 85%, 81%, and 81%. In patients with oral allergy syndrome, sensitivity to profilins or homologues of Bet v 1 was detected in 100% of the cases by all the techniques, except by HRT with rMal d 1, which detected 66% of the cases. CONCLUSIONS: The use of single allergens in the in vitro diagnosis of peach allergy by specific IgE determination, BAT, and CAST offers high specificity and sensitivity, with better results than the HRT.

Understanding patient sensitization profiles in complex pollen areas: a molecular epidemiological study.

BACKGROUND: Allergy diagnosis in patients exposed to multiple pollen species is complex and misdiagnosis is often a cause for unsuccessful specific immunotherapy. OBJECTIVE: We studied the sensitization profile of individual allergens (major, minor and pan-allergens) in pollen-sensitized patients in a region with high exposure to olive pollen by investigating the influence of minor allergens on allergic disease and the association between pan- and minor allergen sensitizations. METHODS: A panel of 13 purified allergens, which included the most relevant allergens in the area, as well as minor olive allergens and pan-allergens, were screened using a high-capacity technology (ADVIA-Centaur) in 891 patients. RESULTS: Olive allergy as measured by specific IgE to Ole e 1 was the leading pollinosis in the area. The minor olive allergens Ole e 7 and Ole e 9 were markers of more severe allergic illness. Profilin sensitization was associated mainly with grass allergy, the second most prevalent pollinosis. Salsola kali pollen allergy was the third most common cause of pollinosis in the area. The prevalence of sensitization to the peach allergen Pru p 3, a nonspecific lipid-transfer protein, was notable. CONCLUSION: Epidemiological analysis by component-resolved diagnosis is a new method, which elucidates the interaction between allergen exposure gradient and patient sensitization. High exposure leads to differential sensitization profiles some of which are associated with more severe allergic conditions. Profilin sensitization, related mainly to grass pollinosis, was a marker of more severe grass pollen sensitization.

Molecular profiles: a new tool to substantiate serum banks for evaluation of potential allergenicity of GMO.

Assessment of the allergenicity of GMOs involves performing a test with a panel of sera obtained from allergic donors. However, there is no clear indication of how to characterize the above-mentioned panel. The patient selection criteria should take into account the geographical location of patients, the intensity and nature of the environmental allergens in the area and the potential cross-reactivity among allergenic molecules. Sera for serum banks, obtained from patients with demonstrated food allergy, should be subjected to a further characterization by screening with a panel of relevant allergenic molecules. A representative panel of these sera should be used in the allergenicity assessment. Finally, the "in vitro" methodologies should have the adequate specificity and sensitivity, and the integrity of the molecules tested should be guaranteed.

Immunoglobulin E recognition patterns to purified Kiwifruit (Actinidinia deliciosa) allergens in patients sensitized to Kiwi with different clinical symptoms.

BACKGROUND: Green kiwifruit allergy is on the rise. However, no surveys testing purified major kiwi allergens have been carried out in a large population, including both kiwi-sensitized [skin prick test (SPT)-positive] and truly kiwi-allergic patients. OBJECTIVE: To isolate major kiwifruit allergens, and to explore their relevance by in vitro and in vivo methods in a large kiwi-sensitized and -allergic population. METHODS: A large group (n=92) of kiwi-sensitized patients with different clinical symptoms were selected, and double-blind, placebo-controlled, food challenges to kiwi were performed in 52 of them. The three major IgE-binding proteins from kiwifruit extracts were isolated and characterized by N-terminal amino acid sequencing and molecular size and glycosylation analysis. The allergenic potency of the three kiwi allergens, and of avocado Pers a 1 as a model allergen associated with the latex-fruit syndrome, was tested by specific IgE quantitation, immunodetection assays and SPTs. RESULTS: The isolated kiwifruit allergens were identified as actinidin Act d 1, glycosylated thaumatin-like Act d 2 and a novel 40 kDa glycoprotein designated as Act d 3.02. Specific IgE to each of the three allergens was found in over 60% of sera from kiwi-sensitized patients, and Act d 1 and Act d 2 induced positive SPT responses in over 50% of the tested patients. A significant link between IgE levels to Act d 1 and Act d 3 and anaphylaxis was uncovered. Avocado Pers a 1 showed an in vitro sensitization prevalence of around 45%, but a low in vivo reactivity. CONCLUSION: Act d 1, Act d 2 and Act d 3 are major allergens in the population studied. Severe symptoms after kiwi ingestion are associated with high IgE levels to Act d 1 and Act d 3.