Fructose biphosphate aldolase: A new cassava allergen.

Background: Food allergy has considerably increased in recent years and this situation has been aggravated mainly by the consumption of more processed and complex foods, since minor or potentially allergenic foods are not required to be labeled. Manihot esculenta (cassava) is a widely consumed food in South America, Africa, and Asia and can be used in the production of flour and starch, as well as several other products. This root can cause allergic reactions with symptoms ranging from mild to severe. Methods: Thus, the aim of this study was the characterization of the immunogenic cassava proteins responsible for sensitizing patients allergic to it. Using a 2D-SDS-PAGE based proteomic approach, six proteins were identified, including Fructose Bisphosphate Aldolase (FBA). Recombinant FBA was produced in Expi293 cells and evaluated by immunoblotting with the serum of 10 individual study subjects. Results: Our results showed six cassava IgE-reactive proteins. From those, recombinant fructose bisphosphate aldolase (FBA) showed a positivity of 80% among tested sera, proving to be a highly sensitizing protein. Conclusion: The recombinant FBA molecule obtained in this study can be important for in vivo diagnostic assays, by producing more accurate results, and for desensitization protocols, in which the use of the isolated molecule produces more precise results by avoiding secondary sensitization. Trial registration: All patients signed a consent form approved by the internal ethics committee CAPPesq, Comissão de Ética para Análise de Projetos de Pesquisa do HC FMUSP (CAAE: 10420619.6.0000.0068).

Eosinophilic Esophagitis: Latent Disease in Patients with Anaphylactic Reaction to Cow's Milk.

BACKGROUND: Food allergy and eosinophilic esophagitis are a substantial and evolving public health issue. Clinicians should know the relationship between these diseases and how one may predispose to the other. This can help minimize misdiagnosis. OBJECTIVE: The objective of this study was to assess esophageal eosinophilia and eosinophilic esophagitis frequency in patients with persistent cow's milk allergy and anaphylaxis manifestations. METHODS: Patients with persistent cow's milk allergy with anaphylaxis manifestations were enrolled from 2012 through 2016 at the São Paulo University Hospital, Brazil. All of them were submitted to endoscopy despite the presence or absence of gastrointestinal symptoms. Demographics data, atopic comorbidities, medication use, endoscopic findings, and esophageal eosinophilia frequency were evaluated. RESULTS: Eighty-nine patients were selected. The median age was 8 years. It was observed that 34 of 89 patients (38.2%; 95% confidence interval [CI]: 28.14%-49.16%) presented with esophageal eosinophilia. Five patients (7.1%) presented proton pump inhibitor-responsive esophageal eosinophilia, and 10 patients (14.2%) presented eosinophilic esophagitis. We found that 29.4% were asymptomatic patients, 23.5% had nonspecific symptoms, 23.5% had persistent typical symptoms, and 23.5% had intermittent typical symptoms. There was an association with inflammatory endoscopy findings in 21 patients (61.7%). CONCLUSIONS: This description demands scientific attention because it is the highest frequency of esophageal eosinophilia yet described in a group of patients with cow's milk allergy presenting with anaphylaxis. Eosinophilic esophagitis is a condition that can coexist "silently" with an IgE-mediated food allergy and is most often underestimated and underdiagnosed.

Relative cost-effectiveness of using an extensively hydrolyzed casein formula in managing infants with cow's milk allergy in Brazil.

OBJECTIVE: To estimate the cost-effectiveness of three alternative dietetic strategies for cow's milk allergy in Brazil: 1) using an extensively hydrolyzed casein formula (eHCF; Nutramigen) as a first-line formula, but switching to an amino acid formula (AAF) if infants remain symptomatic; 2) using an AAF as a first-line formula and then switching to an eHCF after 4 weeks once infants are symptom-free, but switching back to an AAF if infants become symptomatic; and 3) using an AAF as a first-line formula and keeping all infants on that formula. The analysis was conducted from the perspective of the Brazilian public health care system, Sistema Único de Saude. METHODS: Decision modeling was used to estimate the probability of immunoglobulin E (IgE)-mediated and non-IgE-mediated allergic infants developing tolerance to cow's milk by 12 months from starting a formula. The models also estimated the Sistema Único de Saude cost (at 2013/2014 prices) of managing infants over 12 months after starting a formula, as well as the relative cost-effectiveness of each of the dietetic strategies. RESULTS: The probability of developing tolerance to cow's milk by 12 months from starting a formula was higher among infants with either IgE-mediated or non-IgE-mediated allergy who were initially fed with an eHCF, compared with those who were initially fed with an AAF. The total health care cost of initially feeding an eHCF to cow's milk allergic infants was less than that of initially feeding both IgE-mediated and non-IgE-mediated infants with an AAF. CONCLUSION: Within the study's limitations, using an eHCF instead of an AAF for the first-line management of newly-diagnosed infants with cow's milk allergy affords a cost-effective use of publicly funded resources, since it improves the outcome for less cost.

Molecular Diagnosis of Shrimp Allergy: Efficiency of Several Allergens to Predict Clinical Reactivity.

BACKGROUND: The diagnosis of shellfish allergy remains a challenge for clinicians. Several shellfish allergens have been characterized and their IgE epitopes identified. However, the clinical relevance of this sensitization is still not clear. OBJECTIVE: The objective of this study was to identify allergens and epitopes associated with clinical reactivity to shrimp. METHODS: Shrimp-sensitized subjects were recruited and grouped based on the history of shrimp-allergic reactions and challenge outcome. IgE reactivity to recombinant crustacean allergens, and IgE and IgG4 reactivity to peptides were determined. Subjects sensitized to dust mites and/or cockroach without shrimp sensitization or reported allergic reactions, as well as nonatopic individuals, were used as controls. RESULTS: A total of 86 subjects were recruited with a skin prick test to shrimp; 74 reported shrimp-allergic reactions, 58 were allergic (38 positive double-blind placebo-controlled food challenge and 20 recent anaphylaxis), and 16 were tolerant. All subjects without a history of reactions had negative challenges. The individuals with a positive challenge more frequently recognized tropomyosin and sarcoplasmic calcium-binding proteins than those found tolerant by the challenge. Especially a sarcoplasmic-calcium-binding-protein positive test is very likely to result in a positive challenge, though the frequency of recognition is low. Subjects with dust mite and/or cockroach allergy not sensitized to shrimp recognized arginine kinase and hemocyanin. Several epitopes of these allergens may be important in predicting clinical reactivity. CONCLUSION: Tropomyosin and sarcoplasmic-calcium-binding-protein sensitization is associated with clinical reactivity to shrimp. Myosin light chain testing may help in the diagnosis of clinical reactivity. Arginine kinase and hemocyanin appear to be cross-reacting allergens between shrimp and arthropods. Detection of IgE to these allergens and some of their epitopes may be better diagnostic tools in the routine workup of shrimp allergy.

Avaliação da densidade mineral óssea em crianças com dermatite atópica moderada ou grave / Bone mineral density in children with moderate or severe atopic dermatitis

Objetivo: Determinar o impacto da Dermatite Atópica (DA) no estado nutricional e metabolismo ósseo em crianças.Métodos: Quarenta e nove crianças com DA moderada ou grave (4-12 anos) e 48 crianças saudáveis foram avaliadas por z-escore altura/idade, z-escore peso/idade, z-escore IMC, duração e gravidade da doença, uso de Glicocorticoides (GC) tópico e parâmetros ósseos. Conteúdo mineral ósseo (CMO), densidade mineral óssea (DMO) e z-escores foram medidos por absormetria de dupla emissão de raios-X (DXA). Níveis séricos de cálcio, fósforo, fosfatase alcalina, cortisol e telopeptídeo carboxiterminal do colágeno tipo 1 (CTX), e níveis plasmáticos de 25 hidroxivitamina D (25OHD) e hormônio da paratireoide (PTH), foram determinados.Resultados: Crianças com DA apresentaram menor altura para idade quando comparadas às crianças controle (p = 0,007). Menor CMO em coluna lombar [16,5 (6,4) vs. 19,8 (8,3)g, p = 0,027] e fêmur total [12,2 (4,0) vs. 14,2 (5,0)g, p = 0,029] foi encontrado em crianças com DA. Níveis de CTX foram menores em pacientes com DA [1,36 (0,59) vs. 1,67 (0,79)ng/mL, p = 0,026] e tendência a níveis mais baixos de fosfatase alcalina foi observada em crianças com DA [228 (75,3) vs. 255 (70,7) ng/mL, p = 0,074]. Crianças com DA apresentaram níveis mais baixos de cortisol que crianças saudáveis [9,06 (4.8) 10,57 vs. (4,9), p = 0,061], sem diferença significante.Conclusões: Redução em altura para idade, remodelamento ósseo e conteúdo mineral ósseo em crianças com DA moderada ou grave poderia estar associada a fatores incluindo determinantes genéticos, baixa exposição solar, inflamação crônica e uso crônico do GC tópico.

Objective: To determine the impact of atopic dermatitis (AD) on nutritional status and bonemetabolism parameters in children. Methods: Forty-nine children with moderate to severe AD (4-12 years old) and 48 healthy children were assessed using height/age z-scores, weight/agez-scores, body mass index z-scores, disease activity and severity, topical use of glucocorticoids (GC), and bone parameters. Bone mineral content (BMC), bone mineral density (BMD), andz-scores were measured using dual-energy X-ray absorptiometry (DXA). Serum levels ofcalcium, phosphorus, alkaline phosphatase, cortisol, and carboxy terminal telopeptide of type1 collagen (CTX), and plasma levels of 25 hidroxy vitamin D (25OHD) and parathyroid hormone,were determined. Results: AD children presented lower height/age z-score, as compared tocontrols (p = 0.007). Lower BMC at lumbar spine [16.5(6.4) vs. 19.8(8.3)g, p = 0.027] and totalfemur [12.2(4.0) vs. 14.2 (5.0) g, p = 0.029] was found in AD children. CTX levels were lower in ADpatients, as compared to healthy children [1.36(0.59) vs. 1.67(0.79)ng/mL, p = 0.026] and a trendto lower levels of alkaline phosphatase was observed in AD children [228(75.3) vs. 255 (70.7) ng/mL, p = 0.074]. Children with AD presented lower levels of serum cortisol in comparison to thehealthy group [9.06(4.8) vs. 10.57(4.9), p = 0.061], without statistical significance. Conclusions: Reduced height for age, BMD, and bone turnover in children with moderate to severe AD couldbe associated with genetic determinants, insufficient sun exposure, chronic inflammation, andchronic use of topical GC.

Pediatric allergy and immunology in Brazil.

The subspecialty of pediatric allergy and immunology in Brazil is in its early years and progressing steadily. This review highlights the research developed in the past years aiming to show the characteristics of allergic and immunologic diseases in this vast country. Epidemiologic studies demonstrated the high prevalence of asthma in infants, children, and adolescents. Mortality rates and average annual variation of asthma hospitalization have reduced in all pediatric age groups. Indoor aeroallergen exposure is excessively high and contributes to the high rates of allergy sensitization. Prevalence of food allergy has increased to epidemic levels. Foods (35%), insect stings (30%), and drugs (23%) are the main etiological agents of anaphylaxis in children and adolescents. Molecular diagnosis of primary immunodeficiencies (PID) showed a high incidence of fungal infections including paracoccidioidomycosis in X-linked hyper-IgM syndrome, and the occurrence of BCG adverse reactions or other mycobacterial infections in patients with chronic granulomatous disease. Education in pediatric allergy and immunology is deficient for medical students, but residency programs are effective in training internists and pediatricians for the practice of allergy. The field of PID requires further training. Last, this review is a tribute to Prof. Dr. Charles Naspitz, one of the pioneers of our specialty in Brazil.

Measurement of IgE antibodies to shrimp tropomyosin is superior to skin prick testing with commercial extract and measurement of IgE to shrimp for predicting clinically relevant allergic reactions after shrimp ingestion.

BACKGROUND: Shrimp is a frequent cause of food allergy. Tropomyosin is the major allergen in shrimp, and it shares homology to tropomyosins from other crustaceans, dust mites, cockroach, and parasites. OBJECTIVE: The aim of this study was to determine the value of detection of IgE to shrimp tropomyosin in the diagnosis of shrimp allergy. METHODS: We have studied 35 patients with asthma, rhinitis, or both who were sensitized to Dermatophagoides pteronyssinus. All subjects underwent skin prick testing in addition to double-blind, placebo-controlled food challenges (DBPCFC); oral open challenges; or both with shrimp. Measurements of IgE to shrimp and shrimp tropomyosin were carried out by means of CAP and chimeric ELISA, respectively. RESULTS: Oral challenges confirmed the diagnosis of shrimp allergy in 7 patients. IgE measurement to shrimp tropomyosin was positive in 71.4% of the patients with shrimp allergy. Of the 28 patients without shrimp allergy, only 7.1% (2/28) had IgE to shrimp tropomyosin compared with 25% (7/28) who had IgE to shrimp and 35.7% (10/28) who had positive skin prick test responses to shrimp. Sensitivity was similar for all 3 methods (71.4%); in contrast, specificity of IgE to shrimp tropomyosin (92.8%) was greater than that of IgE to shrimp (75%) and skin prick testing (64.2%). With regard to diagnostic efficiency, measurement of IgE to shrimp tropomyosin was superior to measurement of IgE to shrimp and skin prick testing (88.5%, 74.2%, and 65.7%, respectively). CONCLUSION: Use of measurements of IgE to shrimp tropomyosin provided added value to the diagnosis of shrimp allergy.

Reatividade cruzada entre ácaro e camarão: qual o efeito da imunoterapia? / Cross-reactivity between dust mite and shrimp: what is the effect of immunotherapy?

Objetivo: O objetivo desse estudo foi avaliar alterações na resposta clínica e imunológica ao camarão após a imunoterapia com Dermatophagoides pteronyssinus. Métodos: Selecionou-se 35 indivíduos alérgicos a Dermatophagoides pteronyssinus (Der p), os quais foram submetidos a testes cutâneos de leitura imediata. A detecção de IgE específica in vitro foi feita para o ácaro, camarão, e para tropomiosina de camarão. Em todos, avaliou-se reatividade clínica ao camarão através de provocação oral. Dez pacientes foram alocados para o grupo controle, e 25 foram submetidos à imunoterapia alérgeno específica para o ácaro. Os testes cutâneos e a dosagem de IgE sérica específica foram repetidas após a indução da imunoterapia, e após 1 ano do início. A reatividade clínica ao camarão foi reavaliada no final do estudo pela provocação oral. Resultados: No grupo dos pacientes que foram submetidos à imunoterapia, observamos diminuição na reatividade nos testes cutâneos e dosagem de IgE específica para Der p e camarão. Dos 10 pacientes com testes cutâneos positivos para camarão, 4 foram negativos na dosagem após um ano de imunoterapia (p= 0,04). Quanto à dosagem sérica de IgE para camarão, dos 9 positivos no início, 6 ficaram negativos (p= 0,014). Nenhum paciente submetido a imunoterapia desenvolveu nova sensibilização para camarão. Não houve alteração na reatividade clínica ao camarão após imunoterapia. Conclusão: A imunoterapia para Dermatophagoides pteronyssinus foi acompanhada de diminuição da reatividade imunológica para camarão e clinicamente não houve alteração da sensibilidade a camarão.

Objective: The objective of this study was to determine changes in clinical and immunological response to shrimp after immunotherapy with Dermatophagoides pteronyssinus. Methods: We studied 35 allergic subjects to Dermatophagoides pteronyssinus (Der p) submitted to skin tests. The detection of serum specific IgE was performed to mite, shrimp, and tropomyosin from shrimp. In all patients, the clinical reactivity to shrimp was assessed through oral challenge. Ten patients were allocated to the control group, and 25 were submitted to immunotherapy for mite. Skin tests and determination of serum specific IgE were repeated after the induction of limmnunotherapy (3-4 months) and 1 year after of beginning of the treatment. The clinical reactivity to shrimp was assessed again at the end of the study by oral challenge. Results: In the group of patients who were undergoing immunotherapy, we observed decreased reactivity in the skin tests and specific IgE levels to Der p and shrimp. Among the 10 patients with positive skin tests to shrimp, 4 were negative when assessed after one year of immunotherapy (p = 0.04). About serum specific IgE to shrimp, from the 9 positive reactors in the beginning of treatment, 6 became negative (p= 0.014). There was no change in clinical reactivity to shrimp after immunotherapy. Conclusion: The immunotherapy for Dermatophagoides shrimp. pteronyinus was accompanied by decreased immune reactivity to shrimp and c1inically there was no change in sensitivity to shrimp.