Association Between the Seed Storage Proteins 2S Albumin and 11S Globulin and Severe Allergic Reaction After Flaxseed Intake.

BACKGROUND: Given the increased popularity of flaxseed in meals, several cases of allergy to these seeds have been reported. Little is known about the allergens implicated in hypersensitivity reactions to flaxseed. The present study aimed to identify the allergens involved in IgE-mediated reactions in 5 patients with a clinical history of severe systemic symptoms after flaxseed consumption. METHODS: Proteins that were potential allergens with IgE-binding capacity were purified from flaxseed extract using chromatography and identified via MALDI-TOF mass spectrometry. Immunoassays were performed using the 5 allergic patients' sera tested individually and as a pool. RESULTS: Immunoblotting of the flaxseed extract revealed a low-molecular-mass protein (around 13 kDa) in 4 of the 5 patients, while a protein of approximately 55 kDa was detected in 2 patients. The proteins were identified by mass spectrometry as flaxseed 2S albumin, which is included in the WHO/IUIS allergen nomenclature as Lin u 1, and 11S globulin. Inhibition assays revealed in vitro IgE-mediated cross-reactivity between Lin u 1 and peanut and cashew nut proteins, while IgE-mediated recognition of 11S globulin by patients' sera was partially inhibited by several plant-derived sources. CONCLUSIONS: Seed storage proteins from flaxseed were involved in the development of severe symptoms in the 5 patients studied and exhibited cross-reactivity with other allergenic sources. Besides the severity of flaxseed allergy in patients sensitized to 2S albumin, this is the first time that 11S globulin has been identified as a potential allergen. Taking these data into account should ensure a more accurate diagnosis.

Allergen Profile of London Plane Tree Pollen: Clinical and Molecular Pattern in Central Spain.

BACKGROUND AND OBJECTIVES: Platanus acerifolia (London plane tree) is a deciduous tree of the Platanaceae family. Sensitization to this plant varies with geography. Madrid, located in central Spain, has one of the highest London plane tree pollen concentration levels on the Iberian Peninsula. We evaluated both the clinical characteristics and the molecular sensitization pattern of patients with allergy to London plane tree pollen in the region of Madrid. METHODS: Thirty-eight patients allergic to London plane tree pollen were selected according to their clinical symptoms and positive results in skin prick testing and/or specific IgE determination. Serum was collected, and allergen components were evaluated using immunodetection techniques as well as ImmunoCAP. The IgE-binding proteins detected were identified and characterized using mass spectrometry. RESULTS: Analysis of serum samples from allergic patients revealed 9 IgE-binding bands in London plane tree pollen extract. Among these, the 45-kDa protein, which corresponded to Pla a 2, was detected in 76.3% of patients. However, the 18-kDa (Pla a 1) and 9-kDa (Pla a 3) bands were detected in 44.7% and 23.7% of sera, respectively. These results were confirmed using purified proteins. Characterization of the allergen revealed the 27-kDa protein to be glutathione-S-transferase. CONCLUSIONS: The molecular profile of patients sensitized to London plane tree pollen differs from that reported in studies from other locations. In the population we studied, the prevalence of Pla a 2 was higher than that of Pla a 1 and Pla a 3. In addition, the minor allergen previously referred to as Pla a 4 was characterized as glutathione-S-transferase.

Resultados sobre la aplicación cráneo-facial del parche de capsaicina 8 % en una serie de 5 casos / Results of the capsaicin 8% patch in craniofacial application in five clinical cases

Los parches de capsaicina al 8 % son una alternativa de segunda línea para el tratamiento del dolor neuropático periférico. Aunque tiene pocos efectos secundarios, no tiene indicación para el tratamiento cráneo-facial debido a la posible irritación de mucosas por la capsaicina. Sólo hemos encontrado tres publicaciones que refieren la aplicación del parche en estas localizaciones, describiendo 7 casos clínicos. Hemos recogido 4 casos en los que se realizan 5 aplicaciones en total, 3 mujeres (repitiendo aplicación en una de ellas) y 1 hombre, entre 58 y 84 años, con los siguientes diagnósticos: necrosis cáustica en labio inferior tras limpieza dental, neuralgia del trigémino y neuropatía postherpética. Tras comprobar ineficacia de otros tratamientos, se propuso el parche de capsaicina al 8 %, con firma previa de los consentimientos informados de la aplicación de parche en régimen de hospital de día y de tratamiento fuera de ficha técnica. Previamente a la aplicación del parche en la zona cutánea dolorosa, se procedió a realizar protección ocular de ambos ojos con parche oftálmico quirúrgico, y de mucosas oral y nasal con mascarilla facial quirúrgica sellada. La protección se mantuvo durante toda la aplicación del parche y se quitó una vez retirado éste y limpiada la zona de aplicación. Únicamente se reportaron 3 efectos secundarios leves del total de las 5 aplicaciones: un paciente presentó piel eritematosa que cedió espontáneamente, otra paciente refirió sensación de quemazón y dolor que cedió con analgesia endovenosa, y otra paciente explicó dolor leve bien tolerado, que cedió de manera espontánea. En ninguno de los casos se apreciaron efectos secundarios a nivel de mucosas. En cuanto a resultados, dos pacientes notaron mejoría durante uno y dos meses, colocando nuevamente el parche en una de ellas, sin lograr esta segunda vez alivio. Las otras dos pacientes no notaron ningún cambio. El tratamiento con parches de capsaicina 8 % en superficies cráneo-faciales parece tener similar eficacia a su aplicación en otras áreas de la piel. Los efectos secundarios en su aplicación en estas superficies son escasos, al igual que en otras aplicaciones corporales. Creemos que con las medidas de precaución adecuadas en las regiones cráneo-faciales, la utilidad clínica observada del parche de capsaicina 8 % lo sitúa como otra opción de tratamiento para dolor neuropático, sin complicaciones añadidas. No obstante, estudios clínicos con mayor número de pacientes deberían llevarse a cabo para confirmar estos hallazgos (AU)

The capsaicin 8 % patch is a secondary line alternative to neuropathic peripheral pain treatment. Although it has few secondary effects, is not indicated in head and facial treatment due to the possibility of the irritation of mucosa. We have only found three publications related with the patch application in those locations, describing 7 clinical cases. We have analyzed 4 cases in which we have applied 5 patches in total. There were 3 women (repeating the application in one of them) and 1 man, between 58 and 84 years old, with the following diagnosis: caustic necrosis in the inferior lip after dental cleaning, trigeminal neuralgia and post-herpetic neuropathy. Inefficacy of other treatments was confirmed, and after that, the capsaicin 8 % patch was proposed. Informed consent of the application of the patch at day clinic and treatment out of technical data sheet were previously signed. Before the patch was applied to the painful cutaneous area, we proceed with ocular protection of both eyes with surgical ophthalmic patch and oral and nasal mucosa protection with surgical mask hermetically seal. That protection was maintained during the whole application of the patch, and was removed once the capsaicin patch was taken off and the application area was cleaned. There were only 3 mild secondary effects of the total 5 applications: one patient showed erythematic skin that was resolved spontaneously, another patient related burn and pain sensation which was solved with endovenous analgesia. Finally, another patient explained mild pain well tolerated, that was resolved also spontaneously. In no cases there were secondary effects in mucosa. Related with the results, 2 patients felt improvement between one and two months, applying again the patch in one of them, not reaching this time relief in the pain. The other 2 patients did not notice any change. The capsaicin 8 % patch treatment in head and facial areas seems to have similar efficacy as the application in other skin areas. Secondary effects in these surfaces are very low, the same as in other corporal locations. We believe that with the adequate preventive measures in head and facial areas, clinical utility observed with capsaicin 8 % patch places it as another treatment option for neuropathic pain, with no complications added. However, clinical studies with a higher number of patients should carry on to confirm these findings (AU)

Identification of Cucumisin (Cuc m 1), a subtilisin-like endopeptidase, as the major allergen of melon fruit.

BACKGROUND: Allergenic components in melon extracts have not been described in spite of the fact that melon (Cucumis melo) is a frequent allergy-eliciting fruit. The aim of this study was to evaluate allergenic components in melon extract and to report the identification of cucumisin as a major melon allergen. MATERIALS AND METHODS: Sera from 35 patients allergic to melon were selected on the basis of clinical symptoms, skin prick tests and oral challenge test. Allergenic components were detected by sodium dodecyl sulphate polyacrylamide gel electrophoresis and immunoblotting. Molecular characterization of IgE-binding bands was performed by N-terminal amino acid sequencing. RESULTS: More than 10 IgE-binding bands, between 10 and 80 kDa, were identified in melon extract. Out of them, four IgE-binding bands were major allergens: 14 kDa, 36 kDa, 54 kDa and 67 kDa. These major allergens, except 14 kDa band, showed the same N-terminal sequence: T-T-R-S-W-D-F-L. Research conducted with protein databases identified this N-terminal sequence as cucumisin, an alkaline serine protease, which shares structural homology with microbial subtilisin. The molecular mass of the identified bands corresponds with different molecular forms of cucumisin produced during the processing or degradation of the enzyme: 67 kDa native cucumisin, 54 kDa mature cucumisin and 36 kDa NH2-terminal cucumisin fragment. CONCLUSION: Cucumisin (Cuc m 1) and several N-terminal cucumisin fragments are the major allergens of melon. The ubiquitous distribution of this protein family (cucumisin-like proteases) in many plant species and its high structural similarity suggest its potential role as a new panallergen in plant foods.

Allergy to plant-derived fresh foods in a birch- and ragweed-free area.

BACKGROUND: Allergy to plant-derived fresh foods has often been reported in geographical areas where birch or ragweed pollens are frequent and has been attributed to cross-reactivity to pollens. OBJECTIVE: The aim of this study has been to evaluate allergy to plant-derived fresh foods among pollen-allergic patients from a birch and ragweed-free area. METHODS: Ninety-five pollen-allergic patients took part in the study. The study consisted of a questionnaire, skin prick tests and challenge tests. Pollen skin tests to five grasses, eight trees and seven weeds were performed in duplicate. Prick tests (prick by prick) and challenge tests were carried out with the fresh foods. RESULTS: Most patients allergic to pollens were sensitized to grass (Lolium and Phleum; 97.9%), followed by tree (Olea; 82.1%) and weed pollens (Plantago; 64.2%). 35 of the 95 pollen-allergic patients had positive skin test responses to some plant-derived fresh foods, the highest percentage corresponding to several fruits in the Rosaceae family (peach and pear, 26.3%), followed by Cucurbitacea fruits (melon, 13.7%). The 21. 05% of the pollen-allergic patients were allergic to some type of plant-derived fresh food. Peach was the plant-derived fresh food which most frequently elicited allergy symptoms (12.6%), followed by melon (7.36%). The cluster of positive responses to Rosaceae fruits was higher for skin testing than for challenge testing. CONCLUSION: Peach was the most important allergy provoking fruit in a birch and ragweed free-area where apples were consumed at a rate of two times more than peaches and the patients allergic to pollen were principally sensitized to grass pollens.

Pollen allergy in peach-allergic patients: sensitization and cross-reactivity to taxonomically unrelated pollens.

BACKGROUND: Fruit allergy has been attributed to cross-reactive IgE to pollens and has been associated with a particular pollen sensitization. OBJECTIVE: The aim of the study was to evaluate sensitization to several taxonomically unrelated pollens in peach- and pollen-allergic patients and to study cross-reactivity between them. METHODS: One hundred sixty-five patients were evaluated: 70 peach- allergic patients together with 95 pollen-allergic patients (control group). Pollen skin tests in duplicate were performed to 5 grasses, 8 trees, and 7 weeds. Cross-reactivity between peach and taxonomically diverse pollens was determined by radioallergosorbent inhibition and Western blot inhibition tests. Experiments were also carried out after preadsorption of the sera with purified natural profilin. RESULTS: The skin test results revealed that peach-allergic patients frequently reacted to most pollens-grasses, weeds, and trees-even when some of these are not found in our geographic area. There was a statistically significant increase in sensitization frequency to most trees and weeds, with a statistically higher occurrence of asthma (odds ratio 2.98, 95% confidence interval 1.46-6.09). Inhibition test results provided evidence that taxonomically unrelated grasses, weeds, and trees produced various and substantial degrees of inhibition in specific IgE to peach and that the peach extract elicited strong inhibitions to those pollens. Profilin was found to be a relevant cross-reactive antigen in these patients. CONCLUSION: The results of this study provide evidence that peach allergy is linked to sensitization to several taxonomically unrelated pollens. This is attributable to the ubiquitous nature of the IgE binding determinants-such as profilins-between peach and taxonomically unrelated pollens.

A method for quantitation of food biologic activity: results with peach allergen extracts.

OBJECTIVE: The aim of this study was to develop a quantitative skin test assay for measurement of the biologic activity of food allergen extracts, as well as to report the results obtained with a peach extract labeled in food biological units (FBUs). METHODS: We prepared a biologically quantified peach extract. The biologic activity was measured in FBUs by assigning 100 FBU/ml to the biologic activity of the extract that elicited a wheal area with a geometric mean equal to that produced by the prick-prick method by using the food itself in a population of 30 patients allergic to food. We evaluated 265 patients, including 70 patients allergic to peach and 195 control subjects (100 nonatopic subjects and 95 subjects allergic to pollen). The biologically quantified peach extract was used during the study and was compared with four commercial peach extracts, which were labeled in weight per volume. RESULTS: The sensitivity of the nonstandardized commercial peach extracts varied from 4.3% to 74%, with biologic activity being very low in all of them. The sensitivity of the biologically quantified peach extract was 100%, with a 100% concordance between the prick-prick and the skin prick test results. CONCLUSION: These data demonstrate that if peach allergen preparations were biologically standardized in FBUs, the quality of these food extracts used for diagnosis would be improved.

Peach allergy pattern: experience in 70 patients.

We report the clinical characteristics of peach allergy encountered in a population of peach-allergic patients. We evaluated 165 patients. The 70 peach-allergic patients were diagnosed through clinical history, the skin prick-prick test, and open oral challenge and rub tests to peach. As a pollinic control group, 95 pollen-allergic patients were also evaluated. Some 49% of the patients were male and 51% female. The mean age was 20 +/- 8 years. Oral allergy syndrome (86%) was the most common symptom, followed by contact urticaria (61%) and systemic symptoms (26%). Some 67% of the patients were allergic to peach pulp and 36% reported symptoms related to canned peach. Canned peach and pulp symptoms were statistically associated (P < 0.01), and symptoms to canned peach were more frequently reported by patients with systemic symptoms (P < 0.05). On evaluation of the peach-allergic patients' characteristics, three risk factors--allergy to peach pulp, allergy to canned peach, and peach allergy in non-pollen-allergic patients--were found, indicating development of systemic symptoms on eating peach. Most of the peach-allergic patients (81%) also had pollen allergy, which was linked to a higher prevalence of asthma (73%) than in the pollen-allergic patients of the control group (48%); this difference was statistically significant (P < 0.01). Finally, two groups were clearly defined by the seriousness of the peach allergy--the non-pollen-allergic patients were more predisposed to the occurrence of systemic symptoms (> 50%), and the pollen-allergic patients to asthma (> 70%).

Properties and localization of a tyrosine phosphorylated form of hexokinase in mouse sperm.

Mouse sperm possess a phosphotyrosine-containing hexokinase type 1 (HK1) that is associated with the plasma membrane fraction of these cells (Kalab et al., 1994; J. Biol Chem 269:3810-3817). This apparent plasma membrane association appears unique, since somatic HK1 is normally cytoplasmic or bound to the outer mitochondrial membrane via contact sites with a voltage-dependent anion channel (porin) through a porin-binding domain. In male germ cells, three cDNA clones have been described that encode unique HK1 isoforms (HK1-sa, HK1-sb, HK1-sc) that do not contain porin binding domains (Mori et al., 1993: Biol Reprod 49:191-203). This suggests that these proteins might not be localized to the outer mitochondrial membrane and could have alternative functions in germ cells and/or sperm. We demonstrate in the mouse that male germ cells and sperm could potentially express four HK1 isoforms (HK1-sa, HK1-sb, HK1-sc, and the somatic HK1). At the protein level, at least one of the HK1 isoforms becomes phosphorylated on tyrosine residues during spermatogenesis. Treatment of sperm membrane fractions to dissociate the phosphotyrosine-containing HK1 (pY-mHK1) yields results demonstrating that pY-mHK1 has properties of an integral membrane protein. Indirect immunofluorescence using a monoclonal antibody to HK1 demonstrates specific staining both in the head and tail regions of sperm. Surface biotinylation of intact sperm followed by precipitation with either polyclonal HK1 antiserum or with avidin-Sepharose suggests that pY-mHK1 possesses an extracellular domain. These results suggest that mouse sperm contain at least one HK1 isoform that is present on the sperm head, has an extracellular domain, and behaves as an integral membrane protein.

S-adenosyl-L-methionine decarboxylase activity in the rat epididymis: ontogeny and androgenic control.

The authors describe the occurrence of high levels of S-adenosyl-L-methionine decarboxylase (SAMDC) activity in the rat epididymis, and its ontogeny and androgenic control. As early as 15 days of age, SAMDC activity exists, although a peak of activity is observed at 25 days. Bilateral orchidectomy resulted in a decline of epididymal SAMDC activity. However, an androgen-independent fraction, accounting for 34% of total activity, appears to exist in the epididymis. In 45-day-old orchidectomized rats, SAMDC activity was stimulated by testosterone treatment in a dose-dependent manner. However, treatment of 45-day-old intact animals with a high dose of the androgen failed to modify SAMDC activity, indicating that, at this age, the enzyme is maximally stimulated by endogenous androgens. The observed effect of testosterone on castrated rats was completely abolished by concomitant treatment with the antiandrogen flutamide. This compound was ineffective on the androgen-insensitive fraction. To assess the contribution of circulating and luminal androgens to the maintenance of epididymal SAMDC, rats were unilaterally orchidectomized and activity was determined in both epididymides after 7 days. The SAMDC activity was identical in epididymides from both sides, suggesting circulating androgens suffice to maintain normal levels of activity. It was concluded that androgens regulate epididymal SAMDC activity, although an androgen-independent fraction appears to exist.