The allergenic structure of the thaumatin-like protein Ole e 13 is degraded by processing of raw olive fruits.

BACKGROUND: The thaumatin-like protein (TLP) Ole e 13 in raw olive fruit is responsible for occupational allergy in olive oil mill workers. However, these workers do not experience allergic symptoms after ingestion of edible olive. OBJECTIVES: To analyze the presence of IgE-reactive TLP in raw and edible olive fruit and to assess the allergenic potency of both sources. METHODS: The content of TLP in raw and edible olive fruit protein extracts was analyzed using immunoblotting with sera from allergic patients and with olive TLP-specific IgG. The structural and immunological stability of TLP were assayed using immunoblotting after treatment of both raw olive and purified TLP with 0.25 M NaOH solution for 24 hours. Olive pollen extract was investigated by immunoblotting for TLP content. RESULTS: The TLP contained in raw olive fruit was not present in edible olives as a result of maceration before human consumption. No TLP was detected in olive pollen using specific IgG or sera from patients allergic to olive fruit. Sera from patients allergic to olive pollen did not react with purified TLP. CONCLUSIONS: IgE-reactive TLP is not present in edible olive, thus explaining the low number of patients allergic to this highly consumed fruit. Patients allergic to olive pollen are not sensitized toTLP and, therefore, not expected to be at risk of food allergy to olive fruit or TLP plant sources.

The allergenic structure of the thaumatin-like protein ole e 13 is degraded by processing of raw olive fruits

Background: The thaumatin-like protein (TLP) Ole e 13 in raw olive fruit is responsible for occupational allergy in olive oil mill workers. However, these workers do not experience allergic symptoms after ingestion of edible olive. Objectives: To analyze the presence of IgE-reactive TLP in raw and edible olive fruit and to assess the allergenic potency of both sources. Methods: The content of TLP in raw and edible olive fruit protein extracts was analyzed using immunoblotting with sera from allergic patients and with olive TLP-specific IgG. The structural and immunological stability of TLP were assayed using immunoblotting after treatment of both raw olive and purified TLP with 0.25 M NaOH solution for 24 hours. Olive pollen extract was investigated by immunoblotting for TLP content. Results: The TLP contained in raw olive fruit was not present in edible olives as a result of maceration before human consumption. No TLP was detected in olive pollen using specific IgG or sera from patients allergic to olive fruit. Sera from patients allergic to olive pollen did not react with purified TLP. Conclusions: IgE-reactive TLP is not present in edible olive, thus explaining the low number of patients allergic to this highly consumed fruit. Patients allergic to olive pollen are not sensitized to TLP and, therefore, not expected to be at risk of food allergy to olive fruit or TLP plant sources (AU)

Introducción: La aceituna natural contiene una proteína de la familia de las taumatinas (TLP) que es responsable de la alergia ocupacional en trabajadores de molinos de aceite. Sin embargo, éstos no presentan síntomas cuando ingieren aceitunas comestibles. Objetivos: Analizar la presencia de TLP en aceituna natural y comestible, y correlacionar sus niveles con la potencia alergénica de ambos productos. Métodos: El contenido de TLP en los extractos proteicos de las aceitunas fue analizado por inmunotransferencia y tinción con sueros de pacientes alérgicos así como con antisuero específico para TLP de olivo. La estabilidad estructural e inmunológica de la TLP se ensayó mediante inmunotinción después del tratamiento del extracto de aceituna natural y de la TLP purificada con NaOH 0.25 M durante 24 h. También se analizó la presencia de TLP en el polen de olivo por inmunotinción. Resultados: La TLP presente en la aceituna natural no se detecta en la comestible como consecuencia del tratamiento de maceración al que es sometida para obtener el producto apto para el consumo humano. No se observó TLP reactiva en el polen de olivo, ni con anticuerpos específicos ni con sueros de pacientes alérgicos a aceituna. Sueros de pacientes alérgicos al polen de olivo no reaccionan con la TLP purificada de aceituna. Conclusiones: La TLP de olivo no está presente en las aceitunas comestibles lo que explica el escaso número de pacientes alérgicos a la aceituna. Además, los pacientes alérgicos al polen de olivo no están sensibilizados a TLP, por lo que no tendrían riesgo de sufrir alergia alimentaria a aceitunas o a fuentes vegetales de TLPs (AU)

Model predicting survival/exitus after traumatic brain injury: biomarker S100B 24h.

BACKGROUND: The enigma of Traumatic Brain Injury (TBI), reflected in recent scientific literature, is its uncertain consequences, variability of the final prognosis with apparently similar TBI, necessity for peripheral biomarkers, and more specific predictive models. OBJECTIVES: To study the relationship between serum S100B and survival in TBI patients in various serious situations; the S100B level in patients without traumatic pathology or associated tumour, subjected to stressful situations such as neurological intensive care unit (NICU) stay; the possible overestimation caused by extracerebral liberation in TBI patients and associated polytraumatism; the predictive cutoffs to determine the most sensitive and specific chronology; and achieve a predictive prognostic model. METHODS: Patients admitted to the NICU within 6 hours after TBI were selected. We measured: a) clinical: exitus yes/no; age and gender, traumatic mechanism, polytraumatism yes/no, GCS score, unconsciousness duration, amnesia duration, neurological focality, and surgical interventions; b) radiological: CT scan for radiological lesions; c) biochemical: serum SB100B at 6, 24, 48 and 72 hours after TBI and drug abuse detected in the urine; d) GOS on hospital discharge. RESULTS: N: 149 TBI patients, independent of polytraumatism, mean serum S100B at 6, 24, 48, and 72 hours: 2.1, 1.3, 1.2, and 0.6 microg/L, respectively; N: 124 without associated polytraumatism, S100B at 6, 24, 48, and 72 hours: 2.0, 1.4, 1.3, and 0.6 microg/L; N: 50 control I S100B 24 hours: 0.17 microg/L (0.04 - 0.56) and 25 healthy subjects S100B 0.057 microg/L (0.02-0.094). CONCLUSIONS: Significantly higher S100B levels are observed on exitus, with excellent TBI prognosis and evolution performance. Hospital stay in the NICU produces significant increases in S100B compared to healthy subjects, without invalidating it as a biomarker. Polytraumatism associated to TBI does not significantly alter S100B levels. S100B at 24 hours > or = 0.90 microg/L appears to predict unfavourable TBI evolution with a NPV: 94.2% and PPV: 54.9%. We propose a predictive model when we associate S100B at 24 hours with amnesia duration over 30 minutes with a NPV of 85.5% and a PPV of 83.3%.

A deletion variant of the Aspergillus fumigatus ribotoxin Asp f 1 induces an attenuated airway inflammatory response in a mouse model of sensitization.

BACKGROUND: Aspergillus fumigatus is the most prevalent airborne fungal pathogen, and the ribotoxin Asp f 1 is one of its major allergens. Alpha-Sarcin is a natural variant of Asp f 1 produced by the nonpathogenic fungus Aspergillus giganteus. Both proteins show a sequence identity of 87% and almost identical 3-dimensional structures. Alpha-Sarcin delta(7-22) is a deletion mutant that displays reduced immunoglobulin (Ig) E reactivity and is much less cytotoxic than wild-type proteins against human transformed cells. OBJECTIVE: A murine model of sensitization to Asp f 1 was established to test the response elicited by this alpha-sarcin delta(7-22) deletion mutant. METHODS: BALB/c mice were treated intraperitoneally with different mixtures of recombinant wild-type Asp f 1 and/or a suspension of a commercially available A. fumigatus standard extract. Mice were then intranasally challenged with Asp f 1 or alpha-sarcin delta(7-22). Sera were collected for subsequent measurement of Ig levels and histological analysis of the nostrils and lungs. RESULTS: Sensitization to Asp f 1 was successful only when the purified protein was first administered together with the A fumigatus suspension. The model was characterized by elevated levels of total IgE in serum and histological lesions in the lungs and nostrils. These symptoms were less severe when the deletion variant was the protein administered, thus confirming in vivo its lower toxic character. CONCLUSIONS: An easily reproducible mouse model of A fumigatus Asp f 1 sensitization was established. This model revealed alpha-sarcin delta(7-22) to be a potential candidate for immunotherapy.

Long-term improvement of metabolic control without increased risk of hypoglycaemia by intensive insulin regimens in type 1 diabetes patients treated in a regular clinical setting.

AIM: To evaluate if intensive insulin regimen with multiple daily injections (MDI) is successful for treating type 1 diabetes patients over a long period of time in a regular clinical setting. METHOD: This is a prospective, observational seven-year study. Fifty-nine (35 male) type 1 diabetic patients with bad metabolic control (HbA1c> or =9%), aged 31.9 years, range 18-47 were included in the present study. All of them had had at least 5 years of diabetes duration after diagnosis and showed negative responses of serum C-peptide to a standard breakfast. The main control variables are: Metabolic control measured by serum HbA1c values (values < 6.2 % was the treatment objective) and the frequency of hypoglycaemic episodes (episodes/patient-month). RESULTS: Significant decreases in mean+/-SD HbA1c values in this group of patients were observed from the first year of follow-up, with the mean values being: 7.5+/-1.5%, 7.2+/-1.8%, 7.6+/-1.6%, 7.1+/-1.7%, 7+/-1.4, 6.6+/-1.6% and 6.8+/-1.4% for the first, second, third, fourth, fifth, sixth and seventh years of follow-up respectively. Sixteen %, 27.5%, 15.7%, 33.3%, 28.6%, 42% and 33% of the patients reached the treatment objective (HbA1c values<6.2%) for each year of follow-up. Throughout the study period the rate of severe hypoglycaemia (episodes/patient-year) was 0.32+/-0.2 which was not significantly different compared with the value of 0.28+/-0.1 observed the year before the study began. Similarly frequencies of mild/moderate hypoglycaemia episodes (episodes/patient-month) varies between 16.5+/-4 and 21.7+/-5, which are not significantly different from the value of 17.7+/-6 observed the year before the study began. CONCLUSION: Long-term improvement in metabolic control was observed in this group of type 1 diabetes patients with previous bad control, during treatment in a regular clinical setting. A considerable percentage of type 1 diabetic patients with MDI reached the treatment objective in every year of follow-up. Furthermore improvement in metabolic control is not associated with significantly increased frequency of hypoglycaemia episodes.

Determinación de tiroglobulina / Thyroglobulin determination

La tiroglobulina sérica es un marcador tumoral altamente específico y sensible para el seguimiento de pacientes con carcinoma diferenciado de tiroides. La estandarización de la mayoría de inmunoanálisis frente al material de referencia CRM-457 ha reducido, pero no eliminado, la variabilidad entre métodos. Por eso, la monitorización de las concentraciones de tiroglobulina durante el seguimiento de un paciente debe hacerse en el mismo laboratorio y siempre con el mismo método. Cuanto mejor sea la sensibilidad funcional y la precisión interserial del análisis de tiroglobulina, más sensible será éste para detectar recurrencias de forma temprana. Los métodos inmunométricos deben estar protegidos frente al efecto gancho, ya sea con un diseño en 2 etapas o bien procesando las muestras, al menos las de elevada sospecha, de forma directa y diluida. Mención especial requiere la interferencia producida por anticuerpos antitiroglobulina. La inexistencia de una metodología segura para predecir esta interferencia, unido a que cualquier concentración de autoanticuerpos es potencialmente interferente, obliga a acompañar toda determinación de tiroglobulina con la de anticuerpos antitiroglobulina, utilizando métodos de inmunoanálisis lo más sensibles y precisos posible. En presencia de estos autoanticuerpos las concentraciones de tiroglobulina han de interpretarse apropiadamente. Además, existe la posibilidad de que la determinación de tiroglobulina sea interferida por la presencia de anticuerpos heterófilos, o que la tiroglobulina segregada por el tumor resulte inmunológicamente inerte para el análisis utilizado (AU)

Serum thyroglobulin (TG) is a highly specific and sensitive tumoral marker for the follow-up of patients with differentiated thyroid carcinoma. The standardization of most immunoassays against the CRM-457 reference preparation has reduced, but not eliminated, variability among methods. Consequently, the monitoring of TG concentrations during the follow-up of patients should be performed in the same laboratory and always using the same method. The greater the functional sensitivity and the inter-series accuracy of TG analysis, the greater the sensitivity in the early detection of recurrences. Immunometric methods should be protected against the hook effect, whether with a 2 step design or by processing the samples, at least those with high suspicion, directly and in diluted form. Special mention should be made of the interference produced by anti-thyroglobulin antibodies (anti-Tg ab). Because a safe method to predict such interference is lacking and because any level of autoantibody concentration can potentially cause interference, all TG determinations should be accompanied by anti-TG antibody determinations using the most sensitive and accurate immunoassay methods possible. When these autoantibodies are present, TG concentrations should be appropriately interpreted. Moreover, interference with TG determination can be produced by the presence of heterophilic antibodies. In addition, TG segregated by the tumor can be immunologically inert for the assay used (AU)