Fluorocytometric analysis of induced sputum cells in an asthmatic population.

BACKGROUND: Bronchial mucosal inflammation is the major pathogenic process in asthma. In the latest years, induced sputum (IS) examination has become an important non-invasive method of assessing airway inflammation. Flow cytometry has been recently applied to the study of IS though it is not exempt of methodological difficulties. The aim of the present study was to further study if the fluorocytometric analysis of IS could represent a reliable tool to assess the presence of bronchial activated lymphocytes in stable mild asthmatic patients. METHODS: Induced sputa from controls and asthmatic patients were processed in isotonic 3mM dithiothreitol (DTT), a mucolytic agent required for cell dispersion. The individualized cells were then stained with monoclonal antibodies for three-colour flow-cytometric analysis. Total IgE and ECP were measured in serum and in the sputum fluid phase. RESULTS: The cellularity of asthmatic sputa is enriched in eosinophils (mean, 26.63%) with respect to controls, but not in lymphocytes. However, lymphocytes from asthmatics show increased surface expression of activation markers (CD25 in T cells, CD23 in B cells). Surprisingly, no differences were observed in the detected levels of CD54 on IS lymphocytes and eosinophils between asthmatics and non-asthmatics. Furthermore, there was a significantly higher concentration of ECP and total IgE in the sputum from the asthmatic group. CONCLUSION: Fluorocytometric analysis of induced sputum is a reliable non-invasive method for the study of bronchial immune cells. It could provide complementary information on activated cells in the bronchial mucosa even in non-smokers, mild and stable asthmatics and it is reasonable to speculate that it will be useful in monitoring the effect of the treatment in these patients.

Measurement of serum levels of eosinophil cationic protein in the diagnosis of acute gastrointestinal anisakiasis.

Thirty-two patients with abdominal pain and/or intestinal pseudo-obstruction who had consumed raw or undercooked fish in the previous 72 h, were included in a study of anisakiasis, a parasitation of the human gastrointestinal tract by third stage Anisakis simplex larvae. Skin prick test (SPT) against A. simplex were positive in all the patients. High median eosinophil cationic protein (ECP) serum concentrations (> 15 mg/L) at day 0 with normal serum levels at day 30 and a rise of median total and specific IgE against A. simplex at day 30, were observed. We conclude that a raised serum level of ECP in the first 72 h from the onset of symptoms coinciding with a positive SPT against A. simplex and high total and specific immunoglobulin (IgE) in the first month after the parasitation, could be a useful tool in the diagnosis of gastrointestinal anisakiasis, even if the parasite cannot be isolated.

The effect of dithiothreitol on VLA-4 detection in peripheral blood and induced sputum cells.

BACKGROUND: Induced sputum has been shown to be a reliable technique for investigating airway inflammation non-invasively. Flow cytometry could provide useful information in this area. However, the viscosity of the sample entails the use of a mucolytic agent. Dithiothreitol (DDT) is the most frequently used agent although it could affect detection of different inflammatory markers. METHODS: To measure the effect of DDT on the detection of certain adhesion molecules in eosinophils and lymphocytes, sputum was induced from seven non-smoking asthmatic and non-asthmatic subjects treated with 0.1 M DDT. The samples were analyzed by flow cytometry. Whole blood samples from the same subjects were also processed with DTT and analyzed by flow cytometry. RESULTS: Very late activated antigen-4 (VLA-4) levels on eosinophils in intracellular and surface staining were much lower than expected. VLA-4 on lymphocytes was also altered but less so than on eosinophils. VLA-4 levels were also decreased on blood cells after DTT treatment. No abnormalities were found in the detection of CD29 on eosinophils and the beta7-chain in lymphocytes. CONCLUSIONS: Flow cytometry could be used as a complementary method to induced sputum in the investigation of airway inflammation. However, DTT could interfere with the detection of some inflammatory markers, as is the case with VLA-4.

The effect of dithiothreitol on VLA-4 detection in peripheral blood and induced sputum cells

Background: Induced sputum has been shown to be a reliable technique for investigating airway inflammation non-invasively. Flow cytometry could provide useful information in this area. However, the viscosity of the sample entails the use of a mucolytic agent. Dithiothreitol (DDT) is the most frequently used agent although it could affect detection of different inflammatory markers. Methods: To measure the effect of DDT on the detection of certain adhesion molecules in eosinophils and lymphocytes, sputum was induced from seven non-smoking asthmatic and non-asthmatic subjects treated with 0.1 M DDT. The samples were analyzed by flow cytometry. Whole blood samples from the same subjects were also processed with DTT and analyzed by flow cytometry. Results: Very late activated antigen-4 (VLA-4) levels on eosinophils in intracellular and surface staining were much lower than expected. VLA-4 on lymphocytes was also altered but less so than on eosinophils. VLA-4 levels were also decreased on blood cells after DTT treatment. No abnormalities were found in the detection of CD29 on eosinophils and the β7-chain in lymphocytes. Conclusions: Flow cytometry could be used as a complementary method to induced sputum in the investigation of airway inflammation. However, DTT could interfere with the detection of some inflammatory markers, as is the case with VLA-4 (AU)

Antecedentes: El esputo inducido se ha mostrado como una técnica fiable para estudiar de un modo no invasivo, la inflamación de la vía respiratoria en el asma bronquial. La citometría de flujo puede aportar información útil en este campo. No obstante, la viscosidad de la muestra implica la utilización de una gente mucolítico, siendo el Ditiotreitol (DTT) uno de los más usados, a pesar de que puede alterar la expresión de diferentes mediadores inflamatorios. Métodos: Se indujo esputo de 7 sujetos asmáticos y no asmáticos, todos no fumadores. Se trataron las muestras con DTT 0.1M y se analizaron concitometría de flujo. Muestras de sangre de los mismos sujetos igualmente fueron procesadas con DTT 0.1M y analizadas. Resultados: Los niveles de detección de VLA-4 en la superficie y a nivel intracelular en los eosinófilos, fue más baja de lo esperado. La alteración de los niveles de VLA-4 en linfocitos fue menor que en los eosinófilos. En sangre, la detección de VLA-4 disminuyó en las muestras tratadas con DTT. No se observaron anormalidades en la detección de CD29 en eosinófilos ni en la cadena 7 en linfocitos. Conclusiones: La citometría de flujo puede ser usada en el estudio de la inflamación en la vía aérea aunque se debe tener en cuenta que el DTT altera la detección de algunos marcadores, como así se comprueba para el VLA-4 (AU)

Políticas institucionales de prevención de los problemas de salud generados por el consumo de bebidas alcohólicas en España y Europa / Institutional policies in Spain and Europe to prevent alcohol-related health problems

Los problemas relacionados con el consumo de alcohol y las motivaciones que inducen a beber son múltiples y complejas como lo es también su abordaje, que debe sustentarse en una política global, multisectorial y multidisciplinar, con la participación de todas las áreas implicadas: Salud, Educación, Bienestar Social, Empleo, Tráfico, Agricultura, Comercio, Justicia y Hacienda. En España, la descentralización producida en materia de Sanidad e Higiene (art. 148.21 de la Constitución española), conlleva la necesidad de cooperación y coordinación entre las distintas Administraciones Sanitarias. Con dicha finalidad la Ley General de Sanidad previó la creación del Consejo Interterritorial del Sistema Nacional de Salud. Probablemente, la finalización del proceso de trasferencias sanitarias facilitará todo el proceso de coordinación y cooperación, que cada vez se manifiesta como más necesario. En el Sistema Nacional de Salud, la intervención en el campo del alcohol es prioritaria, y así viene recogida en los distintos planes y políticas de salud que a nivel nacional, autonómico y local se están llevando a cabo. Las 17 Comunidades Autónomas han aprobado sus respectivos planes de salud y en todos ellos el alcohol es un área específica de intervención. Las actuaciones y programas sobre alcohol se llevan a cabo en algunas Comunidades desde los Planes Regionales de Drogodependencias y en otras desde el área de Salud Pública. Se recogen los principales objetivos de las distintas actuaciones preventivas contempladas en los distintos planes y políticas de salud, y se presentan las actuaciones que, en el área de prevención de los problemas relacionados con el alcohol, viene desarrollando el Ministerio de Sanidad y Consumo, la Oficina Regional Europea de la Organización Mundial de la Salud y la Unión Europea (AU)

The problems associated with alcohol consumption and the reasons to drink alcohol are multiple and complex, just like the ways to solve them, which must be based on comprehensive, multi-sectorial and multidisciplinary policies, with the collaboration of all the areas involved: Health, Education, Social Welfare, Employment, Traffic, Agriculture, Commerce, Justice and Finance. In Spain, the decentralisation of the Health and Hygiene powers (Art. 148.21 of the Spanish Constitution) requires the continuous co-operation and co-ordination among the Health Authorities of the Autonomous Communities. Consequently, the General Health Law established the Inter-territorial Council of the National Health System. Probably, the completion of the above mentioned decentralisation process will favour the essential co-ordination and co-operation process. Alcohol prevention is a priority within the National Health System, as it is demonstrated by the numerous plans and policies carried out at the national, autonomous and local level. The 17 Autonomous Communities have approved their respective health plans, and all of them include alcohol as a specific intervention area. Some Communities implement alcohol actions and programmes as part of their Regional Drug-dependencies Plans and other Public Health programmes. This chapter explains the main objectives of the various prevention actions included in the different health plans and policies. Besides, we present the actions related with the prevention of alcohol-related problems implemented by the Ministry of Health and Consumer Affairs, the WHO European Region and the European Union (AU)

Anaphylaxis due to hypersensitivity to Anisakis simplex.

Anaphylactic reactions are among the most frequent causes for requests for emergency medical care. It is possible that no clear precipitating factor can be found delaying the onset of adequate treatment and increasing the risk of new episodes of anaphylaxis in the future. Hypersensitivity to Anisakis simplex, an intestinal parasite found in fish, is an unusual cause of anaphylaxis but it should always be borne in mind in countries where a great deal of fish is consumed, especially if it is eaten raw or undercooked. 3 cases of anaphylaxis due to hypersensitivity to A. simplex are described.

[Morbidity of childhood bronchial asthma in the emergency room]. / Estudio de la morbilidad del asma bronquial infantil en un servicio de urgencias.

BACKGROUND: An allergic etiology could be found in more than the 80% of the patients with bronchial asthma. Specific immunotherapy is the only known etiologic treatment, reducing clinical symptoms and requirement of medication. However only a few patients are controlled by an allergist. The aim of the present study was to verify the influence of control and treatment carried out by patients in childhood asthma recruitment that came to the emergency room of our hospital (Hospital Clínico San Carlos) between December 95 and June 96. MATERIAL AND METHOD: Children to 15 years old were included in the study. We analysed: age, sex, clinical record, previous treatment (pharmacological and etiological), basal symptoms, necessity to be admitted to hospital, medical previous control. RESULTS: 163 patients came to emergency room because of asthma exacerbation. They were divided in two groups: less than 3 years (average 17.45 months) and more than 3 years (average 8 years) (Sd: 3.4). Sex: 65% male sex and 35% female sex. 44.17% came during the greater pollinization period in our field, with a statistical significance compared with other periods (p: 0.0023). 81 patients (49.7%) had been previously diagnosed of bronchial asthma. 15% of the patients were controlled by an allergist, with a stable situation in 92% of them. 68.4% of patients who were controlled by a paediatrician and 40% who were controlled in a officially approved centre (p: 0.015) were stable. 55 patients had continuous treatment and 21.8% of them were been treated with immunotherapy. 17.7% of patients without immunotherapy were admitted to hospital. None of the patients on immunotherapy were hospitalised. CONCLUSIONS: It is essential to do an early diagnosis and a combined follow-up by an allergist and a paediatrician to succeed in a good control of the disease from its beginning. In our experience, the children who were controlled by an allergist had a more stable situation and were also more easily controlled. The specific treatment should be initiated in the early stage of the disease being the only factor that modify the immune response and prevents against the development of asthma in children with rhinitis.

Estudio de la morbilidad del asma bronquial infantil en un servicio de urgencias / Study of morbidity of childhood asthma in the emergency department

Introducción: el asma bronquial, en más de un 80 por ciento de los casos, tiene una etiología alérgica, siendo la inmunoterapia el único tratamiento etiológico conocido. A pesar de ello, sólo una minoría de los pacientes asmáticos es controlada por el alergólogo. Nuestro objetivo fue comprobar si en las reagudizaciones del asma bronquial que acudieron a urgencias pediátricas del Hospital Clínico San Carlos de Madrid, entre diciembre de 1995 y junio de 1996, influía el control y tratamiento previo realizado. Material y método: niños hasta 15 años de edad analizando: edad, sexo, antecedentes personales, tratamiento previo realizado (farmacológico y etiológico), situación basal, necesidad de ingreso y control médico previo. Resultados: acudieron a urgencias con crisis de broncoespasmo 163 pacientes, que se dividieron en dos grupos: menores de tres años (edad media 17,45 meses) y mayores de tres años (edad media ocho años) (Sd = 3,4). Sexo: 65 por ciento varones, 35 por ciento mujeres. El 44,17 por ciento acudió durante el período de mayor polinización en nuestra área, dato estadísticamente significativo comparando con el número de pacientes que acudieron en otras épocas del año (p = 0,0023). Habían sido diagnosticados previamente de asma bronquial 81 niños (49,7 por ciento). El 15 por ciento de los pacientes eran controlados por el alergólogo, manteniéndose estables el 92 por ciento, frente al 68,4 por ciento de los que eran controlados por el pediatra y el 40 por ciento de los controlados por los centros concertados, encontrándose una diferencia estadísticamente significativa entre los tres grupos (p = 0,015). De todos los pacientes, 55 realizaban tratamiento de forma habitual y de éstos el 21,8 por ciento lo realizaba con inmunoterapia. Precisaron ingreso hospitalario el 17,7 por ciento de los pacientes que no recibían tratamiento con inmunoterapia y ningún paciente de los que recibía este tratamiento. Conclusiones: es fundamental realizar un diagnóstico precoz y un seguimiento de forma conjunta por el pediatra y por el alergólogo para el buen control de la enfermedad desde sus primeros estadios, ya que, como hemos visto, los niños que eran controlados por el alergólogo presentaban una situación basal más estable y, además, más fácilmente controlable. El tratamiento específico debe iniciarse en las fases tempranas de la enfermedad, ya que modifica la respuesta inmune y previene el desarrollo de asma en niños con rinitis. (AU)

Background: an allergic etiology could be found in more than the 80% of the patients with bronchial asthma. Specific immunotherapy is the only known etiologic treatment, reducing clinical symptoms and requirement of medication. However only a few patients are controlled by an allergist. The aim of the present study was to verify the influence of control and treatment carried out by patients in childhood asthma recruitment that came to the emergency room of our hospital (Hospital Clínico San Carlos) between december 95 and june 96. Material and method: children to 15 years old were included in the study. We analysed: age, sex, clinical record, previous treatment (pharmacological and etiological), basal symptoms, necessity to be admitted to hospital, medical previous control. Results: 163 patients came to emergency room because of asthma exacerbation. They were divided in two groups: less than 3 years (average 17.45 months) and more than 3 years (average 8 years) (Sd: 3.4). Sex: 65% male sex and 35% female sex. 44.17% came during the greater pollinization period in our field, with a statistical significance compared with other periods (p: 0.0023). 81 patients (49.7%) had been previously diagnosed of bronchial asthma. 15% of the patients were controlled by an allergist, with a stable situation in 92% of them. 68.4% of patients who were controlled by a paediatrician and 40% who were controlled in a officially approved centre (p: 0.015) were stable. 55 patients had continuous treatment and 21.8% of them were been treated with immunotherapy. 17.7% of patients without immunotherapy were admitted to hospital. None of the patients on immunotherapy were hospitalised. Conclusions: it is essential to do an early diagnosis and a combined follow-up by an allergist and a paediatrician to succeed in a good control of the disease from its beginning. In our experience, the children who were controlled by an allergist had a more stable situation and were also more easily controlled. The specific treatment should be initiated in the early stage of the disease being the only factor that modify the immune response and prevents against the development of asthma in children with rinitis (AU)

Desensitization to azathioprine.

Azathioprine is an immunosuppressant drug which is an analog to 6-mercaptopurine and has been used in the last 20 years to prevent organ transplant rejection. It has also been used in the treatment of some autoimmunological diseases. We present a case of a 24-year-old woman with systemic lupus erythematosus, suffering from nephritis, who developed angioedema after using azathioprine to control her illness. She had never reported similar episodes. The involvement of the drug was demonstrated by positive oral challenge test without changes in biochemical and complement blood determinations. She reached tolerance to the drug after a desensitization procedure (increasing 5 mg each day to reach 125 mg daily). We are not able to propose the involvement of an IgE-mediated mechanism, but rather a hypersensitive one with a non-dose-dependent effect. These desensitization procedures show great potential as therapeutic safeguards against harmful drugs in some patients. We have not found any other desensitization procedure for this drug.

Occupational asthma caused by champignon flies.

BACKGROUND: Occupational bronchial asthma in mushroom (champignon) workers is unusual, although reports on it appeared in 1938 and 1951; we have not found any others since those dates. Here we report the case of a 52-year-old man who works as a champignon cultivator. He suffered rhinoconjunctivitis and asthma attacks whenever he entered the champignon culture caves. We studied flies as a possible antigen source. We collected these insects from the growing sites in order to identify them, and then prepare an extract; the samples turned out to be of two families of insects of the order Diptera, 98% from the Phoridae family (Brachycera suborder) and 2% from the Sciaridae (Nematocera suborder). METHODS: Skin prick tests, conjunctival provocation tests, serum specific IgE, specific IgE-binding fractions in immunoblotting, and monitoring of PEFR (at work and off work) were performed. RESULTS: IgE-mediated hypersensitivity to these flies was demonstrated by skin prick test, conjunctival provocation test, serum specific IgE, and IgE-binding fractions in immunoblotting. Monitoring of PEFR both at work and off work showed a clear relationship between symptoms, or fall in PEFR, and the workplace. CONCLUSIONS: We report the case of a patient suffering from asthma and rhinoconjunctivitis caused by hypersensitivity to fly proteins.