Role of diesel exhaust particles in the induction of allergic asthma to low doses of soybean.

INTRODUCTION: Exposure to environmental pollutants such as diesel exhaust particles (DEP) increases the risk of asthma and asthma exacerbation. However, the exact mechanisms inducing asthma to low doses of allergens remain poorly understood. The present study aimed to analyse the immunomodulatory effect of the inhalation of DEP in a mouse model exposed to non-asthmagenic doses of soybean hull extract (SHE). MATERIAL AND METHODS: BALB/c ByJ mice were randomly divided into four experimental groups. Two groups received nasal instillations of saline and the other two groups received 3 mg ml-1 SHE during 5 days per week for 3 weeks. One group in each pair also received 150 µg of DEP in the same instillations 3 days per week. SHE-specific IgE levels, oxidative stress, leukocyte pattern and optical projection tomography (OPT) imaging studies were assessed. RESULTS: Inhalation of SHE and/or DEP increased levels of H2O2 in BAL, while coexposure to SHE and DEP increased SHE-specific IgE levels in serum. Inhalation of SHE alone increased eosinophils, B cells, total and resident monocytes and decreased levels of NK cells, while inhalation of DEP increased neutrophils and decreased total monocytes. Regarding dendritic cells (DC), the inhalation of SHE and/or DEP increased the total population, while the inhalation of SHE alone increased Th2-related DCs (CD11b + Ly6C-) and decreased tolerogenic DCs (CD11b-Ly6C-). However, coexposure to SHE and DEP increased oxidative stress-sensitive DCs (CD11b-Ly6C+) and decreased Th1-related DCs (CD11b + Ly6C+). As regards macrophages, inhalation of SHE and DEP decreased total and alveolar populations. DEP deposition in lung tissue did not differ between groups. CONCLUSION: Coexposure to DEP activates the asthmatic response to low doses of soy by triggering the immune response and oxidative stress.

Epidemiology and Risk Factors for Cancer After Lung Transplantation.

Cancer is the third most common cause of death among lung transplant (LT) recipients who survive for more than 1 year. The purpose of this study was to analyze the incidence and risk factors for cancer after LT in a Spanish cohort. The epidemiology and risk factors for cancer were retrospectively analyzed in LT recipients from 2 cities in Spain, Madrid and Barcelona. Of the 1353 LT patients initially included in the study, 125 (9.2%) developed cancer after a mean of 3.7 years. This frequency was 5-fold higher than in the general population. The most prevalent tumors were skin cancer (32%), lymphoproliferative disease (18%), and lung cancer (16.5%). In 4 patients, lung cancer was diagnosed on the day of the operation. The risk of cancer increased with age >55 year (hazard ratio [HR] 2.89 [1.64-5.09]; P < .001), in men (HR 2.8 [1.4-5.6]; P = .004), and in heavy smokers (>20 pack-years) (HR 2.94 [1.64-5.27]; P < .001). Other factors such as sun exposure were not found to be risk factors. In conclusion, prevalence of cancer is high in LT recipients in a Mediterranean country. Skin tumors, lymphoproliferative disease, and lung cancer are the most prevalent cancers. Age, male sex, and smoking were the main risk factors for cancer in this population.

Effect of anti-IgE in occupational asthma caused by exposure to low molecular weight agents.

BACKGROUND: The role of immunoglobulin (Ig)-E in occupational asthma (OA) due to low molecular weight (LMW) agents is not well established compared to classical atopic asthma. In this study, we evaluate whether anti-IgE monoclonal antibody (mAb) has an effect in a mouse model of OA, using persulfate salts. METHODS: On days 1 and 8, BALB/C mice were dermally sensitized with 5% ammonium persulfate (AP) or dimethyl sulfoxide (DMSO). On days 15, 18, and 21, animals were injected intraperitoneally with anti-IgE mAb or PBS 6 hours before challenge with AP or saline. Airway hyper-responsiveness (AHR) using a methacholine test, airway inflammation in bronchoalveolar lavage (BAL) and lung tissue, and total free IgE in serum samples were analyzed 24, 48, and 96 hours after the last challenge. RESULTS: Anti-IgE mAb treatment almost completely neutralized free serum IgE. In AP-sensitized and challenged mice, anti-IgE mAb treatment abolished AHR 24 hour and 48 hour after the last challenge and significantly reduced the total number of eosinophils and neutrophils 48 hour and 96 hour after the last AP challenge compared with nontreated mice. Levels of interleukin (IL)-13 in BAL were also significantly decreased after anti-IgE administration 24 hour and 48 hour after the last AP challenge. Histological analysis of the lung sections from anti-IgE-treated mice revealed normal inflammatory patterns similar to control groups 48 hour after the last challenge. CONCLUSIONS: Anti-IgE-treated mice showed a significant improvement in asthma features related to the AHR and airway inflammation. Anti-IgE mAb has positive effects in OA induced by persulfate salts.

Persistence of respiratory and inflammatory responses after dermal sensitization to persulfate salts in a mouse model of non-atopic asthma.

BACKGROUND: Exposure to ammonium persulfate (AP) has been reported to be the main cause of occupational asthma in hairdressers. The aim of this study is to assess how long the asthmatic response to AP can be induced after dermal sensitization in a mouse model. METHODS: BALB/c mice received dermal applications of AP or dimethylsulfoxide (DMSO) (control) on days 1 and 8. They then received a single nasal instillation (challenge) of AP or saline on days 15, 22, 29, 36, 45, 60 and 90. Respiratory responsiveness to methacholine was measured 24 h after the challenge using a non-specific methacholine provocation test. Pulmonary inflammation was analysed in bronchoalveolar lavage (BAL), and total serum immunoglobulin (Ig) E, IgG1 and IgG2a were measured in serum samples. Histological analysis of lung slides was performed. RESULTS: Mice dermally sensitized and intranasally challenged with AP showed respiratory responsiveness to methacholine as long as 45 days after initial sensitization, as well as increased percentage of neutrophils in BAL compared with the control group. At day 60, dermally sensitized mice still presented bronchial hyperresponsiveness, while the percentage of neutrophils returned to baseline levels similar to those of controls. Total serum IgE increased significantly on day 22 after dermal sensitization. Total serum IgG1 and IgG2a increased from 45 days after dermal sensitization and remained high at 90 days. CONCLUSIONS: Both respiratory responsiveness to methacholine and airway inflammation responses decrease with increasing time between sensitization and challenge. Respiratory responsiveness to methacholine tends to persist longer than inflammation.

[Determining asthma treatment in children by monitoring fractional exhaled nitric oxide, sputum eosinophils and leukotriene B4]. / Monitorización del tratamiento en el asma bronquial mediante determinación de fracción exhalada de óxido nítrico, eosinófilos y leucotrieno B4 en esputo inducido de población infantil.

Sputum eosinophils and exhaled fractional nitric oxide (FENO) are markers of airway inflammation in asthma. Cytokines, cysteinyl-leukotrienes and leukotriene B4 (LTB4) are responsible for this inflammation. The aim of this study is to determine the usefulness of these markers in monitoring asthma treatment in children. FENO, sputum eosinophils, and LTB4 in induced sputum were performed in 10 children (9-15 years old). These determinations were repeated four months later, after the beginning or an increase in the treatment. FENO values tended to decrease (P=.15), pulmonary function tended to improve (P=.10), and sputum eosinophils decreased (P=.003) compared to the first determination. There were no differences in LTB4 concentrations (P=.88). Sputum eosinophils seem to be more precise than FENO in the monitoring of inflammation in asthmatic children.

Biomarkers of pulmonary rejection.

Immunologic complications after lung transplantation (LT) include acute cellular rejection (ACR), antibody-mediated rejection (AMR), and most forms of chronic allograft dysfunction (CAD). ACR is an inflammatory process in which the reaction is mediated by the T-cell population. Most episodes of ACR fully recover with treatment, but repeated bouts are considered to be a risk factor for CAD. Biomarker cytokines interleukin (IL)-10, IL-15, IL-6, CCL5, CCR2 and IFNγ may play significant roles in this complication. Formerly bronchiolitis obliterans syndrome (BOS) or chronic rejection or most forms of CAD were considered to be immunologic complications not amenable therapeutic measures. CAD, the main limitation for long-term survival in LT, is characterized histologically by airway epithelial cell apoptosis and luminal fibrosis in the respiratory bronchioles causing airflow obstruction and, in some cases, lung parenchymal affectations causing restrictive lung disease. Several biomarkers have been studied in CAD, IL-6, IL-8, IL-17, IL-23, IL-13, IFN γ, and TGF ß cytokines, pH, bile acid, and tripsine of gastroesophageal reflux and toll-like receptors of innate immunity. Herein we have reviewed the literature of biomarkers involved in lung rejection.

Soy aeroallergens in thoracic fraction particles (PM10).

BACKGROUND: The aerodynamic diameter of biological particles determines their ability to penetrate the human respiratory system. OBJECTIVE: To assess the content of allergens less than 10 pm in diameter in the particle fraction of airborne dust in order to improve control of exposure to harmful soybean aeroallergens. METHODS: In this study, 98 pairs of particulate matter measuring less than 10 microm in diameter (PM10) and total suspended particulate (TSP) filters were collected in parallel and analyzed for soy aeroallergens by the inhibition enzyme-linked immunosorbent assay. RESULTS: The median levels found were 6 and 22.5 U/m3 for PM10 and TSP filters, respectively. A good correlation was found between soy aeroallergen content in PM10 and TSP filters. The median proportion of soy aeroallergen content in PM10 filters versusTSP filters was 28.6%, and varied widely across different days. CONCLUSIONS: Due to this wide variation between days, it seems that soy aeroallergen content in TSP filters is not a good surrogate of soy allergen content in PM10 filters. Further clinical studies should be conducted to assess differences in the health impact of soy allergen content in PM10 filters and TSP filters.

Soy Aeroallergens in Thoracic Fraction Particles (PM10)

Introducción: El diámetro aerodinámico de las partículas biológicas determina su habilidad para penetrar en el aparato respiratorio. Objetivo: Determinar el contenido de alérgenos de diámetro inferior a 10 μm en la materia particulada del aire, con el fin de mejorar el control de la exposición a los aeroalérgenos nocivos de la soja. Métodos: En este estudio, se muestrearon en paralelo 98 pares de filtros, siendo uno de materia particulada de diámetro <10 μm (PM10) y el otro de partículas suspendidas totales (PST). Mediante un método de ELISA de inhibición se analizó la concentración de aeroalérgenos de soja en todos los filtros. Resultados: La mediana de los niveles encontrados fue de 6 y 22,5 U/m3 para los filtros PM10 y PST, respectivamente. Se encontró una buena correlación entre el contenido de aeroalérgenos de soja en los filtros PM10 y PST. La proporción mediana de aeroalérgenos de soja en los filtros PM10 frente a PST fue de 28,6% y varió ampliamente entre los diferentes días. Conclusiones: Debido a la amplia variación de la proporción de aeroalérgenos de soja en los filtros PM10 frente a PST entre días, parece que el contenido de alérgeno de soja en los filtros PST no es un buen indicador del contenido de dichos alérgenos en los filtros PM10. Por ello, deben ser realizados más ensayos clínicos con el fi n de evaluar si el contenido de alérgenos de soja en los filtros PM10 y PST tienen diferente impacto sobre la salud (AU)

Background: The aerodynamic diameter of biological particles determines their ability to penetrate the human respiratory system. Objective: To assess the content of allergens less than 10 μm in diameter in the particle fraction of airborne dust in order to improve control of exposure to harmful soybean aeroallergens. Methods: In this study, 98 pairs of particulate matter measuring less than 10 μm in diameter (PM10) and total suspended particulate (TSP) filters were collected in parallel and analyzed for soy aeroallergens by the inhibition enzyme-linked immunosorbent assay. Results: The median levels found were 6 and 22.5 U/m3 for PM10 and TSP filters, respectively. A good correlation was found between soy aeroallergen content in PM10 and TSP filters. The median proportion of soy aeroallergen content in PM10 filters versus TSP filters was 28.6%, and varied widely across different days. Conclusions: Due to this wide variation between days, it seems that soy aeroallergen content in TSP filters is not a good surrogate of soy allergen content in PM10 filters. Further clinical studies should be conducted to assess differences in the health impact of soy allergen content in PM10 filters and TSP filters (AU)

Induced sputum cell count and cytokine profile in atopic and non-atopic children with asthma.

BACKGROUND AND AIM: Sputum induction is a semi-invasive technique used to detect and monitor airway inflammation. In this study, the cell profile, and Th1 and Th2 cytokine levels in induced sputum of asthmatic and healthy children (HC) are compared. METHODS: Sputum induction was performed in healthy and asthmatic children by inhalation of hypertonic saline solution. Differential cell count in the specimen obtained was carried out using optic microscopy. IFN-γ, IL-2, IL-10, IL-8, IL-6, IL-4, IL-5, IL-1ß, TNF-α, and IL-12p70 levels were determined in sputum sample supernatants by flow cytometry. RESULTS: Sputum induction was performed in 31 HC and 77 asthmatic children (60 atopic and 17 non-atopic asthma, NAA). Twenty-four samples were obtained in HC and 64 in patients. Median eosinophil count in atopic asthma (AA; 2%) was higher than in NAA (P = 0.02) or HC (P = 0.01). IL-4, IL-5, IFNγ, IL-2, and IL-12p70 concentrations were higher in AA than in NAA or HC. IL-8 was higher in asthmatic children (atopic and non-atopic) than in healthy ones. IL-10 was higher in the healthy group than in the AA group (P = 0.02). CONCLUSIONS: As compared to HC, the inflammatory profile in induced sputum of children with asthma showed an increase in proinflammatory cytokines. Concentrations of IL-10, an anti-inflammatory cytokine, were lower in children with AA than in HC.

Inducción de esputo en niños: desarrollo técnico / Sputum induction in children: technical development

Objetivos: Comparar el rendimiento (total de muestras obtenidas) de nebulizadores ultrasónicos de flujo bajo y alto en la inducción de esputo en niños asmáticos, y valorar los efectos adversos asociados. Pacientes y métodos: Se nebulizó suero salino hipertónico a concentraciones crecientes (3%, 4%, 5%) utilizando nebulizadores ultrasónicos de bajo flujo (Omron NE-U07(R); flujo 1ml/min) y de alto flujo (Omron NE-U12(R); flujo 3 ml/min, y DeVilbiss Ultraneb 3000(R); flujo 2,5ml/min). Resultados: Se realizaron 49 inducciones en 49 pacientes entre 7 y 15 años de edad (en 15 niños se utilizó un nebulizador de bajo flujo y en 34 niños un nebulizador de alto flujo (Omron NEU12(R): 6 casos, DeVilbiss Ultraneb 3000(R): 28 casos). Se obtuvieron 37 muestras. Treinta y seis presentaban<20% de células escamosas y 26 tenían una viabilidad ≥60%. El rendimiento de la prueba fue mayor con los nebulizadores de alto flujo (85,3% de muestras), frente al 53% (p=0,04). El 69% de las muestras obtenidas con los nebulizadores de alto flujo fueron válidas, frente al 62,5% con el de bajo flujo (p=0,7). Con los nebulizadores de alto flujo disminuyó la incidencia de tos (17,6%, p=0,08) y de picor de ojos (0%, p=0,02), respecto al nebulizador de bajo flujo (47% y 20% respectivamente), aunque aumentó el sabor desagradable (82,3%, p<0,001) y la sialorrea (14,7%, p=0,3). Conclusiones: Con los nebulizadores ultrasónicos de alto flujo se consigue un mayor rendimiento de la técnica sin que se observe un aumento de efectos adversos significativos (AU)

Objective: To compare low and high flow nebulizers performance (total of samples) and its side effects on sputum induction in asthmatic children. Patients and methods: Sputum induction was performed by inhalation of a hypertonic saline solution at increasing concentrations (3%, 4% and 5%) using low flow (OMRON NE-U07(R); flow rate 1ml/min), or high flow (OMRON NE-U12(R); flow rate 3ml/min, and DeVilbiss Ultraneb 3000(R); flow rate 2.5ml/min) ultrasonic nebulizers. Results: We performed 49 inductions in 49 patients from 7 to 15 years old (in 15 children we used a low flow nebulizer (Omron NE-U07(R)) and in 34 children a high flow nebulizer (OMRON NEU12(R), 6 patients, and DeVilbiss Ultraneb 3000(R), 28 patients). We obtained 37 samples of which 36 had less than 20% of squamous cells, and 26 had a viability ≥60%. The test performance was higher with high-flow nebulizers, obtaining 85.3% of samples compared to 53% (p=0.04). A total of 69% of samples obtained with the high flow nebulizer were valid, compared to 62.5% (p=0.7) with the low flow nebulizers. With high flow rate nebulizers the incidence of cough (17.6%, p=0.08) and itchy eyes (0%, p=0.02) decreased with the low flow nebulizer (47% and 20% respectively), but bad taste (82.3%, p <0.001) and salivation (14.7%, p=0.3) increased. Conclusions: With high flow rate ultrasonic nebulizers we obtain a higher performance of the technique without an increase in significant side effects (AU)

[Sputum induction in children: Technical development]. / Inducción de esputo en niños: desarrollo técnico.

OBJECTIVE: To compare low and high flow nebulizers performance (total of samples) and its side effects on sputum induction in asthmatic children. PATIENTS AND METHODS: Sputum induction was performed by inhalation of a hypertonic saline solution at increasing concentrations (3%, 4% and 5%) using low flow (OMRON NE-U07; flow rate 1ml/min), or high flow (OMRON NE-U12; flow rate 3ml/min, and DeVilbiss Ultraneb 3000; flow rate 2.5ml/min) ultrasonic nebulizers. RESULTS: We performed 49 inductions in 49 patients from 7 to 15 years old (in 15 children we used a low flow nebulizer (Omron NE-U07) and in 34 children a high flow nebulizer (OMRON NEU12, 6 patients, and DeVilbiss Ultraneb 3000, 28 patients). We obtained 37 samples of which 36 had less than 20% of squamous cells, and 26 had a viability > or =60%. The test performance was higher with high-flow nebulizers, obtaining 85.3% of samples compared to 53% (p=0.04). A total of 69% of samples obtained with the high flow nebulizer were valid, compared to 62.5% (p=0.7) with the low flow nebulizers. With high flow rate nebulizers the incidence of cough (17.6%, p=0.08) and itchy eyes (0%, p=0.02) decreased with the low flow nebulizer (47% and 20% respectively), but bad taste (82.3%, p <0.001) and salivation (14.7%, p=0.3) increased. CONCLUSIONS: With high flow rate ultrasonic nebulizers we obtain a higher performance of the technique without an increase in significant side effects.

Assessment of soy aeroallergen levels in different work environments.

BACKGROUND: Airborne soybean hull proteins are known causes of asthma epidemics around harbours and soy processing plants. Soy flour dust proteins may cause occupational allergy in food and feed industries. OBJECTIVE: To compare enzyme immunoassays (EIAs) for soy hull and soy flour aeroallergens, exposure assessment in various work environments. METHODS: Airborne dust samples (n=324) from soy unloading and/or processing plants, the animal feed industry and pig stables were analysed by two soy flour assays: one assay for measuring complete soy hull proteins and two assays for measuring the purified low-molecular-weight (LMW) soy hull allergens. RESULTS: Immunoblotting confirmed strong differences between antibody specificities and soy preparations. The results of the two soy flour assays and the assay for measuring complete soy hull proteins were highly correlated (r>0.85). The two LMW soy hull assays also showed a strong mutual correlation (r=0.91), but much less correlation with assays for measuring soy flour and complete soy hull. The levels of LMW soy hull proteins were the highest at sites of soybean unloading or processing, while soy flour levels were particularly high in the soy and animal feed industry. CONCLUSIONS: The optimal EIA procedure for soy aeroallergen exposure assessment depends on the type of work environment and the local soy dust composition. Thus, the type of work environment should always be taken into account in future soy allergy studies in order to prevent a possible underestimation of the workers' actual risk of developing soy allergy.

Occupational asthma related to mouse allergen exposure and rhinoconjunctivitis due to collagenase inhalation in a laboratory technician.

We describe the case of a 27-year-old patient working in a research laboratory, who developed occupational asthma to mouse proteins and presented symptoms of rhinoconjunctivitis caused by manipulation of collagenase. Specific inhalation challenge confirmed the diagnosis of occupational asthma to mouse proteins, whereas specific challenge with collagenase only evoked symptoms of rhinitis and conjunctivitis. SDS-PAGE and Western blot analysis for collagenase showed that the patient's IgE antibodies bound specifically to a protein with a molecular weight of 92 kDa. Hence, this was an unusual case of double sensitization. The sensitization to collagenase presented in this report may represent a new occupational disease in technicians working in medical or research laboratories.

An amplified sandwich EIA for the measurement of soy aeroallergens.

BACKGROUND: Soy hull low-molecular-weight (SHLMW) allergens were responsible for the soy asthma epidemics in Barcelona, with one 7.5 kDa protein (Gly m 1) being the main IgE-binding component. The aims of this study were to develop a sensitive sandwich enzyme immunoassay (EIA) using rabbit polyclonal antibodies to measure low levels of SHLMW allergens, and to compare this method with the previously described human IgE EIA-inhibition technique. METHODS: IgG was isolated from serum of rabbits immunized with a chromatographically purified SHLMW extract (SHLMWE). Antibody-binding profiles were compared with those of human IgE anti-soy protein antibodies by Western blot analysis. An amplified sandwich EIA was developed using the purified SHLMWE as a calibration standard. Results were expressed in nanograms per millilitre. To compare the two assays, 54 air samples were analysed by both methods. RESULTS: SDS-PAGE of the SHLMWE revealed four bands of 6, 8, 15 and 17 kDa. Gly m 1 in the SHLMWE was identified by fingerprinting. The detection limit of the assay was 40 pg/mL. The two methods correlated well (r=0.89; P<0.001). The allergen concentration was detected in all 54 (100%) samples by the sandwich EIA but in only 37 (68.5%) by the EIA inhibition. CONCLUSIONS: The amplified sandwich EIA for SHLMW components has a high sensitivity and appeared to be a useful tool for the measurement of airborne SHLMW allergens, even at relatively low concentrations. Moreover, the method uses rabbit antibodies at high dilutions and does not require human sera, with limited availability and quantitative and qualitative pool-to-pool variability.

Weight loss with long-term intermittent treatment with oral oleoyl-estrone in lean Zucker male rats.

To determine whether or not the long-term intermittent treatment with oleoyl-estrone (OE) in rats induces a cumulative weight loss, adult male rats were treated with OE over three alternating 10-day periods, with a 30-day "recovery" period occurring between each period. At the end of the third treatment period, the rats were allowed to recover and were then mated with females. Each treatment period produced a decrease of ca. 7% in body weight with no rebound during the recovery periods, whereas weight changed at the same pace in controls. The greatest difference in body weight was observed during the last days of treatment. OE-treated rats retained their initial protein pools throughout the treatment. Furthermore, treated and control males remained fertile and were able to procreate. Thus, we can conclude that intermittent OE treatment induces a cumulative weight loss in adult male rats.

Technical note: Measurement of total estrone content in foods. Application to dairy products.

Estrone is a powerful growth-inducing hormone that is present in milk, mainly in the form of fatty acid esters, at concentrations that promote growth in experimental animals. We present here a method useful for the measurement of this natural hormone in foods and applied it to several common dairy products. Samples were frozen, finely powdered, and lyophilized then extracted with trichloromethane/methanol; the dry extract was saponified with potassium hydroxide. The free estrone evolved was extracted with ethyl acetate and was used for the estimation of total estrone content through radioimmunoassay. Application of the method to dairy products showed high relative levels of total estrone (essentially acyl-estrone) in milk, in the range of 1 microM, which were halved in skimmed milk. Free estrone levels were much lower, in the nanomolar range. A large proportion of estrone esters was present in all other dairy products, fairly correlated with their fat content. The amount of estrone carried by milk is well within the range, where its intake may exert a physiological response in the sucklings for which it is provided. These growth-inducing and energy expenditure-lowering effects may affect humans ingesting significant amounts of dairy products.