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Biomedical subjects

M J Rodrigo

Publications and source records attributed to M J Rodrigo.

At least 19 recordsLinked to original sources

Association study of proposed candidate genes/regions in a population of Spanish asthmatics.

A number of genes/regions have recently been reported to be linked to asthma or its related phenotypes (i.e. atopy and bronchial hyperresponsiveness), by genetic linkage and allele-sharing methods. We have performed a case-control study comparing the allelic distribution of nine microsatellite markers and two genetic variants in a group of patients attended at emergency room departments because of an acute attack of asthma with respect to an external healthy population of controls. A total of 146 asthmatic subjects and 50 population controls from Barcelona, Spain, were genotyped for nine microsatellite markers from some asthma/atopy candidate genes/regions: the beta-subunit of the high-affinity IgE receptor (Fc epsilonRI-beta) located on chromosome 11; the 5q31-32 candidate region; the T-cell receptor genes, TCR-alpha on chromosome 14 and TCR-beta on chromosome 7. Two genetic variants of the beta-subunit of the high-affinity IgE receptor (Fc epsilonRI-beta) gene were also analyzed. None of the asthmatic or control individuals carried the Ile181Leu variant. There were no significant differences between asthmatic and control subjects neither for the polymorphic markers nor for the other variant of the beta-subunit of the high-affinity IgE receptor (Fc epsilonRI-beta) gene. No association could be observed in this sample of Spanish asthmatics with the genes/regions studied.

Adult↗

Detection of specific antibodies to pigeon serum and bloom antigens by enzyme linked immunosorbent assay in pigeon breeder's disease.

BACKGROUND: Pigeon breeder's disease is an extrinsic allergic alveolitis in the lungs of sensitised people, caused by hypersensitivity reactions to inhaled pigeon antigens. Antigens from different sources of the animal are used for diagnostic purposes, with serum being the most widely used. Bloom is rarely used; very little is known of its antigenicity and diagnostic performance, particularly when used with the enzyme linked immunosorbent assay (ELISA) method, which is the most popular test as it permits measurement of the antibody response. METHODS: To (a) standardise an ELISA for the measurement of specific IgG against pigeon serum and pigeon bloom extract; (b) to establish reference values for specific IgG in 73 non-exposed controls, (c) to show the presence of specific IgG against pigeon serum and bloom in serum samples of 17 patients with bird fancier's lung and 11 asymptomatic fanciers, and (d) to study the similarity of the two antigen sources by cross reactivity experiments. RESULTS: Reference values of specific IgG were defined with the 97.5 percentile (367.9 U/ml for pigeon serum and 953.7 U/ml for pigeon bloom extract). Of symptomatic patients 100% had values higher than the cut off for both antigens. In asymptomatic fanciers values were higher than the cut off for pigeon serum in 45% and bloom extract in 54%. Cross reactivity experiments showed that the two antigens differed in antigenic content although some components may be common to both. CONCLUSION: The ELISA methods used proved to be useful tools for evaluating specific IgG antibody responses against both antigens. The diagnostic performance of both ELISA methods performed with these antigen sources was similar, showing very high sensitivity but moderate specificity. Although some antigenic similarity was found between pigeon serum and bloom extract, cross reactivity studies showed that various antigens seemed to be specific to the bloom extract. However, the antigens responsible for pigeon breeder's disease seem to be present in both antigenic sources.

Adult↗

An amplified ELISA inhibition method for the measurement of airborne soybean allergens.

BACKGROUND: Measurement of the soybean aeroallergen in Barcelona and other cities where soybean is unloaded is of increasing importance in controlling population exposure and evaluating the influence of such exposure on the persistence of asthma symptoms. OBJECTIVE: The aims of the study were: (1) to standardize an amplified ELISA inhibition method for the quantification of soybean aeroallergen and (2) to compare this method to a previously described RAST inhibition method. METHODS AND RESULTS: An amplified competitive ELISA inhibition method with a biotin-streptavidin system was carried out using a pool of sera from soybean-sensitized patients. The results were expressed as U/ml using a low-molecular-mass soybean allergen as reference standard. Reproducibility was calculated by statistically comparing the slope of the regression lines of the standard curve of 4 consecutive assays and by determining the coefficient of variation (CV) of the percent inhibition data for each point of several independent standard curves, each from the same assay (intra-assay) and also from a separate assay (inter-assay). No significant differences in the slopes were obtained by analysis of covariance (ANCOVA) F = 1.04. The CV between assays varied between 4 and 22% (for the assay range used in the reference standard) and was greater than the CV within assays (5-10%). Only values with a CV(%) smaller than 20% were considered acceptable. 78.5% of the samples satisfied this criterion. The RAST inhibition and ELISA inhibition methods were compared by difference plots from the values of 338 air filter eluates. The intraclass correlation coefficient was 0.456 (p < 0.001). After the results of both methods were classified as lower and higher than 165 U/m(3), the kappa index was 0.46 (p < 0.001). CONCLUSIONS: The data obtained in the present study are comparable to those reported from other similar immunoassays. Moreover, despite the difficulty in comparing air-sampling values from different laboratories, the kappa index may be taken to represent fairly good agreement beyond chance between both methods. All these data demonstrate that the present immunoassay is useful for measuring airborne soybean aeroallergens and can also be applied to evaluate the relationship between exposure and the development of asthma symptoms.

Air Pollutants↗

Utility of the antibody response to a conjugated Haemophilus influenzae type B vaccine for diagnosis of primary humoral immunodeficiency.

Antibody response to an Haemophilus influenzae type b (Hib)-conjugated vaccine was studied in 59 healthy adults (mean age: 32 yr) and 22 patients with humoral immunodeficiencies (mean age: 32 yr) to determine its usefulness in the diagnosis of defective antibody formation. Twenty of the healthy adults and nine of the patients were also immunized with a pneumococcal vaccine. Serum specific antibodies were measured by ELISA. Adequate response to both vaccines was defined using the lower limit of the two-tailed 90% probability interval of postimmunization specific IgG of the healthy adults. By using this cutoff, responders were considered to be those with an absolute increase in anti-Hib IgG titers higher than 2.28 microgram/ml, and in anti-Streptococcus pneumoniae IgG higher than 395 arbitrary units/ml. With these criteria, 85% (50 of 59) of the healthy adults responded with anti-Hib IgG and 75% (15 of 20) with anti-pneumococcal IgG. All healthy adults receiving both vaccines responded to at least one. None of the patients with humoral immunodeficiencies responded to either vaccine. Evaluation of the antibody response to both the Hib and pneumococcal vaccines may facilitate the diagnosis of humoral immunodeficiency and selection of patients to receive immunoglobulin therapy.

Adolescent↗

[Occupational asthma due to isocyanates: a study of 21 patients].

BACKGROUND: A descriptive-evolutive study of a series of patients diagnosed with bronchial occupational asthma due to isocyanate exposure. SUBJECTS AND METHODS: Retrospective study of 21 patients diagnosed with bronchial occupational asthma due to isocyanates. Family story, clinical data, functional respiratory study, allergic and laboratory tests were collected. Clinical and functional performance one year after diagnosis was evaluated in patients who had and who had not stopped causal exposure. RESULTS: 17 men and 4 women aged 22 to 59 years were evaluated. In 5 cases (23.8%) family story of atopy was documented and 12 (57.14%) patients were smokers. Seven out of 21 patients were exposed to paint related products at work being this the most common reason. Average time (SD) from the beginning of exposure to development of symptoms was 16.10 (22.5) months. IgE levels were high in 6 (28.57%) and specific IgE to isocyanates was high in 8 (38%), in all cases bronchial hyperreactivity was detected. In 18 patients (85.7%), the study of maxim expiratory flow (MEF) register demonstrated occupational asthma while in the remaining 3 patients a provocation test was performed with positive result. In the annual follow up visit, the 17 patients who avoided casual exposure showed clinical improvement, 7 of them also with functional improvement while in the remaining 10 patients no significant functional change was observed. On the other hand, 3 of the 4 patients who, although protected, underwent exposure, became clinical and functionally worse (p = 0.003, Fisher test). CONCLUSIONS: The etiology of occupational asthma due to isocyanates is not often assessed and usually lately diagnosed. This aspect is worth considering as the illness prognosis seems related to casual agent avoidance.

Adult↗

Isolation and expression analysis of two stress-responsive sucrose-synthase genes from the resurrection plant Craterostigma plantagineum (Hochst.).

The enzyme sucrose synthase (UDP-glucose: D-fructose 2alpha-glucosyltransferase, EC 2.4.1.13) is a key enzyme in carbohydrate metabolism, catalyzing the reversible conversion of sucrose uridine-diphosphate into fructose and UDP-glucose. We report the molecular characterization of two classes of cDNA and genomic clones encoding sucrose synthase from Craterostigma plantagineum Hochst., a resurrection plant in which the turnover of sucrose is considered to have an important role in the unique phenomenon of surviving desiccation. Sucrose-synthase transcript and protein levels are modulated by dehydration and rehydration. In-situ hybridization revealed that transcripts preferentially accumulate in phloem tissues. Promoter analysis underlined a role for class-I sucrose-synthase genes in dehydration stress and in response to cis-abscisic acid. A DNA sequence motif common to class-I sucrose-synthase and sucrose-phosphate-synthase genes was discovered.

Amino Acid Sequence↗

Increased positivity of skin test and allergenic stability of glycerinated soybean hull extracts.

BACKGROUND: Results obtained from previous experiences with soybean hull antigens suggest that the addition of glycerine to the extract increases positivity of the skin test. OBJECTIVE: The purpose of this study was to investigate the effect of this glycerine addition and the influence of different storage temperatures on the potency of soybean hull extracts. METHODS: Twenty-two asthmatic patients admitted to emergency rooms during one of the soybean-related Barcelona asthma epidemics were evaluated 2 years after the last epidemic for: (1) sensitivity to prick test with glycerinated and non-glycerinated extracts, both fresh and stored at 4 degrees C, - 20 degrees C and - 70 degrees C for 30 days, and (2) specific IgE to a fresh hull extract. All extracts were also studied by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: No differences in positivities were detected for prick tests performed the same day of preparation between non-glycerinated (5/22; 22.7%) and glycerinated (4/22; 18.2%) extracts. After 30 days storage, positivities of prick test with non-glycerinated extracts stored at 4 degrees C did not significantly differ from the results obtained on the first day. However, prick tests performed with glycerinated extracts stored for 30 days showed increased positivity for the extracts stored at 4 degrees C (7/22; 31%) (P < 0.05) as well as for those stored at - 20 degrees C and - 70 degrees C (9/22; 40.9%) (P < 0.001). This latter percentage is similar to that of specific immunoglobulin E detected by radioimmunoassay (10/22; 45. 4%). Also, for the glycerinated extracts only, the quantitative response showed an increase in mean diameter of the induration at 30 days at the three temperatures, even though owing to the great standard deviation and limited number of patients, it was only significant for the extract stored at - 20 degrees C (P < 0.01). SDS-PAGE gel densitometry at 30 days demonstrated a loss of protein in the bands with a molecular weight higher than 66.2 kDa for non-glycerinated extracts stored at 4 degrees C. CONCLUSIONS: The results of this study confirm previous observations with soybean hull allergen extracts and indicate that (1) storage of soybean hull extracts with glycerine addition for 30 days at the concentration studied (1:100 w/v) determines a significant increase in the positivity of skin tests, and (2) glycerine addition stabilizes soybean hull extracts at any of the temperatures studied.

Adult↗

Long term outcome of soybean epidemic asthma after an allergen reduction intervention.

BACKGROUND: Asthma outbreaks due to the inhalation of soybean dust released from handling of soybean in the city harbour occurred in Barcelona, Spain from 1981 to 1987. The installation of bag filters in the responsible silo was followed by a substantial reduction of airborne soybean dust released into the atmosphere and the disappearance of asthma outbreaks. A study was undertaken to assess the relevant outcomes in asthma patients affected by soybean epidemic asthma eight years after this environmental intervention. METHODS: A repeat case-control study was performed in 1995 on a population of subjects with epidemic and non-epidemic asthma previously assessed in 1989. The same protocol was used in both surveys to collect data from patients via a questionnaire and respiratory function, skin and laboratory tests were performed under blinded conditions with regard to epidemic and non-epidemic status. Environmental soybean allergen in pollution filters was measured by means of a RAST inhibition technique. RESULTS: During 1995 and 1996 the 24 hour mean airborne levels of soybean allergen on a sample of 39 unloading days (range 31-269 U/m(3)) were systematically below the lowest level ever detected during an epidemic day (1500 U/m(3)). Measurable levels of serum IgE antibodies against soybean were still present in 55% of patients with epidemic asthma compared with 6.0% of those with non-epidemic asthma (p<0.05). These proportions were almost identical to those observed in 1989. The proportion of patients with soybean asthma with symptoms in 1989 who reported the absence of symptoms in 1995 was similar to the control subjects, so most of the relative risks (RRs) of improvement were near to 1. The only statistically significant differences between the two groups were a smaller proportion of patients with epidemic asthma showing improvement in terms of being woken up by attacks of coughing (RR improvement 0.47; 95% CI 0.22 to 0.99) and the need for treatment at the emergency room (RR improvement 0.63; 95% CI 0.41 to 0.96). CONCLUSIONS: Eight years after a large reduction in the levels of airborne soybean allergen half of the former soybean epidemic asthma patients were still sensitised to soybean. These results indicate an initial improvement in soybean epidemic asthma in the two years following the intervention with no further improvement in subsequent years.

Allergens↗

Specific antibody response against the 23-valent pneumococcal vaccine in patients with alpha(1)-antitrypsin deficiency with and without bronchiectasis.

OBJECTIVE: To assess the specific antibody response against polyvalent pneumococcal vaccine in patients with alpha(1)-antitrypsin deficiency (AATD) and respiratory infections. DESIGN AND PARTICIPANTS: We investigated specific IgG, IgG1, and IgG2 antibody responses against the 23-valent antipneumococcal vaccine in 18 patients with AATD phenotype PiZZ, 9 of whom had bronchiectasis and 4 a history of recurrent pneumonia, and compared them with a control group of 40 healthy volunteers. INTERVENTIONS: Blood samples were drawn just prior to and 3 weeks after immunization. MEASUREMENTS AND RESULTS: Quantification of specific IgG and its subclasses was performed by an enzyme-linked immunosorbent assay. For patients with AATD, mean increases in specific antipneumococcal titers were 4.7-fold (25 to 75% quartiles, 2.5- to 6.8-fold) for total IgG, 3.2-fold (1.2- to 4.9-fold) for IgG1, and 2.1-fold (1.8- to 3.7-fold) for IgG2. For the control group, the values were 3.3-fold (1.8- to 5.8-fold) for total IgG, 2. 5-fold (1.9- to 3.4-fold) for IgG1, and 3.1-fold (1.9- to 4.5-fold) for IgG2; differences were not significant. Patients with bronchiectasis showed a tendency toward higher levels of IgG subclasses than both control subjects and patients without bronchiectasis; however, there was a tendency toward lower postvaccination serum levels of specific antipneumococcal IgG, IgG1, and IgG2 in patients with bronchiectasis compared with patients without bronchiectasis, but this trend did not reach statistical significance. Three of the four patients with recurrent pneumonia did not show an appropriate IgG2 response. CONCLUSIONS: These results suggest that, as a group, patients with AATD have a preserved antibody response against pneumococcal polysaccharides. Patients with bronchiectasis show a tendency toward a decreased antibody response, even with increased serum levels of most Ig types. Individuals with an impaired IgG2 response seem to be at increased risk of recurrent pneumonia. Considering the pernicious effect of pulmonary infections on these patients and the preserved antibody response in a majority of them, pneumococcal vaccination should be recommended to patients with AATD.

Adult↗

Blood sample processing effect on eosinophil cationic protein concentration.

BACKGROUND: Asthma is considered to be an inflammatory disease. The most important cell involved in the inflammation is the eosinophil. These cells and their mediators, such as eosinophil cationic protein (ECP), are potential markers of the inflammation's severity. Eosinophil cationic protein may be used for monitoring antiasthma treatment. It is well known that sample processing conditions can affect the ECP blood levels. OBJECTIVE: The aim of this work is to study the effect of temperature, time, and anticoagulants on ECP levels. METHODS: We studied five asthmatic patients and five healthy controls. We obtained three different blood samples from each subject, one with heparin, one with EDTA, and one without anticoagulant. To evaluate the effect of temperature, serum samples were clotted for an hour, one at 0 degree C, one at room temperature, and the other at 37 degrees C. Plasma (heparin and EDTA) samples were treated as follows: one was immediately centrifuged, and two were stored for an hour, one at 0 degree C, and the other at room temperature. Eosinophil cationic protein levels were measured by fluoroimmunoassay (CAP-System ECP FEIA Pharmacia). RESULTS: A higher temperature during blood clotting resulted in a higher ECP concentration. There were no differences between ECP determination in serum samples and plasma samples with heparin, under the same conditions of time and temperature; so clotting may not be necessary for ECP release in vitro. Eosinophil cationic protein was not released in plasma samples with EDTA, neither at 0 degree C nor room temperature. CONCLUSIONS: More studies must be done to clarify the mechanism of the ECP release in vitro.

Adult↗

Characterization of specific immunoglobulin G (IgG) and its subclasses (IgG1 and IgG2) against the 23-valent pneumococcal vaccine in a healthy adult population: proposal for response criteria.

The aim of the study was to standardize an enzyme-linked immunosorbent assay (ELISA) method for the quantification of immunoglobulin G (IgG) and its subclasses (IgG1 and IgG2) against the 23-valent pneumococcal vaccine and to establish the criteria for a normal response to the vaccine. Forty healthy individuals (20 women and 20 men; mean age, 29 years) were studied. All were vaccinated with the 23-valent pneumococcal vaccine; blood samples were drawn just prior to and 3 weeks after immunization. Quantification of specific IgG and its subclasses was performed by an ELISA with the vaccine as the antigen. The linearity of the ELISA method was demonstrated by the similar slopes of the linear regression lines generated from the titration of sera with different antibody concentrations. The specificity of the antibodies against the vaccine was demonstrated by (i) an absorption test with pneumococcal vaccine, (ii) a cross-reactivity experiment with Haemophilus influenzae type b polysaccharide, and (iii) affinity chromatography with protein A-Sepharose. Response to the vaccine was defined by using the lower level of the 90% probability interval (one-tailed) for postimmunization-specific IgG, IgG1, and IgG2. By using this cutoff, responders were considered to be those with an absolute increase in antibody titers higher than 395 arbitrary units/ml for IgG, 0.350 A450 units for IgG1, and 0.314 A450 units for IgG2. Overall, 20 (50%) subjects had IgG, IgG1, and IgG2 responses, 9 (22.5%) had IgG and IgG2 responses, 4 (10%) had IgG1 responses, 3 (7.5%) had IgG and IgG1 responses, and 4 (10%) were nonresponders. Ninety percent of our population responded to the 23-valent pneumococcal vaccine. Up to 10% of healthy individuals may respond to an IgG subclass without significant increases in total IgG titers. The ELISA method that is described may be useful for evaluating the specific antibody response against polysaccharides.

Adult↗

Survival prognostic factors and markers of morbidity in Spanish patients with systemic sclerosis.

OBJECTIVE: To identify survival prognostic factors and markers of morbidity among patients with systemic sclerosis (SSc). PATIENTS AND METHODS: The study included 72 patients diagnosed with SSc. According to the extent of skin involvement, three groups of patients were established: group 1, without sclerosis and with sclerosis of fingers and neck; group 2, with sclerosis of face and distal to elbows and knees; group 3, with generalised sclerosis including the trunk. All patients were included in a study protocol to determine visceral involvement. Cumulative survival after first symptom has been estimated according to the Kaplan-Meier method. The association between a hypothetical prognostic factor and cumulative survival after first symptom was assessed by log rank test. The association between a hypothetical risk factor and the prevalence of severe morbidity was assessed by the odds ratio. Multiple logistic regression models were used to identify the main predictors of severe morbidity. RESULTS: Survival was estimated to be 85% 10 years after first SSc symptom. Survival was higher among SSc patients with skin involvement distal to elbows and knees than among the rest of patients; a forced vital capacity (FVC) on spirometry lower than 70% of expected value was associated with a shorter survival, even after adjustment for diffuse SSc. Skin involvement proximal to elbows or knees was associated with a higher prevalence of severe morbidity (OR = 46.57; p < 0.001). According to a multiple logistic regression, severe morbidity was higher among patients with skin involvement proximal to knees or elbows (OR = 40.92; p < 0.001) or among patients with pulmonary hypertension detected by Doppler echocardiography (OR = 23.66 p < 0.001). CONCLUSIONS: In patients with SSc the extent of skin sclerosis was found to be a determining factor on the prognosis. According to skin sclerosis extent two main subsets of SSc patients with different survival incidence and degree of morbidity could be clearly established: limited SSc, formed by patients with no skin sclerosis or with sclerosis distal to elbows and knees and diffuse SSc, formed by patients with skin sclerosis distal and proximal to elbows and knees. Moreover, lung involvement (FVC < 70% on survival study and pulmonary hypertension on morbidity study) was an important and independent prognostic factor.

Adolescent↗

HLA class II genes in soybean epidemic asthma patients.

From 1981 to 1987, 26 outbreaks of asthma caused by the inhalation of soybean dust, affecting a total of 688 individuals, were detected in Barcelona, Spain. Because only a small proportion of asthmatic individuals living in Barcelona expressed the epidemic phenotype, it is hypothesized that a genetically determined human leukocyte antigen (HLA) Class II factor could have played a role in the susceptible individuals. Accordingly, we studied the distribution of both HLA-DR and HLA-DQ in soybean epidemic asthmatic patients. An analysis of the HLA-DR and HLA-DQ genes for genetic polymorphisms of the beta 1 chain was done with the polymerase chain reaction (PCR) in 78 soybean epidemic asthma patients, and the findings were compared with those for 67 nonepidemic asthmatic individuals and 168 individuals from the general population. An allelic disequilibrium could be established; the risk of epidemic asthma was particularly associated with the DRB1*13 gene (p value corrected for multiple comparisons < 0.02). The association observed for the DRB1*13 gene was stronger in individuals in the lowest tertile for total IgE, with an estimated risk with a 95% confidence interval (CI), of 14.5 (1.6 to 130.8). The combination of two genes from among the DRB1*05-05, DRB1*05-06, and DRB1*06-06 genes was present in epidemic asthmatic subjects only. No association with an HLA-DQB1 allele could be observed. Genetic predisposition could contribute to the response of some asthmatic patients to exposure to soybean dust, having led to their being affected during the epidemics of asthma in Barcelona.

Alleles↗

High prevalence of mollusc shell hypersensitivity pneumonitis in nacre factory workers.

Following the discovery of hypersensitivity pneumonitis caused by the inhalation of mollusc shell dust in two workers from a nacre-button factory, the health status of 26 workers employed in sawing mollusc shells was investigated. The evaluation included the administration of two questionnaires and radiological, functional and immunological assessments of all workers at the outset and 1 year later, when hygienic and therapeutic measures had been taken. Six workers, in whom specific inhalation challenge test was positive, were diagnosed with mollusc shell hypersensitivity pneumonitis, thus yielding a prevalence of 23%. Evidence of diffuse lung disease and systemic symptoms was found in these patients. Nonspecific bronchial hyperreactivity was also found more frequently in patients with mollusc shell hypersensitivity pneumonitis. Specific immunoglobulin G (IgG) level and specific skin testing failed to differentiate patients with mollusc shell hypersensitivity pneumonitis from other exposed workers; whereas, nonspecific skin testing, which was impaired in the patients, did differentiate. Bronchoalveolar lavage and transbronchial biopsy performed in patients with mollusc shell hypersensitivity pneumonitis were consistent with the disease. Removal from an environment containing mollusc shell dust was followed by regression of clinical, radiological and functional changes. The clinical picture of the 20 workers who did not present mollusc shell hypersensitivity pneumonitis remained unchanged, but functional decline was observed despite improvement in the environmental conditions of the factory. This report describes the first series of patients with mollusc shell hypersensitivity pneumonitis studied, and underlines the importance of careful follow-up of workers occupationally-exposed to mollusc shell dust.

Adult↗

IgG subclass deficiencies associated with bronchiectasis.

Only a small number of patients with IgG subclass deficiencies (IgGSD) have been observed to have bronchiectasis. Moreover, in the series of patients with bronchiectasis, IgGSD have not been found at any frequency, and the etiology of bronchiectasis remains unclear in 29 to 49% of cases. Serum concentrations of total IgG, IgA, and IgG subclasses as well as pulmonary function were measured in 65 patients (aged: 10 to 74 yr) with bronchiectasis of unknown etiology. An ELISA test was performed to quantify subclasses 1 through 4 using subclass-specific antihuman monoclonal antibodies. IgG subclass estimation in a healthy population with age-stratified normal ranges was derived from 100 adults, 37 children aged between 10 and 12 yr, and 27 adolescents aged between 13 and 16 yr. Serum concentrations of specific IgG antibodies to Haemophilus influenzae type b capsular polysaccharide (Hib-PRP) were also assayed by an ELISA test in 19 of the patients (10 with IgGSD and nine with non-IgGSD) and in 58 healthy individuals before and 3 wk after immunization with Hib-PRP conjugated to meningococcal outer membrane protein complex (OMPC). Thirty-one patients (48%) had low serum concentrations of one or more IgG subclasses (19 IgG2 deficiencies, 2 IgG3 deficiencies, 3 IgG4 deficiencies, and 7 combined subclass deficiencies). All patients showed increased levels of total IgG, IgG1, and IgA, but this rise was significantly higher in patients without IgGSD. Patients with IgGSD showed impaired antibody response to Hib-PRP compared with patients with non-IgGSD and the control group. IgGSD, particularly IgG2 deficiency, are not an unusual cause of bronchiectasis. Therefore, serum levels of IgG subclasses must be assayed whenever other causes of bronchiectasis have been ruled out.

Adolescent↗

Effect of conditions in obtaining blood samples for ECP testing in children.

Eosinophil Cationic Protein (ECP) is a basic protein found in eosinophil granules. This cell and its mediators are currently considered to be potential indicators of the severity of inflammation in the organism. ECP concentration can be reliably tested using several RIA or ELISA methods. It is well known that the conditions of sample obtention can affect the ECP values in blood. The aim of this study is to establish which parameters affect ECP testing during regular blood sample collection and how they affect it. Blood samples taken for the routine study of five children attended in our department were analysed: four were asthmatic and one child had atopic dermatitis. In the results we observed that ECP was not detected in the blood samples taken with EDTA tripotassium. In both the plasma samples taken with heparin as well as with serum, more ECP was released at a higher temperature. In the release of ECP obtained by coagulation, samples at 37 degrees showed values of between 4 and 20 higher than those obtained for an hour at 0 degrees. There is a considerable variability in the testing of ECP depending on the blood test extraction conditions, the range is bigger in the samples with eosinophils. These results imply the need to define a stricter protocol for obtaining samples than that suggested at present.

Adolescent↗