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M J Rodrigo

Publications and source records attributed to M J Rodrigo.

At least 19 recordsLinked to original sources
PubMed1999

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↗
PubMed1999

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↗
PubMed1999

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↗
PubMed1997

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↗
PubMed1997

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↗
PubMed1997

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↗
PubMed1997

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↗
PubMed1997

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↗
PubMed1996

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↗
PubMed1996

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↗
PubMed1995

[Espartosis. A new type of extrinsic allergic alveolitis among construction plasterers].

The clinical characteristics of three plasterers in contact with esparto and who presented a picture of extrinsic allergic alveolitis which was peculiar because of their being hyperacute and causing hypoxemia. In addition to a review of 5 previously published sporadic cases the clinical picture of this new type of apparently infrequent alveolitis is described. The causal antigen is unknown although contamination by fungus or the addition of raw esparto may be the origin of the disease. After a review of the literature regarding this subject, the authors suggest that, in addition to cases of asthma and chronic airway obstruction, the patients with extrinsic allergic alveolitis be included under the name of espartosis following esparto dust inhalation.

Adult↗
PubMed1995

The value of complement activation products in the assessment of systemic lupus erythematosus flares.

Complement activation products (CAP) have been reported as sensitive markers of disease activity in systemic lupus erythematosus (SLE). We have measured seven parameters of the complement system (C3, C4, factor B, C3a, C4a, iC3b, and the terminal complement complex-TCC-) in 61 SLE patients to study their interrelationship and relative efficacy as diagnostic indicators of lupus activity. Disease activity was judged according to a clinical index (SLEDAI) to be active in 22 and inactive in 39 patients. Subjects with active SLE showed increased levels of C3a, C4a, and TCC compared with those of stable lupus and normal controls, and plasma concentrations of these CAP manifested a positive correlation with disease activity scores. However, values of factor B and iC3b did not correlate with lupus flares. Serum C3 levels were a better reflection of the degree of SLE activity than were C4 levels. The anaphylatoxins were extremely sensitive markers of disease activity but they lacked enough specificity, and iC3b was not at all informative for this purpose. On the whole, TCC concentration was the most useful parameter (77% sensitivity, 80% specificity) to monitor lupus activity, correlating the best with the activity scoring system, and thus offers a better laboratory marker of lupus severity than conventional measurements of complement.

Adolescent↗
PubMed1995

Evolutionary analysis of the picornavirus family.

An exhaustive evolutionary analysis of the picornavirus family has been carried out using the amino acid sequences of several proteins of the viruses including: the capsid proteins (1D, 1B, and 1C) situated at the 5' end of the genome and responsible for the serotype of the viruses, and the viral polymerase (3D), located at the 3' end of the genome. The evolutionary relationships found among the viruses studied support the new classification, recently suggested, in contrast to the classical one, and the existence of a new genus for the picornavirus family. In the new taxonomic organization, five genera form the picornavirus family: (1) aphthoviruses, (2) cardioviruses, (3) hepatoviruses (previously classified as enteroviruses), (4) renteroviruses (which mainly constitute a combination of the previous genera rhinovirus and enterovirus), and (5) a new genus, with a new and unique representative: the echovirus 22. Our analysis also allowed us, for the first time, to propose the most probable sequence of speciation events to have given rise to the current picornavirus family. The bootstrap procedure was used to check the reliability of the phylogenetic trees obtained. The application of the method of the statistical geometry in distance space to internal branches of the tree revealed a high degree of evolutionary "noise," which makes the resolution of some internal branching points difficult.

Capsid↗
PubMed1995

Diagnosis of soybean-induced asthma.

BACKGROUND: The role of soybean dust as a causal agent of asthma has been clearly established since the Barcelona asthma epidemics in the 1980s. The large number of patients who were first seen with asthma symptoms during those epidemics provided an excellent opportunity to study the possibilities of different diagnostic tests. OBJECTIVE: This study was designed to evaluate the usefulness of the skin test and amplified ELISA technique for quantifying specific IgE in the diagnosis of soybean asthma. PATIENTS AND RESULTS: Ninety epidemic asthmatic patients and 95 nonepidemic asthmatic patients were studied 2 years after the last epidemic. Results of prick tests and ELISAs for specific IgE with hull and dust extracts showed a significant difference between the two groups of patients (p < 0.001). Sensitivity and specificity of glycerinated prick test with hull extract in epidemic asthmatic patients were 57.7% and 95.3%, respectively, and ELISA values were 56.6% and 93.7%, respectively. Similar results were obtained with dust extracts. CONCLUSION: Glycerinated skin prick tests and ELISAs with soybean hull and dust extracts have proved effective in the diagnosis of soybean asthma, even 2 years after the epidemics. Taking into account the sensitivity (90.5%) and specificity (93.7%) of ELISA test results for epidemic asthmatic patients found when the epidemic occurred, data from this study suggest that both tests may be very useful for the diagnosis of soybean dust-induced asthma.

Asthma↗
PubMed1995

Relationship between serum IgE and airway responsiveness in adults with asthma.

BACKGROUND: General population studies have shown a relationship between total serum IgE levels and airway responsiveness, but this association has not been documented in patients with asthma. OBJECTIVE: The study assesses the cross-sectional relationship between IgE levels and airway responsiveness in 208 subjects who had had emergency department treatment for asthma at least 2 years earlier. METHODS: All participants completed a standardized respiratory questionnaire and underwent spirometry, allergy skin testing, and a dose-response methacholine challenge test. RESULTS: After adjusting for age and gender, the percentage of patients with asthma and airway responsiveness (provocative concentration causing a 20% fall in forced expiratory volume in 1 second [PC20] < or = 8 mg/ml) increased from 52% in the lower quintile of IgE to 72% in the upper quintile (p < 0.01). After adjusting for age, gender, baseline percent predicted forced expiratory volume in 1 second, and smoking, the association between IgE (both in quintiles and continuous) and PC20 appeared consistent and statistically significant (p < 0.01). This association was stronger in patients who were not given inhaled steroid (odds ratio for twice the concentration of IgE, 1.42; 95% confidence interval, 1.09 and 1.84), than in patients treated with inhaled steroid (odds ratio, 1.10; 95% confidence interval, 0.82 and 1.50). Eosinophilia and skin reactivity were associated with PC20 although to a lesser extent. CONCLUSION: These findings strengthen the role played by IgE in facilitating the development of bronchial responsiveness in patients with asthma.

Administration, Inhalation↗