33rd EUROPEAN CYSTIC FIBROSIS CONFERENCE
16 ‒ 19 JUNE 2010, VALENCIA, SPAIN
©2010 European Cystic Fibrosis Society. All rights reserved. Produced by Elsevier B.V.
16 ‒ 19 JUNE 2010, VALENCIA, SPAIN
©2010 European Cystic Fibrosis Society. All rights reserved. Produced by Elsevier B.V.
1. Genetics - poster presentation
Abstract: 14
Citation: Journal of Cystic Fibrosis 2010; Vol. 9, Supplement 1, page S4
Association of clinical severity of cystic fibrosis (CF) with polymorphism D/I in ACE gene
F.A.L. Marson1, J.D. Ribeiro2, C.S. Bertuzzo1, A.F. Ribeiro2
1 UNICAMP, Genetics, Campinas, Brazil
2 UNICAMP, Pediatrics, Campinas, Brazil
The ACE gene encodes angiotensin-converting enzyme is an enzyme related to pro-inflammatory response. This gene presents a polymorphism called D/I, the D allele is characterized by the deletion of 287pb in intron 16 and responsible for higher expression of the gene.
Objective: To link the polymorphism D/I in ACE gene with the clinical markers of CF (Shwachman score, body mass index-BMI, age at diagnosis, early pulmonary and gastrointestinal symptoms, microorganisms present, spirometry, oxygen saturation).
Method: 181CF patients, 95 (52.6%) males, mean age 15 years. For the analysis of polymorphism D/I was used the technique of PCR. Statistical analysis was performed by SPSS v.10.0. The analysis of quantitative data was performed by analysis of variance for a factor and qualitative data by logistic regression test and Odds Ratio (OR).
Results and Discussion: For patients with allele D, the diagnosis was made on average before the three years of age when compared to individuals homozygous II (OR: 3.07, CI = 1.1-2.6). As for the appearance of the table digestive same occurred (OR: 8.2, CI = 1.4-1.5). As for the Shwachman score, was observed a greater number of patients with genotype DD with scores classified as severe (OR: 6.4, CI = 1.2-34.2).
These results suggest that increased expression of ACE resulting of DD genotype leads to greater lung damage due to inflammation. But the genotype DD seems to protect against chronic infection that can be observed by the presence of greater numbers of patients with genotype II infected by the bacteria Achromobacter xylosoxidans (OR: 4.5, CI = 1.2-17.1; RR: 3.5, CI = 1.3-9.7).
Conclusion: The polymorphism D/I in the ACE gene acts as modifier in CF.
7. Pulmonology - poster presentation
Abstract: 231
Citation: Journal of Cystic Fibrosis 2010; Vol. 9, Supplement 1, page S60
Evaluation of airway resistance through the interrupter technique in cystic fibrosis
P. Marostica1,2, A. Rocha2,3, P. Hommerding3, T. Paim3, M. Donadio3
1 Hospital São Lucas PUCRS, Pediatric Pulmonology, Porto Alegre RS, Brazil
2 Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil
3 Hospital São Lucas da PUCRS, Porto Alegre, Brazil
Few studies have been published on airway resistance measurements using the interrupter technique (IT). We performed a cross-sectional study, evaluating 38 children and adolescents with Cystic Fibrosis (CF), followed at the outpatient CF clinic of Hospital São Lucas from Pontifícia Universidade Católica do Rio Grande do Sul. Airway resistance (Rint) was measured by the IT, followed by spirometry in all patients. Measurements were repeated after inhalation of salbutamol in order to evaluate bronchodilator response.
There was a strong corelation between inverse Rint and forced expiratory volume in one second (FEV1) (r = 0.8, p < 0.001) and fair correlations between the inverse Rint and mid expiratory flow (MEF) (r = 0.74 p < 0.001) and between inverse Rint and body mass index (BMI) (r = 0.62 p < 0.001). The accuracy of bronchodilator esponse by the IT was tested through the ROC (reciever operating curve), comparing results with spirometry bronchodilator response. An area of 0.75 under the curve was obtained, for the cutoff point of -28% of Rint, achieving a sensitivity of 66% and a specificity of 82%.
The findings suggest that Rint shows good correlation with spirometry parameters, although the IT is not sufficiently acurate to replace spirometry in the evaluation of bronchodilator response.
5. Microbiology - poster presentation
Abstract: 104
Citation: Journal of Cystic Fibrosis 2010; Vol. 9, Supplement 1, page S27
Genomovar prevalence of Burkholderia cepacia complex (BCC) in a cystic fibrosis reference center in Brazil
P. Dentini1, C.E. Levy2, C.S. Bertuzzo3, L.C. Bonadia3, A.F. Ribeiro1, J.D. Ribeiro1
1 State University of Campinas, UNICAMP, Department of Pediatrics, Campinas, Brazil
2 State University of Campinas, UNICAMP, Department of Clinical Pathology, Campinas, Brazil
3 State University of Campinas, UNICAMP, Department of Medical Genetics, Campinas, Brazil
Introduction: B. cenocepacia and B. multivorans has been described as the most important BCC genomovars for lung function deterioration in CF patients. The aim of this study was to assess the prevalence and genomovar distribution of BCC among CF patients seen at the outpatient pediatric CF reference center.
Methods: In the last 4 years 2768 samples from respiratory tract of 217 CF patients were obtained (3 cultures/patient/year). The samples were grown in non-selective and selective medium, including the BCSA and phenotypically identified by biochemical tests and the Vitek®II. Subsequently the genomic DNA was extracted and BCC was identified by PCR recA gene amplification, using BCR1 and BCR2 primers.
A second amplification was carried out by nested-PCR method, with BCC genomovars specific primers.
Results and Discussion: BCC prevalence was 20.7% (45/217), 31.1% (14/45) were transient colonization, chronic 26.7% (12/45), 8.9% intermittent (4/45), 26.7% of new cases (12/45) and 6.6% (3/45) were excluded due to patient follow-up loss. Although other studies published in Brazil and other American and European countries have found prevalence of genomovar III - Burkholderia cenocepacia, in this study
the genomovar II - B. multivorans was the most prevalent (26.7%), followed by genomovar I - B. cepacia (24.5%) and genomovar III - B. cenocepacia (8.9%). We could not identify the genomovar from 13 patients and failed to recover the agent for 12 patients. Among patients with chronic colonization was observed the presence of more than one genomovar in 13.3% (6/45). The BCSA selective medium used after 2006 allowed to detect early colonization in 26.7% of patients (12/45).
1. Genetics - poster presentation
Abstract: 15
Citation: Journal of Cystic Fibrosis 2010; Vol. 9, Supplement 1, page S4
Polymorphisms in GCLC and GST(M1, T1 and P1) genes and their performance in clinical severity of cystic fibrosis (CF)
F.A.L. Marson1, J.D. Ribeiro2, C.S. Bertuzzo1, A.F. Ribeiro2
1 UNICAMP, Genetics, Campinas, Brazil
2 UNICAMP, Pediatrics, Campinas, Brazil
For the present study were chosen candidate genes related to modifying the mechanism of action of glutathione (GSH).
Objective: To link the presence of polymorphisms (pol.) of GST and GCLC genes with the degree of seriousness.
Method: 181 patients, 52.6% males, mean age: 15 years. We used the technique of PCR in the genes GSTM1 and T1 for the GSTP1 (pol.313A/G) and GCLC (pol.129C/T and 350A/G) genes was also used restriction enzyme. The data were correlated with: Kanga (EK) and Shwachman (ES) score, BMI, age at diagnosis, onset of symptoms, isolated microorganisms, spirometry and SaO2. Statistical analysis was performed by analysis of variance for a factor, logistic regression test and Odds Ratio (OR).
Results: For the GSTM1, GCLC (pol.129C/T) and GSTP1 gene weren`t statistically significant correlation with clinical markers. For the GSTT1 gene, individuals with at least one allele coding, were classified in the category of low BMI (OR:3.1, CI=1.4-7.1), when GSTM1 and T1 genes were analyzed simultaneously, the same occurred (OR:1.5, CI=1.1-2.7). Patients with M1 and T1 genes with null alleles had the lowest SaO2 (OR:4.3, CI=1.2-20.1) and worst ranking for ES (OR:9.0, CI=1.5-55.1). Regarding the association of bacterial colonization to null allele for T1 and coding to M1 was related to the presence of Pseudomonas aeruginosa mucoid (OR:3.1, CI=1.3-7.6) and non mucoid (OR:4.8, CI=1.7-13.7). For GCLC350(A/G) pol. of the GCLC gene was found a association to genotype A/A with SaO2 (OR:5.8, CI=2.3-14.5) and with worse rating for EK, lower value of FEF25-75% and FEV1/FVC, and more severe regarding FEV1 (OR:4.6, CI=1.3-5.2).
Conclusion: GSTM1, GSTT1 and GCLC (pol.350A/G) genes act as modifiers in CF.
5. Microbiology - poster presentation
Abstract: 133
Citation: Journal of Cystic Fibrosis 2010; Vol. 9, Supplement 1, page S35
Virulence properties of Achromobacter xylosoxidans: cytotoxic activity, interleukin production, biofilm formation and invasion
R.P. Mantovani1, C.E. Levy2, C.Z. Esteves2, J.D. Ribeiro3, A.F. Ribeiro3, D.M. Bernardi3, T. Yano1
1 State University of Campinas, UNICAMP, Biology Institute, Campinas, Brazil
2 State University of Campinas, UNICAMP, Department of Clinical Pathology, Campinas, Brazil
3 State University of Campinas, UNICAMP, Department of Pediatrics, Campinas, Brazil
Achromobacter xylosoxidans has been associated with cystic fibrosis (CF), but its role on human pathogenicity is still not known. Recently we described the cytotoxic activity of cultured supernatants (CS) of A. xylosoxidans to human lung cells (H-460). Among the proteinase studied, lecithinase was the only one detected. Now, the culture supernatants of A. xylosoxidans also showed cytotoxic activity on NCIH292
(human lung mucoepidermoid carcinoma) and this cytotoxic acitivity was not afected after heat treatment (100°C for 20 min), showing it to be thermo stable.
The protein fractions obtained by ultrafiltration (Amiconâ), showed biological activity of the extracts with molecular weigh lower than 5 kDa. This fraction has also stimulated the IL 8 production by these lung cells of 5 folds more than that detected on the negative controls, determined by ELISA (DuoSet, RD Systems, EUA. These data suggests that the thermo stable cytotoxic activity (cytotoxin), is probably the stimulating agent of the proinflamatory cytokine (IL) that may be related to the pathogenicity for lung cells. A. xylosoxidans showed biofilm formation ability on plastic microplates among all the isolates, as verified by flat-bottom microtiter polystyrene plates test. Invasion and intracellular survival in NCI-H292 cells was also observed. The detection of a cytotoxic factor inducing IL 8 production, the bacteria biofilm formation and invasion reinforces the proposal that A. xylosoxidans may present a relevant role in the lung parenchyma destruction in cystic fibrosis patients.
