Analyses of association between PPAR gamma and EPHX1 polymorphisms and susceptibility to COPD in a Hungarian cohort, a case-control study

<p>Abstract</p> <p>Background</p> <p>In addition to smoking, genetic predisposition is believed to play a major role in the pathogenesis of chronic obstructive pulmonary disease (COPD). Genetic association studies of new candidate genes in COPD may lead to improved understanding of the pathogenesis of the disease.</p> <p>Methods</p> <p>Two proposed casual single nucleotide polymorphisms (SNP) <it>(rs1051740, rs2234922) </it>in microsomal epoxide hydrolase (<it>EPHX1</it>) and three SNPs <it>(rs1801282, rs1800571, rs3856806) </it>in peroxisome proliferator-activated receptor gamma (<it>PPARG</it>), a new candidate gene, were genotyped in a case-control study (272 COPD patients and 301 controls subjects) in Hungary. Allele frequencies and genotype distributions were compared between the two cohorts and trend test was also used to evaluate association between SNPs and COPD. To estimate the strength of association, odds ratios (OR) (with 95% CI) were calculated and potential confounding variables were tested in logistic regression analysis. Association between haplotypes and COPD outcome was also assessed.</p> <p>Results</p> <p>The distribution of imputed <it>EPHX1 </it>phenotypes was significantly different between the COPD and the control group (P = 0.041), OR for the slow activity phenotype was 1.639 (95% CI = 1.08- 2.49; P = 0.021) in our study. In logistic regression analysis adjusted for both variants, also age and pack-year, the rare allele of His447His of <it>PPARG </it>showed significant association with COPD outcome (OR = 1.853, 95% CI = 1.09-3.14, P = 0.0218). In haplotype analysis the GC haplotype of <it>PPARG </it>(OR = 0.512, 95% CI = 0.27-0.96, P = 0.035) conferred reduced risk for COPD.</p> <p>Conclusions</p> <p>The "slow" activity-associated genotypes of <it>EPHX1 </it>were associated with increased risk of COPD. The minor His447His allele of <it>PPARG </it>significantly increased; and the haplotype containing the minor Pro12Ala and the major His447His polymorphisms of <it>PPARG </it>decreased the risk of COPD.</p

Liquid phase separation methods for N-glycosylation analysis of glycoproteins of biomedical and biopharmaceutical interest. A critical review

Comprehensive carbohydrate analysis of glycoproteins from human biological samples and biotherapeutics are important from diagnostic and therapeutic points of view. This review summarizes the current state-of-the-art liquid phase separation techniques used in N-glycosylation analysis. The different liquid chromatographic techniques and capillary electrophoresis methods are critically discussed in detail. Miniaturization of these methods is also important to increase throughput and decrease analysis time. The sample preparation and labeling methods for asparagine linked oligosaccharides are also addressed

Impact of education on dimensions of adherence in patients with chronic obstructive pulmonary disease

Objectives To assess the impact of patient education on medication adherence and quality-of-life (QoL) in Hungarian subjects with chronic obstructive pulmonary disease (COPD). Study Design Longitudinal, non-interventional study conducted at three pulmonology outpatient centers in and around Budapest, Hungary. Experimental Subjects visiting the center with COPD were invited to participate in the study. Data collected at baseline included subject demographics, medical history, and responses to the adherence (Morisky Medication Adherence Scale-8 (MMAS-8)) and QoL (EuroQoL-5D-5L (EQ5D), St. George’s Respiratory Questionnaire (SGRQ), and COPD Assessment Tool (CAT)) scales. Subjects were also provided with patient education designed with standardized content. Subjects were asked to provide responses to adherence and QoL scales again at the 3-month follow-up visit. Medication was left unchanged during the course of study. Statistical analysis included independent and paired-samples t-tests, one-way ANOVA, mixed-measures ANOVA and ANCOVA. Results Mean (± standard deviation (SD)) overall adherence score on MMAS-8 scale increased from 6.72 (± 1.46) at baseline to 7.01 (± 1.15) at follow-up (P = .040). A similar increase in mean (± SD) score was observed for question 4 on the MMAS-8 which deals with remembering to carry COPD medication when leaving house (baseline = 0.81 (± 0.40) versus follow-up = 0.89 (± 0.31); P = 0.018). Conclusions Patient education has a positive outcome on medication adherence in subjects with COPD. Further studies will be required to assess if these benefits are translated to patients’ QoL

Capillary electrophoresis analysis of N-glycosylation changes of serum paraproteins in multiple myeloma

Multiple myeloma (MM) is an immedicable malignancy of the human plasma cells producing abnormal antibodies (also referred to as paraproteins) leading to kidney problems and hyperviscosity syndrome. In this paper we report on the N-glycosylation analysis of paraproteins from total human serum as well as the Fc and Fab κ/λ light chain fractions of papain digested immunoglobulins from multiple myeloma patients. Capillary electrophoresis with laser induced fluorescence detection (CE-LIF) was used for the analysis of the N-glycans after endoglycosidase (PNGase F) mediated sugar release and fluorophore labeling (APTS). While characteristic N-glycosylation pattern differences were found between normal control and untreated, treated and remission stage multiple myeloma patient samples at the global serum level, less distinctive changes were observed at the immunoglobulin level. Principal component analysis adequately differentiated the four groups (control and three patient groups) on the basis of total serum N-glycosylation analysis. 12 N-glycan features showed statistically significant differences (p<0.05) among various stages of the disease in comparison to the control at the serum level, while only 6 features were identified with similar significance at the immunoglobulin level, including the analysis of the partitioned Fc fragment as well as the Fabκ and Fabλ chains

Molecular glycopathology by capillary electrophoresis: Analysis of the N-glycome of formalin-fixed paraffin-embedded mouse tissue samples

Capillary electrophoresis with laser-induced fluorescence (CE-LIF) detection was used to analyze endoglycosidase released and fluorophore-labeled N-glycans from formalin-fixed paraffin-embedded (FFPE) mouse tissue samples of lung, brain, heart, spleen, liver, kidney and intestine. The FFPE samples were first deparaffinized followed by solubilization and glycoprotein retrieval. PNGase F mediated release of the N-linked oligosaccharides was followed by labeling with aminopyrene trisulfonate. After CE-LIF glycoprofiling of the FFPE mouse tissues, the N-glycan pool of the lung specimen was subject to further investigation by exoglycosidase array based carbohydrate sequencing. Structural assignment of the oligosaccharides was accomplished by the help of the GUcal software and the associated database, based on the mobility shifts after treatments with the corresponding exoglycosidase reaction mixtures. Sixteen major N-linked carbohydrate structures were sequenced from the mouse lung FFPE tissue glycome and identified, as high mannose (3) neutral biantennary (3) sialylated monoantennary (1) and sialylated bianennary (9) oligosaccharides. Two of these latter ones also possessed alpha(1-3) linked galactose residues

Chronic obstructive pulmonary disease-specific gene expression signatures of alveolar macrophages as well as peripheral blood monocytes overlap and correlate with lung function

Chronic obstructive pulmonary disease (COPD) is a chronic inflammatory disease characterized by progressive airflow limitation and significant extrapulmonary (systemic) effects that lead to co-morbid conditions, though the pathomechanism of COPD is largely undetermined. Alveolar macrophages (AM) derived from peripheral monocytes (MO) appear to play a key role in initiating and/or sustaining disease progression

Monoclonal antibody proteomics: discovery and prevalidation of chronic obstructive pulmonary disease biomarkers in a single step

We define mAb proteomics as the global generation of disease specific antibodies that permit mass screening of biomarkers. An integrated, high-throughput, disease-specific mAb-based biomarker discovery platform has been developed. The approach readily provided new biomarker leads with the focus on large-scale discovery and production of mAb-based, disease-specific clinical assay candidates. The outcome of the biomarker discovery process was a highly specific and sensitive assay, applicable for testing of clinical validation paradigms, like response to treatment or correlation with other clinical parameters. In contrast to MS-based or systems biology-based strategies, our process produced prevalidated clinical assays as the outcome of the discovery process. By re-engineering the biomarker discovery paradigm, the encouraging results presented in this paper clearly demonstrate the efficiency of the mAb proteomics approach, and set the grounds for the next steps of studies, namely, the hunt for candidate biomarkers that respond to drug treatment.6 page(s