Quantitative analysis of the dystrophin gene by real-time PCR

Duchenne and Becker muscular dystrophy (DMD/BMD) are severe X-linked neuromuscular disorders caused by mutations in the dystrophin gene. Our aim was to optimize a quantitative real-time PCR method based on SYBR® Green I chemistry for routine diagnostics of DMD/BMD deletion carriers. Twenty female relatives of DMD/BMD patients with previously detected partial gene deletions were studied. The relative quantity of the target exons was calculated by a comparative threshold cycle method (ΔΔCt). The carrier status of all subjects was successfully determined. The gene dosage ratio for non-carriers was 1.07±0.20, and for carriers 0.56±0.11. This assay proved to be simple, rapid, reliable and cost-effective

Relationship between myocardial viability and improvement in left ventricular function and heart failure symptoms after coronary artery bypass surgery.

BACKGROUND The evaluation of myocardial viability is an important preoperative parameter, predictive of improvement in regional and global left ventricular (LV) function after coronary artery bypass surgery (CABG). However, whether the presence of viability is also associated with relief of heart failure symptoms after revascularization is not always certain. The aims of the study were to define the relationship between extent of viable myocardium and improvement in LV function after CABG and to determine whether preoperative viability testing can predict improvement in heart failure symptoms. METHODS Eighty-five consecutive patients with ischemic cardiomyopathy (mean LVEF 35%) undergoing surgical revascularization were studied with a Tc-99m sestamibi one-day rest/nitrate enhanced myocardial perfusion SPECT imaging (MPI) to assess viability. Regional and global function were measured before and 16 -/+ 6 months after revascularization. We have used the Bull's eye quantitative analysis of MPI scans and 17 segment model of LV function and perfusion evaluation. Heart failure symptoms were graded according to the New York Heart Association (NYHA) criteria, before and 16 -/+ 6 months after revascularization. RESULTS The number of viable segments per patient was directly related to the improvement in LVEF after revascularization (r 0.79, P 4 viable segments representing 24% of the left ventricle yielded the sensitivity of 83% and specificity of 79% respectively for predicting improvement in LVEF. Furthermore, the presence of four or more viable segments predicted improvement in heart failure symptoms after revascularization, with positive and negative predictive values of 79% and 74%, respectively. CONCLUSION The presence of substantial viability (four or more viable segments, 24% of the left ventricle) on myocardial perfusion gated SPECT imaging in patients with ischemic heart failure before CABG surgery has significant correlation with the improvement in LVEF and heart failure symptoms postoperatively

Galectin 3 (LGALS3) Gene Polymorphisms Are Associated with Biochemical Parameters and Primary Disease in Patients with End-Stage Renal Disease in Serbian Population

Galectin 3 plays a significant role in the development of chronic renal failure, particularly end-stage renal disease (ESRD). The aim of our study was to investigate the association between Gal-3 and biochemical parameters and primary disease in ESRD patients, by exploring the polymorphisms LGALS3 rs4644, rs4652, and rs11125. A total of 108 ESRD patients and 38 healthy controls were enrolled in the study. Genotyping of LGALS3 gene rs4644, rs4652, and rs11125 polymorphisms was performed by polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP). By multivariate logistic regression analysis, we found that LGALS3 rs4644 CC and rs4652 AA genotypes were significantly associated with a higher risk for lower hemoglobin, higher level of parathyroid hormone, and also occurrence of diabetes mellitus and arterial hypertension. The CAA haplotype was significantly more common in patients with diabetes, low hemoglobin level, and normal PTH level. It has been observed as well that the ACT haplotype was more common in patients with low glomerular filtration, low PTH, and normal hemoglobin level. We found that the LGALS3 rs4644 and rs4652 gene polymorphism may be involved in the pathogenesis and appearance of complications in ESRD patients and thus could be considered a new genetic risk factor in this population

Analysis of association between polymorphisms of MTHFR, MTHFD1 and RFC1 genes and efficacy and toxicity of methotrexate in rheumatoid arthritis patients

A folate analogue methotrexate (MTX) is the most commonly used disease-modifying drug in the treatment of rheumatoid arthritis. However, the clinical response of RA patients treated with MTX shows interindividual differences and 30% of patients discontinue therapy due to the side effects. In a group of 184 RA patients treated with MTX we have investigated whether polymorphisms in MTHFR (rs1801133, rs1801131), MTHFD1 (rs2236225) and RFC1 (rs144320551) genes may have impact on MTX efficacy and/or adverse drugs effects (ADEs). The efficacy of the MTX therapy has been estimated using the disease activity score in 28 joints (DAS28-ESR) based on EULAR criteria and relative DAS28 values (rDAS28) and all adverse drug events were recorded. Patients were genotyped for selected polymorphism by PCR-RFLP method. According to the EULAR response criteria after 6 months of MTX therapy 146 (79.3%) patients were classified as responders, (17 patients (11.6%) were good and 129 patients (88.4%) were moderate responders) and 38 patients (20.7%) as non-responders. ADEs were observed in 53 (28.8%) patients. The majority of ADEs were mild (36 (19.56%) patients) to moderate (12 (6.25%) patients). Five patients (2.7%) had serious ADEs. Association studies have been conducted between obtained genotypes and the efficacy and toxicity of MTX. We have observed no association between polymorphisms and efficacy or toxicity of MTX in RA patients. [Projekat Ministarstva nauke Republike Srbije, br. 175091