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Rakicevic L.B.,University of Belgrade | Kusic-Tisma J.S.,University of Belgrade | Kovac M.K.,Blood Transfusion Institute of Serbia | Kovac M.K.,University of Belgrade | And 3 more authors.
Scandinavian Journal of Clinical and Laboratory Investigation | Year: 2013

During the last decade genetic factors affecting coumarin therapy have been extensively investigated. The most important genes appear to be CYP2C9 and VKORC1, and different studies have shown that DNA testing can dramatically improve the safety and effectiveness of the therapy. However, the implementation of pharmacogenetic testing in everyday practice is still not a reality. Facilities and ability to get results before the start of therapy are very important. The implementation of specific methodology and equipment for particular type of diagnostics can represent a serious, even impossible, financial hurdle to overcome (especially in developing countries). For this reason, the use of every tool that contributes to rationalization of the existing methods can be a considerable asset. Therefore, we set the goal to rationalize our current DNA sequencing based protocol for analysis of the VKORC1 c.-1639G> A, CYP2C9*2 and CYP2C9*3 variant alleles, in order to obtain shorter and easier procedure. Simplification of the protocol was achieved by setting up multiplex PCR and omitting DNA extraction. This rationalization of the existing DNA sequencing based procedure allows getting results in 12 hours. The new protocol was tested on 118 samples. Obtained results have shown full accordance to those obtained with previous, non-modified protocol. Therefore, given the circumstances, we consider that protocol for pharmocogenetic testing should be made more accessible-both to doctors and patients. It is one of the prerequisites in order to make genotyping prior to the therapy common practice. © 2013 Informa Healthcare. Source

Kotur-Stevuljevic J.,University of Belgrade | Simic-Ogrizovic S.,Clinical Center Serbia | Dopsaj V.,University of Belgrade | Dopsaj V.,Institute for Medical Biochemistry | And 6 more authors.
Clinical Biochemistry | Year: 2012

Background: Atherosclerosis is the main cause of mortality in end stage renal disease (ESRD) patients. Design and methods: Malnutrition, inflammation and diminished paraoxonase activity were used to calculate the sum of risk factors for atherosclerosis development in a cohort of 141 chronic renal disease patients. Kaplan-Meier survival analysis was implemented to assess risk of death. Results: Kaplan-Meier analysis (Log rank = 12.06, P=0.0072) showed higher risk of death with increasing number of risk factors in haemodialysis patients. Conclusions: Malnutrition in combination with inflammation and oxidative stress is associated with higher mortality in patients on long-term haemodialysis. © 2012 The Canadian Society of Clinical Chemists. Source

Becarevic M.,University of Novi Sad | Becarevic M.,Institute for Medical Biochemistry | Ignjatovic S.,Institute for Medical Biochemistry | Ignjatovic S.,University of Belgrade | And 2 more authors.
Clinical Laboratory | Year: 2012

Background: To investigate the association between clinical and serological features of patients with primary antiphospholipid syndrome (PAPS) and TNF-alpha, interleukin 6 (IL-6), and soluble IL-2 receptor (sIL-2R). Methods: ELISA was used for measurement of antibodies (Abs) and TNF-alpha, while IL-6 and sIL-2R were measured by chemiluminescence. Results: PAPS patients with pulmonary emboli showed positive correlation between IgM isotype of anti-annexin A5 antibodies and TNF-alpha (r = 0.894, p = 0.041) and IgM class of anticardiolipin antibodies and sIL-2R (r = 0.900, p = 0.037). In PAPS with cerebrovascular insults, positive correlation was noticed between TNF-alpha and IgG isotype of anticardiolipin (r = 0.624, p = 0.040) and anti-annexin A5 Abs (r = 0.768, p = 0.006). Conclusions: Isotype analysis of antiphospholipid and anti-annexin A5 Abs and investigation of their association with TNF-alpha is important for differentiation of PAPS patients that are prone to further deterioration of arterial and venous thromboses. Source

Becarevic M.,University of Novi Sad | Becarevic M.,Institute for Medical Biochemistry | Stojanovic L.,University of Belgrade | Ignjatovic S.,Institute for Medical Biochemistry | And 3 more authors.
Clinical Rheumatology | Year: 2016

We evaluated the importance of anti-annexin A5 antibodies (aanxA5 Abs) for clinical (thrombosis and/or recurrent pregnancy loss) and serologic (presence of antiphospholipid antibodies: lupus anticoagulant (LA), anticardiolipin (aCL), and anti-β2 glycoprotein I (aβ2GPI) antibodies) features of patients with primary antiphospholipid syndrome (PAPS). Our study included 70 patients with PAPS according to the international consensus criteria for APS. The mean age of the analyzed patients was 45.97 ± 12.72. The disease duration above 5 years was present in 31/70 of patients. Concentrations of analyzed antibodies were measured by ELISA. Cutoff values were set in accordance to the manufacturers’ recommendations. History of recurrent pregnancy loss was associated with double positivity for aanxA5 IgM and LA (χ2 = 4.000, P = 0.046) and triple positivity for aanxA5 IgM + LA + aβ2GPI IgM (χ2 = 4.168, P = 0.041). Venous thromboses were associated with triple positivity for aanxA5 IgM + aCLIgG + aβ2GPI IgM (χ2 = 3.965, P = 0.046). The IgG isotype of aanxA5 Abs was in positive correlation with aCL Abs of the IgG (r = 0.310, P = 0.009) and IgM (r = 0.254, P = 0.034) isotype. The presence of the clinical manifestations of PAPS is increasing with the number of positive conventional aPL and the IgM aanxA5 Abs tests. This new combination of Abs is beneficial even when the number of patients with positivity for aanxA5 Abs is low. This is important in further detection of patients prone to recurrence of thrombotic episodes. © 2016 International League of Associations for Rheumatology (ILAR) Source

Obrenovic R.,Institute for Medical Biochemistry | Petrovic D.,Clinical Center Kragujevac | Majkic-Singh N.,Institute for Medical Biochemistry | Trbojevic-Stankovic J.,Institute for Medical Biochemistry | And 2 more authors.
Clinical Nephrology | Year: 2011

Aim: The aim of this study was to determine the levels of cystatin C, creatinine and creatinine clearance in different trimesters of uncomplicated pregnancy in women with normal kidney function. Subjects and methods: A total of 109 pregnant women was included: Group 1: 38 women (average age 29.63 ± 4.3 y) in the 1st trimester, Group 2: 32 women (average age 33.56 ± 5.95 y) in the 2nd trimester and Group 3: 39 pregnant women (average age 30.1 ± 6.95 y) in the 3rd trimester. Serum cystatin C was determined by the PENIA method (Particle-Enhanced Nephelometric Immuno-Assay), using Behring tests (Behring Diagnostics GmbH, Marburg, Germany). Results were statistically analyzed using the ANOVA. Results: A statistically significant increase in serum cystatin C level was found in the 3rd trimester of pregnancy (0.69 ± 0.16 mg/l vs. 0.78 ± 0.26 mg/l vs. 1.21 ± 0.30 mg/l). Conclusion: It appears that cystatin C is not a reliable marker of kidney function in later stages of pregnancy and that its increase is connected with a combination of several factors, including endotheliasis, hormonal influence and glomerular filtration rate (GFR) alterations. © 2011 Dustri-Verlag Dr. K. Feistle. Source

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