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Asymmetric Dimethylarginine and Its Relation to Atherogenesis
Široká, Romana ; Racek, Jaroslav (advisor) ; Teplan, Vladimír (referee) ; Hyánek, Josef (referee)
ASYMMETRIC DIMETHYLARGININE - COMPARSION OF CHROMATOGRAPHY AND IMMUNOMETRIC METHODS Objective:Asymmetric dimethylarginine (ADMA) is often discussed in connection with hyperhomocysteinemia and its toxic effect on vessel wall. ADMA concentration is usually measured by HPLC (High Performance Liquid Chromatography) after previous derivatisation. Recently, ELISA (Enzym Linked Immuno Assay) methods for ADMA determination were introduced and ELISA kits are commercially available. Method and Result:The aim of the study was to compare HPLC and ELISA methods for ADMA determination. For HPLC determination we used equipments from Thermo separation product (Florida, USA). After solid-phase extraction on polymer cation-exchange column and the following derivatisation with o-phthaldialdehyde the samples were separated using C18 column (mobile phase 8.7% acetonitril, 50 mmol/l phosphate buffer, pH 6.5) and a fluorescence detector. NG-monomethyl-L-arginine was used as an internal standard. ADMA® ELISA kit, based on a competitive principle, was obtained from DLD Diagnostika, Hamburg, Germany. ADMA was measured in EDTA plasma of 40 healthy blood donors and 40 hemodialysis patients with hyperhomocysteinemia. Conclusion: In spite of different principles both methods showed a very good correlation (r = 0.944, p<0.0001). ELISA...
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Cytokines and adipocytokines in surgically ill patients: the role of fat tissue
Teplan, Vladimír ; Gürlich, Robert (advisor) ; Adamec, Miloš (referee) ; Herget, Jan (referee)
Cytokines and adipocytokines have major effect on post-operative prognosis of patients. In obese patients, we find significantly higher expression of proinflammatory mediators. In first part of our project we focus on cytokine expresion after elective abdominal surgery. Abdominal surgery was choosen as model of stress reaction to organism. Our results show, that proinflammatory cytokine expresion was significantly higher in obese patients and especially early elevation of visfatin could be a new marker in diagnosis of postoperative inflammatory complications. Related publications describe methods, design and results of trials. Second part of our poject is focused on patients after kidney transplanation, considering obesity, type of previous dialysis, early postoperative movement a nutrition. Our results show, that there is higher proinflammatory expresion after kidney transplanation with no effect of previous type of dialysis. On the other side early exercise have positive effect on cytokine expresion and decreases ADMA. Related publicatons describe methods, details and results of trials.
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Oxidative and carbonyl stress,microinflammation and cardiovascular risk in patiens with chronic kidney disease
Peiskerová, Martina ; Kalousová, Marta (advisor) ; Teplan, Vladimír (referee) ; Racek, Jaroslav (referee)
Short summary: Background: High cardiovascular risk in patients with chronic kidney disease is partly due to mineral dysbalance, microinflammation and oxidative stress. CKD patients accumulate traditional and non-traditional CV risk factors. FGF23, MMPs and PlGF belong among these non-traditional biomarkers of CV risk. FGF23 is a phosphaturic hormone and inhibitor of calcitriol synthesis. It is associated with vascular calcifications. Matrix-metalloproteinases (e.g. MMP-2, MMP-9) are proteolytic, proinflammatory enzymes, contributing to myocardial remodelation. Placental growth factor (PlGF) is a proangiogenic cytokine that is associated with LV hypertrophy in animal model. Plasmatic FGF23, MMPs and PlGF are elevated in CKD. Aim: We aimed to describe dynamic changes between several novel biomarkers of CV risk (FGF23, MMP-2, MMP-9 and PlGF) in CKD stages 1-5, to describe their mutual correlations and possible association with traditional CV risk markers. We studied possible association of laboratory and echocardiographic parameters in patients with CKD stages 2-4. Methods: In a cross-sectional study we evaluated 80 patiens with CKD 1-5 and 44 healthy controls. In a prospective study we evaluated echocardiographic and laboratory parameters in 62 patients with CKD 2-4 for an average study period of 36±10...
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31P MR Spectroscopy of Human Kidney Grafts Monitoring of MR parameters of human grafts and their comparison with clinical parameters of patients
Vyhnanovská, Pavlína ; Hájek, Milan (advisor) ; Teplan, Vladimír (referee) ; Mechl, Marek (referee)
The aim of this work was to evaluate the possibility of using in vivo phosphorus magnetic resonance spectroscopy (31P MRS) for diagnosis of kidney graft dysfunction after transplantation. In vivo 31P MRS is noninvasive imaging method which make possible to assess energy metabolism. A total of 68 patients with kidney grafts were examined on a 1.5T MR scanner. 31P MRS was performed using the 2D-chemical shift imaging method. The patients were divided into four groups: acute rejection; acute tubular necrosis; late graft dysfunction; and patients with good renal function after transplantation. We measured the signal intensities of phosphomonoesters (PME), inorganic phosphate (Pi), phosphodiesters (PDE) and -, -- adenosine triphosphate (with contribution of -, -adenosine diphosphate) and their ratios. Patients with acute rejection episodes showed significantly elevated PME/ß-ATP, PME/Pi and PDE/Pi signal ratios compared to the control group. The group with acute tubular necrosis has decreased these ratios: PDE/-ATP a PDE/Pi compared to the control group. Patients with late graft dysfunction revealed only significant decrease of PME/Pi ratio. The group of AR patients display higher ratios of PME/Pi and PDE/Pi compared to the group of ATN patients and display higher ratios of PME/Pi compared to the group of LGD....
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Lupus nephritis-novel diagnostic and therapeutic approaches
Pešičková, Satu ; Tesař, Vladimír (advisor) ; Teplan, Vladimír (referee) ; Krejčí, Karel (referee)
Key words: anti-mCRP antibodies, cyclophosphamide, cyclosporine A, lupus nephritis, systemic lupus erythematosus Systemic lupus erythematosus (SLE) is a systemic autoimmune disease. Circulating autoantibodies against the body's own nuclear and cytoplasmic structures and creation of immune complexes play a key role in the pathogenesis of SLE. Antibodies against monomeric C-reactive protein (anti-mCRP) might play role in pathogenesis of lupus nephritis (LN). The aim of this study was to find relation between anti-mCRP and activity of LN and response to therapy. Methods: The study was performed on 57 patients (M/F 0.21, median age 32 years) with LN. In a subanalysis, we focused on 29 patients with newly diagnosed active LN and we followed them up for a median of 5.9 years. Levels of anti-CRP were measured by in house ELISA. Disease activity was measured by SLEDAI. Results: Levels of anti-mCRP were significantly higher in patients with active lupus nephritis (26.78 versus 7.5 AU, p=0.009) and levels of anti-mCRP positively correlated with the activity of SLE as assessed by the SLEDAI score (Spearman's r=0.406, p= 0.002). We found negative prediction of anti-mCRP for worse outcome after two years of standard therapy, OR (95% CI)=13.7 (1.22-770.87); p=0.014. Conclusion: Serum levels of anti-mCRP seem to...
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Study of protein changes in patients with nephrotic syndrome and Anderson-Fabry disease
Vojtová, Lucie ; Zima, Tomáš (advisor) ; Teplan, Vladimír (referee) ; Průša, Richard (referee)
Background: Heavy proteinuria may be caused by either increased glomerulal basement membrane permeability or membrane or podocyte structural damage, and also by impairment of secretion-reabsorption tubular processes. In this study, 60 patients with nephrotic proteinuria and other diagnoses (lupus nephritis, membranous nephropathy, IgA nephropathy, Wegener's granulomatosis) and 20 patients with Anderson-Fabry disease (AFD),which is an X-linked genetic disorder with deficient a-galactosidase A activity, were analysed by the 2D electrophoresis method. The main aim of this work was to investigate possible differences in urine proteins in nephropaties, between healthy controls and AFD patients and to identify abnormal proteins as potential biomarkers of disease. Methods: The urine proteins were devided by isoelectric focusing method using polyacrylamide strips (pH 3-10 linear). The second dimensional SDS electrophoresis was performed in 12 % polyacrylamide gel. The proteins were visualized by silver method and selected proteins were identified by MALDI-TOF MS. The gels were evaluated by Phoretix 2D expression software 2005. Results: We found out that without adding protease inhibitors we can detect proteolysis, with increased quantity of proteins manifested in the area about 10 kDa and decreased quantity...
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Novel biomarkers in patients with renal disease
Zakiyanov, Oskar ; Kalousová, Marta (advisor) ; Teplan, Vladimír (referee) ; Průša, Richard (referee)
Chronic kidney disease (CKD) and acute kidney injury (AKI) are major public health problems. It is important to be able to identify those at high risk of adverse outcome, CKD progression and associated cardiovascular disease. The aim of the thesis was to study novel promising biomarkers, their relationship to kidney function, chronic inflammation and/or cardiovascular risk - placental growth factor (PlGF), pregnancy associated plasma protein A (PAPP-A), matrix metalloproteinase 2 (MMP-2), matrix metalloproteinase 9 (MMP-9), soluble receptor for advanced glycation end products (sRAGE), calcium binding protein S100A12 or extracellular newly identified RAGE binding protein (EN-RAGE), and high mobility group box protein-1 (HMGB-1) in patients with renal diseases including CKD, haemodialysis (HD), AKI patients, and healthy controls for comparison. First study revealed that PlGF is elevated in patients with decreased renal function. Second study demonstrated the association of MMP-2 and PAPP-A with proteinuria in patients with CKD. Moreover, serum MMP-2, MMP-9 and PAPP-A levels significantly differed in patients with various nephropathies. EN-RAGE levels are not elevated in patients with CKD, but are related to inflammatory status. PAPP-A, EN-RAGE and HMGB-1 levels are significantly elevated, but sRAGE and PlGF...
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Novel biomarkers in patients with renal disease
Zakiyanov, Oskar ; Kalousová, Marta (advisor) ; Teplan, Vladimír (referee) ; Průša, Richard (referee)
Chronic kidney disease (CKD) and acute kidney injury (AKI) are major public health problems. It is important to be able to identify those at high risk of adverse outcome, CKD progression and associated cardiovascular disease. The aim of the thesis was to study novel promising biomarkers, their relationship to kidney function, chronic inflammation and/or cardiovascular risk - placental growth factor (PlGF), pregnancy associated plasma protein A (PAPP-A), matrix metalloproteinase 2 (MMP-2), matrix metalloproteinase 9 (MMP-9), soluble receptor for advanced glycation end products (sRAGE), calcium binding protein S100A12 or extracellular newly identified RAGE binding protein (EN-RAGE), and high mobility group box protein-1 (HMGB-1) in patients with renal diseases including CKD, haemodialysis (HD), AKI patients, and healthy controls for comparison. First study revealed that PlGF is elevated in patients with decreased renal function. Second study demonstrated the association of MMP-2 and PAPP-A with proteinuria in patients with CKD. Moreover, serum MMP-2, MMP-9 and PAPP-A levels significantly differed in patients with various nephropathies. EN-RAGE levels are not elevated in patients with CKD, but are related to inflammatory status. PAPP-A, EN-RAGE and HMGB-1 levels are significantly elevated, but sRAGE and PlGF...
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Oxidative and carbonyl stress,microinflammation and cardiovascular risk in patiens with chronic kidney disease
Peiskerová, Martina ; Kalousová, Marta (advisor) ; Teplan, Vladimír (referee) ; Racek, Jaroslav (referee)
Short summary: Background: High cardiovascular risk in patients with chronic kidney disease is partly due to mineral dysbalance, microinflammation and oxidative stress. CKD patients accumulate traditional and non-traditional CV risk factors. FGF23, MMPs and PlGF belong among these non-traditional biomarkers of CV risk. FGF23 is a phosphaturic hormone and inhibitor of calcitriol synthesis. It is associated with vascular calcifications. Matrix-metalloproteinases (e.g. MMP-2, MMP-9) are proteolytic, proinflammatory enzymes, contributing to myocardial remodelation. Placental growth factor (PlGF) is a proangiogenic cytokine that is associated with LV hypertrophy in animal model. Plasmatic FGF23, MMPs and PlGF are elevated in CKD. Aim: We aimed to describe dynamic changes between several novel biomarkers of CV risk (FGF23, MMP-2, MMP-9 and PlGF) in CKD stages 1-5, to describe their mutual correlations and possible association with traditional CV risk markers. We studied possible association of laboratory and echocardiographic parameters in patients with CKD stages 2-4. Methods: In a cross-sectional study we evaluated 80 patiens with CKD 1-5 and 44 healthy controls. In a prospective study we evaluated echocardiographic and laboratory parameters in 62 patients with CKD 2-4 for an average study period of 36±10...
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Pathophysiology of cardiorenal interaction in patients with type 2 diabetes mellitus
Pecková, Marie ; Horáčková, Miroslava (advisor) ; Teplan, Vladimír (referee) ; Linhart, Aleš (referee)
Patients with type 2 diabetes mellitus (DM) suffer often from cardirenal syndrome because of high prevalence of cardiac and renal insufficiency. Functional deterioration of both organs depends on the compensation of DM, arterial hypertension and other metabolic parameters. We asked a question when did the cardiorenal interaction develop and which factors influenced its development. We examined 82 patients with type 2 DM with optimally corrected arterial hypertension, with well compensated diabetic metabolic disorder, without any clinical signs of heart disease and with normal or mild-to-moderate decrease in glomerular filtration rate according to MDRD equation (eGFR). We analysed the associations between eGFR and echocardiographic parameters of left ventricular (LV) diastolic function E' and E/E' on the basis of linear regression and multivariate analysis. According to multivariate analysis eGFR was significantly associated with E' and E/E' either. We assessed the association between E' and GFR by cluster analysis that divided whole cohort into two subgroups on the basis of the value E' = 7.1 cm s-1 . We analysed the associations between the parameters of LV diastolic function, eGFR and other parameters separately in both subgroups. We found out, that cardiorenal interaction (significant association...
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