The Pleiotropic Effects of Atorvastatin on Stable Angina Patients: Evidence by Analysis of High-Density Lipoprotein Size and Subclasses, and Plasma mRNA

Background: High-density lipoproteins (HDL) have atheroprotective biological properties: antioxidative, anti-apoptotic, anti-inflammatory, and they have the efflux capacity of cellular cholesterol. Plasma mRNA analysis can be used to investigate statin pleiotropy in vivo as a new analytical tool for non-invasive assessment of gene expression in vascular beds. The aim of this study was to assess the pleiotropic effects of atorvastatin in stable angina patients with highrisk values (group A) as compared with patients who had borderline and desirable HDL-cholesterol (HDL-C) values (group B). Methods: The atorvastatin therapy (20 mg/day) was given to forty-three patients with stable angina for 10 weeks. We investigated three statin pleiotropy-targeted genes: intercellular adhesion molecule-1, chemokine (C-C motif) ligand 2 and cathepsin S and assessed by gel electrophoresis gradient the effects of atorvastatin on HDL size and subclasses. Results: In group A, after therapy, HDL-C concentration was significantly increased but not in group B. Atorvastatin lowered plasma chemokine (C-C motif) ligand 2 and intercellular adhesion molecule-1 mRNA levels in both groups, but did not change the plasma cathepsin S mRNA levels. In group A only, baseline total bilirubin showed negative cor relations with the genes of cathepsin S (r=-0.506; p = 0.023) and significantly increased after therapy. Conclusions: HDL-C and bilirubin can be promising therapeutic targets in the treatment of cardiovascular diseases. Analysis of cell-free mRNA in plasma might become a useful tool for estimating statin pleiotropy

Glutathione S-transferase A1, M1, P1 and T1 null or low-activity genotypes are associated with enhanced oxidative damage among haemodialysis patients

Background. Increased oxidative stress is a hallmark of end-stage renal disease (ESRD). Glutathione S-transferases (GST) are involved in the detoxification of xenobiotics and protection of oxidative damage. We hypothesized that genetic polymorphism in antioxidant enzymes GSTA1, GSTM1, GSTP1 and GSTT1 is more frequent in ESRD and modulates the degree of oxidative stress in these patients. Methods. GSTA1, GSTM1, GSTP1 and GSTT1 genotypes were determined in 199 ESRD patients and 199 age- and gender-matched controls. Markers of protein and lipid oxidative damage [thiol groups, carbonyl groups, advanced oxidative protein products, nitrotyrosine, malondialdehyde (MDA) and MDA adducts], together with total oxidant status and pro-oxidant antioxidant balance were determined. Results. Individual GST polymorphisms influence vulnerability to both protein and lipid oxidation, with GSTM1-null gene variant having the most pronounced effect. Furthermore, a strong combined effect of null/low-activity GSTM1, GSM, GSTA1 and GSTP1 genotypes in terms of susceptibility towards oxidative and carbonyl stress was found in ESRD patients. When patients were stratified according to GSTM1 and GSTT1, the highest oxidant damage was noted in those with the GSTM1-null/GSTT1-null genotype. The observed effect was even stronger in patients with the third low-activity GSTP1 or GSTA1 genotype. Finally, the level of oxidative and carbonyl stress was most pronounced in the subgroup of patients with all four null or low-activity GSTM1, GSTT1, GSTP1 and GSTA1 genotypes. Conclusions. According to the GST genotype, ESRD patients may be stratified in terms of the level of oxidative and carbonyl stress that might influence cardiovascular prognosis, but could also improve efforts towards individualization of antioxidant treatment

Effects of extracellular calcium on fASIC1 currents

Fish ASIC1 (fASIC1) cloned from Opsanus tau, unlike the rat ASICs, requires Ca2+, in the extracellular preconditioning solution (pH 7.4) to be activated. Here we show that fASIC1 is interacting with Ca2+ in the same way as mammalian ASICs: extracellular Ca2+, is increasing the proportion of channels available for activation by stabilizing the closed state of the channel; in the activation process Ca2+ is released; H+ compete for the binding site of Ca2+ making the gating mechanism both Ca2+ and H+ dependent; H+ stabilizes the desensitized state; Ca2+ blocks the fASIC1 channel; and the affinity of the block is also modulated by H+. The "Ca2+ activation requirement" of fASICI reflects its greater affinity for steady-state desensitization by H+ compared to mammalian ASIC1.nul

Effects of extracellular calcium on fASIC1 currents

Fish ASIC1 (fASIC1) cloned from Opsanus tau, unlike the rat ASICs, requires Ca2+, in the extracellular preconditioning solution (pH 7.4) to be activated. Here we show that fASIC1 is interacting with Ca2+ in the same way as mammalian ASICs: extracellular Ca2+, is increasing the proportion of channels available for activation by stabilizing the closed state of the channel; in the activation process Ca2+ is released; H+ compete for the binding site of Ca2+ making the gating mechanism both Ca2+ and H+ dependent; H+ stabilizes the desensitized state; Ca2+ blocks the fASIC1 channel; and the affinity of the block is also modulated by H+. The "Ca2+ activation requirement" of fASICI reflects its greater affinity for steady-state desensitization by H+ compared to mammalian ASIC1.nul

RFLP and RAPD analysis of maize (Zea mays L.) local populations for identification of variability and duplicate accessions

The genebank in Maize Research Institute "Zemun Polje" maintains the collection of 2178 local populations of maize, characterised and classified using morphological markers. In this work 13 local populations, some of which were suspected to be duplicate accessions, were subjected to RAPD analysis with OPB 1-20 primers, as well as to RFLP analysis with 30 probes in combination with three restriction enzymes. Data thus obtained were used for calculation of Nei's genetic distances among populations and the resulting distance matrix was used for cluster analysis by UPGMA method. Genetic distances of analysed populations ranged between 0.1311 and 0.5075. Genetic distances calculated from RAPD and RFLP data were highly similar, leading to the conclusion that both markers revealed sufficient amount of genetic polymorphism among populations. Both methods showed a high degree of discrimination between populations, so they can be successfully used for polymorphism validation, genetic distance estimation among maize populations and screening of suspected duplicates in maize gene bank

Levels of oxidative stress biomarkers and bone resorption regulators in apical periodontitis lesions infected by Epstein-Barr virus

AimTo investigate whether apical periodontitis lesions infected by Epstein-Barr virus (EBV) exhibit higher levels of oxidative stress biomarkers [8-hydroxydeoxyguanosine (8-OHdG) and oxidized glutathione (GSSG)] and bone resorption regulators [receptor activator of nuclear factor (NF-B) ligand (RANKL) and osteoprotegerin (OPG)] compared to EBV-negative periapical lesions and healthy pulp tissues. MethodologyThe experimental group consisted of 30 EBV-positive and 30 EBV-negative periapical lesions collected in conjunction with apicoectomy. The pulp tissues of 20 impacted third molars were used as healthy controls. The qualitative and quantitative analysis of EBV was performed by nested and real-time polymerase chain reaction (PCR), respectively. The levels of RANKL and OPG were analysed by reverse transcriptase real-time PCR. The levels of 8-OHdG and GSSG were determined by enzyme-linked immunosorbent assay (ELISA). Mann-Whitney U-test and Spearman's correlation were used for statistical analysis. ResultsThe levels of RANKL, OPG, 8-OHdG and GSSG were significantly higher in apical periodontitis lesions compared to healthy pulp controls (P=0.001, P lt 0.001, P lt 0.001 and P lt 0.05, respectively). RANKL and OPG mRNA expression was significantly higher in EBV-positive compared to EBV-negative periapical lesions (P lt 0.05). There was no significant correlation between EBV copy numbers and levels of RANKL, OPG, 8OH-dG and GSSG in apical periodontitis. ConclusionLevels of bone resorption regulators and oxidative stress biomarkers were increased in apical periodontitis compared to healthy pulp tissues. EBV-positive periapical lesions exhibited higher levels of RANKL and OPG compared to EBV-negative periapical lesions. EBV may contribute to progression of apical periodontitis via enhanced production of bone resorption regulators