Genetički polimorfizmi paraoksonaze 1 i podložnost aterogenezi

Introduction. Paraoxonase 1 (PON1) is a multifunctional enzyme associated with high-density lipoprotein particles (HDL). It is a cellular antioxidant that hydrolyses oxidized macromolecules, especially low-density lipoproteins (ox-LDL). Because increased oxidative stress is believed to play a crucial role in the initiation and propagation of atherosclerosis, coding (Q192R and L55M) and promoter (C(-107)T) region polymorphisms of pon1 gene, that are responsible for catalytic efficiency, activity and the level of the enzyme, have been of great interest as a potential markers of susceptibility for atherogenesis. Objective. The aim of the study was to assess possible association between these pon1 gene variants and clinical manifestations of the atherosclerosis and oxidative stress. Methods. A total of 60 angiographically documented patients with manifested atherosclerotic disease and 100 control individuals were analyzed. Genomic DNA was isolated from the peripheral blood cells and genotyping was performed using polymerase chain reaction followed by the restriction fragment length polymorphism (PCR-RFLP) analysis. Results No significant difference in allele and genotype frequencies of all three examined polymorphisms was found between the atherosclerotic patients and healthy controls. The obtained results could not support an association of pon1 gene variants with the oxidative stress and atherogenesis. Conclusion. These polymorphisms cannot be considered risk factors of atherosclerosis in Serbian population. A larger study is required in order to establish possible contribution of pon1 variants to atherosclerosis-related cardiovascular diseases.Uvod. Paraoksonaza 1 (PON1) je multifunkcionalni enzim koji je vezan za lipoproteine visoke gustine (HDL). To je ćelijski antioksidans koji hidrolizuje oksidovane makromolekule, naročito oksidovane lipoproteine niske gustine (ox-LDL). Smatra se da povišeni oksidativni stres igra ključnu ulogu u inicijaciji i propagaciji ateroskleroze, pa su polimorfizmi u kodirajućem (Q192R i L55M) i promotorskom (C(-107)T) regionu gena pon1, koji su odgovorni za katalitičku efikasnost, aktivnost i nivo enzima, od velikog interesa kao potencijalni markeri osetljivosti na aterogenezu. Cilj rada. Cilj ove studije je bio da se ispita moguća povezanost varijanti gena pon1 i kliničkih manifestacija ateroskleroze i oksidativnog stresa. Metode rada. Analizirano je 60 bolesnika s angiografski dokumentovanim manifestacijama ateroskleroze i 100 zdravih ispitanika. Genomska DNK je izolovana iz ćelija periferne krvi, a genotipizacija je urađena primenom reakcije lančane polimeraze, posle koje je urađena analiza dužine restrikcionih fragmenata (tzv. PCR-RFLP analiza). Rezultati. Učestalosti alela i genotipova tri ispitivana polimorfizma nisu pokazale značajne razlike između ispitanika obolelih od ateroskleroze i zdravih osoba. Dobijeni rezultati ne ukazuju na povezanost analiziranih varijanti gena pon1 i oksidativnog stresa i aterogeneze. Zaključak. Ovi polimorfizmi se ne mogu smatrati faktorima rizika za razvoj ateroskleroze u srpskoj populaciji. Potrebna je studija sa većim brojem ispitanika, kako bi se utvrdio mogući doprinos varijanti gena pon1 na nastanak kardiovaskularnih oboljenja u čijoj osnovi je ateroskleroza

Genetički polimorfizam glutation S-transferaze P1 (GSTP1) Ile105Val i osetljivost na aterogenezu kod pacijenata sa dijabetesom tipa 2

One of the characteristics of type 2 diabetes mellitus (T2DM) is the state of persistent oxidative stress (OS) that has been implicated in the pathogenesis of diseases such is atherosclerosis mainly through chronic hyperglycemia that stimulates production of reactive oxygen species (ROS) and increases OS. Glutathione S-transferase P1 (GSTP1) is a member of the cytosolic GST superfamily. It plays an important role in neutralizing OS as an enzyme. Also, it participates in regulation of stress signaling and protects cells against apoptosis via its noncatalytic ligand-binding activity. GSTP1 Ile105Val functional polymorphism influences protein catalytic activity and stability and the aim of this study was to determine whether this gene variation influences susceptibility to atherogenesis in T2DM patients. A total of 240 individuals (140 patients with T2DM, accompanied with clinical manifestations of atherosclerosis, and 100 healthy controls) were included in this study. Genomic DNA was isolated from peripheral blood cells and genotyping was performed using polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP) analysis. We obtained no statistically significant differences in the distribution of alleles and genotypes between cases and controls (P>0.05) but association between Ile/Val (OR=0.6, 95%CI=0.35-1.05, P=0.08) and Val/Val (OR=0.45, 95%CI=0.18-1.11, P=0.08) genotypes and disease approached significance (P=0.08). Our results indicated that a larger study group is needed to establish the true relationship between potentialiy protective allele Val and the disease, and to determine the influence of other GSTP1 polymorphisms on atherogenesis in T2DM patients.Jedna od osobina tipa 2 dijabetes mellitus (T2DM) je stanje perzistentnog oksidativnog stresa (OS) koji je sastavni deo patogeneze bolesti kao što je ateroskleroza i to najčešće putem hronične hiperglikemije koja stimuliše nastanak reaktivnih vrsta kiseonika (tzv ROS) i povećava nivo OS. Glutation S-transferaza P1 (GSTP1) pripada superfamiliji citosolnih GST. Kao enzim, ima važnu ulogu u neutralizaciji OS. Takođe, učestvuje u regulaciji signalnog puta stresa ali i štiti ćelije od apoptoze putem nekatalitičke ligand-vezujuće aktivnosti. Funkcionalni polimorfizam Ile105Val GSTP1 gena utiče na katalitičku aktivnost proteina i na njegovu stabilnost i cilj ove studije je bio da se utvrdi da li ova genska varijanta utiče na osetljivost na aterogenezu kod pacijenata sa T2DM. Ukupno 240 osobe (140 pacijenata sa T2DM i nekom od kliničkih manifestacija ateroskleroze i 100 zdravih kontrola) je bilo uključeno u ovu studiju. Genomska DNK je izolovana iz ćelija periferne krvi a genotipizacija je urađena primenom reakcije lančane polimeraze posle koje je urađena analiza dužine restrikcionih fragmenata (PCR-RFLP analiza). Nismo dobili statistički značajne razlike u raspodeli alela i genotipova između obolelih i kontrola (P>0.05) ali asocijacija između Ile/Val (OR=0.6, 95%CI=0.35-1.05, P=0.08) i Val/Val (OR=0.45, 95%CI=0.18-1.11, P=0.08) genotipova i bolesti se približila značajnosti (P=0.08). Naši rezultaati pokazuju da je potrebna veća grupa ispitanika da bi se utvrdila prava veza između potencijalno protektivnog alela Val i bolesti kao i da bi se utvrdio uticaj drugih polimorfizama gena GSTP1 na aterogenezu kod T2DM pacijenata

Genetic polymorphism of glutathion S-transferase P1 (GSTP1) Ile105Val and susceptibility to atherogenesis in patients with type 2 diabetes mellitus

One of the characteristics of type 2 diabetes mellitus (T2DM) is the state of persistent oxidative stress (OS) that has been implicated in the pathogenesis of diseases such is atherosclerosis mainly through chronic hyperglycemia that stimulates production of reactive oxygen species (ROS) and increases OS. Glutathione S-transferase P1 (GSTP1) is a member of the cytosolic GST superfamily. It plays an important role in neutralizing OS as an enzyme. Also, it participates in regulation of stress signaling and protects cells against apoptosis via its noncatalytic ligand-binding activity. GSTP1 Ile105Val functional polymorphism influences protein catalytic activity and stability and the aim of this study was to determine whether this gene variation influences susceptibility to atherogenesis in T2DM patients. A total of 240 individuals (140 patients with T2DM, accompanied with clinical manifestations of atherosclerosis, and 100 healthy controls) were included in this study. Genomic DNA was isolated from peripheral blood cells and genotyping was performed using polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP) analysis. We obtained no statistically significant differences in the distribution of alleles and genotypes between cases and controls (P>0.05) but association between Ile/Val (OR=0.6, 95%CI=0.35-1.05, P=0.08) and Val/Val (OR=0.45, 95%CI=0.18-1.11, P=0.08) genotypes and disease approached significance (P=0.08). Our results indicated that a larger study group is needed to establish the true relationship between potentialiy protective allele Val and the disease, and to determine the influence of other GSTP1 polymorphisms on atherogenesis in T2DM patients. [Projekat Ministarstva nauke Republike Srbije, br. 175075

Combined GSTM1 and GSTT1 null genotypes are strong risk factors for atherogenesis in a Serbian population

Oxidative stress (OS) plays an important role in atherogenesis and since glutathione S-transferases (GSTs) provide protection against OS, we have tested the hypothesis that deletion polymorphisms in two GSTs (GSTM1 and GSTT1) may affect the risk of developing atherosclerosis. A total of 382 individuals (200 patients with atherosclerosis and 182 healthy controls) were included in this association study. Genomic DNA was isolated from peripheral blood cells or from buccal epithelial cells and genotyping was performed using multiplex-PCR or real-time PCR methods. GSTM1 null genotype was significantly more frequent in atherosclerotic patients than in controls (52.0% vs 34.1%) and individuals with the GSTM1 null genotype had an approximately 2-fold increase in atherosclerosis risk (OR: 2.1, 95%CI=1.39-3.17, P=0.0004). GSTT1 null genotype alone did not show a statistically significant effect on atherosclerosis risk modulation, but the association approached significance (OR: 1.57, 95%CI=0.94-2.64, P=0.08). The combined analysis showed that the presence of both genes had a protective effect against atherosclerosis (OR=0.55, 95%CI=0.37-0.83, P=0.005) while double null genotypes led to a robust atherosclerosis risk increase (OR: 8.14, 95%CI=2.41-27.51, P lt 0.0001). This study demonstrated that the GSTM1 null and combined GSTM1/GSTT1 null genotypes are susceptibility factors for development of atherosclerosis in a Serbian population

Using whole exome sequencing to explore genetic basis of unicuspid aortic valve disease

Normal aortic valve consists of three cusps that develop in the embryonic stage. Unicuspid aortic valve (UAV) is a rare congenital anomaly resulting in only one cusp with estimated prevalence of 0.02% in general population. Aim of this study was to identify genetic variants possibly associated with development of UAV. The study included 17 subjects, namely 5 UAV patients and their healthy family members without UAV disorder. Total DNA was isolated from venous blood samples and whole exomes sequencing (WES) was performed using BGI’s WES protocol. Adapter-trimmed and quality-filtered reads (fastp) were mapped to hg38 reference genome using BWA/SAMtools. VCF files were generated using GATK (BaseRecalibrator, HaplotypeCaller) and annotated with InterVar and AnnoVar tools. Rare heterozygous variants present in UAV patients were found in NOTCH1, TGFB2, MYH6, EGFR, FBN2, C1R, ROBO4 and TBX5, genes associated with development of aortic valves. Among these, most were missense mutations with damaging effects as predicted using in silico tools (SIFT and/or Polyphen). Only mutation in MYH6 p.Ala1130Ser was found in at least two different UAV patients. Also, rare homozygous missense mutation p.Gly577Ser with high damaging potential was found in ADAMTS5 gene. Besides, highly damaging heterozygous missense mutations were detected in gene interacting functional partners (STRING) of genes associated with development of aortic valves: DVL1, THBS1, NOTCH4, ADAMTS3, FBN1, NOTCH2, ADAM17, LRP5, WWTR1, C1S, ANKRD6 and TNNI1, as well as homozygous in ACAN and KNG1. Taken together, malfunctions in ADAMTS5, ACTA2, MYH6, FBN2, AXIN1, CELSR1 or TBX5 networks were found to be common in at least two UAV patients, suggesting existence of genetic basis in UAV disorder, possibly as a result of combined effects of multiple variants.Book of abstract: 4th Belgrade Bioinformatics Conference, June 19-23, 202

Renal Dysfunction Following Elective Endovascular Aortic Aneurysm Repair

Abdominal Aortic Aneurysm (AAA) is a degenerative disease of the aortic wall with potentially fatal complications. Open Repair (OR) was considered the gold standard, until the emergence of Endovascular Aneurysm Repair (EVAR), which is less invasive and equally (if not more) effective. As the popularity of endovascular procedures grows, related complications become more evident, with kidney damage being one of them. Although Acute Kidney Injury (AKI) following EVAR is relatively common, its true incidence is still uncertain. Also, there is insufficient data concerning long-term renal outcomes after EVAR, especially with repeated contrast agent exposure. Despite the lack of firm evidence on the effectiveness of individual strategies, it is evident that prevention of AKI following EVAR requires a multifactorial approach. This review focuses on recent findings based on human studies regarding the current evidence of renal impairment after EVAR, its quantification and strategies for its prevention. © 2019 Bentham Science Publishers.This is the peer-reviewed version of the following article: Isenovic, E., D. Radak, M. Neskovic, and P. Otasevic. "Renal Dysfunction Following Elective Endovascular Aortic Aneurysm Repair." Current vascular pharmacology (2017). [http://dx.doi.org/10.2174/1570161115666171116163203]Published version: [https://vinar.vin.bg.ac.rs/handle/123456789/8038

Genetic polymorphisms of paraoxonase 1 and susceptibility to atherogenesis

Introduction. Paraoxonase 1 (PON1) is a multifunctional enzyme associated with high-density lipoprotein particles (HDL). It is a cellular antioxidant that hydrolyses oxidized macromolecules, especially low-density lipoproteins (ox-LDL). Because increased oxidative stress is believed to play a crucial role in the initiation and propagation of atherosclerosis, coding (Q192R and L55M) and promoter (C(-107)T) region polymorphisms of pon1 gene, that are responsible for catalytic efficiency, activity and the level of the enzyme, have been of great interest as a potential markers of susceptibility for atherogenesis. Objective. The aim of the study was to assess possible association between these pon1 gene variants and clinical manifestations of the atherosclerosis and oxidative stress. Methods. A total of 60 angiographically documented patients with manifested atherosclerotic disease and 100 control individuals were analyzed. Genomic DNA was isolated from the peripheral blood cells and genotyping was performed using polymerase chain reaction followed by the restriction fragment length polymorphism (PCR-RFLP) analysis. Results No significant difference in allele and genotype frequencies of all three examined polymorphisms was found between the atherosclerotic patients and healthy controls. The obtained results could not support an association of pon1 gene variants with the oxidative stress and atherogenesis. Conclusion. These polymorphisms cannot be considered risk factors of atherosclerosis in Serbian population. A larger study is required in order to establish possible contribution of pon1 variants to atherosclerosis-related cardiovascular diseases. [Projekat Ministarstva nauke Republike Srbije, br. 175075

Copper and zinc concentrations in atherosclerotic plaque and serum in relation to lipid metabolism in patients with carotid atherosclerosis

Background/Aim. Some oligoelements are now investigated as possibly having a role in atherosclerosis. The aim of this study was to compare the concentrations of copper and zinc in the serum and carotid plaque and parameters of lipid metabolism in patients with different morphology of carotid atherosclerotic plaque. Methods. Carotid endarterectomy due to the significant atherosclerotic stenosis was performed in 91 patients (mean age 64 ± 7). The control group consisted of 27 patients (mean age 58 ± 9), without carotid atherosclerosis. Atheroscletoric plaques were divided into four morphological groups, according to ultrasonic and intraoperative characteristics. Copper and zinc concentrations in the plaque, carotid artery and serum were measured by atomic absorption spectrophotometry. Results. Serum copper concentrations were statistically significantly higher in the patients with hemorrhagic in comparison to those with calcified plaque (1.2 ± 0.9 μmol/L vs 0.7 ± 0.2 μmol/L, respectively; p = 0.021). Zinc concentrations were statistically significantly lower in plaques of the patients with fibrolipid in comparison to those with calcified plaques (22.1 ± 16.3 μg/g vs 38.4 ± 25.8 μg/g, respectively; p = 0.024). A negative significant correlation was found for zinc and triglycerides in the serum in all the patients (r = -0.52, p = 0.025). In the control group we also demonstrated a positive significant correlation for low-density lipoprotein cholesterol and copper in the serum (r = 0.54, p = 0.04). Conclusion. The data obtained in the current study are consistent with the hypothesis that high copper and lower zinc levels may contribute to atherosclerosis and its sequelae as factors in a multifactorial disease. Further studies are necessary in order to conclude whether high concentration of copper and zinc in the serum could be risk factors for atherosclesrosis

Are de novo acute heart failure and acutely worsened chronic heart failure two subgroups of the same syndrome?

Introduction. Acute heart failure (AHF) is one of the most common diseases in emergency medicine, associated with poor prognosis and high in-hospital and long-term mortality. Objective. To investigate clinical presentation of patients with de novo AHF and acute worsening of chronic heart failure (CHF) and to identify differences in blood levels of biomarkers and echocardiography findings. Methods. This prospective study comprised 64 consecutive patients being grouped according to the onset of the disease into patients with the de novo AHF (45.3%), and patients with acute worsening of CHF (54.7%). Results. Acute congestion (60%) was the most common manifestation of de novo AHF, whereas pulmonary oedema (43.1%) was the most common manifestation of acutely decompensated CHF. Patients with acutely decompensated CHF had significantly higher blood values of creatinine (147.10 vs 113.16 μmol/l; p<0.05), urea (12.63 vs 7.82 mmol/l; p<0.05), BNP (1440.11 vs 712.24 pg/ml; p<001) and NTproBNP (9097.00 vs 2827.70 pg/ml; p<0.01) on admission, and lower values of M-mode left ventricular ejection fraction (LVEF) during hospitalization (49.44% vs 42.94%; p<0.05). The follow-up after one year revealed still significantly higher BNP (365.49 vs 164.02 pg/ ml; p<0.05) and lower average values of both LVEF in patients with acutely worsened CHF (46.62% vs 54.41% and 39.52% vs 47.88%; p<0.05). Conclusion. Considering differences in clinical severity on admission, echocardiography and natriuretic peptide values during hospitalization and after one year follow-up, de novo AHF and acutely worsened CHF are two different subgroups of the same syndrome