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Background:Vitamin D deficiency is repeatedly reported in colorectal cancer (CRC). Since cholesterol and vitamin D share common precursor 7-dehydrocholesterol (7-DHC), it would be important to explore the associations of key vitamin D metabolites and serum lipid parameters in patients with high and low grade CRC. The aim of this study was toanalyze relationships between serum 25(OH)D3,24,25(OH)2D3 and 7-DHC levels and serum lipids inpatients with CRC, and to evaluate their potential for prediction of risk for development of high grade CRC. Methods:We recruited 82 patients CRC and 77 controls.7-DHC, 25(OH)D3 and 24,25(OH)2D3 were quantified by LC-MS/MS methods.Results:7-DHC, 25(OH)D3 and vitamin D metabolic ratio(VDMR) were significantly lower in CRC patients than in control group (P

Insulin-like growth factor 1 (IGF-1) is a regulator of intrauterine growth, and circulating concentrations are reduced in intrauterine growth-restricted fetuses. The aim of our study was to investigate the relationship between IGF-1 levels in newborns and intrauterine growth, expressed as birth weight (BW). The research was designed as a cross-sectional study. The study included 71 premature newborns, gestational age (GA) ≤33 weeks. Quantitative determination of IGF-1 was performed in the 33rd post-menstrual week (pmw) to make the measurements more comparable. We used an enzyme-bound immunosorbent test for quantitative determination of IGF-1. Our results showed the mean IGF-1 level in premature newborns in 33rd pmw to be 23.1±4.56 (range 15.44-39.75) μg/L. There was no difference in IGF-1 values between male (23.1±4.98 μg/L) and female (23.1±4.87 μg/L) newborns. Tere was no significant difference in the average IGF-1 levels between male and female newborns with BW 50th percentile for GA either (p>0.50). Only BW, Inzulinu sličan faktor rasta (IGF-1) je jedan od čimbenika koji utječu na intrauterini rast. Serumske razine IGF-1 su smanjene u fetusima s intrauterinim zastojem rasta. Cilj našega istraživanja bio je ispitati odnos između razine IGF-1 u nedonoščadi i intrauterinog rasta izraženog kao porođajna težina (PT). Istraživanje je provedeno kao presječna studija. U studiju je bilo uključeno 71 nedonošče gestacijske dobi (GD) ≤33 tjedna. Kvantitativno određivanje IGF-1 provedeno je u 33. postmenstruacijskom tjednu (pmt) radi bolje usporedivosti rezultata. Za kvantitativno određivanje IGF-1 rabili smo enzimski imunosorbentni test. Naši rezultati pokazali su da je srednja razina IGF-1 u nedonoščadi u 33. pmt iznosila 23,1±4,56 (raspon 15,44-39,75) μg/L. Nije bilo razlike u vrijednostima IGF-1 između muške (23,1±4,98 μg/L) i ženske (23,1±4,87 μg/L) nedonoščadi. Također nije bilo značajne razlike u srednjim razinama IGF-1 između nedonoščadi s PT 50. percentila za GD (p>0,50). Nedonoščad s niskom PT (

Coronary artery disease (CAD) is one of the most important causes of mortality and morbidity in wide world population. Dyslipidemia, inflammation and oxidative stress may contribute to disruption of endothelium structure and function, atherosclerosis and CAD. Our study was aimed to determine whether Cu/Zn superoxide dismutase (Cu/Zn SOD) and Mn superoxide dismutase (Mn SOD) gene expression could be modulated by oxidative stress in CAD patients.This study included 77 CAD patients and 31 apparently healthy persons. Serum lipid levels, high sensitivity C-reactive protein (hsCRP), total antioxidant status (TAS) and thiobarbituric acid-reacting substances (TBARS) were measured. SOD isoenzymes gene expression was determined in peripheral blood mononuclear cells using quantitative polymerase chain reaction.Mn SOD messenger ribonucleic acid (mRNA) levels were significantly lower in CAD patients than in controls (p=0.011), while Cu/Zn SOD mRNA levels did not change significantly between tested groups (p=0.091). We found significantly lower high-density lipoprotein-cholesterol (HDL-c) (p0.001) and TAS (p0.001) levels and significantly higher hsCRP (p=0.002) and TBARS (p0.001) in CAD patients than in controls. There were significant positive correlations between TAS and Mn SOD mRNA (ρ=0.243, p=0.020) and TAS and Cu/Zn SOD mRNA (r=0.359, p0.001). TBARS negatively correlated only with Cu/Zn SOD mRNA (ρ=-0.215, p=0.040). TAS levels remained independent predictor for Mn SOD mRNA levels (OR=2.995, p=0.034).Results of this study showed that Mn SOD gene expression were decreased in CAD patients compared to controls and can be modulated by non-enzymatic antioxidant status in blood.Koronarna arterijska bolest (KAB) je jedan od najvažnijih uzročnika mortaliteta i morbiditeta u širokoj svetskoj populaciji. Dislipidemija, inflamacija i oksidativni stres mogu doprineti poremecaju strukture i funkcije endotela, nastanku ateroskleroze i KAB. Naša studija je imala za cilj da utvrdi da li ekspresije gena Cu/Zn supsoksid dismutaze (Cu/Zn SOD) i Mn superoksid dismutaze (Mn SOD) mogu da budu promenjene pod uticajem oksidativnog stresa kod pacijenata sa KAB.Ova studija je obuhvatila 77 pacijenata sa KAB i 31 naizgled zdravih osoba kod kojih su određeni nivoi serumskih lipida, visoko osetljivi C-reaktivni protein (hsCRP), ukupni antioksidativni status (TAS) i tiobarbituratna kiselina-reagujuće supstance (TBKRS). Ekspresija gena izoenzima SOD određena je u mononuklearnim ćelijama periferne krvi pomoću metode kvantitativne lančane reakcije polimeraze.Nivoi informacione ribonukleinske kiseline (iRNK) Mn SOD su značajno bili niži kod pacijenata sa KAB nego kod kontrole (p=0,011), dok se nivoi Cu/Zn SOD iRNK nisu značajno razlikovali između ispitanih grupa (p=0,091). Kod pacijenata sa KAB utvrđeni su znatno nižinivoi holesterola u lipoproteinima visoke gustine (HDL-h) (p0,001) i TAS (p0,001) i znatno veće koncentracije hsCRP (p=0,002) i TBKRS (p0,001). Postojale su značajne pozitivne korelacije između TAS i iRNK Mn SOD (ρ=0,243, p=0,020) i TAS i iRNK Cu/Zn SOD (ρ= 0,359, p0,001). TBKRS su bile u negativnoj korelaciji samo sa iRNK Cu/Zn SOD (ρ =-0,215, p= 0,040). TAS je bio nezavisni prediktor za nivoe iRNK Mn SOD (OR=2,995, p=0,034).Rezultati ove studije pokazali su da je ekspresija Mn SOD gena smanjena kod pacijenata sa KAB u poređenju sa kontrolnom grupom i da može biti modulirana neenzimskim antioksidativnim statusom u krvi.

Colorectal cancer (CRC) is a highly prevalent malignancy with multifactorial etiology, which includes metabolic alterations as contributors to disease development. Studies have shown that lipid status disorders are involved in colorectal carcinogenesis. In line with this, previous studies have also suggested that the serum high-density lipoprotein cholesterol (HDL-C) level decreases in patients with CRC, but more recently, the focus of investigations has shifted toward the exploration of qualitative properties of HDL in this malignancy. Herein, a comprehensive overview of available evidences regarding the putative role of HDL in CRC will be presented. We will analyze existing findings regarding alterations of HDL-C levels but also HDL particle structure and distribution in CRC. In addition, changes in HDL functionality in this malignancy will be discussed. Moreover, we will focus on the genetic regulation of HDL metabolism, as well as the involvement of HDL in disturbances of cholesterol trafficking in CRC. Finally, possible therapeutic implications related to HDL will be presented. Given the available evidence, future studies are needed to resolve all raised issues concerning the suggested protective role of HDL in CRC, its presumed function as a biomarker, and eventual therapeutic approaches based on HDL.

Linkage between bis(2-ethylhexyl) phthalate (DEHP), dibutyl phthalate (DBP), and bisphenol A (BPA) co-exposure and type 2 diabetes mellitus (T2DM), as well as ability of multi-strained probiotic to reduce DEHP, DBP and BPA mixture-induced oxidative damage in rat pancreas were investigated. The Comparative Toxicogenomics Database, Cytoscape software and ToppGene Suite were used for data-mining. Animals were sorted into seven groups (n = 6): (1) Control group: corn oil, (2) P: probiotic: Saccharomyces boulardii + Lactobacillus rhamnosus + Lactobacillus plantarum LP 6595 + Lactobacillus plantarum HEAL9; (3) DEHP: 50 mg/kg b.w./day, (4) DBP: 50 mg/kg b.w./day, (5) BPA: 25 mg/kg b.w./day, and (6) MIX: 50 mg/kg b.w./day DEHP + 50 mg/kg b.w/day DBP + 25 mg/kg b.w./day BPA; (7) MIX + P. Rats were sacrificed after 28 days of oral exposure. In silico investigation highlighted 44 DEHP, DBP and BPA mutual genes linked to the T2DM, while apoptosis and oxidative stress were highlighted as the main mechanisms of DEHP, DBP and BPA mixture-linked T2DM. In vivo experiment confirmed the presence of significant changes in redox status parameters (TOS, SOD and SH groups) only in the MIX group, indicating possible additive effects, while probiotic ameliorated mixture-induced redox status changes in rat pancreatic tissue.

Background:Non-cholesterol sterols (NCS) are promising biomarkers for estimation of cholesterol homeostasis properties. In addition, determination of NCS in high-density lipoprotein (HDL) fraction (HDL-NCS) could provide information on cholesterol efflux. However, matrix effects interfere in liquid chromatography–mass spectrometry (LC-MS)analysis of NCS, thereby impairing the method sensitivity.The aims of this study were development, optimization and validation of LC-MS method for quantification of NCS in serum and HDL-NCS. Additionally, matrix effect interfer-ences and methods application in individual serum samples were examined.Methods:HDL precipitating reagent was used for HDL isolation. Matrix effect was examined by comparing different surrogates by simple regression analysis. Validation was conducted according to the FDA-ICH guideline. 20 healthy volunteers were recruited for testing of method application.Results:The observed matrix effect was 30%, and matrix comparison showed that cholesterol was the dominant contributor to the matrix effect. Cholesterol concentration was adjusted by construction of the calibration curve for serum and HDL fraction (5 mmol/L and 2.5 mmol/L,respectively). The intra- and inter- run variabilities for NCSs were 4.7–10.3% for serum NCS and 3.6–13.6% for HDL-NCS and 4.6–9.5% for serum NCSs and 2.5–9.8% for HDL-NCS, respectively. Recovery studies showed satisfactory results for NCSs: 89.8–113.1% for serum NCS and85.3–95.8% for HDL-NCS. Conclusions:The method was successfully developed and optimized. The matrix interference was solved by customising calibration curves for each method and sample type.The measurement of NCS in HDL fraction was proposed for the first time as potentially useful procedure in biomedical researches. Uvod: Neholesterolski steroli (NHS) predstavljaju potencijalne biomarkere homeostaze holesterola. Pored toga, određivanje NHS u HDL frakciji (HDL-NHS) moglo bi da pruži dodatne informacije o efluksu holesterola. Međutim, određivanje NHS metodom tečne hromatografije - masene spektrometrije (LC-MS) je podložno uticaju matriksa usled čega dolazi do sniženja osetljivost metode. Studija je prvenstveno imala za cilj razvoj, optimizaciju i validaciju LC-MS metode za kvantifikaciju NHS u serumu i HDL-NHS. Pored toga, detaljno je ispitan efekat matriksa i metoda primenjena za analizu uzoraka seruma zdravih dobrovoljaca. Metode: Izolacija HDL frakcije je izvršena upotrebom precipitirajućeg reagensa. Efekat matriksa je ispitan poređenjem različitih surogat matriksa regresionom analizom. Validacija je sprovedena u skladu sa FDA-ICH vodičem za validaciju bioanalitičkih metoda. Korišćeni su serumi 20 zdravih dobrovoljaca u cilju kliničke evaluacije metode. Rezultati: Efekat matriksa je iznosio 30% i dalja analiza je pokazala da je holesterol bio glavni uzročnik efekta matriksa. U cilju rešavanja ove interferencije, kalibracione krive za su pripremljene u metanolnom rastvoru holesterola koncentracije 5 mmol/L, a za kvantifikaciju HDL-NHS u metanolnom rastvoru holesterola koncentracije 2,5 mmol/L. Varijacije unutar serije su bile 4,7-10,3% za NHS i 3,6-13,6% za HDL-NHS, a između serija 4,6-9,5% za NHS i 2,5-9,8% za HDL-NHS. Studije prinosa analita su pokazale zadovoljavajuće rezultate: 89,8-113,1% za NHS i 85,3-95,8% za HDL-NHS. Zaključak: Metoda je uspešno optimizovana i validirana. Interferencija usled prisustva holesterola je korigovana upotrebom odgovarajućih kalibracionih krivih za svaku vrstu uzorka. Određivanje HDL-NHS je po prvi put predloženo kao potencijalno korisna procedura u biomedicinskim istraživanjima.

Colorectal cancer (CRC) is a highly prevalent malignancy. Previous studies suggested that cholesterol might play a signficant role in malignant transformation and proliferation. Non-cholesterol sterols (NCS), which are transported by serum lipoproteins alongside cholesterol, are regarded as cholesterol synthesis and absorption markers. Quantification of NCS in serum and HDL fraction (NCSHDL), could provide a better insight into the cholesterol metabolism. The aim of this study was to examine the status of cholesterol synthesis and cholesterol absorption markers in serum and HDL fraction and explore their interrelation in CRC patients. Current study was designed as observational, case-control study. The study included 73 CRC patients and 95 healthy subjects. NCS and NCSHDL concentrations were determined by HPLC-MS/MS. Based on NCS and NCSHDL concentrations, different cholesterol homeostasis indices were calculated. Patients had significantly lower NCS (P

Milk and dairy products exhibit a beneficial nutritional and health effects and represent 20% to 30% of global food consumption. Whey proteins have great anabolic and cardioprotective role. Increased concentration of cholesterol contained in high density lipoproteins (HDL), paraoxonase 1 (PON1) higher activity, and diminished relative proportion of small dense low density lipoproteins (sdLDL) reduce cardiovascular diseases (CVD) risk. Current longitudinal study enrolled 10 healthy volunteers. All participants consumed lyophilized goat whey (LGW) (Koza Nostra®) for two months (3 g per day). Concentrations of basic biochemical parameters were measured by routine methods. Concentration of oxidative stress status parameters were measured spectrophotometrically. Serum PON1 activity was measured kinetically using paraoxon as a substrate. Lipoproteins subfractions were separated by gradient-gel electrophoresis. A significant decrease in urea concentrations (p

Background: Previous studies revealed decreased level of high-density lipoprotein cholesterol (HDL-C) as important factor for development of colorectal cancer (CRC). Quantity and structure of HDL particles depend on activities of lipid transfer proteins lecithin:cholesterol acyltransferase (LCAT) and cholesteryl ester transfer protein (CETP), but this topic is largely unexplored in CRC. The main objective of this study was to investigate activities of LCAT and CETP in patients with CRC. Additionally, we analyzed activity of paraoxonase-1 (PON-1), as a main carrier of HDL-antioxidant function. Materials and methods: Ninety-nine CRC patients and 101 healthy individuals were included. LCAT and CETP activities were assessed by measuring rates of formation and transfer of cholesteryl esters. PON-1 paraoxonase and arylesterase activities were measured. Results: Lower levels of HDL-C (p lt .001) were observed in cohort of patients, alongside with decreased LCAT (p lt .050) and increased CETP activity (p lt .050). Both PON-1 activities were diminished in CRC (p lt .050 and p lt .001 respectively). Univariate logistic regression singled out HDL-C level (OR = 0.218, p lt .001), CETP activity (OR = 1.010, p lt .01) and mass (OR = 0.994, p lt .001) as possible markers of elevated CRC risk. CETP mass maintained its predictive significance when adjusted for traditional risk factors and level of oxidative stress (OR = 0.993, p lt .001; OR = 0.982, p lt .050, respectively). Conclusion: Our results demonstrated increased CETP and decreased LCAT and PON-1 activities in CRC patients. In preliminary analysis CETP mass was identified as potential significant predictor of CRC development, suggesting that alterations in HDL-C levels, alongside with changes in HDL structure might have a role in carcinogenesis.

Background: Obesity-related childhood hypertension is associated with disturbances of serum lipids, but less is known about distribution of lipoprotein subclasses and activities of proteins involved in reverse cholesterol transport in hypertensive obese children. Our objective was to determine low-density lipoprotein (LDL) and high-density lipoprotein (HDL) subclasses distribution and activities of lecithin:cholesterol acyltransferase (LCAT) and cholesteryl ester transfer protein (CETP) in hypertensive and non-hypertensive obese children.Methods: A total of 40 hypertensive and 25 non-hypertensive obese children were enrolled. Lipoprotein subclasses were assessed by polyacrylamide gradient gel electrophoresis. LCAT and CETP activities were determined as a rate of formation and a rate of transfer of cholesteryl esters.Results: Despite of comparable values of serum lipid parameters, a shift toward smaller LDL and HDL subclasses was observed in hypertensive compared to normotensive obese children. Activities of LCAT were similar, but proatherogenic CETP activities were significantly higher in the hypertensive group (p= 0.036). LCAT/net CETP ratio inversely correlated with relative proportion of small, dense LDL particles ( =-0.423; p= 0.025) in the group with hypertension.Conclusions: The results of our study demonstrated a tendency toward altered distribution of lipoprotein subclasses in favor of more proatherogenic particles in childhood hypertension. Also, hypertensive obese children had increased proatherogenic CETP activity.

Cardiovascular complications, as the main cause of mortality in renal patients, are followed with altered lipoproteins composition. Considering that paraoxonase-1 (PON1) is an anti-oxidative enzyme located mainly on HDL particles, the current study has aim to investigate whether failure of kidney function leads to changes in the distribution of PON1 activity between different HDL subclasses.In 77 renal patients (21 chronic kidney disease (CKD) and 56 end stage renal disease (ESRD) patients on dialysis) and 20 healthy subjects PON1 activity on HDLSerum paraoxonase (p0.01) and arylesterase activity (p0.001) of PON1 as well as its concentration (p0.01) were significantly lower in CKD and ESRD patients compared to controls. Relative proportion of HDLThis study showed that altered HDL subclasses distribution, changed PON1 activities on different HDL subclasses as well as diminished anti-oxidative protection could be important factors in atherosclerosis development in CKD and ESRD patients.

Introduction: Cardiovascular complications, as the main cause of mortality in renal patients, are followed with altered lipoproteins composition. Considering that paraoxonase-1 (PON1) is an anti-oxidative enzyme located mainly on HDL particles, the current study has aim to investigate whether failure of kidney function leads to changes in the distribution of PON1 activity between different HDL subclasses. Materials and methods: In 77 renal patients (21 chronic kidney disease (CKD) and 56 end stage renal disease (ESRD) patients on dialysis) and 20 healthy subjects PON1 activity on HDL2 and HDL3 subclasses was determined by zymogram method that combines gradient gel electrophoresis separation of HDL subclasses and measurement of PON1 activity in the same gel. Results: Serum paraoxonase (p lt 0.01) and arylesterase activity (p lt 0.001) of PON1 as well as its concentration (p lt 0.01) were significantly lower in CKD and ESRD patients compared to controls. Relative proportion of HDL3 subclasses was higher in ESRD patients than in healthy participants, while HDL2 subclasses was significantly decreased in CKD (p lt 0.05) and ESRD (p lt 0.001) patients, as compared to controls. Furthermore, control subjects had higher PON1 activity on HDL2 (CKD and ESRD patients p lt 0.001) and HDL3 (CKD p lt 0.05; ESRD patients p lt 0.001) subclasses in comparison with the both patients groups. Also, significant negative correlation was found between paraoxonase activity of PON1 in serum and creatinine concentration (rho = -0.373, p lt 0.01). Conclusions: This study showed that altered HDL subclasses distribution, changed PON1 activities on different HDL subclasses as well as diminished anti-oxidative protection could be important factors in atherosclerosis development in CKD and ESRD patients.

Introduction Cholesterol homeostasis disruption contributes to the development of different pathologies. Non-cholesterol sterols (NCSs) serve as cholesterol synthesis markers (desmosterol and lathosterol), and cholesterol absorption surrogate markers (campesterol, stigmasterol and β-sitosterol). The study aimed to resolve certain new pre-analytical and analytical problems and ensure a reliable and validated method. Materials and methods Method optimization, validation and stability studies were executed in human serum and plasma. Freeze-thaw cycles were done with and without antioxidant. Gas chromatography-mass spectrometer (GC-MS) was used for NCSs confirmation and plasticizer identification, while GC-flame ionization detector (GC-FID) was used for NCSs quantitation. Results Intra- and inter-assay variabilities for all NCSs were 2.75–9.55% and 5.80–7.75% for plasma and 3.10–5.72% and 3.05–10.92% for serum, respectively. Recovery studies showed satisfactory percentage errors for all NCSs: 93.4–105.7% in plasma and 87.5–106.9 in serum. Derivatized samples were stable up to 7 days at − 20 °C and derivatization yield was affected by presence of plasticizers. Fatty acid amids were identified as interfering plastic leachates. Statistically different NCSs concentrations were observed after the 1st freeze-thaw cycle, in antioxidant-free samples, and after the 4th cycle in antioxidant-enriched samples. Conclusions All of the in-house procedures proved to be useful for minimizing the preanalytical and analytical variations, as proven by the validation results.

Objective: The connection of oxidative stress with dyslipidemia creates a newly-emerging atherosclerosis risk factor involved in acute ischemic stroke development. This study analyzed the influence of oxidative stress on structural changes of high-density lipoprotein (HDL) particles connected with modification in protective paraoxonase 1 (PON1) activity. Methods: This study used 185 patients with acute ischemic stroke and 185 apparently healthy controls. Oxidative stress status, PON1 status, lipids and high-sensitivity C-reactive protein (hsCRP) were determined. In isolated HDL lipoprotein fraction we determined selected markers of oxidative stress (malondialdehyde, MDA) and the content of total sulfhydryl (SH) groups. The capability of oxidative and PON1 status parameters to discriminate patients according to survival status was evaluated. Results: Stroke patients had lower HDL-cholesterol than controls and a remarkable fall in PON1 activity (control group-227 U/L, survivors-42 U/L, lethal outcome group-61 U/L, p lt 0.001), along with more prominent inflammation. Pronounced oxidative stress and impaired antioxidative protection was present among patients. HDL fraction analysis revealed a significant decrease of SH groups content (control group vs. patients, p lt 0.05) and increased in MDA content in patients (lethal outcome vs. control group, p lt 0.05). According to logistic regression analysis, the best predictor of disease outcome was oxidative stress marker - prooxidative-antioxidative balance (PAB). Conclusions: Pronounced oxidative stress in this group of acute ischemic stroke patients probably led to HDL structural changes, which could further cause an alteration or decrease of PON1 activity. Evidence of increased prooxidant level associated with decreased protective, antioxidative factors suggests their mutual involvement in this complex pathology.

85th Congress of the European-Atherosclerosis-Society (EAS), 2017.

85th Congress of the European-Atherosclerosis-Society (EAS), 2017.

Background:Cholesterol homeostasis disorders may cause dyslipidemia, atherosclerosis progression and coronary artery disease (CAD) development. Evaluation of non-cholesterol sterols (NCSs) as synthesis and absorption markers, and lipoprotein particles quality may indicate the dyslipidemia early development. This study investigates associations of different cholesterol homeostasis patterns with low-density (LDL) and high-density lipoproteins (HDL) subclasses distribution in statin-treated and statin-untreated CAD patients, and potential use of aforementioned markers for CAD treatment optimization.Methods:The study included 78 CAD patients (47 statin-untreated and 31 statin-treated) and 31 controls (CG). NCSs concentrations were quantified using gas chromatography- flame ionization detection (GC-FID). Lipoprotein subclasses were separated by gradient gel electrophoresis.Results:In patients, cholesterol-synthesis markers were significantly higher comparing to CG. Cholesterol-synthesis markers were inversely associated with LDL size in all groups. For cholesterol homeostasis estimation, each group was divided to good and/or poor synthetizers and/or absorbers according to desmosterol and β-sitosterol median values. In CG, participants with reduced cholesterol absorption, the relative proportion of small, dense LDL was higher in those with increased cholesterol synthesis compared to those with reduced synthesis (pConclusions:The results suggest the existence of different lipoprotein abnormalities according to various patterns of cholesterol homeostasis. Desmosterol/β-sitosterol ratio could be used for estimating individual propensity toward dyslipidemia development and direct the future treatment.

BACKGROUND: . Dyslipidemia is a common feature of chronic kidney disease (CKD). Although it has been observed that the pattern of lipid abnormalities can vary according to the stage of CKD, there is lack of data concerning the distribution of lipoprotein subclasses at various stages of the disease. In addition, association of proatherogenic small, dense low-density lipoprotein (sdLDL) subclasses with markers of inflammation, such is galectin-3, is not sufficiently explored. The aim of this study was to analyze concentrations and relative proportions of sdLDL-cholesterol (sdLDL-C) and galectin-3 in patients with CKD, with respect to the stage of the disease. Also, we sought possible independent associations of galectin-3 and sdLDL-C. Methods. The study involved 100 hemodialysis (HD) and 50 pre-dialysis patients, together with 94 healthy individuals. SdLDL-C was measured by heparin-magnesium precipitation method. Galectin-3 was measured by ELISA technique. Results. Galectin-3 levels were higher in pre-dialysis and HD patients than in the control group (p lt 0.01). The concentration of sdLDL-C was highest in the pre-dialysis group and lowest in HD patients (p lt 0.01). CKD patients with increased galectin-3 concentrations had significantly higher relative proportions of cholesterol in sdLDL (% sdLDL-C) than their counterparts with lower galectin-3 levels (p lt 0.05). Relative proportion of sdLDL-C was shown to be an independent determinant of galectin-3 concentration. Conclusions. Our results demonstrated alterations in concentrations and proportions of sdLDL-C according to the stages of CKD. The observed independent associations of % sdLDL-C and galectin-3 provide further insight into their complex interaction during the progression of atherosclerosis in CKD.

Obstructive sleep apnea (OSA) is associated with dyslipidemia and increased cardiovascular risk. We assessed the effects of apolipoprotein E ( APOE) genotype on low-density lipoprotein (LDL) and high-density lipoprotein (HDL) particle size and lipid subclasses (separated by gradient gel electrophoresis) in patients with OSA. Stable patients (n = 181) prospectively recruited underwent full polysomnography. Both LDL particle size and LDL I proportion were reduced from ∊3∊3 homozygotes to ∊2 carriers and to ∊4 carriers (analysis of variance: P = .024; P = .040, respectively); carriers of the ∊4 allele of the APOE genotype had significantly lower LDL particle size and LDL I proportion compared to ∊3∊3 homozygotes ( P < .05 for both comparisons). Insulin resistance increased from patients with no OSA to those with mild–moderate and to those with severe OSA ( P < .001). In multivariate analysis, LDL size was independently predicted by APOE genotype, male gender, and the presence of metabolic syndrome (MetS; P = .001, P = .020, P = .027, respectively). The HDL particle size was not affected by APOE genotype. Our data demonstrate that both the ∊4 APOE genotype and MetS are independently related to smaller LDL size in patients with OSA.

Congress of the European-Atherosclerosis-Society (EAS), 2016

Unfavorable lipid profile is a major risk factor for cardiovascular disease in renal pathology. In this study, we compared chronic renal patients and healthy controls with different LDL phenotypes (A or B) in respect of various biochemical parameters related to cardiovascular disease. Oxidative stress and anti-oxidative defense parameters [thiobarbituric acid-reacting substances (TBARS), total oxidative status (TOS), total anti-oxidative status (TAS), total protein sulfhydryl (-SH) groups], as well as red blood cell cholesterol distribution were assessed in 40 renal patients and 40 control subjects by standardized assays. LDL particle diameters were determined by polyacrylamide gradient gel electrophoresis. LDL particles are subdivided according to their size into large LDL A phenotype (diameter gt 25.5 nm) and small LDL B phenotype (diameter aecurrency sign25.5 nm). Renal patients with LDL A phenotype had increased oxidative stress (TOS: p lt 0.01, and TBARS: p lt 0.001) and decreased total SH- groups (p lt 0.001) compared to controls with the same LDL phenotype. A notable decrease in hemoglobin-cholesterol adduct was detected in patients with LDL A phenotype (p lt 0.001) and LDL B phenotype (p lt 0.05) compared with appropriate controls. LDL B phenotype was characterized with increased TBARS (p lt 0.05) compared with LDL A phenotype in control group. Increased oxidative stress, decreased anti-oxidative defense followed with unfavorable changes in hemoglobin-cholesterol binding capacity, could have important influence on cardiovascular disease risk in renal patients regardless of LDL phenotype.

Congress of the European-Atherosclerosis-Society (EAS), 2016.

Copeptin is a sensitive and more stable surrogate marker for arginine vasopressin. In this study, we evaluated copeptin levels in carotid endarterectomy (CEA) patients, perioperatively, to determine whether copeptin levels can be related to carotid artery cross clamping (CC) time and to postoperative neurological outcomes. Copeptin, interleukin 6, C-reactive protein, cortisol, and brain natriuretic peptide were measured preoperatively (T1) and 3 hours postoperatively (T3) as well as intraoperatively (T2). We recruited 77 patients. Values of copeptin rose gradually over the observed times: T1 = 7.9 (6.4-9.6), T2 = 12.6 (9.3-16.8), and T3 = 72.3 (49.1-111.2) pmol/L. There was a significant difference for repeated measurement ( P = .000, P = .000, and P = .000). Duration of carotid artery CC during CEA does not affect postoperative copeptin level (CC ≤ 13 minutes: 106.8 ± 93.6 pmol/L, CC > 13 minutes: 96.7 ± 89.1 pmol/L; P = .634). Preoperative copeptin level was significantly higher in patients with ulcerated plaque morphology. Activation of the stress axis in patients undergoing CEA results in copeptin elevation. Duration of CC during CEA does not affect postoperative copeptin levels.

Recent in vitro experiments have indicated that human resistin increases the number of lipoprotein particles secreted by the human hepatocytes and also influences their quality, in terms of generating more proatherogenic lipid particles. The aim of this study is to investigate associations of plasma resistin and peripheral blood mononuclear cells (PBMCs) resistin messenger RNA (mRNA) levels with different prevalence of small, dense low-density lipoprotein particles (sdLDL) in patients with indications for coronary angiography. This study included 65 patients requiring coronary angiography. There were 41 patients without significant stenosis and 24 patients with significant stenosis in at least one major coronary artery. Circulating resistin was measured by enzyme-linked immunosorbent assay; PBMC resistin mRNA was determined by real-time polymerase chain reaction. The LDL and high density lipoprotein subclasses were determined by gradient gel electrophoresis. Plasma resistin (P = 0.031) and PBMCs resistin mRNA (P = 0.004) were significantly higher in patients with proportion of sdLDL particles >= 50%, compared to the group with relative proportion of sdLDL particles lt 50%. Plasma resistin correlated positively with creatinine (r = 0.456, P lt 0.001) and resistin mRNA (r = 0.298, P = 0.014) but negatively with body mass index (r = -0.254, P = 0.034) and total cholesterol (r = -0.286, P = 0.021). Multiple linear regression analysis revealed LDL particle diameter as the only independent predictor of resistin mRNA (R-2 = 0.258; adjR(2) = 0.190). A significant association between resistin, both PBMCs mRNA and plasma protein, and the relative proportion of sdLDL particles in the circulation of coronary artery disease patients has been established, which implies that increased gene expression of resistin in PBMCs and higher resistin concentration in plasma are related to pro-atherogenic LDL particle phenotype.

Congress of the European-Atherosclerosis-Society (EAS), 2016.

83rd EAS Congress, EAS 2015

Objectives: End-stage renal disease (ESRD) is characterized by profound dyslipidemia and enhanced oxidative stress. The patients also show evidence of exhausted and/or deficient anti-oxidative defense enzymes, one of them being glutathione-S-transferase (GST). This study investigates relationship between GST gene polymorphism and low-density lipoprotein (LDL) and high-density lipoprotein (HDL) subclasses in ESRD. Design and methods: GSTM1, T1, and P1 genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism in 160 patients undergoing hemodialysis. LDL and HDL subclasses were separated by gradient gel electrophoresis and biochemical parameters were measured by routine laboratory methods. Results: GSTM1-positive patients had higher proportion of small, dense LDL III particles than those with GSTM1-null genotype (P lt 0.05). Similarly, GSTP1-Ile/Ile patients had higher proportion of LDL III (P lt 0.05), but more HDL 2b and less HDL 3a particles than GSTP1-Ile/Val and Val/Val carriers (P lt 0.05). LDL subclass distribution in smokers with GSTM1-null genotype was shifted towards smaller particles, as compared to GSTM1-positive and GSTM1-null non-smokers. Smokers with GSTP1-Ile/Val and Val/Val genotypes had smaller LDL size than their non-smoking counterparts (P lt 0.05). Both smokers and non-smokers with GSTP1 Ile/Ile genotype had more LDL III particles than non-smokers carrying Val allele. Non-smokers with GSTP1 Ile/Ile genotype had more HDL 2b subclasses than non-smokers with GSTP1-Ile/Val and Val/Val (P lt 0.05), but less HDL 3a particles than smokers with GSTP1-Ile/Val and Val/Val genotypes (P lt 0.05). GSTT1 gene polymorphism had no effect on lipoprotein subclass distributions. Conclusions: Our results demonstrate significant associations between low activity GST genotypes and proatherogenic lipoprotein particles in hemodialysis patients which might further increase their cardiovascular disease risk.

Pregnancy is associated with alterations in low-density lipoprotein (LDL) and high-density lipoprotein (HDL) subclasses, but the exact pattern of these variations remains controversial. This study investigates longitudinal changes of plasma LDL and HDL particles distributions during the course of normal pregnancy, as well as associations of maternal LDL and HDL subclasses distributions before delivery with parameters of newborn size. Blood samples were collected from 41 healthy pregnant women throughout entire pregnancy, before delivery and 7 weeks postpartum. LDL and HDL subclasses were determined by gradient gel electrophoresis, while other biochemical parameters were measured by standard laboratory methods. During gestation LDL size significantly decreased (P lt 0.001), due to reduction in relative proportion of LDL I (P lt 0.01) and increase of LDL II (P lt 0.001) and IIIA (P lt 0.05) subclasses. In the same time, HDL size and proportions of HDL 2a particles significantly decreased (P lt 0.001), with concomitant increase of HDL 3b and 3c subclasses (P lt 0.05). Observed alterations were associated with changes in serum triglyceride levels. Rearrangement in LDL subclasses distribution during gestation was transient, while postpartum HDL subclasses distribution remained shifted toward smaller particles. Higher proportion of LDL IVB in maternal plasma before delivery was an independent predictor of smaller birth weights and lengths, while higher proportions of LDL IVB and HDL 2a subclasses were independent determinants of newborns' smaller head circumferences. Routine gestational and prenatal care in otherwise normal pregnancy could be complemented with evaluation of LDL and HDL particles distribution in order to ensure an adequate size of the newborn.

Context.—Systemic inflammatory diseases are associated with proatherogenic lipoprotein profile, but there is a lack of information regarding overall distributions of lipoprotein subclasses in sarcoidosis. Objective.—To investigate whether patients with sarcoidosis have altered distributions of plasma low-density lipoprotein (LDL) and high-density lipoprotein (HDL) particles. Design.—Seventy-seven patients with biopsy-proven sarcoidosis (29 with acute and 48 with chronic sarcoidosis) treated with corticosteroids and 77 age- and sex-matched controls were included in the study. Low-density lipoprotein and HDL subclasses were determined by gradient gel electrophoresis, while inflammatory markers and lipid parameters were measured by standard laboratory methods. Results.—Compared to controls, patients had fewer LDL I subclasses (P < .001), but more LDL II and III (P < .001) subclasses. This pattern was evident in both acute and chronic disease groups. Patients also had smaller HDL size (P < .001) and higher proportions of HDL 2a (P = .006) and 3a particles (P = .004). Patients with chronic sarcoidosis had smaller LDL size than those with acute disease (P = .02) and higher proportions of HDL 3a subclasses (P = .04) than controls. In acute sarcoidosis, relative proportions of LDL and HDL particles were associated with levels of inflammatory markers, whereas in chronic disease an association with concentrations of serum lipid parameters was found. Conclusions.—The obtained results demonstrate adverse lipoprotein subfraction profile in sarcoidosis with sustained alterations during disease course. Evaluation of LDL and HDL particles may be helpful in identifying patients with higher cardiovascular risk, at least for prolonged corticosteroid therapy due to chronic disease course.

Carotid endarterectomy (CEA) is a surgical procedure performed to reduce the incidence of embolic and thrombotic stroke. Although only a preventive procedure, CEA carries the risk of perioperative complications. There is constant searching for an optimal anesthetic technique. There are pros and cons for both anesthetic techniques used: regional (RA) and general anesthesia (GA). A large number of studies have compared RA and GA techniques in CEA surgery patients. The primary outcome was the proportion of patients with stroke, myocardial infarction, or death. However, neither the GALA trial nor the pooled analysis was adequately powered to reliably detect an effect of type of anesthesia on mortality. It may therefore be appropriate to consider other additional parameters (stress response, incidence of postoperative delirium and cognitive impairment, functional recovery, total surgery time, intensive care unit requirement, hospital stay, hospital costs and patients satisfaction) when comparing the outcomes of the two techniques.Although, the debate continues as to whether regional anesthesia or general anesthesia is safer, the choice of anesthetic technique is a complex decision and surgical teams should be able to offer both RA and GA. The individual approach is the ideal choice and should be determined at the discretion of the surgeon, anesthetist and patient depending on the clinical situation and own preferences.