Hypomethylation of LINE-1 but not Alu in lymphocyte subsets of systemic lupus erythematosus patients

BackgroundT lymphocytes from SLE patients have a global decrease in the 5-methylcytosine content. Previous studies have identified hypomethylation in the promoter of several genes but there is limited study in the interspersed repetitive sequences (IRSs).MethodsWe examined and compared the methylation levels of long interspersed nuclear element 1 s (LINE-1) and Alu elements in normal and SLE CD4+ T lymphocytes, CD8+ T lymphocytes and B lymphocytes by the combined bisulfite restriction analysis-interspersed repetitive sequences (COBRA-IRS).ResultsHypomethylation of LINE-1 but not Alu was found in CD4+ T lymphocytes, CD8+ T lymphocytes, and B lymphocytes of SLE patient (P = 0.005, 0.002, and 0.007, respectively). Moreover, when the SLE patients were divided into active and inactive groups, LINE-1 hypomethylation was more significantly distinguished in both CD4+ and CD8+ T lymphocytes of patients from the active SLE group when compared to the controls. Surprisingly, Alu hypomethylation was also observed in CD8+ T lymphocytes from the inactive SLE group when compared to the normal controls (P = 0.0056).ConclusionsThe hypomethylation in each lymphocyte subset of SLE was IRSs specific, mainly found in LINE-1 rather than Alu.     

Folate and choline metabolism gene variants and development of uterine cervical carcinoma

ObjectiveIt has been reported that aberrant DNA methylation can be associated with HPV infection and cervical tumorigenesis. The aim of this study was to evaluate the possibility that polymorphic variants of genes that may affect DNA methylation status are associated with the risk of cervical cancer in the Polish population.Design and methodEmploying PCR-RFLPs and HRM analyses, we examined the prevalence of BHMT Arg239Gln (rs3733890), MTR Asp919Gly (rs1805087), MTHFR Ala222Val (rs1801133), MTHFD1 Arg653Gln (rs2236225) and MTRR Ile22Met (rs1801394) genotypes and alleles in patients with advanced cervical cancer (n = 124) and controls (n = 168).ResultsThe odds ratio (OR) for BHMT Gln/Gln genotype was 0.433 (95% CI = 0.1780–1.054; p = 0.0602). The OR for patients having the BHMT Arg/Gln or Gln/Gln genotypes was 0.579 (95% CI = 0.3622–0.924; p = 0.0216). We also observed a significantly higher frequency of the BHMT 239Gln allele in controls than in patients, p = 0.0165. The genotype and allele frequencies of the MTR Asp919Gly, MTHFR Ala222Val, MTHFD1 Arg653Gln and MTRR Ile22Met gene variants did not display statistical differences between patients with cervical cancer and controls. We also did not find a significant association between the distribution of any genotypes or alleles and cancer characteristics.ConclusionOur results might suggest the protective role of the BHMT 239Gln variant in cervical cancer incidence.     

Elevated levels of myeloperoxidase, white blood cell count and 3-chlorotyrosine in Taiwanese patients with acute myocardial infarction

ObjectivesInflammation, a major risk factor for acute myocardial infarction (AMI), is associated with leukocytic activation, secretion of myeloperoxidase (MPO) and generation of the oxidative damage marker, 3-chlorotyrosine (3-Cl-Tyr). To study their association with AMI and their value in diagnosis of AMI, white blood cell (WBC) count, plasma MPO, plasma 3-Cl-Tyr, and conventional risk factors such as cardiac troponin I and CK-MB were examined in AMI patients during the onset of chest pain.MethodsAfter obtaining informed consent, blood samples were collected from 77 AMI patients during the onset of chest pain and from 53 normal controls. The samples were analyzed for WBC count using SE-9000 automated analyzer. Plasma MPO was measured by an enzyme-linked immunosorbent assay. Plasma levels of 3-Cl-Tyr, a product of MPO, were analyzed by HPLC coupled with Coularray electrochemical detection.ResultsThe WBC, plasma MPO and 3-Cl-Tyr levels were significantly elevated in AMI patients than in normal controls (p < 0.001). The levels of WBC, MPO and 3-Cl-Tyr alone were strongly associated with the prevalence of AMI. Plasma MPO was correlated with 3-Cl-Tyr (r = 0.389, p < 0.01) and WBC counts (r = 0.405, p < 0.01) respectively. The ROC curve analyses suggested that MPO had the best specificity and sensitivity among these oxidative stress-related markers.ConclusionPlasma MPO value should be considered as a better marker for early diagnosis of AMI, as compared with WBC count or 3-Cl-Tyr.     

Usefulness of glycated albumin as an indicator of glycemic control status in patients with hemolytic anemia

BackgroundIn patients with hemolytic anemia (HA), glycated hemoglobin (HbA_1C) presents lower values in relation to glycemia because the lifespan of erythrocytes is shortened, whereas glycated albumin (GA) is not affected. In the present study, we examined the usefulness of GA as an indicator of glycemic control status in patients with HA.MethodsWe enrolled 21 patients with HA. A total of 202 patients with type 2 diabetes mellitus (T2DM) without complications were used as controls.ResultsWe identified a significant correlation between GA and HbA_1C in the patients with HA. However, in a comparison between the patients with HA and those with T2DM, the regression line showed a leftward shift in the former group. There was a significant positive correlation between hemoglobin (Hb) and HbA_1C in the patients with HA (R = 0.541, p = 0.025), although there was no significant correlation between Hb and GA. There was an inverse correlation between Hb levels and GA/HbA_1C ratio (R = ? 0.710, p = 0.001). The measured HbA_1C levels were lower than the HbA_1C levels estimated from mean plasma glucose levels, whereas the GA/3 levels were close to the estimated HbA_1C levels.ConclusionsGA is a useful indicator of glycemic control status in patients with HA.     

Variation of serum C-reactive protein (CRP) over time in pediatric cancer patients with febrile illness and its relevance to identified pathogen

ObjectiveTo evaluate the correlation of serum CRP with clinical and laboratory parameters proven to be related to the cause of infection in pediatric cancer patients.MethodsWe studied prospectively for a 12-month period, 37 pediatric cancer patients, who presented with 70 episodes of febrile illness (38 bacterial and 13 viral infections).At fever's onset and 48 h later, infection indices, such as CRP, WBC, ANC were measured in the peripheral blood. Moreover we calculated the change rate of CRP over 48 h [CRP/t = (CRP48h ? initial CRP) / t (t = 2 days)]. Cultures of biological fluids, PCR and antibody detection of infectious agents were also obtained.ResultsWhen comparing patients with viral vs. bacterial infections, mean CRP levels on admission (11.0 vs. 33.1 mg/L, p = 0.005) and at 48 h (13.4 vs. 71.9 mg/L, p = 0.0007), and CRP/t (0.9 vs. 18.8 mg/L/day, p = 0.030) were significantly lower in the group with viral infection.At 48 h - follow-up, patients with positive culture had higher CRP levels (57.3 vs. 43.3 mg/L, p = 0.048) and higher CRP/t (15.9 vs. 7.7 mg/L/day, p = 0.025), compared to those without proven infection. CRP/t at 48 h was correlated with both the fever duration (r = 0.27, p = 0.027) and maximum temperature (Tmax) during the febrile episode (r = 0.30, p = 0.013).ConclusionsSingle CRP values on fever initiation can differentiate between viral and bacterial infections in febrile pediatric cancer patients. Moreover the change rate of CRP over time (CRP/t) is offered as a prognostic index of bacterial infection and a marker of the total duration of fever and Tmax.     

Diagnostic accuracy, reproducibility and robustness of fibrosis blood tests in chronic hepatitis C: a meta-analysis with individual data

ObjectivesTo evaluate the diagnostic accuracy of liver fibrosis tests and its influencing factors in a meta-analysis with individual data.Design and methodsFour independent centers provided four blood tests and Metavir staging from 825 patients with chronic hepatitis C.ResultsFibroMeter AUROC (0.840) for significant fibrosis was superior to those of Fibrotest (0.803, p = 0.049), APRI (0.789, p = 0.001) and Hepascore (0.781, p < 0.001). The misclassification rate was lower for FibroMeter (23%) than for Fibrotest and Hepascore (both 28%, p < 0.001). The variation in the diagnostic cut-offs of tests among centers, reflecting the overall reproducibility, was: FibroMeter: 4.2%, APRI: 24.0%, Fibrotest: 24.2%, Hepascore: 35.0%. Accordingly, the proportion of patients diagnosed with significant fibrosis changed: FibroMeter: 0.8%, Hepascore: 2.4% (p = 0.02 vs FibroMeter), Fibrotest: 5.8% (p < 10^? 3), APRI: 18.2% (p < 10^? 3).ConclusionsThis study on clinical applicability shows significant differences in diagnostic accuracy, inter-center reproducibility, and robustness of biomarkers to changes in population characteristics between blood tests.     

Synergistic effects of the polymorphisms in the PAI-1 and IL-6 genes with smoking in determining their associated risk with coronary artery disease

ObjectivesTo assess the relationship between IL-6 and PAI-1 polymorphisms and coronary artery disease (CAD) and to observe the interactions between these polymorphic variants and smoking in the CAD risk.Design and methodThe study population consisted of 178 patients with angiographically documented CAD and 202 blood donors. The analyses of genetic polymorphisms were performed using the PCR-RFLP method.ResultsThe frequency of PAI-1 5G allele was higher in the entire CAD group than in control group (p = 0.04, OR = 1.35). Also the 5G allele carriers (4G5G + 5G5G) were more frequent in patients than in controls (p = 0.03, OR = 1.93). The number of women carrying 5G allele was again significantly higher among patients (OR = 10.95 p = 0.0075). The IL-6 C allele frequency was higher only in the CAD male subgroup (p = 0.035, OR = 1.44). We found synergistic and cumulative effects between specific genotype patterns and smoking in determining the risk of CAD, especially between PAI-1(4G5G + 5G5G)+IL-6(CC) and smoking (SIM = 4.18 and p = 0.0005, OR = 9.20, respectively).ConclusionsThere are synergistic and cumulative effects of 5G allele of PAI-1 polymorphism and C allele of IL-6 polymorphism with smoking in determining their associated risk with CAD.     

Cystatin C as a predictor of all-cause mortality and myocardial infarction in patients with non-ST-elevation acute coronary syndrome

ObjectivesTo investigate the predictive value of cystatin C among patients diagnosed with non-ST-elevation acute coronary syndrome (nSTE-ACS).Design and methodsAdmission serum samples from 245 nSTE-ACS patients were measured with a novel cystatin C immunoassay based on a dry-reagent, double monoclonal design. Creatinine concentrations, estimated glomerular filtration rates (eGFR) and one-year follow-up data were available for these patients.ResultsDuring the follow-up period, 34 (14%) of patients had myocardial infarction (MI) and 25 (11%) died. Increased serum cystatin C was an independent predictor of all-cause mortality and combined events (all-cause mortality and MI) after adjustment to non-biomarker baseline factors, hazard ratio (HR) 2.19 (per increase of 1 tertile; 95% Cl 1.28–3.78, p = 0.0046) and 1.75 (1.22–2.51, p = 0.0024), respectively. Corresponding values for eGFR were 2.56 (1.43–4.59, p = 0.0016) and 1.76 (1.23–2.53, p = 0.0022), respectively. Creatinine was not an independent predictor of endpoints (p > 0.05).ConclusionsCystatin C was associated with an increased risk of death and combined events in patients with nSTE-ACS.     

Heritability of genetic variants of resistin gene in patients with coronary artery disease: a family-based study

ObjectiveThe resistin gene (RETN) ?420 C > G and + 299 G > A polymorphism was investigated in a case-control study from forty complex Pakistani families with coronary artery disease (CAD) history. Heritability of the susceptible/variant alleles was investigated from parent–offspring trios in these families.MethodResistin levels were determined from 239 individuals by enzyme-linked immunosorbent assay. Genotyping was performed using polymerase chain reaction and restriction fragment length polymorphism.ResultsElevated resistin levels were observed from CAD cases vs. controls (p < 0.0001). The RETN ? 420 C > G and + 299 G > A polymorphism was more prevalent in cases vs. controls (p < 0.0001). The transmission disequilibrium test revealed a significant association of the ? 420 and + 299 polymorphism with CAD (χ² = 34.4, p < 0.0001 and χ² = 31.6, p < 0.0001, respectively).ConclusionElevated resistin, and the RETN ?420 C > G and + 299 G > A polymorphism may contribute to familial CAD. The ? 420 and + 299 variant alleles are transmitted more frequently from parent to affected offspring. This is the first report on the association of the RETN + 299 G > A polymorphism with CAD.     

Association of exonic variants of Klotho with metabolic syndrome in Asian Indians

BackgroundKlotho, an anti-aging gene, is a functional candidate for metabolic syndrome. We conducted a cross-sectional study to evaluate the association of the genetic variants of Klotho with metabolic syndrome and surrogates of insulin resistance in Asian Indians.MethodsWe recruited 428 clinically normal subjects for the study. Genotyping was done by polymerase chain reaction and restriction fragment length polymorphism.ResultsSignificant and borderline associations of the KL-VS (OR = 15.88 [95%CI, 2.56–98.70], p = 0.003) and C1818T (OR = 0.28 [95%CI, 0.07–1.07], p = 0.063) variants of the Klotho gene, respectively, were observed with metabolic syndrome. The association of the KL-VS variant with metabolic syndrome could be linked to its observed influence on high blood glucose (OR = 6.92 [95% CI = 1.75–27.44], p = 0.006), high blood pressure (OR = 5.21 [95%CI = 1.00–38.43], p = 0.046), insulin resistance (OR = 3.59, [95%CI = 1.01–12.79], p = 0.048) and trend towards its association with hypertriglyceridemia (OR = 3.69 [95%CI = 0.92–14.77], p = 0.065).ConclusionsThe genetic variants of Klotho might predict risk for metabolic syndrome and insulin resistance in Asian Indians. However, larger studies in other ethnic populations are warranted to determine the role of these gene variants in the etiology of metabolic syndrome.     

Coenzyme Q10 levels are low and associated with increased mortality in post-cardiac arrest patients

AimSurvival after cardiac arrest (CA) is limited by the profound neurologic insult from ischemia–reperfusion injury. Therapeutic options are limited. Previous data suggest a benefit of coenzyme Q₁₀ (CoQ₁₀) in post-arrest patients. We hypothesized that plasma CoQ₁₀ levels would be low after CA and associated with poorer outcomes.MethodsProspective observational study of post-arrest patients presenting to a tertiary care center. CoQ₁₀ levels were drawn 24 h after return of spontaneous circulation (ROSC) and compared to healthy controls. Levels of inflammatory cytokines and biomarkers were analyzed. Primary endpoints were survival to discharge and neurologic status at time of discharge.Results23 CA subjects and 16 healthy controls were enrolled. CoQ₁₀ levels in CA patients (0.28 μmol L^?1, inter-quartile range (IQR): 0.22–0.39) were significantly lower than in controls (0.75 μmol L^?1, IQR: 0.61–1.08, p < 0.0001). The mean CoQ₁₀ level in CA patients who died was significantly lower than in those who survived (0.27 vs 0.47 μmol L^?1, p = 0.007). There was a significant difference in median CoQ₁₀ level between patients with a good vs poor neurological outcome (0.49 μmol L^?1, IQR: 0.30–0.67 vs 0.27 μmol L^?1, IQR: 0.21–0.30, p = 0.02). CoQ₁₀ was a statistically significant predictor of poor neurologic outcome (adjusted p = 0.02) and in-hospital mortality (adjusted p = 0.026).ConclusionCoQ₁₀ levels are low in human subjects with ROSC after cardiac arrest as compared to healthy controls. CoQ₁₀ levels were lower in those who died, as well as in those with a poor neurologic outcome.     

Plasma ceruloplasmin as a biomarker for obesity: a proteomic approach

ObjectivesThis study aimed to investigate new biomarkers of obesity particularly in relation with inflammation-associated proteins using protein differential display techniques.Design and methodsComparison of protein expression in plasma between non-obese (n = 109, body mass index, BMI < 25 kg/m²) and obese (n = 32, BMI ≥ 25 kg/m²) groups was carried out using two-dimensional gel electrophoresis (2-DE) analysis. ELISA was also performed for validation.ResultsAmong six differentially expressed protein spots, ceruloplasmin (Cp) and fibrinogen were over-expressed in obese group. Plasma Cp levels were significantly higher in obese group than non-obese group (34.0 ± 8.6 vs. 41.3 ± 12.7 mg/dL, p < 0.001) and positively correlated with age (r = 0.253, p < 0.005), BMI (r = 0.265, p < 0.001) and hsCRP (r = 0.385, p < 0.001). In stepwise multiple linear regression analysis, plasma Cp along with hsCRP were found predictors for obesity (adjusted β-coefficient = 0.266, p < 0.01).ConclusionElevated plasma Cp levels were significantly associated with obesity, which may be suggested to be a marker of obesity.     

Mitochondrial depolarisation and oxidative stress in rheumatoid arthritis patients

ObjectivesTo investigate mitochondrial membrane integrity, lipid peroxidation and cytotoxicity in peripheral lymphocytes (PL) from rheumatoid arthritis (RA) patients.Design and methodsSouth African black RA patients (HIV^?) were recruited into the study. Mitochondrial membrane potential (Δψ_m) was analysed in PL using the JC-1 dye distribution assay and flow cytometry. Correlations between Δψ_m and clinical parameters were tested for statistical significance. Cytotoxicity (LDH) and lipid peroxidation (thiobarbituric acid reactive substances (TBARS)) was also determined.ResultsOur findings show significantly elevated levels of cytotoxicity (p = 0.0029) and lipid peroxidation (p = 0.0030) in RA. A significantly higher percentage of circulating PL contained depolarised mitochondria (p = 0.0003) which correlated with disease activity and C-reactive protein levels in patients. Collapse of Δψ_m also negatively correlated to absolute lymphocyte counts (r = ? 0.4041; p = 0.0197).ConclusionThese findings suggest a possible role for mitochondrial membrane alterations in the pathology of RA.     

HOMA-IR and non-HDL-C as predictors of high cholesteryl ester transfer protein activity in patients at risk for type 2 diabetes

Background and aimsMetabolic syndrome (MS) and type 2 diabetes are highly associated with an abnormal lipoprotein profile, which may be generated and accentuated by high cholesteryl ester transfer protein (CETP) activity. Given the difficulty in measuring CETP activity, the aim was to identify simple biochemical predictors of high CETP activity.Design and methodsEighty five subjects at risk for type 2 diabetes were classified according to the presence of MS. Lipoprotein profile, HOMA-IR and endogenous CETP activity were evaluated.ResultsAs expected, MS patients presented higher concentration of glucose, insulin, triglycerides and non-HDL-C and lower HDL-C levels. Moreover, MS patients exhibited increased HOMA-IR and CETP activity. Employing a ROC curve for MS, high CETP activity was defined as > 250% ml^? 1 h^? 1. The predictive variables of high CETP were non-HDL-C ≥ 160 mg/dl (OR = 11.1;95%IC = 3.3–38.2;p < 0.001) and HOMA-IR > 2.1 (OR = 4.4;95%IC = 1.3–14.8;p < 0.05).ConclusionsHigh non-HDL-C and insulin resistance were predictors for increased CETP activity which measurement is not accessible for clinical laboratories.     

Pro-inflammatory and atherogenic circulating factors in non-alcoholic fatty liver disease associated to metabolic syndrome

BackgroundIt is not elucidated if liver fat deposits associated to metabolic syndrome (MS) aggravate the atherogenic state. We evaluated, in MS patients, if the presence of non-alcoholic hepatic steatosis (HS) determines differences in inflammatory markers and VLDL characteristics.MethodsSeventy-five patients with MS were divided into 2 groups depending on the presence or absence of HS, assessed by ultrasound. Lipid profile, free fatty acids (FFA), VLDL composition, adiponectin, tumor necrosis factor-alpha (TNF-α), high sensitivity C-reactive protein (hs-CRP), and soluble adhesion molecules (sVCAM-1 and sICAM-1) were measured.ResultsHS patients presented increased triglycerides levels, HOMA-IR and FFA. Patients with HS showed a reduction in adiponectin (p = 0.04) and increase in hs-CRP (p = 0.02), independently of insulin-resistance (IR). FFA correlated positively with TNF-α (p = 0.04) and inversely with adiponectin (p = 0.01). hs-CRP correlated with all inflammatory markers, independently of IR: TNF-α (r = 0.34, p = 0.02), sVCAM-1 (r = 0.29 p = 0.03), sICAM-1 (r = 0.56, p = 0.01), adiponectin (r = ?0.34, p = 0.04). HS patients presented higher VLDL mass and number of particles. Adiponectin correlated with VLDL cholesterol content (r = ?0.47, p = 0.04), independently of IR. VLDL, once secreted, would suffer from changes, becoming more atherogenic.ConclusionsSimple HS would play an important role increasing cardiovascular risk, independently of IR. hs-CRP may represent a useful biomarker of this condition.     

Expression analysis and clinical utility of L-Dopa decarboxylase (DDC) in prostate cancer

BackgroundL-Dopa decarboxylase (DDC) is a pyridoxal 5′-phosphate-dependent enzyme that was found to be involved in many malignancies. The aim of this study was to investigate the mRNA expression levels of DDC in prostate tissues and to evaluate its clinical utility in prostate cancer (CaP).MethodsTotal RNA was isolated from 118 tissue specimens from benign prostate hyperplasia (BPH) and CaP patients and a highly sensitive quantitative real-time RT-PCR (qRT-PCR) method for DDC mRNA quantification has been developed using the SYBR Green^® chemistry. LNCaP prostate cancer cell line was used as a calibrator and GAPDH as a housekeeping gene.ResultsDDC was found to be overexpressed, at the mRNA level, in the specimens from prostate cancer patients, in comparison to those from benign prostate hyperplasia patients (p < 0.001). Logistic regression and ROC analysis have demonstrated that the DDC expression has significant discriminatory value between CaP and BPH (p < 0.001). DDC expression status was compared with other established prognostic factors, in prostate cancer. High expression levels of DDC were found more frequently in high Gleason's score tumors (p = 0.022) as well as in advanced stage patients (p = 0.032).ConclusionsOur data reveal the potential of DDC expression, at the mRNA level, as a novel biomarker in prostate cancer.     

CLSI EP17-A protocol: a useful tool for better understanding the low end performance of total prostate-specific antigen assays

BackgroundClinical Laboratory and Standards Institute (CLSI) published EP17-A guideline, recommending new definitions for low end performances: Limit of Blank (LoB), Limit of Detection (LoD) and Limit of Quantitation (LoQ). The aim of this study was to determine LoB, LoD and LoQ by applying EP17-A to Hybritech and World Health Organization (WHO) calibrated Access Total PSA assays, and verify the correlation between results generated by the same reagent with both calibrations, particularly at low end concentrations.MethodsAccording to EP17-A, serum pools of anonymous routine patient samples residuals were analyzed on a UniCelDxI800 with the chemiluminescent Access®Hybritech®TotalPSA assay.ResultsLoB: 0.0046 μg/L Hybritech, 0.005 μg/L WHO calibration; LoD: 0.014 μg/L Hybritech, 0.015 μg/L WHO; LoQ at 20% coefficient of variation (CV%) 0.0414 μg/L Hybritech, 0.034 μg/L WHO. Regression Hybritech y = 0.2398× + 4.2017 (R² = 0.9515); WHO y = 0.2248× + 3.4335 (R² = 0.9596) with no statistical difference. Comparison between Hybritech and WHO at low PSA levels indicated an excellent Pearson's and intraclass correlation (r = 0.999, p < 0.001; ICC = 0.974, p < 0.001).ConclusionsOur results show that the Access Total PSA assay is suitable for prostate cancer recurrence and PSA velocity evaluation; Hybritech and WHO calibrated values can both be used for clinical purposes even at low levels.     

Clinical significance of Stratifin,ERα and PR promoter methylation in tumor and serum DNA in Indian breast cancer patients

Objectives:The objective of this study was to determine the concordance of promoter methylation of stratifin, ERα and PR in tumor and circulating DNA in breast cancer patients and their association with clinicopathological parameters and disease prognosis.Design and methods:Methylation specific PCR were carried out to investigate the promoter methylation status of stratifin, ERα and PR in tumor and circulating DNA in 100 breast cancer patients in a prospective study. The effect of promoter methylation on protein expression was evaluated by immunohistochemistry.Results:Significant association was observed between promoter methylation of stratifin in tumors (61%) and paired sera (56%) (r = 0.78; p ≤ 0.001). Loss of stratifin expression was observed in 47% tumors and was associated with poor overall survival (p = 0.05). Significant correlation was observed between methylation status of ERα with PRB (p < 0.0001, OR = 20.8, 95% CI = 7.4–58.0) and stratifin (p = 0.003, OR = 2.0, 95% CI = 0.8–4.4).Conclusion:This study underscores the potential utility of serum DNA methylation of these genes as surrogate for tumor DNA methylation as a promising tool for cancer diagnosis.     

Predominance of large VLDL particles in metabolic syndrome, detected by size exclusion liquid chromatography

ObjectiveTo study size heterogeneity of triglyceride rich lipoproteins (TRL) in metabolic syndrome (MS).Design and methodsThirty MS patients and 14 healthy subjects were included. In fasting serum we measured: lipid profile, free fatty acids (FFA) and adiponectin; TRL were isolated (d < 1.006 g/mL) and analysis by size exclusion HPLC followed by UV detection was performed; each subfraction was expressed as percentage of total TRL.ResultsMS patients, even those with normal triglycerides, presented higher proportion of very large VLDL (90 nm diameter) and large VLDL (60 nm) and slightly lower of typical VLDL (37 nm) (p < 0.04); increased FFA (p = 0.04) and lower adiponectin (p = 0.001). FFA correlated with large VLDL% (r = 0.58; p = 0.003), independently of insulin-resistance and waist. Furthermore, the lower the adiponectin, the greater the predominance of large VLDL (r = ? 0.40; p = 0.04).ConclusionMS was associated with large VLDL, described as more atherogenic beyond triglyceride levels. Size exclusion HPLC would represent a useful tool for assessing subfractions' lipoprotein profile.