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.     

Does non-alcoholic fatty liver impair alterations of plasma lipoproteins and associated factors in metabolic syndrome?

BackgroundHepatic steatosis (HS) is closely associated to metabolic syndrome (MS). Both, VLDL-triglyceride oversecretion and intrahepatic deposits, can take place. We evaluated VLDL characteristics, CETP, hepatic lipase (HL), IDL and small dense LDL (sdLDL), in patients with HS associated to MS.MethodsWe studied 3 groups matched by age and sex: 25 MS patients with HS (diagnosed by ultrasonography), 25 MS patients without HS and 25 healthy controls. Main measurements were: lipid profile, free fatty acids, VLDL composition, VLDL size by HPLC, CETP and HL activities, IDL-cholesterol and sdLDL-cholesterol.ResultsPatients with HS presented higher triglyceride levels, HOMA-IR and free fatty acids, VLDL mass and VLDL-apoB (p < 0.05). No differences in VLDL composition were observed. MS groups presented higher proportion of large VLDL than controls (p < 0.05). HS group showed higher CETP than controls (p = 0.01) and almost higher than MS without HS (p = 0.06). CETP correlated with VLDL-cholesterol content, r = 0.48, p < 0.005. The increase in sdLDL-cholesterol correlated with CETP (r = 0.47) and HL (r = 0.56), independent of insulin resistance (p < 0.003).ConclusionDespite intrahepatic fat, patients with HS secreted higher number of VLDL particles. CETP would have a remodeling action on VLDL in circulation, enriching it in cholesterol and also favoring, together with HL, the formation of sdLDL.     

Comparative study between anion-exchange HPLC and homogeneous assay methods in regard to the accuracy of high- and low-density lipoprotein cholesterol measurement

Objectives:A convenient method based on anion-exchange HPLC was recently developed to determine cholesterol levels of lipoproteins (HDL, LDL, IDL, VLDL, and chylomicron). The present study was performed to compare this HPLC method to homogenous assay in regard to measurement accuracy of HDL and LDL cholesterol.Design and methods:Serum samples (n = 105), including three samples from cholestasis patients, were measured by homogenous assay with Cholestest-LDL and CholestestN-HDL (Daiichi Chemicals, Tokyo) and by HPLC as reported previously (J Lipid Res 2003; 44: 1404–12).Results:The homogenous assay for HDL cholesterol correlated strongly with the HPLC method for HDL cholesterol (r = 0.976). Two samples from cholestasis patients could not be measured by homogenous assay but were measured by HPLC. The homogenous assay for LDL cholesterol correlated modestly with the HPLC method for LDL cholesterol (r = 0.823). Three outlier samples, from cholestasis patients with serum cholesterol levels > 17 mmol/L, were observed in this correlation analysis. Homogenous assay data showed that these LDL cholesterol levels were 15.2–34.7 mmol/L. However, HPLC data showed that these LDL cholesterol levels were 3.6–8.2 mmol/L, and that the major lipoprotein fractions were VLDL and IDL. The difference in LDL cholesterol levels (homogenous assay data minus HPLC data) was positively correlated with VLDL cholesterol levels.Conclusions:When measuring samples from cholestasis patients, homogenous assay may give inaccurate results. In contrast, the HPLC method is likely to be capable of accurately measuring HDL and LDL cholesterol levels without the involving VLDL.     

The relationship between insulin-like growth factor-1 and metabolic syndrome, independent of adiponectin

BackgroundInsulin-like growth factor-1 (IGF-1) is associated with obesity and aging, and was recently linked to metabolic syndrome (MetS) and insulin resistance. However, little is known about the relationship between IGF-1 and adiponectin (adiponectin), another marker of MetS.MethodsWe measured the plasma IGF-1 and adiponectin levels of 3099 subjects (1869 males, 55.9 ± 10.8 y). We applied the Korean-modified International Diabetes Foundation (k-IDF) criteria for determination of, and risk assessment for, MetS.ResultsK-IDF criteria-based MetS occurred in 37.0% (n = 1146) of patients. IGF-1 (91.5 vs. 97.3 ng/ml, p < 0.001) and adiponectin (3.95 vs. 4.23 μg/ml, p < 0.001) were significantly lower in MetS patients than without MetS. Lower IGF-1 was associated with increasing numbers of MetS abnormalities, independent of adiponectin (p for trend < 0.001, F = 12.615, p < 0.001 in ANCOVA). MetS prevalence in individuals with both high IGF-1 and adiponectin levels (6.7%, n = 206) was significantly lower than in other groups. Both high IGF-1 and adiponectin group was associated with reduced MetS risk after adjusting for other confounding factors (OR 0.694, 95% CI 0.493–0.977, p = 0.036).ConclusionsIGF-1 was associated with MetS independent of adiponectin in our study. The independent relationship between IGF-1 and MetS provides insight into the pathophysiologic mechanisms of MetS.     

Measurement of mycophenolic acid and its glucuronide using a novel rapid liquid chromatography-electrospray ionization tandem mass spectrometry assay

ObjectivesMycophenolic acid (MPA), the active metabolite of the ester prodrug mycophenolate mofetil is an immunosuppressant which selectively inhibits inosine-monophosphate dehydrogenase. The requirement for therapeutic drug monitoring shown in previous studies raises the necessity of acquiring accurate and sensitive methods to measure MPA and also its metabolite mycophenolic acid glucuronide (MPAG).Design and methodsWe developed a robust, rapid, sensitive and highly specific HPLC–electrospray ionization mass spectrometry method to assay MPA and its metabolite MPAG in human plasma and serum. Ion suppression was investigated by a post column infusion experiment.ResultsDetermination of MPA and MPAG were performed during a 3.0-min run time. Multiple calibration curves for the analysis of MPA and MPAG exhibited consistent linearity and reproducibility in the range of 0.05 to 100 mg/L (r > 0.999) and 6 to 400 mg/L r > 0.998, respectively. Limits of detection were 0.009 mg/L for MPA and 4.5 mg/L for MPAG and lower limits of quantification were 0.011 mg/L for MPA and 4.9 mg/L for MPAG.Interassay imprecision was < 6.0% for both substances. Mean recovery was 48.9% (range 43.3–60.0%) for MPA and 112.2% (range 95.0–127.0%) for MPAG. Agreement was relatively good for MPA (n = 122) between the presented method and a validated ELISA method (Viva analyzer, Siemens Medicals Solutions Diagnostics, NY). The Passing–Bablok regression line was: EMIT = 0.91 (LC–MS/MS) + 0.17 [mg/L]; r = 0.97.ConclusionsThis simple, robust and interference-free LC–MS/MS assay allows the rapid and accurate determination of MPA and MPAG in human plasma and other body fluids.     

The ratio of serum leptin to adiponectin provides adjunctive information to the risk of metabolic syndrome beyond the homeostasis model assessment insulin resistance: the Korean Genomic Rural Cohort Study

BackgroundLeptin and adiponectin are adipokines, shown to have opposing functions for fat metabolism and development of metabolic syndrome. We determined if the ratio of serum leptin to adiponectin (L/A ratio) adjunctively contributes to the risk of metabolic syndrome beyond the homeostasis model assessment of insulin resistance (HOMA-IR).MethodsThis study included 1532 men and 1856 women, aged 40–70 y assessed in the Korean Genomic Rural Cohort Study from 2005 to 2008. The serum concentrations of adiponectin and leptin were measured by radioimmunoassay. Area under the receiver operating characteristic curve (AUROC) analyses were used to describe the ability of L/A ratio and HOMA-IR to differentiate between subjects with and without metabolic syndrome.ResultsThere were no significant differences in the ability of L/A ratio and HOMA-IR to predict metabolic syndrome (AUROC of L/A ratio vs. HOMA-IR, 0.771 vs. 0.774, p = 0.8006 for men; 0.677 vs. 0.691, p = 0.3088 for women). There was a significant adjunctive contribution by the L/A ratio, beyond that of HOMA-IR, to the risk of metabolic syndrome in men (p < 0.0001 with 0.028 increased AUROC) and women (p = 0.025 with 0.017 increased AUROC).ConclusionsThe L/A ratio provides significant adjunctive information to the risk of metabolic syndrome beyond HOMA-IR alone. The L/A ratio could be a good surrogate marker to assess metabolic syndrome.     

Lymphocytic enzymes and lipid peroxidation in patients with metabolic syndrome

ObjectivesMetabolic syndrome (MetS) is considered a state of chronic inflammation. This study aimed to ascertain selected parameters of purinergic and cholinergic systems related to glucose metabolism and inflammation, as well as _γ-glutamyltransferase (GGT) and N-acetyl-b-glucosaminidase (NAG) activities and lipoperoxidation in lymphocytes of patients with MetS.Design and methodsThe adenosine deaminase (ADA), dipeptidyl peptidase IV (DPP-IV), acetylcholinesterase (AChE), GGT and NAG activities, as well as thiobarbituric acid reactive substances (TBARS) levels were investigated in lymphocytes of patients with MetS (n = 38) and healthy volunteers (n = 41). We also evaluated the insulin levels, anthropometric measurements and routine biochemical analyses.ResultsADA (p < 0.05), DPP-IV and AChE (p < 0.0001) activities were higher in patients with MetS when compared to the control group. Furthermore, we observed correlations between ADA and DPP-IV activities (p = 0.0002; r = 0.5945), TBARS levels and ADA (p = 0.0021; r = 0.5172) and DPP-IV activities (p = 0.0022; r = 0.5010).ConclusionsOur findings showed that MetS might cause tissue distress that disturbed lymphocytic ADA, DPP-IV and AChE activities in response to inflammatory stimuli. These alterations evidence clinical abnormalities, since these enzymatic systems are able to regulate several aspects of adipose tissue function and inflammatory state of MetS and could be used successfully both for preventing and for halting the progression of MetS.     

Carotid intima media thickness is related positively to plasma pre ß-high density lipoproteins in non-diabetic subjects

BackgroundLipid-poor or lipid-free high density lipoprotein (HDL) particles, designated pre ß-HDL, stimulate removal of cell-derived cholesterol to the extracellular compartment, which is an initial step in the reverse cholesterol transport pathway. Pre ß-HDL levels may be elevated in subjects with established cardiovascular disease. We determined the relationship of carotid intima media thickness (IMT), a marker of subclinical atherosclerosis, with pre ß-HDL in subjects without clinically manifest cardiovascular disease.MethodsIMT and plasma pre ß-HDL, assayed by crossed immuno-electrophoresis, were determined in 70 non-diabetic subjects (aged 56 ± 9 years; non-smokers only; 27 women).ResultsIMT was correlated positively with pre ß-HDL, both expressed as plasma apolipoprotein (apo) A-I concentration (r = 0.271, p = 0.023) and as% of apo A-I (r = 0.341, p = 0.004). In contrast, IMT was correlated inversely with HDL cholesterol (r = ? 0.253, p = 0.035). IMT was also related positively to pre ß-HDL after adjustment for age, sex, systolic blood pressure (in apoA-I concentration, ß = 0.203, p = 0.043; in% of plasma apoA-I, ß = 0.235, p = 0.023). IMT remained associated with pre ß-HDL after additional adjustment for either body mass index, plasma glucose, cholesterol, triglycerides, HDL cholesterol, apoA-I and apoB.ConclusionSubclinical atherosclerosis may relate to higher plasma pre ß-HDL independently of apoA-I and HDL cholesterol levels.     

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.     

Urinary calcium response to high dose vitamin D3 with calcium supplementation in patients with multiple sclerosis

ObjectiveTo characterize the effect of vitamin D₃ intake on urinary calcium:creatinine ratios across predefined ranges of serum 25(OH)D.DesignPatients with multiple sclerosis (n = 25) received escalating doses of vitamin D₃ (4000–40,000 IU/d) with calcium (1200 mg/d).ResultsUrinary calcium:creatinine was driven by increased 25(OH)D when concentrations were < 75 nmol/L (r = 0.424, p = 0.009) and > 200 nmol/L (r = 0.281, p = 0.01), but no relationship existed when 25(OH)D concentrations were 76–200 nmol/L.ConclusionsA “safe”, physiological range of 25(OH)D concentrations is 75–200 nmol/L.     

Plasma adiponectin as an independent predictor of early death after acute intracerebral hemorrhage

BackgroundHyperadiponectinemia or hypoadiponectinemia is associated with different diseases. There is a paucity of data on circulating plasma adiponectin concentrations in human intracerebral hemorrhage (ICH). We investigated the plasma adiponectin concentrations in patients with intracerebral hemorrhage, and analyzed the correlation of adiponectin with the severity of brain injury and early mortality after ICH.MethodsThirty controls and 86 patients with acute ICH were included. Plasma samples were obtained on admission and at days 1, 2, 3, 5, and 7 after ICH. Their concentrations were measured by enzyme-linked immunosorbent assay.ResultsAfter ICH, plasma adiponectin level of the patients increased immediately within 6 h, peaked within 24 h, plateaued at day 2, and decreased gradually thereafter. It was substantially higher than that in the controls in a period of 7 days. A multivariate analysis showed plasma adiponectin level was an independent predictor for 1-week mortality (odds ratio, 1.199; 95% CI: 1.035–1.389; P = 0.015) and that it was associated with Glasgow coma scale (GCS) score (t = ? 3.596, P = 0.001) and plasma C-reactive protein level (t = 4.194, P < 0.001). A receiver operating characteristic curve identified that a plasma adiponectin level > 16.4 μg/ml predicted the 1-week mortality of patients with a sensitivity of 65.6% and a specificity of 90.7% (AUC, 0.789; 95% CI: 0.688–0.870). The predictive value of adiponectin concentration was significantly lower than that of GCS score (P = 0.007) and hematoma volume (P = 0.022). Adiponectin could not improve the predictive values of GCS score (P = 0.317) and hematoma volume (P = 0.226).ConclusionsAdiponectin is an independent indicator of early death and may play an anti-inflammatory role after intracerebral hemorrhage.     

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.     

Plasma proprotein convertase subtilisin–kexin type 9 is predominantly related to intermediate density lipoproteins

ObjectivesProprotein convertase subtilisin–kexin type 9 (PCSK9) is a key regulator of low density lipoprotein (LDL) receptor processing, but the PCSK9 pathway may also be implicated in the metabolism of triglyceride-rich lipoproteins. Here we determined the relationship of plasma PCSK9 with very low density lipoprotein (VLDL) and LDL subfractions.Design and methodsThe relationship of plasma PCSK9 (sandwich enzyme-linked immunosorbent assay) with 3 very low density lipoprotein (VLDL) and 3 low density lipoprotein (LDL) subfractions (nuclear magnetic resonance spectroscopy) was determined in 52 subjects (30 women).ResultsIn age- and sex-adjusted analysis plasma PCSK9 was correlated positively with total cholesterol, non-high density lipoprotein cholesterol and LDL cholesterol (r = 0.516 to 0.547, all p < 0.001), as well as with triglycerides (r = 0.286, p = 0.044). PCSK9 was correlated with the VLDL particle concentration (r = 0.336, p = 0.017) and with the LDL particle concentration (r = 0.362, p = 0.010), but only the relationship with the LDL particle concentration remained significant in multivariable linear regression analysis. In an analysis which included the 3 LDL subfractions, PCSK9 was independently related to intermediate density lipoproteins (IDL) (p < 0.001), but not to other LDL subfractions.ConclusionsThis study suggests that plasma PCSK9 predominantly relates to IDL, a triglyceride-rich LDL subfraction. The PCSK9 pathway may affect plasma triglycerides via effects on the metabolism of triglyceride-rich LDL particles.     

Effects of a single dose of oral iron on hepcidin concentrations in human urine and serum analyzed by a robust LC-MS/MS method

BackgroundThe measurement of serum hepcidin, a peptide hormone that regulates iron metabolism, is clinically important to the understanding of iron homeostasis in health and disease. To date, the quantification of serum hepcidin levels by conventional immunological detection methods has proven problematic due to challenges in obtaining high quality antibodies which demonstrate good reproducibility. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) has been employed recently for more sensitive quantification of hepcidin; however, this method has high background levels and therefore less than optimal specificity.MethodsIn order to increase the specificity of the mass spectrometry based assay, we developed a robust, ultra-performance liquid-chromatography-tandem mass spectrometry (UPLC-MS/MS) protocol using multiple selected reaction monitoring (mSRM) for quantification of hepcidin levels in urine and serum of human subjects. With this assay, we assessed levels of hepcidin before and for up to 8 h after oral ingestion of ferrous sulfate in ten adult human subjects without known disease.ResultsThe linear response of hepcidin quantitation on each instrument was measured, and the correlation coefficients of these calibrations were r² = 0.9512 ± 0.0202 (n = 5) for urine and r² = 0.9709 ± 0.0291 (n = 5) for serum [r² = mean ± SD]. Compared to baseline, the levels of urinary hepcidin between 2–4 h and 4–8 h of both women and men showed significant increases with p < 0.05 and p < 0.001, respectively. The levels of serum hepcidin between 4 h and 8 h in both women and men showed significant increases, compared with baseline values, with both p < 0.01. Interestingly, we also observed some degree of oscillation of levels, occurring at later time points.ConclusionsWe have developed and validated a new method for measuring hepcidin concentrations in human serum and urine and used it to demonstrate early increases with iron supplement in both urinary and serum levels of hepcidin, which return to baseline levels, except in urine samples from men.     

The alarm secretory leukocyte protease inhibitor increases with progressive metabolic dysfunction

BackgroundSecretory leukocyte protease inhibitor (SLPI) is an alarm antiprotease secreted by neutrophils and mucous membranes that potently inhibits the inflammatory cascade; however, the role of SLPI in human disease remains largely unknown. We hypothesized that SLPI is related to chronic low-grade inflammatory diseases, such as metabolic syndrome (MS) or type-2 diabetes (T2DM).MethodsWe examined associations between circulating SLPI (ELISA) and quantitative traits of MS (ATPIII criteria) in 261 Caucasian men with various degrees of metabolic dysfunction. Subjects had neither MS nor T2DM (n = 140), either diagnosis (n = 44) or both diagnoses (n = 77).ResultsCirculating SLPI increased with progressive metabolic dysfunction, with a mean increase of 4.4 ng/ml (95% IC 2.4 to 6.3 ng/ml; p < 0.001) for each unit increase in the criteria used to define MS. Circulating SLPI showed independent associations with uric acid [β = 5.1 (95% CI 3.4 to 6.7), p < 0.00001], serum lipids, pulse pressure and inflammatory markers.ConclusionsCirculating SLPI increases with progressive metabolic dysfunction and is related to metabolic and inflammatory parameters in men.     

Serum vascular adhesion protein-1 is higher in subjects with early stages of chronic kidney disease

ObjectivesAn increased level of serum vascular adhesion protein-1 (VAP-1) has been found in patients with diabetes mellitus and vascular disorders. This study examined whether serum VAP-1 levels are associated with chronic kidney disease (CKD).Design and methodsWe included 262 subjects aged 30 and above with fasting plasma glucose level < 7 mmol/L checked within 1 year. First morning urine specimens were collected. Microalbuminuria was defined if urinary albumin-to-creatinine ratio ≥ 30 μg/mg creatinine. The glomerular filtration rate (GFR) was estimated. CKD stages were defined according to the suggestions of the National Kidney Foundation. Serum VAP-1 levels were analyzed by immunofluorometric assay.ResultsSerum VAP-1 levels were positively associated with the urinary albumin-to-creatinine ratio ( r = 0.29, p < 0.0001) and negatively associated with estimated GFR (r = ?0.24, p =  0.0001). Subjects with CKD stage 2 (N =  51) and stage 3 (N =  91) had significantly higher levels of serum VAP-1 than those without CKD (p =  0.0003 and p =  0.035, adjusted for age and gender, respectively). A high serum VAP-1 level was associated with the presence of CKD (OR 1.63 for 1 SD increase of VAP-1, p =  0.018), adjusting for age, sex, and smoking. Ordered logit models revealed that high serum VAP-1 levels correlated with advanced stages of CKD.ConclusionsSerum levels of VAP-1 are associated with the severity of kidney damage or stages of kidney disease. The true mechanism which links the serum VAP-1 and CKD remains to be elucidated in further studies.     

Lack of association between paraoxonase 1 Q192R polymorphism and multiple sclerosis in relapse phase: A case-control study

ObjectivesThe aim of this study was to estimate the serum activity of paraoxonase 1(PON1) and assess the distribution of PON1 polymorphisms in MS patients in the relapse phase.Design and methodsPON1 and arylesterase (ARE) serum activities were measured in two equal groups (each group 63 cases) of relapsing–remitting MS patients and healthy individuals.ResultsMean values for serum PON1 and ARE activities were 90.3 ± 63.4 and 182.1 ± 128.7 IU/L for patients and 99.9 ± 73.3 and 190.8 ± 150.3 IU/L for controls. Those values were not statistically significant (p = 0.242 and p = 0.378), respectively. Comparing genotype distributions and allele frequencies in both groups for PON1 Q192R and PON L55M polymorphisms did not show any statistical difference.ConclusionIn a selected group of MS patients in relapsing phase no statistically significant difference in PON1 and ARE activities was detected but the mean values for the serum enzyme activities were lower in MS patients.     

Improved measurement of LINE-1 sequence methylation for cancer detection

BackgroundMethylation of long interspersed nuclear element-1 (LINE-1) sequences varies among normal cells and it is often decreased in cancer genomes and white blood cells (WBC) of cancer patients. Current measurement techniques of genome-wide level are inadequate because LINE-1 methylation is distinctive at each locus. Here, we improved the detection of cancer by combining information of LINE-1 methylation pattern and level.MethodsCombined bisulfite restriction analysis (COBRA) of LINE-1, COBRA LINE-1, was used to test cancer cell lines, two oral rinse cohorts, and WBC from normal and cancer patients. COBRA LINE-1 separated LINE-1 sequences into 4 products depending on the methylation statuses of 2 CpG dinucleotides, as follows: 2 unmethylated CpGs (^uC^uC), partial methylation (^mC^uC), 1 methylated CpG (^mC), and 1 unmethylated CpG (^uC).ResultsThe association between ^mC^uC and ^uC^uC was directly correlated in normal cells (r = 0.4895, p = 0.0009) but inversely correlated in cancer (r = ? 0.8979, p = 0.0002). Oral rinse AUC values of ^uC^uC were 0.763 and 0.926 and methylation levels were 0.707 and 0.621, respectively. ^uC^uC, but not overall methylation level, differentiated cancer WBC from normal (p = 0.0082 and p = 0.4830, respectively).ConclusionLINE-1 partial methylation represents hypomethylation in normal cells but hypermethylation in cancer cells. This information improves LINE-1 methylation detection in cancer.     

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.     

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.