standardized methods. Brachial blood pressure was measured in the left arm of the supine subjects, after 5 min of quiet rest, with a digital electronic tensiometer. A minimum of three blood pressure readings were taken on three separate occasions at least 2 weeks apart, and the medians of these three values were used. A 75 g oral glucose tolerance test was performed with sampling for plasma glucose. Liver ultrasonography was performed in all participants by the same trained operator, who was blinded to participants’ details, using a Toshiba Aplio 50 ultrasound apparatus equipped with a 3.5-MHz linear transducer. Longitudinal, sub costal, ascending, and oblique scans were performed. The ultrasonographic criteria used to diagnose fatty liver included liver and kidney echo discrepancy, the presence of an increased liver echogenicity or “bright liver”, poor echo penetration into the deep portion of the liver, and vascular blurring either singly or in combination. The protocol was approved by the local ethical committee, and written informed consent was obtained from all participants in accordance with principles of Helsinki Declaration. automated technique based on a Creatinine Jaffe compensated ‘method for serum and plasma implemented in an auto-analyzer. Serum uric acid was measured by the URICASE/POD method implemented in an auto-analyzer. Albumin concentration was determined with a Alb2 kit on a Cobas C6000 analyzer. High sensitivity Creactive protein levels were measured PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19645691 by automated instrument. An automated nephelometric technology using the BNTM II System analyzer was employed to measure plasma fibrinogen concentrations. Plasma insulin concentration was measured with a chemiluminescence-based assay, and total serum IGF-1 concentrations were determined by chemiluminescent immunoassay. Definitions Glucose tolerance status was diagnosed according to the American Diabetes Association criteria: normal glucose tolerance when fasting plasma glucose was,5.6 mmol/l and 2 h post-load,7.8 mmol/l, isolated impaired fasting glucose when FPG was 5.66.9 mmol/l and 2 h postload,7.8 mmol/l, impaired glucose tolerance when FPG was 6.9 mmol/l and 2-h get Debio-1347 post-load was 7.811.0 mmol/l, type 2 diabetes when FPG was $7.0 mmol/l and/or 2 h post-load was $11.1 mmol/l. The NAFLD fibrosis score was calculated according to the following formula: 21.675 + 0.0373 x age + 0.0943 x BMI +1.13 x IFG or diabetes + 0.99 x AST/ALT ratio 20.013 x platelet 2 0.66 x albumin. Two cutoff points were used to divide the subjects in three groups: low risk of fibrosis, intermediate risk of fibrosis, and high risk of fibrosis . The homeostasis model assessment index of insulin resistance was calculated as fasting insulin 6 fasting glucose/ 22.5. Estimated glomerular filtration rate was calculated by using the CKD-EPI equation: eGFR = 141 x mina x max21.209 x 0.993Age x 1.018, where Scr is serum creatinine, k is 0.7 for females and 0.9 for males, a is 2 0.329 for females and 20.411 for males, min indicates the minimum of Scr/k or 1, and max indicates the maximum of Scr/k or 1. CKD was defined as eGFR,60 ml/min/1.73 m2. Metabolic syndrome was defined as having three or more of the following criteria: waist circumference.102 cm in men and.88 cm in women, triglycerides.1.69 mmol/l or on treatment for elevated triglycerides, HDL,1.04 mmol/l in men and, 1.3 mmol/l in women or on treatment for reduced HDL, blood pressure.130/85 mmHg or on antihypertensive treatment, fasting glucose $5.6 m

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