Immulite 2000 system

C-reactive protein (CRP). Wide-range CRP was measured in fresh serum samples by a commercial latex-enhanced immunoturbidimetry method on an Advia 2400 Clinical Chemistry System (Siemens). Reference values with the method were 0–0.5 mg/dL. The measurement was available for 1499 individuals [45 (link)].
Erythrocyte sedimentation rate (ESR). ESR was measured in blood samples in K3EDTA tubes (Becton Dickinson, Franklin Lakes, NJ, USA) with a TEST-1 automatic apparatus (Alifax, Padova, Italy). The TEST-1 apparatus was validated against the Westergren reference method according to the criteria of the International Council for Standardization in Hematology. Reference values are 0–20 mm/h for men and 0–30 mm/h for women. ESR determination was available for 1472 participants [37 (link)].
Serum cytokines. Serum concentrations of interleukin (IL)-6, IL-8, TNF-alpha, and soluble IL-2 receptor (sIL-2R) were determined in fresh serum samples using a chemiluminescent immunoassay (IMMULITE 2000 System, Siemens Health Care Diagnostics, Barcelona, Spain). Results were available for 1499 individuals [46 (link)].

Serum total-PSA (tPSA) and free-PSA (fPSA) were measured using the Immulite^® 2000 system (Siemens Healthcare Diagnostics srl, Siemens, Milan, Italy). For PCA3 analyses, urine samples were collected after digital rectal examination (DRE) using a Progensa™ PCA3 (Gen-Probe, San Diego, CA, USA) assay following the manufacturer’s instructions. PCA3 mRNA and PSA mRNA values were determined. The PCA3 score was determined by calculating the PCA3 mRNA/PSA mRNA × 1000 ratio. Urinary creatinine was measured using a Cobas 6000 (c501) analyser (Roche Diagnostics S.p.a., Monza, Italy).

Fasting glucose levels were routine measurements by the Department of Clinical Biochemistry. Fasting plasma levels of insulin were measured using Immulite 2000 System (Siemens, NY, USA) and insulin resistance was calculated using the homeostasis model assessment (HOMA-IR) [24 (link)]. Metabolic syndrome was defined according to the 2009 consensus definition for high risk populations [25 (link)]. The assessment of metabolic syndrome was based on the presence of at least three of the five following criteria: elevated waist circumference (≥94 cm, as recommended for high risk populations); elevated triglycerides (≥1.7 mmol/L); reduced HDL-C (<1.0 mmol/L); elevated blood pressure (systolic ≥130 or diastolic ≥85 mm Hg, or anti-hypertensive treatment); elevated fasting glucose (≥100 mg/dL).

Routine blood tests, including complete blood count (CBC), liver function tests, serum creatinine, serum cholesterol and triglycerides were performed.
Serum AFP was evaluated by chemiluminescent immunometric technique on the Immulite 2000 system (Siemens Medical Solutions Diagnostics, Los Angeles, California, USA). The eGFR was estimated based on the following formula: $eGFR=186\times {\left(\frac{Creatinine}{88.4}\right)}^{-1.154}\times {Age}^{-0.203}\times (1.210\hspace{0.5em}if\hspace{0.5em}black)\times (0.742\hspace{0.5em}if\hspace{0.5em}female)$ (Levey et al., 1999 (link)). Quantitative assessments of serum AGEs were measured by ELISA kit (MyBioSource, San Diego, CA 92195-3308, USA, Cat No. MBS267540). The homeostasis model assessment (HOMA) method (Matthews et al., 1985 (link)) was evaluated as follows: Insulin resistance (HOMA-IR) = $\frac{fasting\hspace{0.5em}serum\hspace{0.5em}insulin\hspace{0.5em}\left(FSI\right)\times fasting\hspace{0.5em}plasma\hspace{0.5em}glucose\hspace{0.5em}\left(FPG\right)}{405}$