Lipodystrophy was evaluated according to this categorization in the face, arms, legs, buttocks, abdomen, neck and breasts. The sum of the values corresponding to each corporal zone indicated the degree of lipodystrophy: nonexistent (0), slight (1–6), moderate (7–12) and severe (13–18). In this study we included only moderate and severe cases in order to avoid an overlap between the LD+ and LD− subsets. The LD+ group comprised 26 patients with pure lipoatrophy and 106 patients with Z-IETD-FMK molecular weight the mixed type. No cases of pure lipohypertrophy were recorded.

With respect to severity, 109 had moderate and 23 had severe lipodystrophy. After an overnight fast, 20 mL of blood obtained from a peripheral vein was collected in Vacutainer™ (Becton Dickinson Immunocytometry Systems, San Jose, CA, USA) ethylenediaminetetraacetic acid (EDTA) tubes. Five millilitres of whole blood was used to determine the CD4 T-cell count. Five hundred microlitres was used for DNA isolation with a MagNa Pure LC Instrument (Roche Diagnostics, selleck kinase inhibitor Basel, Switzerland). Plasma and serum were obtained by centrifugation at 3500 g for 15 min at 4 °C and stored at −80 °C until use. HIV-1 infection and plasma HIV-1 viral load were assessed as described elsewhere [14]. The CD4 T-cell count was determined using a flow cytometer FAC Scan (Becton Dickinson Immunocytometry Systems). Data acquired were analysed using the multiset program

(Becton Dickinson Immunocytometry Systems). Plasma glucose, total cholesterol, HDL cholesterol and triglycerides were determined in an ADVIA 1200 (Siemens AG, Munich, Germany) auto-analyser using standard enzyme methods. Low-density lipoprotein (LDL) cholesterol was calculated using

the Friedewald formula [16]. Fasting plasma insulin was measured Etoposide using a specific immunoradiometric assay (Medgenix Diagnostics, Fleunes, Belgium) in which proinsulin did not cross-react. The intra- and inter-assay coefficients of variation (CVs) were 6% and 7%, respectively. The homeostasis model assessment of insulin resistance (HOMA-IR) as a marker for insulin resistance was calculated according to the formula [fasting glucose (in millimoles per litre) × fasting insulin (in microunits per millilitre)/22.5] [17]. Soluble tumour necrosis factor receptor 1 (sTNF-R1) and sTNF-R2 were assessed as previously described [18]. Adiponectin levels were measured using a standardized radioimmunoassay kit from Linco Research (Linco Research Inc., St. Charles, MO, USA). The kit has a sensitivity of 1 ng/mL. The intra- and inter-assay CVs were 8% and 12%, respectively. Plasma FABP-4 was measured using the Human Adipocyte FABP ELISA (BioVendor Laboratory Medicine, Palackeho, Czech Republic). The sensitivity was 0.1 ng/mL. The intra- and inter-assay CVs were 5.2% and 3.8%, respectively. The leptin concentration in plasma was determined with a Human Leptin ELISA kit (Assaypro, St Charles, MO, USA); the lowest detectable level was 0.15 pg/mL with an intra-assay CV of 4.