At week 24, all participants were offered 6 months of uridine and pravastatin. Oral PD98059 solubility dmso uridine supplementation was provided as Nucle-omaxX® (Pharma Trade Healthcare, Spanga, Sweden), a dietary supplement with a high
content (17%; 36 g per sachet) and availability of uridine. The uridine dose was based on the findings of previous studies showing efficacy of uridine supplementation for lipoatrophy at this dose [13] and rapid entry of exogenous uridine from plasma into cells where uridine pools turn over with a half-life of 13–18 h [19]. Participants could adapt their uridine dose to one sachet daily (for 30 days per month) for significant gastrointestinal intolerance to uridine three times a day (tid), as diarrhoea is a known side-effect of uridine [20]. Clinical and biological assessments were performed at randomization, week 4, week 12 and week 24. Anthropometric parameters (weight, umbilical waist circumference and maximum hip circumference) were measured at each visit. Height was recorded at baseline. Blood was collected for
fasting total cholesterol, high-density lipoprotein (HDL) cholesterol, triglycerides, glucose and insulin, as well as safety measures (hepatic transaminases, creatinine, electrolytes and full blood count). Patients randomized to receive uridine had a uridine plasma concentration measurement performed at baseline, week 1 and week LDK378 supplier 24. All uridine plasma levels were quantified using high-performance liquid chromatography (HPLC) with ultraviolet detection at a wavelength of 262 nm; the range of detection was 0.25–10 μg/mL and the coefficient of variation <10%. Plasma was extracted using acetonitrile to precipitate plasma proteins. The extract was centrifuged at 15 000 g for 5 min to separate the supernatant from the precipitate. The supernatant was evaporated to dryness at 50°C and Methane monooxygenase the residue suspended in the mobile phase. An aliquot of the resuspended
fluid was injected onto the HPLC column. Separation was performed on a Phenomenex Aqua (Torrance, California, USA) column (250 × 4.6 mm) with a mobile phase of water containing phosphate buffer. Quantitative HIV-1 RNA (viral load) was measured using a Roche COBAS TaqMan HIV-1 test (COBAS AmpliPrep; Roche Diagnostic, Basel, Switzerland) at baseline and at weeks 4, 12 and 24. Adherence was assessed by pill count and empty pack return at all Australian sites by a study pharmacist. Body composition was quantified at baseline, week 12 and week 24 by dual-energy X-ray absorptiometry (DEXA). DEXA scans were performed on a GE Lunar Prodigy machine (General Electric Health Care, Madison, WI) using software version 7.51 (enCORE GE Lunar Platform, General Electric). Cross-validation between sites was carried out using a body composition phantom.