Carey Special Immunology Unit at University Hospitals Case Medica

Carey Special Immunology Unit at University Hospitals Case Medical Center in Cleveland, OH. All individuals provided written informed consent to participate in the HIV Metabolic Research Center trials and also to have their blood stored for use in future HIV-related metabolic research. This study was approved by the University Hospital Case Medical Center Institutional Review Board with a waiver for further informed consent. All data collected, demographics, HIV and cardiovascular characteristics, laboratory values and stored samples were obtained on the date on which FMD was performed. The primary outcome

of this study was endothelial function determined using FMD of the brachial artery. Secondary outcomes of interest included markers of inflammation [interleukin-6 (IL-6), soluble tumour necrosis factor receptors I and II (sTNFR-I and -II), high-sensitivity C-reactive protein (hs-CRP), soluble intercellular adhesion molecule-1 (sICAM-1) and soluble

vascular cell adhesion molecule-1 (sVCAM-1)], coagulation (D-dimer and fibrinogen), oxidative stress (F2-isoprostanes), lipoprotein levels and insulin resistance estimated using the homeostasis model assessment of insulin resistance (HOMA-IR). Endothelial function was evaluated by measuring FMD of the brachial artery with ultrasound [15] as previously described [16]. Participants were instructed to come learn more fasting, to not take anti-hypertensive medications and not to use tobacco or caffeine-containing products for 12 h before the study. All studies were performed by a single technologist (CW) using a Phillips iU22 Ultrasound and an L10-7 MHz linear array transducer (Phillips Healthcare, Bothell, WA, USA) and a 5-min occlusion time. Images were read using Brachial Artery Analyzer software (Medical Imaging Applications LLC, Coralville, IA, USA), a semi-automated, border-interfacing program. For FMD determination, brachial artery diameters before and after confirmed reactive hyperaemia were measured in triplicate and averaged from a 1-cm segment of the artery. FMD is expressed

as a percentage change from baseline brachial artery diameter to brachial artery diameter after reactive hyperaemia. Plasma from each participant Clomifene was previously stored at −70°C immediately after processing. Stored samples were then batched and tested for the markers of inflammation, coagulation and oxidative stress outlined above. IL-6, sTNFR-I and -II, sICAM-1 and sVCAM-1 were determined by quantitative sandwich enzyme-linked immunosorbent assays (ELISAs) (R&D Systems, Minneapolis, MN, USA). Interassay variability was 2.02–15.36%, 3.66–5.77%, 2.13–3.79%, 3.43–7.37% and 4.76–8.77%, respectively. hs-CRP and fibrinogen were determined using particle enhanced immunonephelometric assays on a BNII nephelometer (Siemens, Indianapolis, IN, USA). Interassay variability was 3.01–6.46% and 3.42–7.59%, respectively.

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