Hscrp

We measured biomarkers of metabolic functioning [hemoglobin A1C (HbA1C) and adiponectnin], and inflammation [high-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-a)]. HbA1C was measured by turbidimetric inhibition immunoassay (Roche Diagnostics, Indianapolis, IN). Adiponectin was measured by enzyme-linked immunosorbent assay (ELISA) (ALPCO Diagnostics Inc, Salem, NH). hs-CRP was measured using particle enhanced turbidimetric assay (Roche Diagnostics, Indianapolis, IN). IL-6 was measured by paramagnetic particle, chemiluminescent immunoassay (Beckman Coulter, Fullerton, CA). TNF-a was measured using quantitative sandwich enzyme immunoassay (R&D Systems Inc., Minneapolis, MN).

The study procedures conducted after informed consent and assent included a physical examination of heart rate and blood pressure measurement, weight and height measurement, and a CXR. If any evidence of CVD was found in the physical examination or CXR, subjects were excluded from the study. If the CXR was normal, the subject would then have blood drawn for complete blood count, fasting lipid profiles, hs-CRP (hs-CRP, Roche Diagnostics GmbH, Mannheim, Germany), and NT-pro-BNP (Elecsys proBNP, Roche Diagnostics GmbH, Mannheim, Germany). For the HIV-infected subjects, CD4 and HIV-1 RNA were also included. The subjects then underwent an echocardiogram to assess cardiac anatomy and function. The cIMT measurement was performed right after the echocardiogram. The case record forms were filled in using data extracted from the medical records, which included demographic data, medical history including previous illnesses and hospitalizations, and HIV-related treatment.

At the time of CPET, participants underwent history and physical examination including assessment of body mass index (BMI) and fasting blood draw. Blood specimens were immediately processed and stored at −80°C. Plasma N-terminal pro-B type natriuretic peptide was assayed using an electrochemiluminescence immunoassay (Roche, NT-proBNP, intra-assay coefficient of variation 2.4–3.8%) and high-sensitivity C-reactive protein was ascertained using an immunoturbidimetric assay (Roche, hsCRP, intra-assay coefficient of variation 0.4–8.4%). Medical records were reviewed in detail to ascertain medical history. Resting spirometry was performed. Echocardiography within 1 year of exercise testing was reviewed, with abstraction of LVEF, the presence of left atrial enlargement ascertained via left atrial diameter measures, LV hypertrophy, and diastolic dysfunction (Supplemental Table 1). All available medical records were reviewed with the following clinical outcomes adjudicated through 10/29/2018: first cardiovascular (CV) hospitalization (defined as any hospitalization for principal cardiovascular cause, including arrhythmia, acute coronary syndrome, coronary revascularization, heart failure) and all-cause mortality, which was ascertained using the social security death index and electronic hospital records.

Blood samples were tested for serum ferritin (Roche Cobas Integra 400 Plus analyser) and highly sensitive C-reactive protein (hsCRP; Roche Diagnostics, Indianapolis, IN, USA). The detection limit of CRP was 0.1 mg/L, and a value of 0.001 was assigned to observations below the detection limit, as recommended by the Biomarkers Reflecting Inflammation and Nutritional Determinants of Anaemia (BRINDA) working group [19 ].