Hair concentrations of antiretrovirals are associated with virologic outcomes in cohorts of HIV-positive individuals but have never before been examined in a clinical trial. We show for the first time the predictive utility of hair antiretroviral concentrations in a large HIV treatment-naïve trial, AIDS Clinical Trials Group (ACTG) protocol A5257.

Rationale Assays to quantify antiretrovirals in hair samples are increasingly used to monitor adherence and exposure in both HIV prevention and treatment studies. Atazanavir (ATV) is a protease inhibitor used in combination antiretroviral therapy (ART). We developed and validated a liquid chromatography/tandem mass spectrometry (LC/MS/MS)-based method to quantify ATV in human hair, per the NIH Division of AIDS Clinical Pharmacology Quality Assurance (CPQA) program and the FDA bioanalytical method validation guidelines. Methods ATV was extracted from hair using optimized methods and the extracts were injected onto a BDS C-18 column (5 μm, 4.6 × 100 mm), followed by isocratic elution via a mobile phase composed of 55% acetonitrile, 45% water, 0.15% acetic acid, and 4 mM ammonium acetate, at a flow rate of 0.8 mL/min prior to analysis by MS/MS. Levels were quantified using positive electrospray ionization by multiple reaction monitoring (MRM) for the transitions MH+ m/z 705.3 to m/z 168.0 and MH+ m/z 710.2 to m/z 168.0 for ATV and ATV-d5 (internal standard), respectively. Results Our assay demonstrated a linear standard curve (r = 0.99) over the concentration range of 0.0500 ng ATV/mg hair to 20.0 ng/mg hair. The inter- and intraday accuracy of ATV quality control (QC) samples was −1.33 to 4.00% and precision (% coefficient of variation (%CV)) was 1.75 to 6.31%. The %CV for ATV levels in hair samples from highly adherent patients (incurred samples) was less than 10%. No significant endogenous peaks or crosstalk were observed in the specificity test with other HIV drugs. The overall extraction efficiency of ATV from incurred hair samples was greater than 95%. Conclusions This highly sensitive, highly specific and validated assay can be considered for therapeutic drug monitoring for HIV-infected patients on ATV-based ART.

Affordable novel strategies are needed to treat COVID-19 cases complicated by respiratory compromise in resource limited settings. We report a mixed-methods pre-post assessment of 1) the useability of CPAP/O2 helmet non-invasive ventilation (NIV) to treat COVID-19, at ~1% the cost of mechanical ventilation; 2) the effectiveness of a train-the-trainer practice facilitation intervention; and 3) whether use of CPAP/O2 helmet NIV was associated with increased COVID-19 infection among healthcare workers. At baseline, eight COVID-19 treatment centers in Nigeria (CircumVent network) received CPAP/O2 helmet systems, and were instructed on its use. After five months, clinicians within the CircumVent netwok participated in a 2-day train-the-trainers educational intervention. The physicians completed i) standardized forms on patient demographics, clinical course, and outcomes for patients seen in the treatment centers; ii) standardized surveys of feasibility and acceptability of use of CPAP/O2 helmet systems; and iii) in-depth-interviews to explore facilitators and barriers to implementation of CPAP/O2 helmet NIV. Physicians described the CPAP/O2 helmet ventilator as easy to use and they felt comfortable training their staff on its use. They rated CPAP/O2 helmet NIV as feasible, acceptable, and appropriate (mean score of 4.0, 3.8, and 3.9 out of 5, respectively, on standardized scales). Case report forms for 546 patients with suspected and/or confirmed COVID-19 infection were obtained between May 2020 and November 2021. Of these, 69% (n=376) were treated before the training; and 29.7% (n=162) were treated with CPAP/O2 helmet ventilation. CPAP/O2 helmet NIV was well-tolerated by patients, with 12% reporting claustrophobia, and 2% reporting loose- or tight-fitting helmets. Although patient outcomes improved among CPAP/O2 helmet users overall, this was not associated with training (P=0.2). This finding persisted after adjustment for disease severity at presentation. Serosurvey of 282 health workers across treatment centers revealed that 40% (n=112) were seropositive for SARS-CoV-2. Seropositivity was significantly associated with direct contact with COVID-19 patients and limited access to PPE and hand hygiene during aerosol generating procedures (P = 0.02), but not use of CPAP/O2 helmet (Ps ≥ 0.2). In conclusion, physicians effectively used CPAP/O2 helmet NIV systems to treat COVID-19 patients in Nigeria without need for practice facilliation of their training and without increased risk of infection among healthcare workers. The use of CPAP/O2 helmet NIV could be an important strategy for treating individuals with COVID-19 infection and other disease conditions complicated by respiratory distress, particularly in settings were resources such mechanical ventilation are limited.