Publications

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.

OBJECTIVE

To assess feasibility, safety, and patient satisfaction of same-day discharge (SD) following peripheral arterial interventions.

BACKGROUND

Although diagnostic angiography is routinely performed as a same-day procedure, same-day percutaneous trans-luminal angioplasty is less common. Because there is very low incidence of peri-procedural complications after 4 hr, discharge after this window is possible provided the patient is able to ambulate and has necessary social support. To-date, safety and patient satisfaction related to SD has not been studied systematically in this population.

METHOD

After providing informed consent, patients undergoing out-patient peripheral arterial interventions in a single institution between 2011 and 2015 were randomized to usual care (overnight stay, OS) or SD following successful interventions. Patient satisfaction, complications, and readmission status was ascertained by blinded telephone interviewers at 48-72 hr and 10 days post-procedure.

RESULTS

A total of 24 patients consented. Of these, 5 (21.7%) failed screening, leaving 19 patients for randomization to control (n = 10) and experimental group (n = 9) conditions. The SD group experienced zero complications, however their Likert scale rating scores were significantly lower than OS for perceived level of safety (P = 0.02) and likelihood of having the procedure again (P = 0.004).

CONCLUSION

This small, single-center randomized study found that among carefully selected peripheral arterial interventions, SD may be feasible and safe. However, patient satisfaction and perceived safety were significantly lower among SD compared to the OS condition. Larger prospective studies are warranted to confirm these findings.

Jamaican men who have sex with men (MSM) have experienced widespread stigma and discrimination. Much of the research on Jamaican MSM has focused on HIV risk behaviors. We examined the social and romantic relationships of Jamaican MSM and how these factors fostered a sense of community in an antihomosexual environment. Qualitative in-depth interviews were conducted with 30 MSM ages 18 to 29 years. Women and familial matriarchal figures were more likely to accept someone identified as homosexual and provide protection against homophobia. Jamaican MSM affirmed their identity by providing emotional support and safe spaces, which aided in building a sense of community. Relationships with friends and intimate partners were a source of love and validation as opposed to simply sexual gratification. The social and romantic relationships of Jamaican MSM transcended the social boundaries of homophobia, affirmed sexual identity and orientation, and served as facilitators across most general societal and cultural interactions.

BACKGROUND

High-risk human papillomavirus (hrHPV) causes virtually all cervical cancers. Trans masculine (TM) people (those assigned female at birth who identify with a gender other than female) have low uptake of conventional cervical cancer screening. Self-collected hrHPV DNA testing has high levels of acceptability among cisgender (non-transgender) females and may support increased cervical cancer screening uptake in TM individuals.

OBJECTIVE

To assess the test performance and acceptability of self-collected vaginal specimens in comparison to provider-collected cervical swabs for hrHPV DNA detection in TM individuals ages 21-64 years.

METHODS

Between March 2015-September 2016, 150 TM participants with a cervix (mean age = 27.5 years; SD = 5.7) completed a one-time study visit comprised of a self-report survey, self-collected vaginal HPV DNA swab, clinician-administered cervical HPV swab, and brief interview on acceptability of clinical procedures. Participants were randomized to complete either self- or provider-collection first to minimize ordering effects. Self- and provider-collected samples were tested for 13 hrHPV DNA types using a DNA Hybridization Assay. The primary outcome variable was the concordance (kappa statistic) and performance (sensitivity, specificity) of self-collected vaginal HPV DNA specimens versus provider-collected cervical HPV swabs as the gold standard.

RESULTS

Of the 131 participants completing both the self- and provider-collected HPV tests, 21 cases of hrHPV were detected by the provider cervical swab (gold standard; 16.0% hrHPV prevalence); 15 of these cases were accurately detected by the self-collected vaginal swab (71.4% concordance) (Kappa = 0.75, 95% Confidence Interval [CI]: 0.59, 0.92; p<0.001). Compared to the provider-collected cervical hrHPV DNA sample (gold standard), the self-collected vaginal hrHPV DNA test demonstrated a sensitivity of 71.4% (95% CI: 0.52, 0.91; p = 0.0495) and specificity of 98.2% (95% CI: 0.96, 1.00; p<0.0001). Over 90% of participants endorsed a preference for the self-collected vaginal swab over provider-collected cervical swab.

CONCLUSION

Self-collected vaginal swabs are highly acceptable to TM as a means to test for hrHPV DNA. Test performance of this self-collection method for hrHPV detection in TM is consistent with previous studies in cisgender females. Self-collected vaginal swab testing for hrHPV DNA represents a reasonable and patient-centered strategy for primary cervical cancer screening in TM patients unwilling to undergo provider collection of specimens via speculum exam.

Infection with M. tuberculosis is associated with inconsistent and incomplete elimination of the bacteria, despite development of antigen-specific T cell responses. One mechanism employed by M. tuberculosis is to limit availability of antigen for activation of CD4 T cells. We examined the utility of systemic administration of epitope peptides to activate pre-existing T cells in mice infected with M. tuberculosis. We found that systemic peptide administration: 1) selectively activates T cells specific for the epitope peptide; 2) loads MHC class II on lung macrophages and dendritic cells; 3) activates CD4 T cells in the lung parenchyma; 4) has little antimycobacterial activity. Further studies revealed that CD4 T cells in lung lesions are distant from the infected cells, suggesting that, even if they can be activated, the positioning of CD4 T cells and their direct interactions with infected cells may be limiting determinants of immunity in TB.

Antigen-specific CD4 and CD8 T cells are important components of the immune response to , yet little information is currently known regarding how the breadth, specificity, phenotype, and function of -specific T cells correlate with infection outcome in humans. To facilitate evaluation of human -specific T cell responses targeting multiple different Ags, we sought to develop a high throughput and reproducible T cell response spectrum assay requiring low blood sample volumes. We describe here the optimization and standardization of a microtiter plate-based, diluted whole blood stimulation assay utilizing overlapping peptide pools corresponding to a functionally diverse panel of 60 Ags. Using IFN-γ production as a readout of Ag specificity, the assay can be conducted using 50 μl of blood per test condition and can be expanded to accommodate additional Ags. We evaluated the intra- and interassay variability, and implemented testing of the assay in diverse cohorts of -unexposed healthy adults, foreign-born adults with latent infection residing in the United States, and tuberculosis household contacts with latent infection in a tuberculosis-endemic setting in Kenya. The -specific T cell response spectrum assay further enhances the immunological toolkit available for evaluating -specific T cell responses across different states of infection, and can be readily implemented in resource-limited settings. Moreover, application of the assay to longitudinal cohorts will facilitate evaluation of treatment- or vaccine-induced changes in the breadth and specificity of Ag-specific T cell responses, as well as identification of -specific T cell responses associated with infection outcomes.