Improving the clinic-laboratory-interface in the context of HIV diagnosis, treatment, and monitoring.

PURPOSE OF REVIEW: Laboratory support is central to clinical medicine. The present review focuses on the health systems strengthening needs of the clinic-laboratory-interface (CLI) in developing countries within the context of HIV diagnosis, treatment, and monitoring. RECENT FINDINGS: Although significant improvements have been made in reducing error rates in the analytical phase of pathology services within laboratories, there remain substantial deficits in both the preanalytic and postanalytic phases of the entire value chain that require strengthening. Recent interventions have largely aimed at improving the CLI focus through technological advancements. Although these are important, addressing CLI deficiencies within the context of the World Health Organization's health systems strengthening building blocks has been relatively neglected, and data are very limited in resource-limited countries. Broad health systems strengthening in the CLI is required. The present study reviews the key elements of the CLI associated with poor laboratory functioning, and proposes specific areas for improvement. The present study discuss case studies in South Africa and India, as well as reviewing data supporting interventions in public health clinics to improve the CLI. SUMMARY: The CLI is a neglected area of the laboratory value chain. Reducing errors and increasing efficiencies will improve the laboratory service to patients.

Subtype C gp140 Vaccine Boosts Immune Responses Primed by the South African AIDS Vaccine Initiative DNA-C2 and MVA-C HIV Vaccines after More than a 2-Year Gap.

A phase I safety and immunogenicity study investigated South African AIDS Vaccine Initiative (SAAVI) HIV-1 subtype C (HIV-1C) DNA vaccine encoding Gag-RT-Tat-Nef and gp150, boosted with modified vaccinia Ankara (MVA) expressing matched antigens. Following the finding of partial protective efficacy in the RV144 HIV vaccine efficacy trial, a protein boost with HIV-1 subtype C V2-deleted gp140 with MF59 was added to the regimen. A total of 48 participants (12 U.S. participants and 36 Republic of South Africa [RSA] participants) were randomized to receive 3 intramuscular (i.m.) doses of SAAVI DNA-C2 of 4 mg (months 0, 1, and 2) and 2 i.m. doses of SAAVI MVA-C of 1.45 × 10(9) PFU (months 4 and 5) (n = 40) or of a placebo (n = 8). Approximately 2 years after vaccination, 27 participants were rerandomized to receive gp140/MF59 at 100 µg or placebo, as 2 i.m. injections, 3 months apart. The vaccine regimen was safe and well tolerated. After the DNA-MVA regimen, CD4(+) T-cell and CD8(+) T-cell responses occurred in 74% and 32% of the participants, respectively. The protein boost increased CD4(+) T-cell responses to 87% of the subjects. All participants developed tier 1 HIV-1C neutralizing antibody responses as well as durable Env binding antibodies that recognized linear V3 and C5 peptides. The HIV-1 subtype C DNA-MVA vaccine regimen showed promising cellular immunogenicity. Boosting with gp140/MF59 enhanced levels of binding and neutralizing antibodies as well as CD4(+) T-cell responses to HIV-1 envelope. (This study has been registered at ClinicalTrials.gov under registration no. NCT00574600 and NCT01423825.).

Implementation of HIV and Tuberculosis Diagnostics: The Importance of Context.

BACKGROUND: Novel diagnostics have been widely applied across human immunodeficiency virus (HIV) and tuberculosis prevention and treatment programs. To achieve the greatest impact, HIV and tuberculosis diagnostic programs must carefully plan and implement within the context of a specific healthcare system and the laboratory capacity. METHODS: A workshop was convened in Cape Town in September 2014. Participants included experts from laboratory and clinical practices, officials from ministries of health, and representatives from industry. RESULTS: The article summarizes best practices, challenges, and lessons learned from implementation experiences across sub-Saharan Africa for (1) building laboratory programs within the context of a healthcare system; (2) utilizing experience of clinicians and healthcare partners in planning and implementing the right diagnostic; and (3) evaluating the effects of new diagnostics on the healthcare system and on patient health outcomes. CONCLUSIONS: The successful implementation of HIV and tuberculosis diagnostics in resource-limited settings relies on careful consideration of each specific context.

Optimization of a polyglutamine aggregation inhibitor peptide (QBP1) using a thioflavin T fluorescence assay.

Polyglutamine protein aggregates are a hallmark of several neurodegenerative diseases, including Huntington's disease, and increasing evidence suggests that reducing or inhibiting aggregation produces a therapeutic benefit in animal models of disease. Part of the challenge in designing compounds that interfere with protein aggregation is having a sensitive and consistent in vitro assay that allows for efficient screening and lead optimization. Here we describe a simplified polyglutamine assay that uses a soluble, pathological-length polyglutamine construct (62 glutamines [Q62]) fused to glutathione-S-transferase (GST) and measure aggregate formation with fluorescence generated by thioflavin T binding. Controlled release of Q62 from GST using proteolytic cleavage resulted in time-dependent aggregate formation that was not observed for a non-pathological-length GST-Q19 construct. Cleavage of the polyglutamine domain from GST increased the rate of Q62 aggregation from days to hours, significantly decreasing the time for compound analysis. Controlled aggregate formation combined with the fluorescence sensitivity of the dye thioflavin T allowed us to screen a series of peptide analogs for lead optimization of a previously identified peptide aggregation inhibitor, QBP1. QBP1 analogs showed the greatest inhibitory potency when added prior to Q62 aggregate initiation, suggesting that the mechanism of inhibition was interference with early formed aggregates that were not detectable by ultraviolet or dye binding. The assay detected activities that differed by three orders of magnitudes with Z' = 0.56, which is suitable for high-throughput screening and allowed us to do lead optimization of QBP1 analogs for pharmacophore model building.

Investigation of HIV in amniotic fluid from HIV-infected pregnant women at full term.

BACKGROUND: In the absence of interventions and breast-feeding, the in utero transmission rate of human immunodeficiency virus (HIV) is estimated to be 10%-15%, and the role that amniotic fluid (AF) plays in this is unclear. OBJECTIVES: Levels of cytomegalovirus (CMV) in AF and levels of HIV-1 in AF, maternal blood, and fetal cord blood were assessed.Study design. We enrolled 23 HIV-1-positive women with healthy, singleton pregnancies who underwent elective cesarean section (CS) at full term. The Roche Amplicor HIV-1 Monitor test (version 1.5) was used for determination of maternal plasma VLs. The NASBA Nuclisens assay was used for determination of VLs in other samples. To determine the feasibility of detecting viral infections in AF, CMV polymerase chain reaction DNA extraction was performed on the AF samples by use of the QIAamp DNA kit. RESULTS: HIV-1 RNA was not detected in either AF or fetal cord blood. CMV was detected in 4 AF samples. Maternal CD4(+) T cell counts were 158-654 cells/mL (mean, 405 cells/mL). The maternal plasma VLs ranged from below detectable limits to 169,990 copies/mL (mean, 33,700 copies/mL). CONCLUSIONS: In the absence of medical complications and before labor, AF collected during elective CS from women who had received either zidovudine or nevirapine during late-stage pregnancy was free of HIV.

Molecular characterization of duck hepatitis B virus isolates from South African ducks.

The objective of the study was to characterize the genome of duck hepatitis B virus (DHBV) isolates from South African Pekin ducks. Duck serum and liver samples were collected from two commercial duck farms from geographically distinct regions of South Africa. In total, 498 duck serum samples were tested for the presence of DHBV DNA using either sub-genomic or full-length polymerase chain reaction (PCR) assays. The overall prevalence of DHBV infection in South African ducks was 47%. In addition, 30% of 59 liver tissues tested were DHBV DNA-positive. Six randomly selected serum or liver samples were used to clone and sequence the genomes of the South African DHBV strains. All six isolates had DHBV genomes of 3,021 nucleotides with three characteristic overlapping reading frames encoding the polymerase, surface and core gene products. No X-like gene with a traditional start codon was found. Following phylogenetic analysis, the South African DHBV isolates clustered with DHBV isolates from other "Western" countries, including United States of America, Canada, Germany and India. On translation of the open reading frames, the South African isolates were found to share signature amino acids in the polymerase and surface genes with the "Western" country isolates as opposed to those of Chinese DHBV isolates.

Pharmacists' opinions regarding level of involvement in emergency preparedness and response.

OBJECTIVE: To assess opinions of pharmacist-members of the American Pharmacists Association Academy of Pharmacy Practice and Management (APhA-APPM) regarding the appropriate level of pharmacists' involvement in emergency preparedness and response activities and to determine whether opinions differed according to demographic characteristics. DESIGN: Cross-sectional, descriptive, Web-based survey. SETTING: United States. PARTICIPANTS: Five hundred eighteen APhA-APPM member-pharmacists. MAIN OUTCOME MEASURES: Responses to survey questions. RESULTS: Respondents to our survey indicated that pharmacists should have a high level of involvement in emergency preparedness and response activities. Traditional pharmacy practice activities (such as medication preparation and dispensing) and patient education were the most highly supported roles for pharmacists. Newer activities such as surveillance, vaccine administration, and mobilization were also strongly supported. Demographic characteristics, such as age, sex, degree, state of residence, practice setting, and employment setting, did not influence respondents' opinions. The only characteristic that influenced pharmacist opinions was previous participation in local and/or state emergency preparedness and response activities. Compared with other respondents, pharmacists who participated in these activities gave higher ratings to these possible roles for pharmacists: surveillance, triage/evaluation, community planning and preparation, mobilization, and training of others. CONCLUSION: Pharmacist-members of APhA-APPM who responded to this survey believe that participating in public health activities related to emergency preparedness and response is important for members of the pharmacy profession.