Patient trust and positive attitudes maximize non-communicable diseases management in rural Tanzania.

The objectives of this study were to identify difficulties and their related contexts non-communicable disease (NCD) patients in rural Tanzania experienced, examine how patients managed the situation by seeking better treatment of the diseases, and propose a realistic approach for optimizing disease management with long-term perspectives in resource-limited settings, based on views of patients (PTs), health-care providers (HPs), and health volunteers (HVs). Nine focus group discussions were performed with 56 participants of PTs, HPs, and HVs in three district hospitals in the Dodoma region. Their views and self-care practices were extracted, and the verbatim data were analyzed to derive codes and categories. The types of NCDs reported by the PTs were hypertension (HT), diabetes mellitus (DM), and HT/DM comorbidity. Reported barriers to disease management included discontinuation of treatment due to various factors and a lack of positive messages regarding disease management in NCD care. The following points were addressed in relation to the improved management of NCDs: (i) positive attitudes and coping skills, (ii) support from family members, (iii) good communication between PTs and HPs, and (iv) trustworthy relationships with HVs. The findings suggest that to gain the trust of PTs in optimizing disease control in overstretched health-care systems, patient support systems should be strengthened by empowering positive attitudes.

Non-communicable diseases (NCDs) are the leading cause of death globally. NCDs are common in low- and middle-income countries and their prevalence has been growing more prominent. In Tanzania, one-third of all deaths are NCD-related. This study aims to identify the factors that may lead to the improved management of NCDs in rural Tanzania based on actual situations in patients' daily lives. We conducted focus group discussions with three different groups (patients with hypertension and/or diabetes mellitus [PTs], health volunteers [HVs], and health-care providers [HPs]). The results revealed that PTs faced various barriers such as treatment discontinuation and a lack of positive messages regarding disease management in NCD care. However, the following points were indicated by the participants for the improved management of NCDs: (i) positive attitudes and coping skills, (ii) support from family members, (iii) good communication between PTs and HPs, and (iv) trustworthy relationships with HVs. Thus, to gain the trust of PTs in optimizing disease control and complications in overstretched health-care systems, patient support systems need to be strengthened by adopting a community empowerment approach, delivering supportive messages, and building reliable relationships.

A chromosome 5q31.1 locus associates with tuberculin skin test reactivity in HIV-positive individuals from tuberculosis hyper-endemic regions in east Africa.

One in three people has been infected with Mycobacterium tuberculosis (MTB), and the risk for MTB infection in HIV-infected individuals is even higher. We hypothesized that HIV-positive individuals living in tuberculosis-endemic regions who do not get infected by Mycobacterium tuberculosis are genetically resistant. Using an "experiment of nature" design that proved successful in our previous work, we performed a genome-wide association study of tuberculin skin test positivity using 469 HIV-positive patients from prospective study cohorts of tuberculosis from Tanzania and Uganda to identify genetic loci associated with MTB infection in the context of HIV-infection. Among these individuals, 244 tested were tuberculin skin test (TST) positive either at enrollment or during the >8 year follow up, while 225 were not. We identified a genome-wide significant association between a dominant model of rs877356 and binary TST status in the combined cohort (Odds ratio = 0.2671, p = 1.22x10-8). Association was replicated with similar significance when examining TST induration as a continuous trait. The variant lies in the 5q31.1 region, 57kb downstream from IL9. Two-locus analyses of association of variants near rs877356 showed a haplotype comprised of rs877356 and an IL9 missense variant, rs2069885, had the most significant association (p = 1.59x10-12). We also replicated previously linked loci on chromosomes 2, 5, and 11. IL9 is a cytokine produced by mast cells and TH2 cells during inflammatory responses, providing a possible link between airway inflammation and protection from MTB infection. Our results indicate that studying uninfected, HIV-positive participants with extensive exposure increases the power to detect associations in complex infectious disease.

A Locus at 5q33.3 Confers Resistance to Tuberculosis in Highly Susceptible Individuals.

Immunosuppression resulting from HIV infection increases the risk of progression to active tuberculosis (TB) both in individuals newly exposed to Mycobacterium tuberculosis (MTB) and in those with latent infections. We hypothesized that HIV-positive individuals who do not develop TB, despite living in areas where it is hyperendemic, provide a model of natural resistance. We performed a genome-wide association study of TB resistance by using 581 HIV-positive Ugandans and Tanzanians enrolled in prospective cohort studies of TB; 267 of these individuals developed active TB, and 314 did not. A common variant, rs4921437 at 5q33.3, was significantly associated with TB (odds ratio = 0.37, p = 2.11 × 10(-8)). This variant lies within a genomic region that includes IL12B and is embedded in an H3K27Ac histone mark. The locus also displays consistent patterns of linkage disequilibrium across African populations and has signals of strong selection in populations from equatorial Africa. Along with prior studies demonstrating that therapy with IL-12 (the cytokine encoded in part by IL12B, associated with longer survival following MTB infection in mice deficient in CD4 T cells), our results suggest that this pathway might be an excellent target for the development of new modalities for treating TB, especially for HIV-positive individuals. Our results also indicate that studying extreme disease resistance in the face of extensive exposure can increase the power to detect associations in complex infectious disease.

Client satisfaction with family planning services in the area of high unmet need: evidence from Tanzania Service Provision Assessment Survey, 2014-2015.

BACKGROUND: Client satisfaction has been found to be an important factor for the uptake and continuation of family planning services. This study aimed to examine the current status of and factors associated with client's satisfaction with family planning services in Tanzania, which has a high unmet need for family planning. METHODS: The study used data from the Tanzania Service Provision Assessment survey of 2014-2015. A facility was classified as having high service readiness for FP if it scored at least 67.7% on a composite score based on three domains (staff training and guidelines, basic diagnostic equipment, and basic medicines), following criteria developed by the World Health Organization. The exit interview questionnaire was used to collect information from women about their level of satisfaction, whether "very satisfied," "more or less satisfied," or not satisfied with the services received. The response was dichotomized into "Yes" if the woman reported being very satisfied with services received otherwise coded as "No". Unadjusted and adjusted logistic regression models were used to assess the association between the client satisfaction and covariate variables; service readiness, facility type, managing authority, location, management meetings, supervision, provider's sex, and working experience, clients' age and education. All analyses were weighted to correct for non-response, disproportionate and complex sampling by using the "SVY" command in Stata 14. RESULTS: Out of the 1188 facilities included in the survey, 427 (35.9%) provided family planning services. A total of 1746 women participated in observations and exit interviews. Few (22%) facilities had a high readiness to provide family planning services. While most facilities had the recommended equipment available, only 42% stocked contraceptives (e.g. oral pills, injectable contraceptives and/or condoms). Further, trained staff and clinical guidelines were present in only 30% of services. Nevertheless, the majority (91%) of clients reported that they were satisfied with services. In the multivariate analysis, a high service readiness score [AOR = 2.5, 95% CI; 1.1-6.0], receiving services from private facilities [AOR = 2.3, 95% CI; 1.1-5.0], and being in the age group 20 to 29 years [AOR = 0.3, 95% CI; 0.1-0.7] were all significantly associated with clients' satisfaction with family planning services. CONCLUSION: There is a high level of client satisfaction with family planning services in Tanzania. Maintaining and exceeding this level will require improvements in the provision of staff training and the availability of contraceptives in existing services.

Auditory Impairments in HIV-Infected Children.

OBJECTIVES: In a cross-sectional study of human immunodeficiency virus (HIV)-infected adults, the authors showed lower distortion product otoacoustic emissions (DPOAEs) in HIV+ individuals compared with controls as well as findings consistent with a central auditory processing deficit in HIV+ adults on antiretroviral therapy. The authors hypothesized that HIV+ children would also have a higher prevalence of abnormal central and peripheral hearing test results compared with HIV- controls. DESIGN: Pure-tone thresholds, DPOAEs, and tympanometry were performed on 244 subjects (131 HIV+ and 113 HIV- subjects). Thirty-five of the HIV+, and 3 of the HIV- subjects had a history of tuberculosis treatment. Gap detection results were available for 18 HIV- and 44 HIV+ children. Auditory brainstem response results were available for 72 HIV- and 72 HIV+ children. Data from ears with abnormal tympanograms were excluded. RESULTS: HIV+ subjects were significantly more likely to have abnormal tympanograms, histories of ear drainage, tuberculosis, or dizziness. All audiometric results were compared between groups using a two-way ANOVA with HIV status and ear drainage history as grouping variables. Mean audiometric thresholds, gap detection thresholds, and auditory brainstem response latencies did not differ between groups, although the HIV+ group had a higher proportion of individuals with a hearing loss >25 dB HL in the better ear. The HIV+ group had reduced DPOAE levels (p < 0.05) at multiple frequencies compared with HIV- subjects. No relationships were found between treatment regimens or delay in starting treatment and audiological parameters. CONCLUSIONS: As expected, children with HIV+ were more likely to have a history of ear drainage, and to have abnormal tympanograms. Similar to the adult findings, the HIV+ group did not show significantly reduced audiometric thresholds, but did have significantly lower DPOAE magnitudes. These data suggest that (1) HIV+ children often have middle ear damage which complicates understanding the direct effects of HIV on the hearing system, and (2) even when corrected for confounders DPOAEs were lower in the HIV+ group. Previous studies suggest ototoxicity from antiretroviral drugs is an unlikely cause of the reduced DPOAE magnitudes. Other possibilities include effects on efferent pathways connecting to outer hair cells or a direct effect of HIV on the cochlea.

Auditory impairments in HIV-infected individuals in Tanzania.

OBJECTIVES: Abnormal hearing tests have been noted in human immunodeficiency virus (HIV)-infected patients in several studies, but the nature of the hearing deficit has not been clearly defined. The authors performed a cross-sectional study of both HIV+ and HIV- individuals in Tanzania by using an audiological test battery. The authors hypothesized that HIV+ adults would have a higher prevalence of abnormal central and peripheral hearing test results compared with HIV- controls. In addition, they anticipated that the prevalence of abnormal hearing assessments would increase with antiretroviral therapy (ART) use and treatment for tuberculosis (TB). DESIGN: Pure-tone thresholds, distortion product otoacoustic emissions (DPOAEs), tympanometry, and a gap-detection test were performed using a laptop-based hearing testing system on 751 subjects (100 HIV- in the United States, plus 651 in Dar es Salaam, Tanzania, including 449 HIV+ [130 ART- and 319 ART+], and 202 HIV-, subjects. No U.S. subjects had a history of TB treatment. In Tanzania, 204 of the HIV+ and 23 of the HIV- subjects had a history of TB treatment. Subjects completed a video and audio questionnaire about their hearing, as well as a health history questionnaire. RESULTS: HIV+ subjects had reduced DPOAE levels compared with HIV- subjects, but their hearing thresholds, tympanometry results, and gap-detection thresholds were similar. Within the HIV+ group, those on ART reported significantly greater difficulties understanding speech in noise, and were significantly more likely to report that they had difficulty understanding speech than the ART- group. The ART+ group had a significantly higher mean gap-detection threshold compared with the ART- group. No effects of TB treatment were seen. CONCLUSIONS: The fact that the ART+/ART- groups did not differ in measures of peripheral hearing ability (DPOAEs, thresholds), or middle ear measures (tympanometry), but that the ART+ group had significantly more trouble understanding speech and had higher gap-detection thresholds indicates a central processing deficit. These data suggest that: (1) hearing deficits in HIV+ individuals could be a CNS side effect of HIV infection, (2) certain ART regimens might produce CNS side effects that manifest themselves as hearing difficulties, and/or (3) some ART regimens may treat CNS HIV inadequately, perhaps due to insufficient CNS drug levels, which is reflected as a central hearing deficit. Monitoring of central hearing parameters could be used to track central effects of either HIV or ART.

Recurrent tuberculosis risk among HIV-infected adults in Tanzania with prior active tuberculosis.

BACKGROUND: Active tuberculosis is common among human immunodeficiency virus (HIV)-infected persons living in tuberculosis-endemic areas, but the hazard of subsequent tuberculosis disease has not been quantified in a single prospective cohort. METHODS: Among HIV-infected, BCG-immunized adults with CD4 counts ≥200 cells/µL who received placebo in the DarDar tuberculosis vaccine trial in Tanzania, we compared the prospective risk of active tuberculosis between subjects who did and who did not report prior active tuberculosis. All subjects with a positive tuberculin skin test without prior active tuberculosis were offered isoniazid preventive treatment. Definite or probable tuberculosis was diagnosed during active follow-up using rigorous published criteria. RESULTS: We diagnosed 52 cases of definite and 92 cases of definite/probable tuberculosis among 979 subjects during a median follow-up of 3.2 years. Among the 80 subjects who reported prior active tuberculosis, 11 (13.8%) subsequently developed definite tuberculosis and 17 (21.3%) developed definite/probable tuberculosis, compared with 41 (4.6%) and 75 (8.3%), respectively, of 899 subjects without prior active tuberculosis (definite tuberculosis risk ratio [RR], 3.01; 95% confidence interval [CI], 1.61-5.63, P < .001; definite/probable tuberculosis RR, 2.55; 95% CI, 1.59-4.09, P < .001). In a Cox regression model adjusting for age, CD4 count, and isoniazid receipt, subjects with prior active tuberculosis had substantially greater hazard of subsequent definite tuberculosis (hazard radio [HR], 3.69; 95% CI, 1.79-7.63, P < .001) and definite/probable tuberculosis (HR, 2.78; 95% CI, 1.58-4.87, P < .001). CONCLUSIONS: Compared to subjects without prior tuberculosis, the hazard of active tuberculosis is increased 3-fold among HIV-infected adults with prior active tuberculosis. Clinical Trials Registration. NCT0052195.

Pure-tone audiometric threshold assessment with in-ear monitoring of noise levels.

OBJECTIVE: Our objective was to obtain reliable threshold measurements without a sound booth by using a passive noise-attenuating hearing protector combined with in-ear 1/3-octave band noise measurements to verify the ear canal was suitably quiet. DESIGN: We deployed laptop-based hearing testing systems to Tanzania as part of a study of HIV infection and hearing. An in-ear probe containing a microphone was used under the hearing protector for both the in-ear noise measurements and threshold audiometry. The 1/3-octave band noise spectrum from the microphone was displayed on the operator's screen with acceptable levels in grey and unacceptable levels in red. Operators attempted to make all bars grey, but focused on achieving grey bars at 2000 Hz and above. STUDY SAMPLE: 624 adults and 197 children provided 3381 in-ear octave band measurements. Repeated measurements from 144 individuals who returned for testing on three separate occasions were also analysed. RESULTS: In-ear noise levels exceeded the maximum permissible ambient noise levels (MPANL) for ears not covered, but not the dB SPL levels corresponding to 0 dB HL between 2000-4000 Hz. In-ear noise measurements were repeatable over time. CONCLUSIONS: Reliable audiometry can be performed using a passive noise-attenuating hearing protector and in-ear noise measurements.

Reaching Patients With Noncommunicable Diseases in Rural Tanzania Using Mobile Devices and Community Trust: Qualitative Study.

BACKGROUND: A health service using mobile devices, mobile health (mHealth), has been widely applied to programs focusing on maternal and child health and communicable diseases in sub-Saharan African countries. However, mHealth apps for noncommunicable disease (NCD) services remain limited. OBJECTIVE: This study aimed to explore the acceptability and potential usability of SMS text messaging for patients and health care providers for the management of NCDs as part of an implementation research in rural Tanzania. METHODS: Nine focus group discussions were conducted with 56 participants (21 community health workers [CHWs], 17 patients, and 18 health care professionals [HPs]) in 3 districts in the Dodoma region, Tanzania. The interview guides were prepared in Swahili, and each session was recorded, transcribed, and translated into English. The focus group discussions consisted of the following topics: (1) perceptions of the participants about the possible use of mobile devices and SMS text messages as an mHealth platform in community health services; and (2) experiences of mobile device use in health activities or receiving health services via a mobile phone in the past. RESULTS: CHWs and HPs reported having familiarity using mobile devices to provide health services, especially for reaching or tracing patients in remote settings; however, patients with NCDs were less familiar with the use of mobile devices compared with the other groups. Hesitation to receive health services via SMS text messaging was seen in the patient group, as they wondered who would send health advice to them. Some patients expected services beyond what mHealth could do, such as aiding in recovery from a disease or sending notifications about the availability of prescription medications. CHWs showed interest in using text messaging to provide health services in the community; however, the concerns raised by CHWs included the cost of using their own mobile devices. Moreover, they demanded training about NCD management before engaging in such an activity. CONCLUSIONS: This study explored views and experiences regarding the possible installation of an mHealth intervention for managing NCDs in rural Tanzania. Although HPs and CHWs had experience using mobile devices to provide health services in non-NCD projects, only a few patients (3/17, 17%) had heard about the use of mobile devices to receive health services. To improve the suitability and acceptability of the intervention design for patients with NCDs, their trust must be earned. Involving CHWs in the intervention is recommended because they have already been appointed in the community and already know how to communicate effectively with patients in the area.

DAR-901 vaccine for the prevention of infection with Mycobacterium tuberculosis among BCG-immunized adolescents in Tanzania: A randomized controlled, double-blind phase 2b trial.

BACKGROUND: SRL172 prevented disease due to Mycobacterium tuberculosis in a Phase 3 trial. DAR-901 represents a scalable manufacturing process for SRL172. We sought to determine if DAR-901 would prevent infection with M. tuberculosis among BCG-primed adolescents age 13-15 years in Tanzania. METHODS: Adolescents with a negative T- SPOT.TBR interferon gamma release assay (IGRA) were randomized 1:1 to three intradermal injections of DAR-901 or saline placebo at 0, 2 and 4 months. Repeat IGRAs were performed at 2 months, and at 1, 2, and 3 years. The primary efficacy outcome was time to new TB infection (IGRA conversion to positive); the secondary outcome was time to persistent TB infection (IGRA conversion with repeat positive IGRA). RESULTS: Among 936 participants screened 667 were eligible and randomized to their first dose of vaccine or placebo (safety cohort). At 2 months, 625 participants remained IGRA-negative and were scheduled for the additional two doses (efficacy cohort). DAR-901 was safe and well-tolerated. One DAR-901 recipient developed a vaccine site abscess. Neither the primary nor secondary endpoints differed between the two treatment arms (p = 0.90 and p = 0.20, respectively). DAR-901 IGRA converters had median responses to ESAT-6 of 50.1 spot-forming cells (SFCs) vs. 19.6 SFCs in placebo IGRA converters (p = 0.03). CONCLUSIONS: A three-dose series of 1 mg DAR-901 was safe and well-tolerated but did not prevent initial or persistent IGRA conversion. DAR-901 recipients with IGRA conversion demonstrated enhanced immune responses to ESAT-6. Since protection against disease may require different immunologic responses than protection against infection a trial of DAR-901 to prevent TB disease is warranted. TRIAL REGISTRATION: The trial is registered at ClinicalTrials.gov as NCT02712424.

Hearing complaints in HIV infection originate in the brain not the ear.

OBJECTIVE: Evidence suggests damage to brain auditory pathways, rather than inner ear damage, underlies the hearing difficulties HIV+ individuals report. But, antiretroviral therapy (ART) may affect the hearing system and also lead to hearing complaints. DESIGN: Longitudinal study of HIV+ and HIV- individuals in Dar es Salaam Tanzania. A subset of this cohort started ART while in the study allowing the effects of ART to be studied directly. METHODS: The ability to hear quiet sounds (pure-tone audiometry), cochlear outer hair cell function [distortion-product otoacoustic emissions (DPOAEs)], and gaps-in-noise detection thresholds (a central auditory processing test) were assessed at each visit. Visits were scheduled for 6-month intervals, but the number and spacing of visits varied. In the group that started ART while in the study, 107 HIV+ individuals had audiometric thresholds, 98 had DPOAEs, and 98 had gap measurements suitable for analysis. Data were analyzed using a linear mixed model with time and starting ART as fixed effects and individual participant repeated measures as random effects. RESULTS: Starting ART did not affect audiometric or gap detection thresholds. The slope of the DPOAE amplitude vs. time relationship was more negative after starting ART but did not differ from the HIV- group. Gap thresholds were higher in the HIV+ group. CONCLUSION: ART did not affect audiometric thresholds significantly suggesting common ART drugs are not major ototoxins. The gap detection results from the study show effects on central auditory processing in HIV+ individuals, supporting the origin of HIV-related hearing complaints in the central auditory system.

Sialic acid levels in breast milk from HIV-positive Tanzanian women and impact of maternal diet.

OBJECTIVE: To quantify total sialic acid in milk from HIV-positive Tanzanian mothers and to determine the impact of maternal diet on milk sialic acid levels. DESIGN: Milk samples were analyzed from 74 HIV-positive, Tanzanian women enrolled in a randomized, controlled clinical study of a dietary macronutrient supplement. Women were provided with a daily protein-calorie supplement and a micronutrient supplement or micronutrient supplement only during the last trimester of pregnancy and up to the first 6 months of breastfeeding. METHODS: Milk samples were collected at approximately 2 weeks and at least 3 months postpartum and assayed for total sialic acid. Milk sialic acid was assessed relative to maternal macronutrient intake, age, BMI, CD4+ cell count and infant birth weight. RESULTS: The mean concentration of milk sialic acid was highest in the first 2 weeks postpartum (6.89 ±â€Š2.79 mmol/l) and declined rapidly by 3 months (2.49 ±â€Š0.60 mmol/l). Sialic acid content in milk was similar between both treatment arms of the study, and did not correlate with maternal macronutrient intake. No correlation was found between maternal age, BMI, CD4+ cell count or infant birth weight and total milk sialic acid concentration. CONCLUSION: Milk sialic acid levels in HIV-positive, Tanzanian women without malnutrition are comparable with reported values for women of European descent and show a similar temporal decline during early lactation. These findings suggest that total milk sialic acid is maintained despite macronutrient deficiencies in maternal diet and support a conserved role for milk sialic acid in neonatal development.

Pre- and post-natal macronutrient supplementation for HIV-positive women in Tanzania: Effects on infant birth weight and HIV transmission.

OBJECTIVE: To determine if a protein-calorie supplement (PCS) plus a micronutrient supplement (MNS) improves outcomes for HIV-infected lactating women and their infants. DESIGN: Randomized, controlled trial. SETTING: Dar es Salaam, Tanzania. SUBJECTS, PARTICIPANTS: Pregnant HIV-infected women enrolled in PMTCT programs who intended to breastfeed for 6 months. INTERVENTION: Randomization 1:1 to administration of a PCS plus MNS versus MNS alone among 96 eligible women beginning in the third trimester and continuing for 6 months of breast-feeding. MAIN OUTCOME MEASURE(S): Primary: infant weight at 3 months. Secondary: maternal BMI at 6 months. RESULTS: PCS resulted in significant increases in daily energy intake compared to MNS at all time points (range of differences: +388-719 Kcal); and increases in daily protein intake (range of differences: +22-33 gm). Infant birth weight (excluding twins) was higher in the PCS than MNS groups: 3.30 kg vs 3.04 kg (p = 0.04). Infant weight at 3 months did not differ between PCS and MNS groups: 5.63 kg vs 5.99 kg (p = 0.07). Maternal BMI at 6 months did not differ between PCS and MNS groups: 24.3 vs 23.8 kg/m2 (p = 0.68). HIV transmission occurred in 0 infants in the PCS group vs 4 in the MNS group (p = 0.03). CONCLUSIONS: In comparison to MNS the PCS + MNS intervention was well tolerated, increased maternal energy and protein intake, and increased infant birth weight, but not weight at 3 months or maternal BMI at 6 months. Reduced infant HIV transmission in the PCS + MNS group was observed. TRIAL REGISTRATION: Clinical Trials.Gov NCT01461863.

Fine epitope signature of antibody neutralization breadth at the HIV-1 envelope CD4-binding site.

Major advances in donor identification, antigen probe design, and experimental methods to clone pathogen-specific antibodies have led to an exponential growth in the number of newly characterized broadly neutralizing antibodies (bnAbs) that recognize the HIV-1 envelope glycoprotein. Characterization of these bnAbs has defined new epitopes and novel modes of recognition that can result in potent neutralization of HIV-1. However, the translation of envelope recognition profiles in biophysical assays into an understanding of in vivo activity has lagged behind, and identification of subjects and mAbs with potent antiviral activity has remained reliant on empirical evaluation of neutralization potency and breadth. To begin to address this discrepancy between recombinant protein recognition and virus neutralization, we studied the fine epitope specificity of a panel of CD4-binding site (CD4bs) antibodies to define the molecular recognition features of functionally potent humoral responses targeting the HIV-1 envelope site bound by CD4. Whereas previous studies have used neutralization data and machine-learning methods to provide epitope maps, here, this approach was reversed, demonstrating that simple binding assays of fine epitope specificity can prospectively identify broadly neutralizing CD4bs-specific mAbs. Building on this result, we show that epitope mapping and prediction of neutralization breadth can also be accomplished in the assessment of polyclonal serum responses. Thus, this study identifies a set of CD4bs bnAb signature amino acid residues and demonstrates that sensitivity to mutations at signature positions is sufficient to predict neutralization breadth of polyclonal sera with a high degree of accuracy across cohorts and across clades.

Deficiencies of macronutrient intake among HIV-positive breastfeeding women in Dar es Salaam, Tanzania.

We compared macronutrient intake, food insecurity, and anthropometrics in breastfeeding women: 40 HIV-positive women not yet on antiretroviral therapy and 40 HIV-negative women. Calculated deficits at 2 weeks were 517 kcal per day for HIV-positive women vs 87 kcal per day surplus for HIV-negative women (P = 0.01) and 29 g protein per day for HIV-positive women vs 16 g protein per day for HIV-negative women (P = 0.04). Food insecurity scores were 11.3 for HIV-positive women vs 7.8 for HIV-negative women (P < 0.01). Enhanced dietary education together with macronutrient supplementation may be required to improve health outcomes in HIV-positive women and their infants.

Maintenance of Sensitivity of the T-SPOT.TB Assay after Overnight Storage of Blood Samples, Dar es Salaam, Tanzania.

Background. T-SPOT.TB is an interferon gamma release assay for detecting Mycobacterium tuberculosis infection. The requirement to process within 8 hours is constraining, deters use, and leads to invalid results. Addition of T Cell Xtend reagent may allow delayed processing, but has not been extensively field tested. Design. Consecutive AFB smear positive adult tuberculosis patients were prospectively recruited in Dar es Salaam, Tanzania. Patients provided a medical history, 1-3 sputum samples for culture and 1 blood sample which was transported to the laboratory under temperature-controlled conditions. After overnight storage, 25 µL of T Cell Xtend reagent was added per mL of blood, and the sample was tested using T-SPOT.TB. Results. 143 patients were enrolled: 57 patients were excluded because temperature control was not maintained, 19 patients were excluded due to red blood cell contamination, and one did not provide a sputum sample for culture. Among 66 evaluable patients, overall agreement between T-SPOT.TB and culture was 95.4% (95%CI; 87.1-99.0%) with Kappa value 0.548. Sensitivity of T-SPOT.TB when using T Cell Xtend reagent was 96.8% (95%CI; 88.8-99.6%). Conclusions. When T Cell Xtend reagent is added to specimens held overnight at recommended temperatures, T-SPOT.TB is as sensitive as the standard assay in patients with tuberculosis.