Predictors of growth and body composition in HIV-infected children beginning or changing antiretroviral therapy.

OBJECTIVES: The aim of the study was to describe growth and body composition changes in HIV-positive children after they had initiated or changed antiretroviral therapy (ART) and to correlate these with viral, immune and treatment parameters. METHODS: Ninety-seven prepubertal HIV-positive children were observed over 48 weeks upon beginning or changing ART. Anthropometry and bioelectrical impedance analysis results were compared with results from the National Health and Nutrition Examination Survey 1999-2002 (NHANES) to generate z-scores and with results for HIV-exposed, uninfected children from the Women and Infants Transmission Study (WITS). Multivariate analysis was used to evaluate associations between growth and body composition and disease parameters. RESULTS: All baseline lean and fat mass measures were below those of controls from NHANES. Weight, height and fat free mass (FFM) index (FFM/height(2)) z-scores increased over time (P = 0.004, 0.037 and 0.027, respectively) and the waist:height ratio z-score decreased (P = 0.045), but body mass index and per cent body fat z-scores did not change. Measures did not increase more than in uninfected WITS controls. In multivariate analysis, baseline height, mid-thigh circumference and FFM z-scores related to CD4 percentage (P = 0.029, P = 0.008 and 0.020, respectively) and change in FFM and FFM index z-scores to CD4 percentage increase (P = 0.010 and 0.011, respectively). Compared with WITS controls, baseline differences in height and mid-thigh muscle circumference were also associated with CD4 percentage. Case-control differences in change in both subscapular skinfold (SSF) thickness and the SSF:triceps skinfold ratio were inversely associated with viral suppression. No measures related to ART class(es) at baseline or over time. CONCLUSIONS: In these HIV-positive children, beginning or changing ART was associated with improved growth and lean body mass (LBM), as indicated by FFM index. Height and LBM related to CD4 percentage at baseline and over time. Altered fat distribution and greater central adiposity were associated with detectable virus but not ART class(es) received.

Bacterial safety of flash-heated and unheated expressed breastmilk during storage.

Heat-treated breastmilk is one infant-feeding option recommended by the WHO to reduce mother-to-child transmission of HIV in developing countries. Flash-heat, a simple pasteurization method that a mother could perform in her home, has been shown to inactivate cell-free HIV-1. Since heating may affect the naturally occurring antimicrobial properties found in breastmilk, storing heated breastmilk may present a safety issue in resource-poor settings due to lack of refrigeration and potential contamination. To address this, we investigated the ability of flash-heat to eliminate bacteria and to prevent growth over time compared with unheated breastmilk. We collected breastmilk samples from 38 HIV positive mothers in South Africa and aliquoted them to flash-heated and unheated controls. Samples were stored at room temperature for 0, 2, 6 and 8 h and then plated and incubated for 24 h at 37 degrees C in CO(2). We performed total colony counts and identified Escherichia coli, Staphylocuccus aureus and Group A and Group B streptococci. Unheated samples had a significantly higher number of samples positive for bacterial growth at each time point (p < 0.0001), as well as mean colony-forming units (CFU)/ml in those samples that were positive at each time point (p < 0.0001). In addition, unheated samples had a significantly higher rate of bacterial propagation over time than flash-heated samples when comparing log values of CFU/ml across 0-8 h (p < 0.005). No pathogenic growth was observed in the flash-heated samples, while the unheated samples showed growth of E. coli (n = 1) and S. aureus (n = 6). Our data suggest that storage of flash-heated breastmilk is safe at room temperature for up to 8 h.

Effects of lipolysis or heat treatment on HIV-1 provirus in breast milk.

BACKGROUND: Transmission of HIV-1 infection through breastfeeding is associated with integrated DNA (provirus) in milk cells. Reduction of HIV-1 DNA in milk may lessen infectivity. PURPOSE: To investigate efficacy of two methods available in developing countries to reduce HIV-1 proviral DNA in breast milk. METHODS: Methods simulated field conditions; milk was heated by bringing it to a boil, for instance, over a cooking fire, and lipolysis was done at room temperature. Four HIV-positive pregnant women were recruited for this pilot study, instructed to feed formula exclusively, and to stimulate milk production using pumping. Milk was collected twice weekly for 3 weeks and analyzed qualitatively for HIV-1 proviral DNA by polymerase chain reaction at three stages: 1) fresh, 2) after standing for 6 hours, and 3) after having been brought to the boiling point. RESULTS: Seventeen samples from 4 mothers were analyzed. Fifteen of 17 fresh samples (88%) had measurable HIV-1 proviral DNA despite all mothers' having had low or undetectable plasma viral loads. Lipolysis (standing at room temperature) for 6 hours did not destroy proviral DNA: 6 of 7 samples (86%) tested positive for DNA after lipolysis. No samples of milk (n = 8) brought to a boil were positive for HIV-1 proviral DNA (p <.0001). CONCLUSIONS: This preliminary evidence suggests that inherent lipolytic activity of fresh breast milk is inadequate for destruction of HIV-1; bringing breast milk to a boil may result in decreased HIV-1 infectivity; and breast milk cell-associated HIV-1 may not reflect plasma viral load. Nutritional value or possible bacterial contamination of milk treated in this manner was not assessed.

Plasma glutathione concentrations in non-infected infants born from HIV-infected mothers: developmental profile.

Glutathione (GSH) is the primary antioxidant in humans. Oxidative cellular injury is postulated to be centrally involved in diverse processes including aging, cancer, cardiovascular disease, and Human Immunodeficiency Virus (HIV) disease progression. Normal plasma GSH concentrations have been well characterized in healthy children and adults, but not during infant development. The objectives of this study were to: a) measure plasma GSH concentrations in non-infected infants born from HIV-infected mothers, to b) assess the developmental variations with age and gender, and c) evaluate for possible associations with growth, anemia, and other maternal and infant variables. One hundred and seventy (170) plasma samples from 44 HIV-uninfected infants (birth to 18 mos.) born to HIV-infected mothers from the Women and Infant Transmission Study (Puerto Rico site) were analyzed. The total plasma GSH geometric mean concentration for all samples analyzed was 1.94 (1.06) mumoles/L. A developmental effect of age was seen with lower concentrations in younger infants (0-2 months) than in older infants 4-18 months. There was no significant effect of gender, anemia, zidovudine exposure, maternal age, maternal CD4 cell percent, or infant growth, although a trend towards increasing GSH concentration was seen with increasing weight for height z-score. These findings have multiple clinical ramifications including prediction of capacity to detoxify oxidants at different ages, and partial explanation for the increased viral loads seen in HIV-infected infants.

Seroreversion in human immunodeficiency virus-exposed but uninfected infants.

The goal of this study was to describe seroreversion (SR) in a cohort of human immunodeficiency virus-exposed but uninfected infants. Groups of patients who seroreverted very early or late were examined for salient clinical and immunologic characteristics of the mother or infant. The mean time (+/- s.d.) to seroreversion by enzyme-linked immunoabsorbent assay (ELISA) was 50.1 +/- 14.8 weeks, or 11.6 months (n = 84); the range of times to antibody loss by ELISA was 17.9 to 82.0 weeks. The mean time to seroreversion by Western blot was 68.3 +/- 12.6 weeks, or 15.8 months (n = 51), with a range of 44.9 to 94.1 weeks. Initial anti-human immunodeficiency virus titer as measured by cord blood ELISA optical density (OD) was found to relate significantly to mean time to seroreversion. No relationship to time to seroreversion was demonstrated for gestational age, maternal or neonatal serum immunoglobulin concentrations, maternal CD4 cell counts, maternal alcohol consumption, infantile diarrhea or failure to thrive. The lengthy time to seroreversion seen here demonstrates the 1994 revised Centers for Disease Control and Prevention definition of human immunodeficiency virus infection (based on seropositivity by both ELISA and confirmatory tests persisting beyond 18 months of age) to be accurate in our population. We recommend Western blot testing be used as confirmation for positive ELISAs only after 18 months of age.

Non-neurogenic mechanism for reserpine-induced release of catecholamines from the adrenal medulla of neonatal rats: possible modulation by opiate receptors.

Neonatal rats do not have functional splanchnic nerve connections to the adrenal medulla until approximately one week of postnatal age, yet they are able to respond to some drugs or stresses by releasing adrenal catecholamines. Reserpine (5 mg/kg s.c.) resulted in significant loss (20-40%) of neonatal catecholamines within 4 h; unlike the acute effects of reserpine in the adult, depletion was not prevented by pretreatment with a nicotinic blocking agent, demonstrating that the effect in the neonate is non-neurogenic. Depletion did not result simply from inhibition of catecholamine storage by reserpine, but rather, the non-neurogenic depletion represented net movement of the amines from the storage granules into the extracellular space, as evaluated by the subcellular distribution of catecholamines. These results suggest that the non-neurogenic mechanism represents release of catecholamines. The immature response mechanism disappeared by 11 days of age and was replaced by a completely neurogenic release, demonstrating that the special mechanism is lost within several days of the onset of functional innervation of the adrenal. Experiments also were carried out to test whether endogenous opioids and/or opiate receptors are involved in the non-neurogenic mechanism. Naloxone (5 mg/kg s.c.) potentiated the depletion of catecholamines by reserpine, while methadone (2.5 mg/kg s.c.) inhibited the non-neurogenic response. In contrast, no potentiation of release by naloxone or inhibition by methadone was seen in adult rats. Thus, opiate receptors may modulate only the immature secretory mechanism.