Adverse childhood experiences and mental health conditions among multiracial adolescents.

OBJECTIVE: The objective of this study was to verify the factor structure of the household dysfunction type of ACE using data from the National Survey of Children's Health (NSCH), and then examine whether household dysfunction (measured as a latent construct) was associated with mental health conditions among multiracial adolescents. DESIGN: We used cross-sectional data collected in 2016 from caregivers who completed the NSCH and analyzed data from a subpopulation of adolescents (12-17) who reported more than one race (n = 1,231). Mplus 8.4 was used to conduct confirmatory factor analysis and probit models from a structural equation modeling framework. RESULTS: Results from this study indicated that the household dysfunction type of ACE, as a latent construct, had good model fit and was significantly associated with depression [standardized coefficient [B] = .50, 95% confidence interval [CI] .36, .65], anxiety [B = .61, 95% CI .48, .73], behavior problems [B = .58, 95% CI .44, .72], and ADHD [B = .54, 95% CI .38, .69] for multiracial adolescents. CONCLUSIONS: Household dysfunction may result in adolescents being separated (physically or emotionally) from their caregivers, which may hinder adolescents' ability to establish or maintain one of the most important relationships needed to promote racial/ethnic identity development and mental health. Implications for advancements in theory and NSCH are presented.

Demographic and Pathogens of Domestic, Free-Roaming Pets and the Implications for Wild Carnivores and Human Health in the San Luis Region of Costa Rica.

Habitat loss and degradation, restricted ranges, prey exploitation, and poaching are important factors for the decline of several wild carnivore populations and additional stress from infectious agents is an increasing concern. Given the rapid growth of human populations in some regions like Costa Rica, pathogens introduced, sustained, and transmitted by domestic carnivores may be particularly important. To better understand the significance of domestic carnivore pathogens for wildlife, we determine the prevalence of infection and possible mechanisms for contact between the two groups. The demographics, role in the household, and pathogens of pet dogs and cats was studied during three annual spay/neuter clinics in San Luis, Costa Rica. Most dogs were owned primarily as pets and guard animals, but ~10% were used for hunting. Cats were owned primarily as pets and for pest control. Both roamed freely outdoors. We detected high prevalences of some pathogens (e.g., carnivore protoparvovirus 1 and Toxoplasma gondii). Some pathogens are known to persist in the environment, which increases the probability of exposure to wild carnivores. This study demonstrated that domestic pets in San Luis, home to a number of protected and endangered wildlife species, are infected with pathogens to which these wild species are potentially susceptible. Additionally, results from our questionnaire support the potential for domestic and wild animal contact, which may result in disease spillover.

Can Conformity-Based Volumetric Modulated Arc Therapy Improve Dosimetry and Speed of Delivery in Radiation Therapy to Lumbosacral Spine Compared with Conventional Techniques?

PURPOSE: Volumetric modulated arc therapy (VMAT) has generally been perceived as too time and resource intensive for palliative radiation therapy mainly because of the need for extensive organs at risk contouring. Dose-limiting conformity-based objectives can be automatically generated and are commonly used to conform isodoses closely around the target volumes during inverse planning. The aim of this study was to determine if conformity-based objectives can be used to create VMAT plans for lumbosacral spine palliative radiation therapy without organs at risk contours, which will improve conformity, dose homogeneity, and speed of delivery compared with standard forward planning approaches. METHODS: A total of 25 patients were retrospectively replanned using three different planning techniques: (1) anterior-posterior parallel opposed pair (POP); (2) single isocenter anterior-posterior half-beam block junctioned to three fields, posterior and two laterals (JUNC); and (3) VMAT single arc. Treatment volume included L1-S5 vertebrae prescribed to 20 Gy in five fractions. Conformality index, homogeneity index, contour, planning, and treatment time were compared for each technique. RESULTS: Planning target volume V95 ≥95% was maintained for all 75 replans. VMAT was superior to POP and JUNC in terms of conformality (POP 2.0 vs. JUNC 1.8 vs. VMAT 1.2; P < .01) and homogeneity (POP 1.1 vs. JUNC 1.1 vs. VMAT 1.0; P < .01). Planning times for POP were the lowest (3.2 minutes). VMAT and POP had similar delivery times (1.5 minutes), which were approximately half the JUNC delivery time (3.2 minutes). CONCLUSIONS: Conformity-based VMAT was dosimetrically superior to conventional field-based planning and reduced delivery time. This reduction in normal tissue dose as well as reduced time spent on the treatment couch can potentially improve the quality of life in palliative patients receiving radiotherapy to the lumbosacral spine.

Deep sequence analysis of HIV adaptation following vertical transmission reveals the impact of immune pressure on the evolution of HIV.

Human immunodeficiency virus (HIV) can adapt to an individual's T cell immune response via genomic mutations that affect antigen recognition and impact disease outcome. These viral adaptations are specific to the host's human leucocyte antigen (HLA) alleles, as these molecules determine which peptides are presented to T cells. As HLA molecules are highly polymorphic at the population level, horizontal transmission events are most commonly between HLA-mismatched donor/recipient pairs, representing new immune selection environments for the transmitted virus. In this study, we utilised a deep sequencing approach to determine the HIV quasispecies in 26 mother-to-child transmission pairs where the potential for founder viruses to be pre-adapted is high due to the pairs being haplo-identical at HLA loci. This scenario allowed the assessment of specific HIV adaptations following transmission in either a non-selective immune environment, due to recipient HLA mismatched to original selecting HLA, or a selective immune environment, mediated by matched donor/recipient HLA. We show that the pattern of reversion or fixation of HIV adaptations following transmission provides insight into the replicative cost, and likely compensatory networks, associated with specific adaptations in vivo. Furthermore, although transmitted viruses were commonly heavily pre-adapted to the child's HLA genotype, we found evidence of de novo post-transmission adaptation, representing new epitopes targeted by the child's T cell response. High-resolution analysis of HIV adaptation is relevant when considering vaccine and cure strategies for individuals exposed to adapted viruses via transmission or reactivated from reservoirs.

Magnetically-enabled biomarker extraction and delivery system: towards integrated ASSURED diagnostic tools.

Diagnosis of asymptomatic malaria poses a great challenge to global disease elimination efforts. Healthcare infrastructure in rural settings cannot support existing state-of-the-art tools necessary to diagnose asymptomatic malaria infections. Instead, lateral flow immunoassays (LFAs) are widely used as a diagnostic tool in malaria endemic areas. While LFAs are simple and easy to use, they are unable to detect low levels of parasite infection. We have developed a field deployable Magnetically-enabled Biomarker Extraction And Delivery System (mBEADS) that significantly improves limits of detection for several commercially available LFAs. Integration of mBEADS with leading commercial Plasmodium falciparum malaria LFAs improves detection limits to encompass an estimated 95% of the disease reservoir. This user-centered mBEADS platform makes significant improvements to a previously cumbersome malaria biomarker enrichment strategy by improving reagent stability, decreasing the processing time 10-fold, and reducing the assay cost 10-fold. The resulting mBEADS process adds just three minutes and less than $0.25 to the total cost of a single LFA, thus balancing sensitivity and practicality to align with the World Health Organization's ASSURED criteria for point-of-care (POC) testing.

Vision Therapy for Binocular Dysfunction Post Brain Injury.

PURPOSE: To prospectively evaluate the effectiveness of home-based computer vergence therapy for the treatment of binocular vision disorders in adults at least 3 months after an acquired brain injury. METHODS: Eligibility criteria included presence of binocular dysfunction characterized by receded near point of convergence (≥6 cm break), insufficient positive fusional vergence at near (failing Sheard's criterion or <15△ blur or break), insufficient negative fusional vergence at near (<12△ blur or break), and/or reduced vergence facility at near (<15 cycles per minute with 12△BO/3△BI). Participants were prescribed 12 weeks of home-based computer vergence therapy. Phoria (cover test), negative fusional vergence, positive fusional vergence, near point of convergence, vergence facility, and symptoms (convergence insufficiency symptom survey [CISS]) were assessed at baseline and after 4, 8, and 12 weeks of prescribed therapy. ANOVA was used to evaluate change in each measure. Percentage successful was also determined. RESULTS: Nineteen participants were enrolled (mean age 45.4 ± 12.9 years); six participants were lost to follow-up. Baseline findings were orthophoria at distance, 7.2△ exophoria at near, near point of convergence break = 17.5 cm, near point of convergence recovery = 21.8 cm, negative fusional vergence = 12.3△, positive fusional vergence blur = 8.4△, vergence facility = 3.9 cycles per minute, and CISS = 32.1. ANOVA showed a statistically significant improvement for near point of convergence break (p = 0.002) and recovery (p < 0.001), positive fusional vergence blur (p < 0.0001), break (p < 0.0001), and recovery (p < 0.0001), negative fusional vergence blur (p = 0.037), break (p = 0.003), and recovery (p = 0.006), vergence facility (p < 0.0001), and CISS (p = 0.0001). The percentage of patients who were classified as "successful" or "improved" was 69% for near point of convergence (<6 cm or decrease of ≥4 cm), 77% for positive fusional vergence (>15△ and passing Sheard's criterion or increase of ≥10△), 77% for negative fusional vergence (≥12△ or increase of ≥6△), 62% for positive fusional vergence and near point of convergence composite, and 92% for vergence facility (15 cycles per minute or increase of 3 cycles per minute). CONCLUSIONS: The majority of participants who completed the study experienced meaningful improvements in signs and symptoms.

Antiretroviral therapy reduces the magnitude and T cell receptor repertoire diversity of HIV-specific T cell responses without changing T cell clonotype dominance.

After initiation of antiretroviral therapy (ART), HIV loads and frequencies of HIV epitope-specific immune responses decrease. A diverse virus-specific T cell receptor (TCR) repertoire allows the host to respond to viral epitope diversity, but the effect of antigen reduction as a result of ART on the TCR repertoire of epitope-specific CD8(+) T cell populations has not been well defined. We determined the TCR repertoires of 14 HIV-specific CD8(+) T cell responses from 8 HIV-positive individuals before and after initiation of ART. We used multiparameter flow cytometry to measure the distribution of memory T cell subsets and the surface expression of PD-1 on T cell populations and T cell clonotypes within epitope-specific responses from these individuals. Post-ART, we noted decreases in the frequency of circulating epitope-specific T cells (P = 0.02), decreases in the number of T-cell clonotypes found within epitope-specific T cell receptor repertoires (P = 0.024), and an overall reduction in the amino acid diversity within these responses (P < 0.0001). Despite this narrowing of the T cell response to HIV, the overall hierarchy of dominant T cell receptor clonotypes remained stable compared to that pre-ART. CD8(+) T cells underwent redistributions in memory phenotypes and a reduction in CD38 and PD-1 expression post-ART. Despite extensive remodeling at the structural and phenotypic levels, PD-1 was expressed at higher levels on dominant clonotypes within epitope-specific responses before and after initiation of ART. These data suggest that the antigen burden may maintain TCR diversity and that dominant clonotypes are sensitive to antigen even after dramatic reductions after initiation of ART.

Dominant clonotypes within HIV-specific T cell responses are programmed death-1high and CD127low and display reduced variant cross-reactivity.

HIV epitope-specific T cell responses are often comprised of clonotypic expansions with distinct functional properties. In HIV(+) individuals, we measured programmed death-1 (PD-1) and IL-7Rα expression, MHC class I tetramer binding, cytokine production, and proliferation profiles of dominant and subdominant TCR clonotypes to evaluate the relationship between the composition of the HIV-specific T cell repertoire and clonotypic phenotype and function. Dominant clonotypes are characterized by higher PD-1 expression and lower C127 expression compared with subdominant clonotypes, and TCR avidity positively correlates with PD-1 expression. At low peptide concentrations, dominant clonotypes fail to survive in culture. In response to stimulation with peptides representing variant epitopes, subdominant clonotypes produce higher relative levels of cytokines and display greater capacity for cross-recognition compared with dominant clonotypes. These data indicate that dominant clonotypes within HIV-specific T cell responses display a phenotype consistent with ongoing exposure to cognate viral epitopes and suggest that cross-reactive, subdominant clonotypes may retain greater capacity to suppress replication of viral variants as well as to survive in the absence of strong antigenic signaling.

Clonal expansion and TCR-independent differentiation shape the HIV-specific CD8+ effector-memory T-cell repertoire in vivo.

Flexibility of the HIV-specific T-cell receptor repertoire is a hallmark of HIV-1 infection. Altered differentiation of HIV-specific CD45RO(+)/CCR7(-) (TemRO) CD8(+) effector-memory T cells into CD45RA(+)/CCR7(-) (TemRA) CD8(+) effector-memory T cells as well as increased expression of the senescence marker CD57 has been frequently observed HIV-1 infection, but the structural relationship between clonal expansion and T-cell differentiation has not been defined. In this study, we demonstrate that HIV-specific clonotypes have differing degrees of TemRA differentiation but always maintain a significant proportion of TemRO-phenotype cells. These data indicate that structural constraints of the TCR/peptide major histocompatibility complex interaction play a central role in the TemRA differentiation of HIV-specific CD8(+) T cells in chronic HIV-1 infection. Clonotypes with a predominantly TemRA phenotype had a substantial fraction of cells without expression of CD57; and in contrast to the high clonotypic variability of TemRA differentiation, expression of CD57 was highly correlated among T-cell clonotypes within epitope-specific responses, indicating TCR-independent expression of CD57 in vivo. Our data highlight the importance of the structural composition of the TCR repertoire for the effector-memory differentiation of the immune response in chronic viral infections and suggest that TCR-dependent and -independent homeostasis shapes the pathogen-specific effector-memory repertoire in vivo.

Maternal transmission of human immunodeficiency virus escape mutations subverts HLA-B57 immunodominance but facilitates viral control in the haploidentical infant.

Expression of HLA-B57 is associated with restricted replication of human immunodeficiency virus (HIV), but the mechanism for its protective effect remains unknown. If this advantage depends upon CD8 T-cell recognition of B57-restricted epitopes, mother-to-child transmission of escape mutations within these epitopes could nullify its protective effect. However, if the B57 advantage is largely mediated by selection for fitness-attenuating viral mutations within B57-restricted epitopes, such as T242N in TW10-Gag, then the transmission of such mutations could facilitate viral control in the haploidentical infant. We assessed the consequences of B57-associated mutations on replication capacity, viral control, and clinical outcome after vertical transmission in 13 mother-child pairs. We found that expression of HLA-B57 was associated with exceptional control of HIV during infancy, even when mutations within TW10 and most other B57-restricted epitopes were transmitted, subverting the natural immunodominance of HLA-B57. In contrast, most B57-negative infants born to B57-positive mothers progressed rapidly to AIDS. The presence of T242N led to a reproducible reduction in viral fitness, as demonstrated by in vitro assays using NL4-3 constructs encoding p24 sequences from individual mothers and infants. Associated compensatory mutations within p24-Gag were observed to reverse this impairment and to influence the propensity of T242N to revert after transmission to B57-negative hosts. Moreover, primary failure to control viremia was observed in one infant to whom multiple compensatory mutations were transmitted along with T242N. These parallel in vivo and in vitro data suggest that HLA-B57 confers its advantage primarily by driving and maintaining a fitness-attenuating mutation in p24-Gag.

Fluctuations of functionally distinct CD8+ T-cell clonotypes demonstrate flexibility of the HIV-specific TCR repertoire.

T-cell receptor (TCR) diversity of virus-specific CD8+ T cells likely helps prevent escape mutations in chronic viral infections. To understand the dynamics of the virus-specific T cells in more detail, we followed the evolution of the TCR repertoire specific for a dominant HLA-B*08-restricted epitope in Nef (FLKEKGGL) in a cohort of subjects infected with HIV. Epitope-specific CD8+ T cells used structurally diverse TCR repertoires, with different TCRbeta variable regions and with high amino acid diversity within antigen recognition sites. In a longitudinal study, distinct Vbeta populations within the HIV-specific TCR repertoire expanded simultaneously with changes in plasma viremia, whereas other Vbeta populations remained stable or even decreased. Despite antigenic variation in some subjects, all subjects had the consensus sequence present during the study period. Functional analysis of distinct Vbeta populations revealed differences in HIV-specific IFN-gamma secretion ex vivo as well as differences in tetramer binding, indicating functional heterogeneity among these populations. This contrasts with findings in a subject on antiretroviral therapy with suppression of viremia to less than 50 copies/mL, where we observed long-term persistence of a single clonotype. Our findings illustrate the flexibility of a heterogeneous HIV-1-specific CD8+ TCR repertoire in subjects with partial control of viremia.