A highly reproducible quantitative viral outgrowth assay for the measurement of the replication-competent latent HIV-1 reservoir

Cure of Human Immunodeficiency Virus (HIV) infection remains elusive due to the persistence of HIV in a latent reservoir. Strategies to eradicate latent infection can only be evaluated with robust, sensitive and specific assays to quantitate reactivatable latent virus. We have taken the standard peripheral blood mononuclear cell (PBMC) based viral outgrowth methodology and from it created a logistically simpler and more highly reproducible assay to quantify replication-competent latent HIV in resting CD4$^{+}$ T cells, both increasing accuracy and decreasing cost and labour. Purification of resting CD4$^{+}$ T cells from whole PBMC is expedited and achieved in 3 hours, less than half the time of conventional protocols. Our indicator cell line, SupT1-CCR5 cells (a clonal cell line expressing CD4, CXCR4 and CCR5) provides a readily available standardised readout. Reproducibility compares favourably to other published assays but with reduced cost, labour and assay heterogeneity without compromising sensitivity.This work was submitted on behalf ofthe UK Biomedical Research Centres CHERUB cooperative and was supported by the National Institutes for Health Research (NIHR, Cambridge Biomedical Research Centres), Medical Research Council (MR/N02043X/1) and the Academy of Medical Sciences

A unified model for age-velocity dispersion relations in Local Group galaxies: Disentangling ISM turbulence and latent dynamical heating

We analyze age-velocity dispersion relations (AVRs) from kinematics of individual stars in eight Local Group galaxies ranging in mass from Carina ($M_{*} \sim 10^{6}$) to M31 ($M_{*} \sim 10^{11}$). Observationally the $\sigma$ vs. stellar age trends can be interpreted as dynamical heating of the stars by GMCs, bars/spiral arms, or merging subhalos; alternatively the stars could have simply been born out of a more turbulent ISM at high redshift and retain that larger velocity dispersion till present day - consistent with recent IFU studies. To ascertain the dominant mechanism and better understand the impact of instabilities and feedback, we develop models based on observed SFHs of these Local Group galaxies in order to create an evolutionary formalism which describes the ISM velocity dispersion due to a galaxy's evolving gas fraction. These empirical models relax the common assumption that the stars are born from gas which has constant velocity dispersion at all redshifts. Using only the observed SFHs as input, the ISM velocity dispersion and a mid-plane scattering model fits the observed AVRs of low mass galaxies without fine tuning. Higher mass galaxies above $M_{vir} > 10^{11}$ need a larger contribution from latent dynamical heating processes (for example minor mergers), in excess of the ISM model. Using the SFHs we also find that supernovae feedback does not appear to be a dominant driver of the gas velocity dispersion compared to gravitational instabilities - at least for dispersions $\sigma \gtrsim 25$ km/s. Together our results point to stars being born with a velocity dispersion close to that of the gas at the time of their formation, with latent dynamical heating operating with a galaxy mass-dependent efficiency. These semi-empirical relations may help constrain the efficiency of feedback and its impact on the physics of disk settling in galaxy formation simulations

On the increase in thermal diffusivity caused by the perovskite to post-perovskite phase transition and its implications for mantle dynamics

The thermal diffusivity (κ) of perovskite and post-perovskite CaIrO3 has been measured, at elevated pressure and temperatures up to 600 °C, using the X-radiographic Ångström method. At high temperatures we find that the thermal diffusivity of post-perovskite is slightly below twice that of isochemical perovskite over the temperature range investigated. Assuming a similar effect occurs in MgSiO3 post-perovskite, the effect of the contrasting thermal transport properties between perovskite and post-perovskite on mantle dynamics has been investigated using simple two-dimensional convection models. These show a reduction in extent and increase in depth of post-perovskite lenses, as well as increased core–mantle-boundary heat-flux, broader upwellings and more vigorous downwellings when compared to the reference, constant a, case

Magnetic structure and field-dependent magnetic phase diagram of Ni2In-type PrCuSi

The magnetic structure of the ternary equiatomic intermetallic compound PrCuSi is investigated using neutron powder diffraction experiments in 0 T as well as in external magnetic fields up to 2 T. The PrCuSi compound crystallizes in the hexagonal Ni2In-type structure, in the space group P63/mmc. In this structure, cationic ordering of Cu and Si takes place. The antiferromagnetic phase transition in the Pr sublattice takes place at K in 0 T. Under an external magnetic field of 2 T, a field-induced ferromagnetic phase is observed. Magnetoelastic coupling is evidenced by an increase in the unit cell volume. Clear signatures of a mixed antiferromagnetic and ferromagnetic phase in weak, intermediate fields, 0.4–0.8 T, are obtained from the present study. Using the present set of experimental data, we construct the H  −  T phase diagram of PrCuSi

‘Do i care?’ young adults' recalled experiences of early adolescent overweight and obesity: a qualitative study

<p>Objective: Individual behaviour change to reduce obesity requires awareness of, and concern about, weight. This paper therefore describes how young adults, known to have been overweight or obese during early adolescence, recalled early adolescent weight-related awareness and concerns. Associations between recalled concerns and weight-, health- and peer-related survey responses collected during adolescence are also examined.</p> <p>Design: Qualitative semi-structured interviews with young adults; data compared with responses to self-report questionnaires obtained in adolescence.</p> <p>Participants: A total of 35 participants, purposively sub-sampled at age 24 from a longitudinal study of a school year cohort, previously surveyed at ages 11, 13 and 15. Physical measures during previous surveys allowed identification of participants with a body mass index (BMI) indicative of overweight or obesity (based on British 1990 growth reference) during early adolescence. Overall, 26 had been obese, of whom 11 had BMI99.6th centile, whereas 9 had been overweight (BMI=95th–97.9th centile).</p> <p>Measures: Qualitative interview responses describing teenage life, with prompts for school-, social- and health-related concerns. Early adolescent self-report questionnaire data on weight-worries, self-esteem, friends and victimisation (closed questions).</p> <p>Results: Most, but not all recalled having been aware of their overweight. None referred to themselves as having been obese. None recalled weight-related health worries. Recollection of early adolescent obesity varied from major concerns impacting on much of an individual's life to almost no concern, with little relation to actual severity of overweight. Recalled concerns were not clearly patterned by gender, but young adult males recalling concerns had previously reported more worries about weight, lower self-esteem, fewer friends and more victimisation in early adolescence; no such pattern was seen among females. Conclusion: The popular image of the unhappy overweight teenager was not borne out. Many obese adolescents, although well aware of their overweight recalled neither major dissatisfaction nor concern. Weight-reduction behaviours are unlikely in such circumstances.</p&gt

Cytotoxic polyfunctionality maturation of cytomegalovirus-pp65-specific CD4 + and CD8 + T-cell responses in older adults positively correlates with response size

Cytomegalovirus (CMV) infection is one of the most common persistent viral infections in humans worldwide and is epidemiologically associated with many adverse health consequences during aging. Previous studies yielded conflicting results regarding whether large, CMV-specific T-cell expansions maintain their function during human aging. In the current study, we examined the in vitro CMV-pp65-reactive T-cell response by comprehensively studying five effector functions (i.e., interleukin-2, tumor necrosis factor-α, interferon-γ, perforin, and CD107a expression) in 76 seropositive individuals aged 70 years or older. Two data-driven, polyfunctionality panels (IL-2-associated and cytotoxicity-associated) derived from effector function co-expression patterns were used to analyze the results. We found that, CMV-pp65-reactive CD8 + and CD4 + T cells contained similar polyfunctional subsets, and the level of polyfunctionality was related to the size of antigen-specific response. In both CD8 + and CD4 + cells, polyfunctional cells with high cytotoxic potential accounted for a larger proportion of the total response as the total response size increased. Notably, a higher serum CMV-IgG level was positively associated with a larger T-cell response size and a higher level of cytotoxic polyfunctionality. These findings indicate that CMV-pp65-specific CD4 + and CD8 + T cell undergo simultaneous cytotoxic polyfunctionality maturation during aging

Treatment of Highly Drug-Resistant Pulmonary Tuberculosis

BACKGROUND Patients with highly drug-resistant forms of tuberculosis have limited treatment options and historically have had poor outcomes. METHODS In an open-label, single-group study in which follow-up is ongoing at three South African sites, we investigated treatment with three oral drugs — bedaquiline, pretomanid, and linezolid — that have bactericidal activity against tuberculosis and to which there is little preexisting resistance. We evaluated the safety and efficacy of the drug combination for 26 weeks in patients with extensively drug-resistant tuberculosis and patients with multidrug-resistant tuberculosis that was not responsive to treatment or for which a second-line regimen had been discontinued because of side effects. The primary end point was the incidence of an unfavorable outcome, defined as treatment failure (bacteriologic or clinical) or relapse during follow-up, which continued until 6 months after the end of treatment. Patients were classified as having a favorable outcome at 6 months if they had resolution of clinical disease, a negative culture status, and had not already been classified as having had an unfavorable outcome. Other efficacy end points and safety were also evaluated. RESULTS A total of 109 patients were enrolled in the study and were included in the evaluation of efficacy and safety end points. At 6 months after the end of treatment in the intention-to-treat analysis, 11 patients (10%) had an unfavorable outcome and 98 patients (90%; 95% confidence interval, 83 to 95) had a favorable outcome. The 11 unfavorable outcomes were 7 deaths (6 during treatment and 1 from an unknown cause during follow-up), 1 withdrawal of consent during treatment, 2 relapses during follow-up, and 1 loss to follow-up. The expected linezolid toxic effects of peripheral neuropathy (occurring in 81% of patients) and myelosuppression (48%), although common, were manageable, often leading to dose reductions or interruptions in treatment with linezolid. CONCLUSIONS The combination of bedaquiline, pretomanid, and linezolid led to a favorable outcome at 6 months after the end of therapy in a high percentage of patients with highly drug-resistant forms of tuberculosis; some associated toxic effects were observed. (Funded by the TB Alliance and others; ClinicalTrials.gov number, NCT02333799. opens in new tab.

Arduous implementation: Does the Normalisation Process Model explain why it's so difficult to embed decision support technologies for patients in routine clinical practice

Background: decision support technologies (DSTs, also known as decision aids) help patients and professionals take part in collaborative decision-making processes. Trials have shown favorable impacts on patient knowledge, satisfaction, decisional conflict and confidence. However, they have not become routinely embedded in health care settings. Few studies have approached this issue using a theoretical framework. We explained problems of implementing DSTs using the Normalization Process Model, a conceptual model that focuses attention on how complex interventions become routinely embedded in practice.Methods: the Normalization Process Model was used as the basis of conceptual analysis of the outcomes of previous primary research and reviews. Using a virtual working environment we applied the model and its main concepts to examine: the 'workability' of DSTs in professional-patient interactions; how DSTs affect knowledge relations between their users; how DSTs impact on users' skills and performance; and the impact of DSTs on the allocation of organizational resources.Results: conceptual analysis using the Normalization Process Model provided insight on implementation problems for DSTs in routine settings. Current research focuses mainly on the interactional workability of these technologies, but factors related to divisions of labor and health care, and the organizational contexts in which DSTs are used, are poorly described and understood.Conclusion: the model successfully provided a framework for helping to identify factors that promote and inhibit the implementation of DSTs in healthcare and gave us insights into factors influencing the introduction of new technologies into contexts where negotiations are characterized by asymmetries of power and knowledge. Future research and development on the deployment of DSTs needs to take a more holistic approach and give emphasis to the structural conditions and social norms in which these technologies are enacte