Finding the missing and unknown: Novel educational approaches to warming up cold cases.

In recent years, students in police academies and higher education institutions around the world have worked together to analyse cold cases including long-term missing persons cases in collaboration with investigators and prosecutors. In 2020, three European organisations, the Police Expert Network on Missing Persons (PEN-MP), AMBER Alert Europe and Locate International, succeeded in connecting these educational organisations enabling them to work collectively on cases and conduct cold case analyses (CCA) across international borders. The International Cold Case Analysis Project (ICCAP) learning objectives were to 1) collect the necessary information about the victim, 2) reconstruct the crime, and 3) investigate trace control. In a learning objective-based evaluation using Computer-Assisted Web Interviewing, 76 participating students from the German and International ICCAP teams were asked to complete a pre- and post-review questionnaire to self-assess their personal competence development. Participants reported significant increases in competence in all evaluated areas, thus demonstrating that authentic and relevant collaborations can enrich the learning environment, promote the use of professional skills, and provide significant knowledge exchange opportunities between academia and industry. Drawing on case studies of cold case missing persons' investigations and unidentified found remains, this article shares how university academics, students and community volunteers can work together nationally and internationally to find out what has happened to missing people and how we can more effectively identify the previously unidentified. In so doing, we share the expertise required to progress these cold cases and provide recommendations to support other institutions and organisations in adopting this innovative approach.

Innovation from the inside: Collaborating for school readiness.

This paper will critically reflect on a service evaluation project that was undertaken within Sussex Community NHS Foundation Trust in 2014/15. The project sought to provide a new way of working that supported health visitors (HVs) and school nurses (SNs) in developing effective collaborative and partnership working practices in order to meet the health needs and improve the health outcomes of children aged four to five years in preparation and readiness for school. HVs and SNs are well placed to work with families and provide the early interventions and health support required to support school readiness, e.g. behaviour, sleep, eating and continence advice. Historically, within Susssex Community NHS Foundation Trust, this public health approach has been taken on by the SN service. However, problems were identified locally with this model due to several factors including reduced staffing and confusion regarding transition of care from HVs to SNs. In response, a new way of working was considered locally to ensure the best possible service for families.

Integrating in silico and in vitro analysis of peptide binding affinity to HLA-Cw*0102: a bioinformatic approach to the prediction of new epitopes.

BACKGROUND: Predictive models of peptide-Major Histocompatibility Complex (MHC) binding affinity are important components of modern computational immunovaccinology. Here, we describe the development and deployment of a reliable peptide-binding prediction method for a previously poorly-characterized human MHC class I allele, HLA-Cw*0102. METHODOLOGY/FINDINGS: Using an in-house, flow cytometry-based MHC stabilization assay we generated novel peptide binding data, from which we derived a precise two-dimensional quantitative structure-activity relationship (2D-QSAR) binding model. This allowed us to explore the peptide specificity of HLA-Cw*0102 molecule in detail. We used this model to design peptides optimized for HLA-Cw*0102-binding. Experimental analysis showed these peptides to have high binding affinities for the HLA-Cw*0102 molecule. As a functional validation of our approach, we also predicted HLA-Cw*0102-binding peptides within the HIV-1 genome, identifying a set of potent binding peptides. The most affine of these binding peptides was subsequently determined to be an epitope recognized in a subset of HLA-Cw*0102-positive individuals chronically infected with HIV-1. CONCLUSIONS/SIGNIFICANCE: A functionally-validated in silico-in vitro approach to the reliable and efficient prediction of peptide binding to a previously uncharacterized human MHC allele HLA-Cw*0102 was developed. This technique is generally applicable to all T cell epitope identification problems in immunology and vaccinology.

Is 1 alanine transaminase >200 IU enough to define an alanine transaminase flare in HIV-infected populations? A new definition derived from a large cohort study.

OBJECTIVES: Recent studies have suggested that highly active antiretroviral therapy may lead to rises in alanine transaminase (ALT) among HIV-infected patients. However, the definition of an ALT flare is arbitrary and the extent to which such increases represent normal fluctuations has not been explored. METHODS: Using data from untreated, hepatitis B virus/hepatitis C virus-negative, HIV-infected patients, we derived a definition for an ALT flare by exploring a series of ALT thresholds (from 100 to 200 IU/L). The resulting definition (2 consecutive ALTs > 200 measured >2 weeks apart) was applied to all patients in the UK Collaborative HIV Cohort (CHIC) Study, and Poisson regression was used to identify factors associated with ALT flares. RESULTS: Five hundred and twenty six of 12,206 eligible patients (4.3%) had > or =1 ALT flare, resulting in a total of 615 episodes of ALT flares. The overall rate of an ALT flare was 1.19 (95% confidence interval: 1.10 to 1.28) per 100 person-years. Higher risk of ALT flare was associated with lower CD4 counts, detectable viral loads, being under follow-up in earlier calendar years, prior clinical AIDS, receipt of nevirapine either with didanosine/stavudine or without didanosine/stavudine, receipt of ritonavir, detectable anti-hepatitis C virus, and detectable hepatitis B surface antigen. CONCLUSIONS: Associations between known risk factors may be under/over estimated if using single values, that is, 1 ALT > 200, to define ALT flares. We recommend studies to use a more stringent measure and suggest our derived definition of an ALT flare.

Kinetics of expansion of epitope-specific T cell responses during primary HIV-1 infection.

Multiple lines of evidence support a role for CD8(+) T cells in control of acute/early HIV replication; however, features of the primary HIV-specific CD8(+) T cell response that may impact on the efficiency of containment of early viral replication remain poorly defined. In this study, we performed a novel, comprehensive analysis of the kinetics of expansion of components of the HIV-specific CD8(+) T cell response in 21 acutely infected individuals. Epitope-specific T cell responses expanded asynchronously during primary infection in all subjects. The most rapidly expanded responses peaked as early as 5 days following symptomatic presentation and were typically of very limited epitope breadth. Responses of additional specificities expanded and contracted in subsequent waves, resulting in successive shifts in the epitope immunodominance hierarchy over time. Sequence variation and escape were temporally associated with the decline in magnitude of only a subset of T cell responses, suggesting that other factors such as Ag load and T cell exhaustion may play a role in driving the contraction of HIV-specific T cell responses. These observations document the preferential expansion of CD8(+) T cells recognizing a subset of epitopes during the viral burst in acute HIV-1 infection and suggest that the nature of the initial, very rapidly expanded T cell response may influence the efficiency with which viral replication is contained in acute/early HIV infection.

A prospective study of delayed sleep phase syndrome in patients with severe resistant obsessive-compulsive disorder.

There have been relatively few studies examining sleep in patients with obsessive-compulsive disorder (OCD) and these have produced contradictory findings. A recent retrospective study identified a possible association between OCD and a circadian rhythm sleep disorder known as delayed sleep phase syndrome (DSPS). Patients with this pattern of sleeping go to bed and get up much later than normal. They are unable to shift their sleep to an earlier time and, as a result, suffer considerable disruption to social and occupational functioning. In this study, we examined the sleep of patients with OCD prospectively. We aimed to establish the frequency of DSPS in this population and any associated clinical or demographic factors which might be implicated in its aetiology.

New assessment tools that measure sleep vital signs: the SleepMed Insomnia Index and the Sleep Matrix.

Insomnia is the leading sleep disorder in the US; however, diagnosis is often problematic. This pilot study assessed the clinical value of a novel diagnostic insomnia questionnaire. The SleepMed Insomnia Index (SMI) was administered to 543 consecutive patients and 50 normal control subjects during a pilot study. Mean SMI scores were assessed based on subsequent sleep-related diagnoses. The SMI scores for patients with sleep-related disorders were significantly higher than those for the control group (p < 0.001) and highest for the 90 patients comprising the insomnia group. Analysis of the SMI scores from the 90 insomnia patients indicates a high degree of reliability (Cronbach's alpha: 0.7). These data support our clinical experience with this diagnostic tool which indicates a strong likelihood of disrupted nighttime sleep in patients with high SMI scores. Following further validation, the SMI may prove to be a valuable tool for evaluating sleep disorders, specifically as an aid in the diagnosis of insomnia. The Sleep Matrix is a visual tool that quantifies a sleep complaint by combining scores from the Epworth Sleepiness Scale (ESS) and the SMI. The SMI measures an insomnia component while the ESS is an accepted measure of daytime sleepiness. The Sleep Matrix visually displays the complexity of the sleep complaint in an effort to differentiate insomnia with differing etiologies from other sleep disorders and measure treatment outcomes. To pilot test the Sleep Matrix, the tool was administered to 90 patients with insomnia and to 22 normal controls. Plots from the insomnia patients were concentrated into the "insomnia zone" while scores from the normal controls were located in the "normal zone" located in the lower left quadrant. Additional research using the Sleep Matrix could provide data that the tool could be utilized to visually aid the clinician in the diagnosis of unknown sleep complaints.

HIV-1 epitope-specific CD8+ T cell responses strongly associated with delayed disease progression cross-recognize epitope variants efficiently.

The ability of HIV-1-specific CD8(+) T cell responses to recognize epitope variants resulting from viral sequence variation in vivo may affect the ease with which HIV-1 can escape T cell control and impact on the rate of disease progression in HIV-1-infected humans. Here, we studied the functional cross-reactivity of CD8 responses to HIV-1 epitopes restricted by HLA class I alleles associated with differential prognosis of infection. We show that the epitope-specific responses exhibiting the most efficient cross-recognition of amino acid-substituted variants were those strongly associated with delayed progression to disease. Not all epitopes restricted by the same HLA class I allele showed similar variant cross-recognition efficiency, consistent with the hypothesis that the reported associations between particular HLA class I alleles and rate of disease progression may be due to the quality of responses to certain "critical" epitopes. Irrespective of their efficiency of functional cross-recognition, CD8(+) T cells of all HIV-1 epitope specificities examined showed focused TCR usage. Furthermore, interpatient variability in variant cross-reactivity correlated well with use of different dominant TCR Vbeta families, suggesting that flexibility is not conferred by the overall clonal breadth of the response but instead by properties of the dominant TCR(s) used for epitope recognition. A better understanding of the features of T cell responses associated with long-term control of viral replication should facilitate rational vaccine design.