Search

Human cytomegalovirus (HCMV) is carried lifelong by ∼80 % of adults worldwide, generating distinct disease syndromes in transplant recipients, people with HIV (PWH) and neonates. Amino acids 15-23 encoded by the HCMV gene UL40 match positions 3-11 of HLA-A and HLA-C, and constitute a "signal peptide" able to stabilise cell surface HLA-E as a restriction element and a ligand of NKG2A and NKG2C. We present next generation sequencing of UL40 amplified from 15 Australian renal transplant recipients (RTR), six healthy adults and four neonates, and 21 Indonesian PWH. We found no groupwise associations between the presence of multiple sequences and HCMV burden (highest in PWH) or HCMV-associated symptoms in neonates. Homology between UL40 and corresponding HLA-C and HLA-A peptides in 11 RTR revealed perfect matches with HLA-C in three individuals, all carrying HCMV encoding only VMAPRTLIL - a peptide previously associated with viremia. However indices of the burden of HCMV did not segregate in our cohort.

Around 80% of adults worldwide carry human cytomegaloviris (HCMV). The HCMV gene UL18 is a homolog of HLA class I genes and encodes a protein with high affinity for the NK and T-cell cytotoxicity inhibitor LIR-1. UL18 was deep sequenced from blood, saliva or urine from Indonesian people with HIV (PWH) (n = 28), Australian renal transplant recipients (RTR) (n = 21), healthy adults (n = 7) and neonates (n = 4). 95% of samples contained more than one variant of HCMV UL18, as defined by carriage of nonsynonymous variations. When aligned with immunological markers of the host’s burden of HCMV, the S318N variation associated with high levels of antibody reactive with HCMV lysate in PWH over 12 months on antiretroviral therapy. The A107T variation associated with HCMV antibody levels and inflammatory biomarkers in PWH at early timepoints. Variants D32G, D248N, V250A and E252D aligned with elevated HCMV antibody levels in RTR, while M191K, E196Q and F165L were associated with HCMV-reactive T-cells and proportions of Vδ2− γδ T-cells—populations linked with high burdens of HCMV. We conclude that UL18 is a highly variable gene, where variation may alter the persistent burden of HCMV and/or the host response to that burden.

Around 80% of adults worldwide carry human cytomegaloviris (HCMV). The HCMV gene

Cytomegalovirus (CMV) has been implicated in vascular pathologies and may warrant inclusion in cardiovascular predictive algorithms. We addressed this in healthy older adults and renal transplant recipients (RTR) as they retain a high burden of CMV. RTR (n = 45) stable more than 2 years after transplantation and 58 age-matched healthy adults were assessed. Plasma inflammatory biomarkers (soluble isoform of the interferon-β receptor [sIFNAR2], soluble tumour necrosis factorreceptor-1 [sTNFR1], soluble cluster of differentiation 14 [sCD14], C reactive protein, P-selectin, intracellular cell adhesion molecule-1, vascular cell adhesion molecule-1), and measures of CMV burden (antibodies, saliva CMV DNA, and interferon γ responses to CMV) were assessed in 2014 and evaluated in 2017 as predictors of vascular health-defined using flow-mediated dilatation (FMD), pulse wave velocity (PWV), and augmentation indices (Aix@ 75). Linear regression models adjusted for age, sex, and body mass index (BMI) were optimized to identify risk factors. In 2017, RTR had inferior vascular health marked by impaired FMD and PWV. Detectable CMV DNA (P = .02) was associated with impaired FMD, whilst CMV glycoprotein B (gB) antibody attenuated this effect (P = .03) (adjusted R2 = .42). In healthy adults, the optimal model for predicting FMD (R2 =.22) incorporated high P-selectin (P = .03) and low ICAM-1 (P = .03) levels with no significant impact of CMV. Elevated sIFNAR2 (P = .04) and gB antibody (P = .06) levels predicted increasing Aix@ 75 (poor vascular health) in healthy adults (R2 = .4), whilst optimal models for RTR (R2 = .37) linked low sIFNAR2 and CMV IE-1 antibody levels with lower Aix@ 75 (better vascular health). CMV IE-1 antibody was also protective in relation to PWV in healthy adults (R2 = .55). Overall, measures of active CMV replication were more predictive of impaired FMD in RTR than standard biomarkers, but increased CMV gB antibodies may be protective.

Human cytomegalovirus (HCMV) is a beta-herpesvirus carried by ~80% of adults worldwide. Acute infections are often asymptomatic in healthy individuals but generate diverse syndromes in neonates, renal transplant recipients (RTR), and people with HIV (PWH). The HCMV gene UL111a encodes a homolog of human interleukin-10 (IL-10) that interacts with the human IL-10 receptor. Deep sequencing technologies were used to sequence UL111a directly from 59 clinical samples from Indonesian PWH and Australian RTR, healthy adults, and neonates. Overall, 93% of samples contained more than one variant of HCMV, as defined by at least one nonsynonymous variation. Carriage of these variants differed between neonates and adults, Australians and Indonesians, and between saliva and blood leukocytes. The variant alleles of N41D and S71Y occurred together in Australian RTR and were associated with higher T-cell responses to HCMV pp65. The variant P122S was associated with lower levels of antibodies reactive with a lysate of HCMV-infected fibroblasts. L174F was associated with increased levels of antibodies reactive with HCMV lysate, immediate-early 1 (IE-1), and glycoprotein B (gB) in Australian RTR and Indonesians PWH, suggesting a higher viral burden. We conclude that variants of UL111a are common in all populations and may influence systemic responses to HCMV.

Human cytomegalovirus (HCMV) is a beta-herpesvirus carried by ∼80% of the world’s population. Acute infections are asymptomatic in healthy individuals but generate diverse syndromes in neonates, solid organ transplant recipients, and HIV-infected individuals. The HCMV gene US28 encodes a homolog of a human chemokine receptor that is able to bind several chemokines and HIV gp120. Deep sequencing technologies were used to sequence US28 directly from 60 clinical samples from Indonesian HIV patients and Australian renal transplant recipients, healthy adults, and neonates. Molecular modeling approaches were used to predict whether nine nonsynonymous mutations in US28 may alter protein binding to a panel of six chemokines and two variants of HIV gp120. Ninety-two percent of samples contained more than one variant of HCMV, as defined by at least one nonsynonymous mutation. Carriage of these variants differed between neonates and adults, Australian and Indonesian samples, and saliva samples and blood leukocytes. Two nonsynonymous mutations (N170D and R267K) were associated with increased levels of immediate early protein 1 (IE-1) and glycoprotein B (gB) HCMV-reactive antibodies, suggesting a higher viral burden. Seven of the nine mutations were predicted to alter binding of at least one ligand. Overall, HCMV variants are common in all populations and have the potential to affect US28 interactions with human chemokines and/or gp120 and alter responses to the virus. The findings relied on deep sequencing technologies applied directly to clinical samples, so the variants exist in vivo. IMPORTANCE Human cytomegalovirus (HCMV) is a common viral pathogen of solid organ transplant recipients, neonates, and HIV-infected individuals. HCMV encodes homologs of several host genes with the potential to influence viral persistence and/or pathogenesis. Here, we present deep sequencing of an HCMV chemokine receptor homolog, US28, acquired directly from clinical specimens. Carriage of these variants differed between patient groups and was associated with different levels of circulating HCMV-reactive antibodies. These features are consistent with a role for US28 in HCMV persistence and pathogenesis. This was supported by in silico analyses of the variant sequences demonstrating altered ligand-binding profiles. The data delineate a novel approach to understanding the pathogenesis of HCMV and may impact the development of an effective vaccine.

The majority of adults in the world (around 83%) carry antibodies reactive with HCMV and are thought to retain inactive or latent infections lifelong. The virus is transmitted via saliva, so infection events are likely to be common. Indeed, it is hard to imagine a life without exposure to HCMV. From 45 seronegative individuals (13 renal transplant recipients, 32 healthy adults), we present seven cases who had detectable HCMV DNA in their blood and/or saliva, or a CMV-encoded homologue of IL-10 (vIL-10) in their plasma. One case displayed NK cells characteristic of CMV infection before her HCMV DNA became undetectable. In other cases, the infection may persist with seroconversion blocked by vIL-10. Future research should seek mechanisms that can prevent an individual from seroconverting despite a persistent HCMV infection, as HCMV vaccines may not work well in such people.

Human cytomegalovirus (HCMV) infections are common following renal transplantation and may have long-lasting effects. HCMV can be measured directly by viral DNA or indirectly via host immune responses. HCMV-encoded microRNA (miRNA) may alter the pathobiology of HCMV infections and contribute to the progression of HCMV disease. HCMV-encoded miRNAs can be detected in blood but have not been sought in saliva. We investigated saliva samples from 32 renal transplant recipients (RTR) and 12 seropositive healthy controls for whom immunological data was available. Five HCMV-encoded miRNAs (miR-UL112-5p, miR-US5-2-3p, miR-UL36, miR-US25-2-3p and miR-UL22A) were sought using primer probe assays. HCMV miRNA species were detected in saliva from 15 RTR and 3 healthy controls, with miR-US5-2-3p most commonly detected. The presence of HCMV miRNAs associated with increased T-cell responses to HCMV IE-1 in RTR, suggesting a link with frequent reactivations of HCMV.

Background Cytomegalovirus (CMV) has been implicated in accelerated cardiovascular changes and may warrant inclusion in predictive algorithms. This is addressed here in healthy older adults and in renal transplant recipients (RTR) stable on therapy as they retain a high burden of CMV. Methods RTR (n=45) stable >2 years after transplantation and 58 age-matched healthy adults were recruited in 2014 and returned in 2017. Venous blood samples and saliva were collected, frozen and stored. Plasma proteins linked with inflammation [soluble interferon a receptor 2 (sIFNaR2), sTNFR1, sCD14, C-reactive protein (CRP)], vasculopathy [p-selectin, intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1)], and metrics of CMV burden [antibodies reactive with CMV lysate, gB or IE-1 (by ELISA), CMV DNA in saliva (by PCR) and T-cell IFNg responses to CMV antigens (by ELISpot)] were assessed in 2014. In 2017, vascular endpoint measurements included brachial artery flow mediated dilatation (FMD), pulse wave analysis [augmented index corrected to heart rate of 75 beats per minute (Aix@75)] and pulse wave velocity (PWV). Results In 2017, RTR had lower FMD % (p Conclusions Overall measures of a high burden of CMV, predicted a low FMD marking poor peripheral vascular health in RTR several years after assessment, with more potency than was achieved with plasma markers of systemic and vascular inflammation, which was observed in healthy controls. CMV antibodies can predict future measures of arterial stiffness such as Aix@75 and PWV in healthy controls, but need further investigation in larger cohorts.

Cytomegalovirus (CMV) is a highly prevalent virus and a common cause of morbidity in solid organ transplant patients. It is also known for its long-lasting imprint on the immune system, expanding populations of highly differentiated T cells and natural killer (NK) cells with novel phenotypes. However, it is unclear whether these cells mark success or failure in the management of an active infection. We assessed CMV reactivation in 54 renal transplant recipients (RTRs) by measuring CMV DNA in plasma samples. Function and phenotype of T cells and NK cells were then assessed in seven RTR with detectable CMV DNA. The patient with highest CMV viral load (P1) displayed increased NK cell function and abundant highly differentiated T cells. We compare P1 with the other six patients and review possible scenarios of cross-regulation between NK cells and T cells.

Human cytomegalovirus (CMV) is often transmitted through saliva. The salivary gland is a site of CMV replication and saliva can be used to diagnose congenital CMV infections. CMV replication is monitored in whole blood or plasma in renal transplant recipients (RTR) and associates with clinical disease. However, these assays may not detect replication in the salivary gland and there is little data linking detection in saliva with systemic infection and clinical sequelae. RTR (n = 82) were recruited &gt, 2 years after transplantation. An in-house quantitative PCR assay was used to detect CMV UL54 in saliva samples. CMV DNA was sought in plasma using a commercial assay. Vascular health was predicted using flow mediated dilatation (FMD) and plasma biomarkers. CMV-reactive antibodies were quantified by ELISA and circulating CMV-specific T-cells by an interferon-&gamma, ELISpot assay. V&delta, 2&minus, &gamma, &delta, T-cells were detected using multicolor flow cytometry reflecting population expansion after CMV infection. The presence of CMV DNA in saliva and plasma associated with plasma levels of antibodies reactive with CMV gB and with populations of circulating V&delta, T -cells (p &lt, 0.01). T-cells reactive to CMV immediate early (IE)-1 protein were generally lower in patients with CMV DNA in saliva or plasma, but the level of significance varied (p = 0.02&ndash, 0.16). Additionally, CMV DNA in saliva or plasma associated weakly with impaired FMD (p = 0.06&ndash, 0.09). The data suggest that CMV detected in saliva reflects systemic infections in adult RTR.

Alzheimer's disease (AD) is an incurable neurodegenerative condition resulting in progressive cognitive decline. Pathological features include Aβ plaques, neurofibrillary tangles, neuroinflammation and neuronal death. Purinergic receptors 7 and 4 (P2X7R and P2X4R) and calcium/calmodulin-dependent kinase kinase 2 (CaMKK2) are implicated in neuronal death. We used immunohistochemistry to investigate the distribution of these proteins in neurones from frontal cortex of donors (n = 3/group; aged 79–83 years) who died with and without AD. Neurones were identified morphologically and immunoperoxidase staining was achieved using commercial antibodies. Immunoreactive neurones were counted for each protein by 2–3 raters blinded to the diagnoses. We observed no differences in percentages of P2X7R, P2X4R or CaMKK2 positive neurones (p = 0.2–0.99), but sections from individuals with AD had marginally fewer neurones (p = 0.10). Hence P2X7R, P2X4R or CaMKK2 appear to be expressed in neurones from older donors, but expression does not associate with AD.

Altered T cell profiles have been linked with metrics of persistent cytomegalovirus (CMV) infections in healthy aging and older HIV patients stable on antiretroviral therapy (ART). In this study, we use CMV DNA to identify active infections, and levels of CMV-reactive antibody to assess the persistent burden of CMV in a longitudinal study of 78 young adult patients beginning ART in Jakarta, Indonesia, with200 CD4 T cells/μL. CMV antibodies, inflammatory markers (C-reactive protein [CRP], soluble interferon-α/β receptor) and T cell phenotypes were assessed before ART (V0) and after 1, 3, 6, and 12 months (V1-V12). CMV DNA was detected in 41 patients (52%) at V0, irrespective of CD4 T cell counts, gender, age, or plasma HIV RNA. CMV DNA+ patients had higher levels of antibody reactive with CMV Immediate Early 1 (IE-1) at V0 and V12 (p = 0.04), and with CMV lysate at V12 (p = 0.01). Detectable CMV DNA did not align with inflammatory markers, but associated with lower CD4/CD8 ratios until V3. CMV antibody levels correlated inversely with proportions of naive CD4 and CD8 T cells, and directly with proportions of CD57

Renal transplant recipients (RTR) display high burdens of cytomegalovirus (CMV) and accelerated cardiovascular change. NK cells can control CMV and may contribute to vascular pathologies. Polymorphisms in genes encoding the inhibitory receptor LILRB1 and its ligand HLA-G, and the activating receptor NKG2C may illuminate the role of NK cells in vascular health and CMV immunity. We assessed 81 healthy adults and 82 RTR >2 years after transplantation. RTR had higher humoral and T-cell responses to CMV, and impaired vascular health. A 14bp indel in HLA-G associated with increased flow-mediated dilatation of the brachial artery. The T allele of LILRB1 rs1061680 associated with increased carotid intimal media thickness (cIMT) in RTR and controls. A 16 kb deletion encompassing the NKG2C gene associated with lower cIMT values and higher humoral and T-cell responses to CMV. Hence all polymorphisms tested had small but discernable effects on vascular health. The NKG2C deletion may act via CMV.

A set of cases concerning child injury is described in which there are several human factors/ergonomics (HFE) issues. Each panelist describes an injury or death of a child with a brief overview of the events that occurred. Major HFE issues are presented and discussed using the framework of the hazard-control hierarchy of designing out, guarding against, and warning about hazards. Consideration is not only given to children but also caretakers in the design of useable and safe products. A secondary purpose of the panel is to discuss interest in forming a special interest or technical group on children’s HFE issues.

Infant sling carriers are hands-free, fabric carrying devices used to secure an upright or reclined infant to a caregiver. With traditional, unstructured slings, safety depends solely on the wearer’s use and knowledge of how to wrap an infant onto the caregiver, since safety is not passively designed into the product. In recent years, semi-structured slings with seemingly simple instructions began being marketed to make slings easier to use. However, infants younger than 4 months of age are suffocating in both unstructured and semi-structured slings. This paper describes research undertaken to identify factors contributing to suffocation, explains why efforts to improve product usability instead created new suffocation hazards, why better warnings and instructions fail to adequately address these hazards, and thus why the government and industry should adopt more effective options to address this deadly hazard.

This alternative format session synthesizes the experiences of five, diverse, forensic human factors practitioners who have actively participated in widely varying voluntary committees for standards and codes and found similar benefits of performing this work. Although such work is typically non-compensated, actively participating on voluntary standards committees can benefit society by improving safety standards and codes and simultaneously benefit the forensic human factors professional through contacts and networking, increased knowledge, technical expertise, and professional credibility. A strong need exists for more human factors input on voluntary committees. It is the authors’ hope that sharing their individual experiences will persuade other forensic human factors practitioners to similarly join in this mutually beneficial volunteer work.

While voluntary standards can improve the safety of consumer products by setting minimum safety requirements, such standards are rarely all encompassing and should not be relied upon as the sole criteria for product safety. A product can meet the technical requirements of a voluntary standard yet still pose serious hazards. This paper explores the limitations of voluntary standards through specific examples and recommends additional actions and resources to conduct a thorough product safety evaluation.

A research project to evaluate and improve warnings concerning unexpected missiling of carbonated beverage caps is described. The methodologies employed, which have wider application, included the use of commonly accepted evaluative criteria, observation of bottle opening behavior, directed interviews, evaluative feedback from interviewees, iterative testing of modified labeling, and symbol comprehension testing.

Welfare reform and related changes have the potential to significantly affect states' financing of child welfare services both directly and indirectly. While the actual effects of these changes are not yet known, there have been no baseline data on states financing practices upon which to measure change. This article seeks to fill this data gap, reporting the results of a survey that collected information from state child welfare administrators on the amount states spend on child welfare services, the sources of these funds, and the amount expended on different types of services. Total spending in State Fiscal Year 1996 exceeded $14.4 billion. The majority of both federal and state child welfare funds supported children in out-of-home care. States relied heavily on federal Medicaid, Emergency Assistance, and Social Services Block Grant funds to pay for child welfare services though states were found to vary significantly in their financing practices. Implications of these findings for welfare reform implementation are discussed.

This paper presents the results of two studies which comprehension tested safety symbols in published ANSI and ISO voluntary standards. Study 1 tested safety symbols from the ANSI Z535.3 Criteria for Safety Symbols standard. Study 2 tested additional safety symbols from the ANSI Z535.3 standard as well as additional symbols from the ISO 7010 safety symbols standard. Overall, comprehension of the symbols in both studies was relatively poor. Only two of the twelve symbols tested in Study 1 and five of the fourteen symbols tested in Study 2 met ANSI Z535.3 acceptance criteria. Adding red to hazard alerting symbols to depict heat improved comprehension, but adding color and determinants to information symbols did not. Implications of these results for future revisions of the ANSI Z535.3 standard are discussed.

The 1997 revision to the ANSI Z535.3 Criteria for Safety Symbols standard refined and expanded the structure of the original 1991 version. The revision contains improved symbol creation guidelines, comprehension test procedures, symbol surround shape and color guidance, and symbol examples. This paper explains the revisions of interest, the role of human factors analyses in driving the standard's evolution, and the reasons why symbols created and tested according to the revised criteria are expected to be more effective in terms of consistency, legibility, and most importantly, comprehension. Research areas for future revisions are also identified.

Improvements in the 1997 revision of the ANSI Z535.4 Product Safety Signs and Labels standard were based largely on human factors considerations. The design guidelines for safety messages have been revised, affecting their content, order, grammatical style, and physical appearance. There are also revisions concerning the signal word panel, the use of pictorials, and multi-lingual, and multi-hazard warnings. Warnings designed according to the revised criteria are expected to be more recognizable, legible, readable, and understandable, and therefore, more effective. This paper explains the revisions of interest and the role of human factors in the standard's evolution. Potential liability issues related to the standard and suggestions regarding future revisions are also discussed.

The Federal Small Parts regulation banned toys with small parts intended for children less than three years old because of the hazard of choking. However, choking incidents continued to occur because children had access to older children's toys. To address this problem, Congress recently passed an Act requiring choking hazard labels for balloons, small balls, marbles, and toys containing small parts intended for children between three and six years old. This paper discusses the analysis of choking incidents, the development of labels, the events leading to passage of the Act requiring labels for toys, and a critique of the final labels. Balloons and toys most likely to be given to children less than three will now be labeled with an explicit warning about choking. However, these labels would have been more effective had they followed standardized labeling format. This deficiency has negative implications for other labels.