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Ambient fine particulate matter (PM2.5) pollution is a major health risk. Networks of low-cost sensors (LCS) are increasingly being used to understand local-scale air pollution variation. However, measurements from LCS have uncertainties that can act as a potential barrier to effective decision making. LCS data thus need adequate calibration to obtain good quality PM2.5 estimates. In order to develop calibration factors, one or more LCS are typically co-located with reference monitors for short or long periods of time. A calibration model is then developed that characterizes the relationships between the raw output of the LCS and measurements from the reference monitors. This calibration model is then typically transferred from the co-located sensors to other sensors in the network. Calibration models tend to be evaluated based on their performance only at co-location sites. It is often implicitly assumed that the conditions at the relatively sparse co-location sites are representative of the LCS network overall and that the calibration model developed is not overfitted to the co-location sites. Little work has explicitly evaluated how transferable calibration models developed at co-location sites are to the rest of an LCS network, even after appropriate cross-validation. Further, few studies have evaluated the sensitivity of key LCS use cases, such as hotspot detection, to the calibration model applied. Finally, there has been a dearth of research on how the duration of co-location (short-term or long-term) can impact these results. This paper attempts to fill these gaps using data from a dense network of LCS monitors in Denver deployed through the city's “Love My Air” program. It offers a series of transferability metrics for calibration models that can be used in other LCS networks and some suggestions as to which calibration model would be most useful for achieving different end goals.

A total of 20 urban neighbourhood-scale eddy covariance flux tower datasets are made openly available after being harmonized to create a 50 site–year collection with broad diversity in climate and urban surface characteristics. Variables needed as inputs for land surface models (incoming radiation, temperature, humidity, air pressure, wind and precipitation) are quality controlled, gap-filled and prepended with 10 years of reanalysis-derived local data, enabling an extended spin up to equilibrate models with local climate conditions. For both gap filling and spin up, ERA5 reanalysis meteorological data are bias corrected using tower-based observations, accounting for diurnal, seasonal and local urban effects not modelled in ERA5. The bias correction methods developed perform well compared to methods used in other datasets (e.g. WFDE5 or FLUXNET2015). Other variables (turbulent and upwelling radiation fluxes) are harmonized and quality controlled without gap filling. Site description metadata include local land cover fractions (buildings, roads, trees, grass etc.), building height and morphology, aerodynamic roughness estimates, population density and satellite imagery. This open collection can help extend our understanding of urban environmental processes through observational synthesis studies or in the evaluation of land surface environmental models in a wide range of urban settings. These data can be accessed from https://doi.org/10.5281/zenodo.7104984 (Lipson et al., 2022).

Abstract Background Mastitis is a common reason new mothers visit their general practitioner (GP). In Australia, the Therapeutic Guidelines: Antibiotic provides practical advice to GPs managing a range of infections, including mastitis. It is not known if Australian GPs prescribe antibiotics and order investigations as recommended for the management of mastitis. Methods A convergent mixed methods design integrated quantitative analysis of a general practice dataset with analysis of interviews with GPs. Using the large-scale primary care dataset, MedicineInsight, (2021–2022), antibiotics prescribed and investigations ordered for mastitis encounters were extracted. Mastitis encounters were identified by searching ‘Encounter reason’, ‘Test reason’ and ‘Prescription reason’ free text field for the term ‘mastitis’; ‘granulomatous mastitis’ was excluded. Clinical encounters for mastitis occurring within 14 days of a previous mastitis encounter were defined as belonging to the same treatment episode. Semi-structured interviews were conducted with 14 Australian GPs using Zoom or telephone in 2021–2022, and analysed thematically. The Pillar Integration Process was used to develop a joint display table; qualitative codes and themes were matched with the quantitative items to illustrate similarities/contrasts in findings. Results During an encounter for mastitis, 3122 (91.7%) women received a prescription for an oral antibiotic; most commonly di/flucloxacillin ([59.4%]) or cefalexin (937 [27.5%]). Investigations recorded ultrasound in 303 (8.9%), blood tests (full blood examination [FBE]: 170 [5.0%]; C-reactive protein [CRP]: 71 [2.1%]; erythrocyte sedimentation rate [ESR]: 34 [1.0%]) and breast milk or nipple swab cultures in approximately 1% of encounters. Analysis using pillar integration showed consistency between quantitative and qualitative data regarding mastitis management. The following themes were identified: - GPs support continued breastfeeding. - Antibiotics are central to GPs' management. - Antibiotics are mostly prescribed according to Therapeutic Guidelines. - Analgesia is a gap in the Therapeutic Guidelines. - Low use of breast milk culture. Conclusions Prescribing antibiotics for mastitis remains central to Australian GPs’ management of mastitis. Interview data clarified that GPs were aware that antibiotics might not be needed in all cases of mastitis and that delayed prescribing was not uncommon. Overall, GPs followed principles of antibiotic stewardship, however there is a need to train GPs about when to consider ordering investigations.

Abstract Background There is a severe shortage of corneas for donation, globally, for transplantation and research purposes. One group of individuals who could potentially be donors are those who die within the inpatient palliative care unit. The aim of the study was to understand clinician and patient perceptions of corneal donations and discussion of donation in palliative care units. Methods A qualitative design was utilised with data collected through semi-structured interviews and analysed using qualitative content analysis. A total of 46 interviews were undertaken involving inpatient palliative care unit patients (19) and clinicians (27) in three major inpatient palliative care units in South Australia. Results Very few patient participants reported being asked about corneal donations during their time in palliative care. Most inpatient palliative care unit clinicians did not raise the topic as they felt other areas of care took precedence. Inpatient palliative care unit patients thought if inpatient palliative care unit clinicians did not raise the topic, then it was not important. There were some differences between patient and clinician views, such as preference about who raises the possibility of donation and when the discussion might occur. Conclusions Findings suggest that patients are receptive to discussing corneal donations, but clinicians are not initiating these. This is a missed opportunity for donors and potential recipients. We recommend that clinicians routinely discuss eye donation as part of palliative care.

Objective To examine longitudinal trends in clinical management of lactational mastitis in women attending general practice.Design Open cohort study.Setting Australian general practice using data from MedicineInsight.Participants Women aged 18 to 44 years with one or more clinical encounters for lactational mastitis between January 2011 and July 2022.Primary and secondary outcome measures The primary outcome measure was the proportion of prescribed oral antibiotics based on the antibiotic type. Secondary outcome measures were the proportion of women prescribed other medications (eg, antifungals, lactation suppressants) or ordered selected clinical investigations including breast ultrasound, blood test, breast milk culture, nipple swab culture or breast aspirate. Outcomes were examined based on the calendar year and individual- or clinical practice-level characteristics.Results Among 25 002 women who had one or more clinical encounters related to mastitis, 90.9% were prescribed oral antibiotics. While the proportion of women prescribed an oral antibiotic remained consistent from 2011 to 2022 (91.1% vs 92.5%), there were changes in the proportion receiving prescriptions for di/flucloxacillin (46.1% vs 60.4%) and cefalexin (38.6% vs 26.5%). Fewer than 12% of women were clinically investigated for their mastitis encounter, most commonly a breast ultrasound (7.1%), followed by a selected blood test (3.8%). Requests for breast milk cultures, nipple swab cultures or breast aspirates occurred in less than 1.1% of individuals. Significant increases were evident with respect to ordering of all clinical investigations, with rates at least doubling between 2011 and 2022 (6.6% vs 14.7%). Large variability in clinical management was evident according to both individual- (eg, concessional status) and clinical practice-level characteristics (eg, remoteness).Conclusions Australian general practitioners commonly prescribe oral antibiotics to women with mastitis and largely in line with clinical guidelines. Their use of clinical investigations as part of mastitis management has increased over the last decade.

Abstract High-precision searches for an electric dipole moment of the neutron (nEDM) require stable and uniform magnetic field environments. We present the recent achievements of degaussing and equilibrating the magnetically shielded room (MSR) for the n2EDM experiment at the Paul Scherrer Institute. We present the final degaussing configuration that will be used for n2EDM after numerous studies. The optimized procedure results in a residual magnetic field that has been reduced by a factor of two. The ultra-low field is achieved with the full magnetic-field-coil system, and a large vacuum vessel installed, both in the MSR. In the inner volume of $$\sim 1.4~\hbox {m}^3$$ ∼ 1.4 m 3 , the field is now more uniform and below 300 pT. In addition, the procedure is faster and dissipates less heat into the magnetic environment, which in turn, reduces its thermal relaxation time from $$12~\text {h}$$ 12 h down to $$1.5~\text {h}$$ 1.5 h .

Background:. Carpal tunnel syndrome affects up to 6% of the general population, and surgical intervention is often required to ameliorate symptoms. Osteoarthritis (OA) is a common condition that often coexists with carpal tunnel syndrome. We hypothesized that patients with preexisting OA use more healthcare resources after carpal tunnel release (CTR) than patients without arthritis. Methods:. This was a retrospective cohort study at a single academic center between January 1, 2018 and November 1, 2021. Patients who underwent CTR were included. Preoperative carpal tunnel symptoms, diagnostic tests, medications, and concomitant OA were abstracted. Hand, wrist, and basal joint arthritis were specified. The primary outcome was healthcare utilization represented by duration and frequency of hand clinic and occupational therapy (OT) follow-up. In total, 312 hands were included. Multivariable analysis was performed. Results:. The average duration of hand clinic follow-up among patients without arthritis was 25.3 days compared with 87.1 days for patients with any arthritis (P = 0.0375) and 172 days for patients with wrist arthritis (P = 0.012). The average number of postoperative surgeon visits was increased in patients with hand arthritis, with an average of 2.3 visits versus 1.34 visits for patients without arthritis (P = 0.003). Both the number of OT visits and the duration of OT follow-up did not differ between cohorts. Conclusion:. After CTR, patients with preexisting OA use more healthcare resources than patients without OA.

We applied an integrative framework to illuminate and discuss the complexities of exurbanization in Macon County, North Carolina. The case of Macon County, North Carolina, highlights the complexity involved in addressing issues of exurbanization in the Southern Appalachian region. Exurbanization, the process by which urban residents move into rural areas in search of unique natural amenities and idealized lifestyles, can often have a dramatic impact on the local economy, culture, and environment. Within Macon County, complex debates and tensions among multiple stakeholders struggle to address local residential development. How can better problem definition benefit rural communities in addressing exurbanization pressures and effects? We asserted that a key factor in the shortcomings of previous solutions was the shortsightedness inherent in policy that attempts to treat individual symptoms without being able to adequately characterize the underlying problem. The goal of the integrative framework is to initiate an iterative process of transparent negotiation, which recognizes a range of potential choices to be considered and to embrace the social complexities that can at times overwhelm scholars and practitioners, inviting simplification and polarization of the issues. This new and emerging framework offers a novel way of approaching conservation and development issues where other frameworks have failed. It helps acknowledge the difficult choices, i.e., trade-offs, that have to be made in a material process like exurbanization. Trade-offs will be necessary in any negotiation related to conservation. Therefore, conflict surrounding specific values, e.g., cultural, financial, or ecological, must be acknowledged upfront to move deeper into issues of plurality. Given the complexity, understanding how the process of exurbanization is being played out within Macon County provided not only an opportunity to demonstrate the functionality of an integrative approach, but also a call for further study of exurbanization dynamics.

Healthy dietary patterns rich in flavonoids may benefit cognitive performance over time. Among socioeconomically disadvantaged groups, the association between flavonoid intake and measures of cognition is unclear. This study sought to identify associations between flavonoid intake and cognitive performance among Healthy Aging in Neighborhoods of Diversity across the Life Span (HANDLS) study participants (n = 1947) across three study visits. Flavonoid intakes were assessed via two 24-h dietary recalls. Cognitive performance was assessed via the Trail Making Test (TMT)-A and TMT-B, which provide measures of attention and executive function, respectively. Mixed effects linear regression was used to model TMT scores over three study visits against visit 1 (v1) flavonoid intake, time (years from v1), and the interaction between v1 flavonoid intake and time, capturing both the cross-sectional association between flavonoid intake and time at v1 as well as the longitudinal association between v1 flavonoid intake and the change in TMT scores over time. Prior to adjustment, inverse cross-sectional associations at v1 were observed between (1) anthocyanidin intake and TMT-A scores for the overall sample and (2) total flavonoid, anthocyanidin, flavan-3-ol, flavone, and flavonol intake and TMT-B scores for the overall sample and among White adults. Only the association between anthocyanidin intake and TMT-B at v1 among White adults persisted after adjustment (for demographic characteristics such as age). One possible explanation for the few significant associations is universally low flavonoid intakes resulting from the consumption of an unhealthy dietary pattern.

Reactivation of latent Human Cytomegalovirus (HCMV) in CD34+ hematopoietic progenitor cells (HPCs) is closely linked to hematopoiesis. Viral latency requires maintenance of the progenitor cell quiescence, while reactivation initiates following mobilization of HPCs to the periphery and differentiation into CD14+ macrophages. Early growth response gene 1 (EGR-1) is a transcription factor activated by Epidermal growth factor receptor (EGFR) signaling that is essential for the maintenance of CD34+ HPC self-renewal in the bone marrow niche. Down-regulation of EGR-1 results in mobilization and differentiation of CD34+ HPC from the bone marrow to the periphery. In the current study we demonstrate that the transcription factor EGR-1 is directly targeted for down-regulation by HCMV miR-US22 that results in decreased proliferation of CD34+ HPCs and a decrease in total hematopoietic colony formation. We also show that an HCMV miR-US22 mutant fails to reactivate in CD34+ HPCs, indicating that expression of EGR-1 inhibits viral reactivation. Since EGR-1 promotes CD34+ HPC self-renewal in the bone marrow niche, HCMV miR-US22 down-regulation of EGR-1 is a necessary step to block HPC self-renewal and proliferation to induce a cellular differentiation pathway necessary to promote reactivation of virus.

Background: Patients from residential aged care facilities are commonly exposed to inappropriate polypharmacy. Unplanned inpatient admissions can provide an opportunity for review of complex medical regimens and deprescribing of inappropriate or nonbeneficial medications. The aim of this study was to assess the efficacy, safety and sustainability of in-hospital deprescribing. Methods: We followed a prospective, multi-centre, cohort study design, with enrolment of 106 medical inpatients age 75 years and older (mean age was 88.8 years) who were exposed to polypharmacy prior to admission and with a planned discharge to a nursing home for permanent placement. Descriptive statistics were calculated for relevant variables. The Short Form-8 (SF-8) health survey was used to assess changes in health-related quality of life (HRQOL) at 90-day follow up, in comparison with SF-8 results at day 30. Results: Deprescribing occurred in most, but not all patients. There were no differences between the groups in principal diagnosis, Charlson index, number of medications on admission or number of Beers list medications on admission. At 90 days, mortality and readmissions were similar, though the deprescribed group had significantly higher odds of better emotional wellbeing than the nondeprescribed group [odds ratio (OR) = 5.08, 95% confidence interval (CI): 1.93, 13.39; p = 0.001]. In the deprescribing group, 31% of the patients still alive at 90 days had medications restarted in primary care. One-year mortality rates were similar. Conclusions: Deprescribing medications during an unplanned hospital admission was not associated with mortality, readmissions, or overall HRQOL.

Human cytomegalovirus (HCMV) infection is a serious complication in hematopoietic stem cell transplant (HSCT) recipients due to virus-induced myelosuppression and impairment of stem cell engraftment. Despite the clear clinical link between myelosuppression and HCMV infection, little is known about the mechanism(s) by which the virus inhibits normal hematopoiesis because of the strict species specificity and the lack of surrogate animal models. In this study, we developed a novel humanized mouse model system that recapitulates the HCMV-mediated engraftment failure after hematopoietic cell transplantation. We observed significant alterations in the hematopoietic populations in peripheral lymphoid tissues following engraftment of a subset of HCMV+ CD34+ hematopoietic progenitor cells (HPCs) within the transplant, suggesting that a small proportion of HCMV-infected CD34+ HPCs can profoundly affect HPC differentiation in the bone marrow microenvironment. This model will be instrumental to gain insight into the fundamental mechanisms of HCMV myelosuppression after HSCT and provides a platform to assess novel treatment strategies.

Cushing’s syndrome (CS) due to ectopic adrenocorticotrophic hormone (ACTH) is associated with a variety of tumours most of which arise in the thorax or abdomen. Prostate carcinoma is a rare but important cause of rapidly progressive CS. To report a case of severe CS due to ACTH production from prostate neuroendocrine carcinoma and summarise previous published cases. A 71-year-old male presented with profound hypokalaemia, oedema and new onset hypertension. The patient reported two weeks of weight gain, muscle weakness, labile mood and insomnia. CS due to ectopic ACTH production was confirmed with failure to suppress cortisol levels following low- and high-dose dexamethasone suppression tests in the presence of a markedly elevated ACTH and a normal pituitary MRI. Computed tomography demonstrated an enlarged prostate with features of malignancy, confirmed by MRI. Subsequent prostatic biopsy confirmed neuroendocrine carcinoma of small cell type and conventional adenocarcinoma of the prostate. Adrenal steroidogenesis blockade was commenced using ketoconazole and metyrapone. Complete biochemical control of CS and evidence of disease regression on imaging occurred after four cycles of chemotherapy with carboplatin and etoposide. By the sixth cycle, the patient demonstrated radiological progression followed by recurrence of CS and died nine months after initial presentation. Prostate neuroendocrine carcinoma is a rare cause of CS that can be rapidly fatal, and early aggressive treatment of the CS is important. In CS where the cause of EAS is unable to be identified, a pelvic source should be considered and imaging of the pelvis carefully reviewed.

The FCST (Female-Female Competition Stress Test) was constructed to assess stress from adolescent female-female competition. Such stress is particularly relevant to the Reproductive Suppression Hypothesis that has been suggested as one possible explanation for the development of anorexic type behavior in young women. A series of items generated an initial test in the first phase of development. In the second phase, three studies were conducted to assess reliability and validity. Female undergraduates retrospectively rated the female-female competition stress they had experienced as adolescents, and their responses were factor-analyzed. In a second group of subjects, FCST scores were correlated with a measure of anxiety. In the third study, three samples provided prototypicality ratings of the test items to determine both the agreement among raters and which items were not sufficiently prototypical of female-female competition stress to be retained. In the final phase, the responses of adolescent girls to the FCST and several measures of body image were factor analyzed, showing the utility of the FCST.

Background: Older adults prefer to stay in their homes for as long as possible but are often unaware of the resources in their community to help them to remain in their home. Access to resources may be important among older adults, representing a critical area for intervention. Objectives: The study aim was to evaluate the feasibility of the Health Empowerment Intervention (HEI) and to explore the impact of the HEI on the theoretical mediating variables of health empowerment and purposeful participation in goal attainment and the outcome variable of well-being with home-bound older adults. Method: Fifty-nine eligible homebound older adults were randomly assigned to the intervention group or the comparison group. The HEI consisted of 6 weekly visits, whereas the comparison group received a weekly newsletter for 6 weeks. Participants were measured at baseline, after the 6-week protocol, and at 12 weeks. Data were analyzed using descriptive statistics, t test, Χ² and analysis of covariance. Results: There was a significant difference between groups in education. The participants in the intervention found the sessions to be helpful in recognizing resources. In addition, participants in the intervention group had significantly higher scores in the mediator purposes participation in goal attainment, F(2. 83) = 3.71, p = .03. There was no significant main effect for the mediator healih empowerment; however, the intervention group increased in the subscale personal growth from baseline to 12 weeks, F(1, 83) = 3,88, p = .05. Discussion: This randomized control trial provided initial support for the hypothesis that homebound older adults receiving the HEI would find the intervention acceptable and have significantly improved health empowerment, purposeful participation in goal attainment, and well-being than an atentional comparison group receiving a weekly newsletter. [ABSTRACT FROM AUTHOR]

Recruiting older adults to participate in intervention research is essential for advancing the science in this field. Developing a relevant recruitment plan responsive to the unique needs of the population before beginning a project is critical to the success of a research study. This article describes our experiences in the process of recruiting homebound older adults to test a community-based health empowerment intervention. In our study, the trust and partnership that existed between the research team and Community Action Agency facilitated the role of the home-delivered meal drivers as a trusted and untapped resource for study recruitment. Researchers can benefit from thinking creatively and developing meaningful partnerships when conducting research with older adults. [ABSTRACT FROM AUTHOR]

B cell differentiation and humoral immune responses are markedly suppressed by the persistent environmental contaminant, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The suppression of humoral immune responses by TCDD occurs by direct actions on the B cell and involves activation of the aryl hydrocarbon receptor. Transcriptional regulation of paired box gene 5 (Pax5), an important regulator of B cell differentiation, is altered by TCDD in concordance with the suppression of B cell differentiation and humoral immunoglobulin M response. We hypothesized that TCDD treatment leads to dysregulation of Pax5 transcription by interfering with the basic B cell differentiation mechanisms and aimed to determine the effects of TCDD on upstream regulators of Pax5. A critical regulator of B cell differentiation, B lymphocyte–induced maturation protein-1 (Blimp-1) acts as a transcriptional repressor of Pax5. In lipopolysaccharide (LPS)-activated murine B cell lymphoma, CH12.LX, Blimp-1 messenger RNA, and DNA-binding activity within the Pax5 promoter were suppressed by TCDD. Furthermore, LPS activation of CH12.LX cells upregulated DNA-binding activity of activator protein 1 (AP-1) at three responsive element–like motifs within the Blimp-1 promoter. TCDD treatment of LPS-activated CH12.LX cells suppressed AP-1 binding to these motifs between 24 and 72 h, in concordance with the suppression of Blimp-1 by TCDD. A more comprehensive analysis at 72 h demonstrated that the suppression of AP-1 binding within the Blimp-1 promoter by TCDD was concentration dependent. In summary, our findings link the TCDD-mediated suppression of Blimp-1 through AP-1 to the dysregulation of Pax5, which ultimately leads to the suppression of B cell differentiation and humoral immune responses. [ABSTRACT FROM AUTHOR]

Abstract We present a novel Active Magnetic Shield (AMS), designed and implemented for the n2EDM experiment at the Paul Scherrer Institute. The experiment will perform a high-sensitivity search for the electric dipole moment of the neutron. Magnetic-field stability and control is of key importance for n2EDM. A large, cubic, 5 m side length, magnetically shielded room (MSR) provides a passive, quasi-static shielding-factor of about $$10^5$$ 10 5 for its inner sensitive volume. The AMS consists of a system of eight complex, feedback-controlled compensation coils constructed on an irregular grid spanned on a volume of less than 1000 m $$^3$$ 3 around the MSR. The AMS is designed to provide a stable and uniform magnetic-field environment around the MSR, while being reasonably compact. The system can compensate static and variable magnetic fields up to $$\pm \, 50 \, {\upmu }\hbox {T}$$ ± 50 μ T (homogeneous components) and $$\pm \, 5 \, {\upmu }\hbox {T/m}$$ ± 5 μ T/m (first-order gradients), suppressing them to a few $${\upmu }\hbox {T}$$ μ T in the sub-Hertz frequency range. The presented design concept and implementation of the AMS fulfills the requirements of the n2EDM experiment and can be useful for other applications, where magnetically silent environments are important and spatial constraints inhibit simpler geometrical solutions.

Human cytomegalovirus (HCMV) encodes multiple putative G protein-coupled receptors (GPCRs). US28 functions as a viral chemokine receptor and is expressed during both latent and lytic phases of virus infection. US28 actively promotes cellular migration, transformation, and plays a major role in mediating viral latency and reactivation; however, knowledge about the interaction partners involved in these processes is still incomplete. Herein, we utilized a proximity-dependent biotinylating enzyme (TurboID) to characterize the US28 interactome when expressed in isolation, and during both latent (CD34+ hematopoietic progenitor cells) and lytic (fibroblasts) HCMV infection. Our analyses indicate that the US28 signalosome converges with RhoA and EGFR signal transduction pathways, sharing multiple mediators that are major actors in processes such as cellular proliferation and differentiation. Integral members of the US28 signaling complex were validated in functional assays by immunoblot and small-molecule inhibitors. Importantly, we identified RhoGEFs as key US28 signaling intermediaries. In vitro latency and reactivation assays utilizing primary CD34+ hematopoietic progenitor cells (HPCs) treated with the small-molecule inhibitors Rhosin or Y16 indicated that US28 -RhoGEF interactions are required for efficient viral reactivation. These findings were recapitulated in vivo using a humanized mouse model where inhibition of RhoGEFs resulted in a failure of the virus to reactivate. Together, our data identifies multiple new proteins in the US28 interactome that play major roles in viral latency and reactivation, highlights the utility of proximity-sensor labeling to characterize protein interactomes, and provides insight into targets for the development of novel anti-HCMV therapeutics.

The human betaherpesviruses including human cytomegalovirus (HCMV), human herpesvirus (HHV)-6a and HHV-6b, and HHV-7 infect and establish latency in CD34+ hematopoietic stem and progenitor cells (HPCs). The diverse repertoire of HPCs in humans and the complex interactions between these viruses and host HPCs regulate the viral lifecycle, including latency. Precise manipulation of host and viral factors contribute to preferential maintenance of the viral genome, increased host cell survival, and specific manipulation of the cellular environment including suppression of neighboring cells and immune control. The dynamic control of these processes by the virus regulate inter- and intra-host signals critical to the establishment of chronic infection. Regulation occurs through direct viral protein interactions and cellular signaling, miRNA regulation, and viral mimics of cellular receptors and ligands, all leading to control of cell proliferation, survival, and differentiation. Hematopoietic stem cells have unique biological properties and the tandem control of virus and host make this a unique environment for chronic herpesvirus infection in the bone marrow. This review highlights the elegant complexities of the betaherpesvirus latency and HPC virus-host interactions.

Suppression of humoral immune responses by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been well established to require the aryl hydrocarbon receptor; however, the downstream mechanisms for this immunotoxic response remain poorly understood. Based on evidence demonstrating that primary hepatocytes pretreated with interferon-gamma (IFN-γ) exhibited decreased induction of cytochrome P450 1A1 (CYP1A1) by TCDD, and that serum factors alter the sensitivity of the in vitro T-cell–dependent IgM antibody forming cell (AFC) response, it was hypothesized that IFN-γ attenuates suppression of humoral immune responses by TCDD. In fact, concomitant addition of IFN-γ (100 U/ml) produced a concentration-related attenuation of TCDD-mediated suppression of the anti-sheep erythrocyte (anti-sRBC) IgM AFC response. Time-of-addition studies performed by adding 100 U/ml IFN-γ at 0, 1, 2, 4, 12, 24, 48, and 72 h post-TCDD showed that suppression of the AFC response was prevented only when IFN-γ was added within 2 h of TCDD treatment. mRNA levels of the IgM components, immunoglobulin κ light chain, immunoglobulin μ heavy chain, and immunoglobulin J-chain were significantly decreased by TCDD treatment, an effect that was completely reversed by IFN-γ (100 U/ml) cotreatment. Further studies showed that IFN-α, IFN-β, and IFN-γ significantly attenuate TCDD-induced increases in CYP1A1 mRNA levels to varying degrees, but concentrations as high as 1000 U/ml of type I IFNs did not reverse the effect of TCDD on the anti-sRBC IgM AFC response. In summary, IFN-γ prevents TCDD-mediated suppression of the IgM AFC response in a concentration- and time-related manner by altering transcriptional effects associated with TCDD treatment. [ABSTRACT FROM AUTHOR]

Ultra-low-mass axions are a viable dark matter candidate and may form a coherently oscillating classical field. Nuclear spins in experiments on Earth might couple to this oscillating axion dark-matter field, when propagating on Earth's trajectory through our Galaxy. This spin coupling resembles an oscillating pseudo-magnetic field which modulates the spin precession of nuclear spins. Here we report on the null result of a demonstration experiment searching for a frequency modulation of the free spin-precession signal of \magHg in a 1 $\mu$T magnetic field. Our search covers the axion mass range $10^{-16}$ eV $\lesssim m_a \lesssim 10^{-13}$ eV and achieves a peak sensitivity to the axion-nucleon coupling of $g_{aNN} \approx 3.5 \times 10^{-6}$ GeV$^{-1}$.

Background: Therapeutic angiogenesis aims to induce new blood vessel growth in ischemic tissues; however, previous clinical trials have had limited success. Studies of uterine angiogenesis revealed a specialized subset of natural killer (NK) cells, called uterine NK (uNK) cells, which have unique proangiogenic abilities. Methods: We show that uNK cells in mice express ephrin-B2, a regulator of angiogenesis, to induce tubule formation in an ex vivo coculture tubule formation assay. We next induced the expression of ephrin-B2 by splenic NK (sNK) cells harvested from male mice. Results: We showed that induced NK (iNK) cells can also instruct endothelial cells to form tubules using ephrin-B2. Conclusions: We concluded that Ephrin-B2 is a marker of proangiogenic uNK cells and that a proangiogenic phenotype characterized by ephrin-B2 can be induced in sNK cells to induce therapeutic angiogenesis. : Clinical significance: Peripheral arterial disease threatens puts another 2 million limbs at risk for amputation annually; however, the clinical trials aimed at inducing angiogenesis within ischemic limbs have yielded disappointing results. New molecular therapies should be developed to address this. Like chimeric antigen receptor T cell therapy for melanoma, engineering a patient’s own cells into a population of proangiogenic cells represents a novel strategy to induce angiogenesis. Previous studies have shown that induction of a proangiogenic phenotype in natural killer cells was able to improve perfusion in the placentas of mice. Dissecting the molecular mechanisms behind these phenomena will elucidate the specific molecular targets required, and streamline the cell engineering protocol. We show that induction on natural killer cells of ephrin-B2, a proangiogenic cell membrane receptor that controls angiogenesis throughout life, induces a proangiogenic phenotype in these cells, which is the first step to a cell-based therapy for critical ischemia.

Abstract Background Home death is one of the key performance indicators of the quality of palliative care service delivery. Such a measure has direct implications on everyone involved at the end of life of a dying patient, including a patient’s carers and healthcare professionals. There are no studies that focus on the views of the team of integrated inpatient and community palliative care service staff on the issue of preference of place of death of their patients. This study addresses that gap. Methods Thirty-eight participants from five disciplines in two South Australian (SA) public hospitals working within a multidisciplinary inpatient and community integrated specialist palliative care service, participated in audio-recorded focus groups and one-on-one interviews. Data were transcribed and thematically analysed. Results Two major and five minor themes were identified. The first theme focused on the role of healthcare professionals in decisions regarding place of death, and consisted of two minor themes, that healthcare professionals act to: a) mediate conversations between patient and carer; and b) adjust expectations and facilitate informed choice. The second theme, healthcare professionals’ perspectives on the preference of place of death, comprised three minor themes, identifying: a) the characteristics of the preferred place of death; b) home as a romanticised place of death; and c) the implications of idealising home death. Conclusion Healthcare professionals support and actively influence the decision-making of patients and family regarding preference of place of death whilst acting to protect the relationship between the patient and their family/carer. Further, according to healthcare professionals, home is neither always the most preferred nor the ideal place for death. Therefore, branding home death as the ideal and hospital death as a failure sets up families/carers to feel guilty if a home death is not achieved and undermines the need for and appropriateness of death in institutionalised settings.

Abstract We present the design of a next-generation experiment, n2EDM, currently under construction at the ultracold neutron source at the Paul Scherrer Institute (PSI) with the aim of carrying out a high-precision search for an electric dipole moment of the neutron. The project builds on experience gained with the previous apparatus operated at PSI until 2017, and is expected to deliver an order of magnitude better sensitivity with provision for further substantial improvements. An overview is of the experimental method and setup is given, the sensitivity requirements for the apparatus are derived, and its technical design is described.

Abstract Background Current international consensus is that ‘early’ referral to palliative care services improves cancer patient and family carer outcomes. In practice, however, these referrals are not routine. An approach which directly addresses identified barriers to early integration of palliative care is required. This protocol details a trial of a standardized model of early palliative care (Care Plus) introduced at key defined, disease-specific times or transition points in the illness for people with cancer. Introduced as a ‘whole of system’ practice change for identified advanced cancers, the key outcomes of interest are population health service use change. The aims of the study are to examine the effect of Care Plus implementation on (1) acute hospitalisation days in the last 3 months of life; (2) timeliness of access to palliative care; (3) quality and (4) costs of end of life care; and (5) the acceptability of services for people with advanced cancer. Methods Multi-site stepped wedge implementation trial testing usual care (control) versus Care Plus (practice change). The design stipulates ‘control’ periods when usual care is observed, and the process of implementing Care Plus which includes phases of planning, engagement, practice change and evaluation. During the practice change phase, all patients with targeted advanced cancers reaching the transition point will, by default, receive Care Plus. Health service utilization and unit costs before and after implementation will be collated from hospital records, and state and national health service administrative datasets. Qualitative data from patients, consumers and clinicians before and after practice change will be gathered through interviews and focus groups. Discussion The study outcomes will detail the impact and acceptability of the standardized integration of palliative care as a practice change, including recommendations for ongoing sustainability and broader implementation. Trial registration Australian New Zealand Clinical Trials Registry ACTRN 12619001703190 . Registered 04 December 2019.

Innate-like B cells (ILBs) are a heterogeneous population B cells which participate in innate and adaptive immune responses. This diverse subset of B cells is characterized by the expression of CD5 and has been shown to secrete high levels of immunoglobulin M (IgM) in the absence of infection or vaccination. Further, CD5+ ILBs have been shown to express high basal levels of lymphocyte specific protein tyrosine kinase (LCK) and programmed cell death protein-1 (PD-1), which are particularly sensitive to stimulation by interferon gamma (IFNγ). Previous studies have demonstrated that activation of the aryl hydrocarbon receptor (AHR), a cytosolic ligand-activated transcription factor, results in suppressed IgM responses and is dependent on LCK. A recent study showed that CD5+ ILBs are particularly sensitive to AHR activation as evidenced by a significant suppression of the IgM response compared to CD5- B cells, which were refractory. Therefore, the objective of this study was to further investigate the role of LCK and PD-1 signaling in AHR-mediated suppression of CD5+ ILBs. In addition, studies were conducted to establish whether IFNγ alters the levels of LCK and PD-1 in CD5+ ILBs. We found that AHR activation led to a significant upregulation of total LCK and PD-1 proteins in CD5+ ILBs, which correlated with suppression of IgM. Interestingly, treatment with recombinant IFNγ reduced LCK protein levels and reversed AHR-mediated IgM suppression in CD5+ ILBs in a similar manner as LCK inhibitors. Collectively, these results support a critical role for LCK and PD-1 in AHR-mediated suppression of the IgM response in human CD5+ ILBs.

Abstract The Diabetes and Antenatal Milk Expressing (DAME) randomised controlled trial (RCT) was conducted in 2011–2015, at six sites in Melbourne, Australia to explore the effect of advising women with diabetes in pregnancy to express breast milk from 36 weeks gestation. Infants whose mothers were randomised to express in pregnancy were more likely to be exclusively breast milk fed during their hospital stay, and there was no evidence of harm. This paper explores women's views and experiences of antenatal expressing. In this two‐arm RCT, 635 women with diabetes in pregnancy who were otherwise of low medical risk were randomised at 36–37 weeks gestation to usual care (not expressing, n = 316), or the intervention, where women were advised to hand express for 10 min twice daily until birth (n = 319). Semistructured face‐to‐face interviews were conducted with 10 women who expressed antenatally. They were asked about their experiences of antenatal expressing, including how they felt about the overall experience, the amount of breast milk they expressed, making time to express, and their experience of breastfeeding. Thematic analysis of the in‐depth interviews identified six themes: (1) learning and adapting expressing, (2) feelings and sensations associated with expressing, (3) support, (4) dis/empowerment, (5) health, and (6) the value of breast milk. Women had both positive and negative experiences of antenatal expressing. If health professionals are advising antenatal expressing to women, it is important they understand the range of outcomes and experiences.

ABSTRACT Human cytomegalovirus (HCMV) is a highly prevalent beta-herpesvirus and a significant cause of morbidity and mortality following hematopoietic and solid organ transplant, as well as the leading viral cause of congenital abnormalities. A key feature of the pathogenesis of HCMV is the ability of the virus to establish a latent infection in hematopoietic progenitor and myeloid lineage cells. The study of HCMV latency has been hampered by difficulties in obtaining and culturing primary cells, as well as an inability to quantitatively measure reactivating virus, but recent advances in both in vitro and in vivo models of HCMV latency and reactivation have led to a greater understanding of the interplay between host and virus. Key differences in established model systems have also led to controversy surrounding the role of viral gene products in latency establishment, maintenance, and reactivation. This review will discuss the details and challenges of various models including hematopoietic progenitor cells, monocytes, cell lines, and humanized mice. We highlight the utility and functional differences between these models and the necessary experimental design required to define latency and reactivation, which will help to generate a more complete picture of HCMV infection of myeloid-lineage cells.

Restrictions, social isolation, and uncertainty related to the global COVID-19 pandemic have disrupted the ways that parents and children maintain family routines, health, and wellbeing. Companion animals (pets) can be a critical source of comfort during traumatic experiences, although changes to family routines, such as those caused by COVID-19, can also bring about challenges like managing undesirable pet behaviours or pet-human interactions. We aimed to examine the relationship between pet attachment and mental health for both parents and their children during the COVID-19 pandemic in Australia. A total of 1,034 parents living with a child under 18 years and a cat or dog completed an online cross-sectional survey between July and October 2020. Path analysis using multivariate linear regression was conducted to examine associations between objective COVID-19 impacts, subjective worry about COVID-19, human-pet attachment, and mental health. After adjusting for core demographic factors, stronger pet-child attachment was associated with greater child anxiety (parent-reported, p < .001). Parent-pet attachment was not associated with self-reported psychological distress (p = .42), however, parents who reported a strong emotional closeness with their pet reported greater psychological distress (p = .002). Findings highlight the role of pets during times of change and uncertainty. It is possible that families are turning to animals as a source of comfort, during a time when traditional social supports are less accessible. Alternatively, strong pet attachment is likely to reflect high levels of empathy, which might increase vulnerability to psychological distress. Longitudinal evidence is required to delineate the mechanisms underpinning pet attachment and mental health.

We report on a search for a new, short-range, spin-dependent interaction using a modified version of the experimental apparatus used to measure the permanent neutron electric dipole moment at the Paul Scherrer Institute. This interaction, which could be mediated by axion-like particles, concerned the unpolarized nucleons (protons and neutrons) near the material surfaces of the apparatus and polarized ultracold neutrons stored in vacuum. The dominant systematic uncertainty resulting from magnetic-field gradients was controlled to an unprecedented level of approximately 4 pT cm ^−1 using an array of optically-pumped cesium vapor magnetometers and magnetic-field maps independently recorded using a dedicated measurement device. No signature of a theoretically predicted new interaction was found, and we set a new limit on the product of the scalar and the pseudoscalar couplings $g_sg_p\lambda^2 \lt 8.3 \times 10^{-28}\,$ m ^2 (95% C.L.) in a range of 5 µ m $\lt \lambda \lt 25$ mm for the monopole–dipole interaction. This new result confirms and improves our previous limit by a factor of 2.7 and provides the current tightest limit obtained with free neutrons.

The POLISH2 optical polarimeter has been in operation at the Lick Observatory 3 m Shane telescope since 2011, and it was commissioned at the Gemini North 8 m in 2016. This instrument primarily targets exoplanets, asteroids, and the Crab Pulsar, but it has also been used for a wide variety of planetary, galactic, and supernova science. POLISH2's photoelastic modulators, employed instead of rotating wave plates or ferroelectric liquid crystal modulators, offer the unprecedented ability to achieve sensitivity and accuracy of order 1 ppm (0.0001%), which are difficult to obtain with conventional polarimeters. Additionally, POLISH2 simultaneously measures the intensity (Stokes I ), linear polarization (Stokes Q and U ), and circular polarization (Stokes V ), which fully describe the polarization state of incident light. We document our laboratory and on-sky calibration methodology and our archival on-sky database, and we demonstrate the conclusive detection of circular polarization of certain objects.

Abstract Background Subcutaneous penile modifications (SPMs) are more prevalent in Southeast Asian culture and have been growing in popularity in Western culture. SPMs are often made of domino tiles, or other available pieces of plastic, shaved into a desired shape and placed in unsterile conditions. Previous literature indicates a high risk of infection and the need for surgical removal. Case presentations Seven patients presented to the emergency department in the Southwest border region with complications from SPMs. All the patients complained of pain, four presented with signs of infection, and four SPMs required removal in the emergency department. Removal consisted of a dorsal penile nerve block and making an incision over the SPM to remove the foreign body. Three of the patients had their SPMs done during a previous incarceration under unsterile conditions. Conclusions SPMs appear to be growing in popularity among Western culture, and emergency department health care providers should be aware of trends in body modifications as well as potential complications. The conditions in which SPMs are often placed pose a high risk for infection. In some cases, placement and/or removal of SPMs pose a risk of damage to the corpora, arteries, and nerves of the penis. In the absence of overt bleeding, or suggestion of neurologic injury, dorsally placed superficial foreign bodies of the penis may be amenable to emergency department removal.

Vegetable and fruit consumption plays an important role in lowering the risk of obesity, heart disease, diabetes, stroke and some cancers. Yet, only 20% of the population eats the daily recommended amount of produce, and 10% of children do not consume any vegetables or fruit on a daily basis. To increase vegetable and fruit intake across the population, which can extend lives and reduce health care expenditures while also enhancing demand for California agricultural products, the UC Nutrition Policy Institute has conducted research to guide public policy and institutional and community programs. The institute has studied produce consumption among children and families in child care settings, emergency food systems, schools and federal nutrition assistance programs. Conducting research to support policymakers in deciding on effective nutrition programs and policies involves partnering with community organizations and decision-makers from the outset, collecting qualitative as well as quantitative data, and sharing research findings with those who can act upon them.

Background: Virtual reality (VR) using head-mounted displays (HMDs) has demonstrated to be an effective tool for treating various somatic and psychological symptoms. Technological advances and increased affordability of VR technology provide an interesting option for delivering psychological interventions to patients in palliative care. The primary aim of this systematic review was to synthesise the available research on the use of VR for enhancing psychological and somatic outcomes for palliative care patients. Secondary aims included assessing general satisfaction and overall usability. Method: A pre-registered systematic literature search was conducted according to PRISMA guidelines using OVID Emcare, Cochrane Library, Embase, Medline, PsycINFO, and PubMed Care Search: Palliative Care Knowledge Network. Peer-reviewed experimental, quasi-experimental, observational, case, and feasibility studies consisting of single or multiple VR sessions using HMDs that reported psychological and/or somatic outcomes were included. Results: Eight studies published between 2019 and 2021 were included, representing 138 patients. While the reported quantitative psychological and somatic outcomes were ambiguous, the qualitative outcomes were largely positive. Participants were generally satisfied with VR, and most studies reported the VR interventions as usable, feasible, and acceptable. Conclusions: VR shows promise in palliative care and generally addresses a range of symptoms with few adverse effects. Future research should consist of adequately powered RCTs evaluating dosage and focusing on providing meaningful activities to enhance outcomes further.

Xenobiotic-mediated activation of the aryl hydrocarbon receptor (AHR) is immunotoxic in a number of immune cell types, with the B cell being a well-established sensitive target. Recent advances have provided evidence that the B cell repertoire is a heterogeneous population, with subpopulations exhibiting vastly different cellular and functional phenotypes. Recent work from our laboratory identified the T cell specific kinase lck as being differentially regulated by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), which is a potent activator of AHR. While LCK is primarily expressed in T cells, a subset of CD5+ B cells also express LCK. CD5 positivity describes a broad class of B lymphocytes termed innate-like B cells (ILBs) that are critical mediators of innate immunity through constitutive secretion of polyvalent natural immunoglobulin M (IgM). We hypothesized that CD5+ ILBs may be sensitive to AHR-mediated immunotoxicity. Indeed, when CD5+ B cells were isolated from the CD19+ pool and treated with TCDD, they showed increased suppression of the CD40 ligand-induced IgM response compared to CD5- B cells. Further, characterization of the CD5+ population indicated increased basal expression of AHR, AHR repressor (AHRR), and cytochrome p450 family 1 member a1 (CYP1A1). Indeed the levels of AHR-mediated suppression of the IgM response from individual donors strongly correlated with the percentage of the B cell pool that was CD5+, suggesting that CD5+ B cells are more sensitive to AHR-mediated impairment. Together these data highlight the sensitive nature of CD5+ ILBs to AHR activation and provide insight into mechanisms associated with AHR activation in human B cells.

Abstract There are numerous reports in the medical literature documenting urethral foreign bodies with nearly all cases found in men and often associated with underlying psychiatric disorders, autoerotic stimulation, and/or intoxication. Patients most commonly presented with localized penile pain, hematuria, dysuria, and occasionally obstruction. Although endoscopic removal by a urologist is often the first‐line treatment, this report describes evaluation and management considerations and presents a novel extraction technique that may allow emergency physicians in consultation with urology to perform removal of some unusual urethral foreign bodies in the emergency department. We report a novel extraction technique using a pediatric foley catheter under ultrasound guidance that has been applied during multiple encounters with 2 individuals who have each presented multiple times at a single emergency department (ED) for evaluation because of urethral foreign body insertion. The foreign body materials have ranged from small pieces of rubber to cellophane to styrofoam and most commonly plastic utensils. Urologic extraction may be required in some cases, but ED removal can be considered. A final discussion of the creation of a multidisciplinary care plan to address resource use concerns also is described.

Each herpesvirus family establishes latency in a unique cell type. Since herpesviruses genomes are maintained as episomes, the viruses need to devise mechanisms to retain the latent genome during cell division.

Human cytomegalovirus (HCMV) causes serious disease in immunocompromised individuals and is a significant problem during transplantation. The virus can establish a latent infection in CD34 +

This scoping review examines the strength of evidence for the effectiveness of public policy-led place-based initiatives designed to improve outcomes for disadvantaged children, their families and the communities in which they live. Study designs and methods for evaluating such place-based initiatives were assessed, along with the contexts in which initiatives were implemented and evaluated. Thirty-two reports relating to 12 initiatives were included. Eleven initiatives used a quasi-experimental evaluation to assess impact, although there were considerable design variations within this. The remaining initiative used a pre- and post- evaluation design. Place-based initiatives by definition aim to improve multiple and interrelated outcomes. We examined initiatives to determine what outcomes were measured and coded them within the five domains of pregnancy and birth, child, parent, family and community. Across the 83 outcomes reported in the 11 studies with a comparison group, 30 (36.4%) demonstrated a positive outcome, and all but one initiative demonstrated a positive outcome in at least one outcome measure. Of the six studies that examined outcomes more than once post baseline, 10 from 38 outcomes (26.3%) demonstrated positive sustained results. Many initiatives were affected by external factors such as policy and funding changes, with unknown impact on their effectiveness. Despite the growth of place-based initiatives to improve outcomes for disadvantaged children, the evidence for their effectiveness remains inconclusive.

(Series Information) European Papers - A Journal on Law and Integration, 2018 3(1), 183-202 | Article | (Table of Contents) I. Introduction. - II. The Europeanisation programme. - III. The United Kingdom opt-in. - IV. The legislative background. - IV.1. The Brussels instruments. - IV.2. The Rome instruments. - V. Brexit. - V.1. Brexit: the political background. - V.2. The European Union (Withdrawal) Bill. - VI. The position of Scotland within the UK in the matter of private international law. - VI.1. The European Union (Withdrawal) Bill and the Scottish devolution settlement. - VII. The prospective regime: what should be the UK's aim? - VII.1. Unilateralism. - VII.2. Divergence. - VIII. A possible compromise solution: a bespoke UK/EU27 agreement. - VIII.1. The future role and jurisdiction of the CJEU. - IX. An alternative possible compromise solution: a Lugano II template. - IX.1. The role of the CJEU in the Lugano regime. - X. Failing UK/EU27 agreement... - XI. The Hague Conference on Private International Law. | (Abstract) Professors Crawford and Carruthers comment, from a British perspective, on the possible effects of Brexit upon European civil justice harmonisation measures, with particular reference to the Brussels I Recast, Brussels II bis, Rome I and Rome II Regulations.

Abstract The strict species specificity of Human Cytomegalovirus (HCMV) has impeded our understanding of antiviral adaptive immune responses in the context of a human immune system. We have previously shown that HCMV infection of human hematopoietic progenitor cells engrafted in immune deficient mice (huNSG) results in viral latency that can be reactivated following G-CSF treatment. In this study, we characterized the functional human adaptive immune responses in HCMV latently-infected huBLT (humanized Bone marrow-Liver-Thymus) mice. Following infection, huBLT mice generate human effector and central memory CD4+ and CD8+ T-cell responses reactive to peptides corresponding to both IE and pp65 proteins. Additionally, both HCMV specific IgM and IgG B-cell responses with the ability to neutralize virus were detected. These results indicate that the HCMV huBLT mouse model may provide a valuable tool to study viral latency and reactivation as well as evaluate HCMV vaccines and immune responses in the context of a functional human immune system.

ABSTRACT Human cytomegalovirus (HCMV) infection of CD34+ hematopoietic progenitor cells (CD34+ HPCs) provides a critical reservoir of virus in stem cell transplant patients, and viral reactivation remains a significant cause of morbidity and mortality. The HCMV chemokine receptor US28 is implicated in the regulation of viral latency and reactivation. To explore the role of US28 signaling in latency and reactivation, we analyzed protein tyrosine kinase signaling in CD34+ HPCs expressing US28. US28-ligand signaling in CD34+ HPCs induced changes in key regulators of cellular activation and differentiation. In vitro latency and reactivation assays utilizing CD34+ HPCs indicated that US28 was required for viral reactivation but not latency establishment or maintenance. Similarly, humanized NSG mice (huNSG) infected with TB40E-GFP-US28stop failed to reactivate upon treatment with granulocyte-colony-stimulating factor, but viral genome levels were maintained. Interestingly, HCMV-mediated changes in hematopoiesis during latency in vivo and in vitro was also dependent upon US28, as US28 directly promoted differentiation toward the myeloid lineage. To determine whether US28 constitutive activity and/or ligand-binding activity were required for latency and reactivation, we infected both huNSG mice and CD34+ HPCs in vitro with HCMV TB40E-GFP containing the US28-R129A mutation (no CA) or Y16F mutation (no ligand binding). TB40E-GFP-US28-R129A was maintained during latency and exhibited normal reactivation kinetics. In contrast, TB40E-GFP-US28-Y16F exhibited high levels of viral genome during latency and reactivation, indicating that the virus did not establish latency. These data indicate that US28 is necessary for viral reactivation and ligand binding activity is required for viral latency, highlighting the complex role of US28 during HCMV latency and reactivation. IMPORTANCE Human cytomegalovirus (HCMV) can establish latency following infection of CD34+ hematopoietic progenitor cells (HPCs), and reactivation from latency is a significant cause of viral disease and accelerated graft failure in bone marrow and solid-organ transplant patients. The precise molecular mechanisms of HCMV infection in HPCs are not well defined; however, select viral gene products are known to regulate aspects of latency and reactivation. The HCMV-encoded chemokine receptor US28, which binds multiple CC chemokines as well as CX3CR1, is expressed both during latent and lytic phases of the virus life cycle and plays a role in latency and reactivation. However, the specific timing of US28 expression and the role of ligand binding in these processes are not well defined. In this report, we determined that US28 is required for reactivation but not for maintaining latency. However, when present during latency, US28 ligand binding activity is critical to maintaining the virus in a quiescent state. We attribute the regulation of both latency and reactivation to the role of US28 in promoting myeloid lineage cell differentiation. These data highlight the dynamic and multifunctional nature of US28 during HCMV latency and reactivation.

ABSTRACT The ability of human cytomegalovirus (HCMV) to reactivate from latent infection of hematopoietic progenitor cells (HPCs) is intimately linked to cellular differentiation. HCMV encodes UL7 that our group has shown is secreted from infected cells and induces angiogenesis. In this study, we show that UL7 is a ligand for Fms-like tyrosine kinase 3 receptor (Flt-3R), a well-known critical factor in HPC differentiation. We observed that UL7 directly binds Flt-3R and induces downstream signaling cascades, including phosphatidylinositol 3-kinase (PI3K)/Akt and mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathways. Importantly, we show that UL7 protein induces differentiation of both CD34+ HPCs and CD14+ monocytes. Last, we show that an HCMV mutant lacking UL7 fails to reactivate in CD34+ HPCs in vitro as well as in humanized mice. These observations define the first virally encoded differentiation factor with significant implications not only for HCMV reactivation but also for alteration of the hematopoietic compartment in transplant patients. IMPORTANCE Human cytomegalovirus (HCMV) remains a significant cause of morbidity and mortality in allogeneic hematopoietic stem cell transplant recipients. CD34+ hematopoietic progenitor cells (HPCs) represent a critical reservoir of latent HCMV in the transplant population, thereby providing a source of virus for dissemination to visceral organs. HCMV reactivation has been linked to HPC/myeloid cellular differentiation; however, the mechanisms involved in these events are poorly understood at the molecular level. In this study, we show that a viral protein is a ligand for Fms-like tyrosine kinase 3 receptor (Flt-3R) and that the binding of HCMV UL7 to the Flt-3R triggers HPC and monocyte differentiation. Moreover, the loss of UL7 prevents viral reactivation in HPCs in vitro as well as in humanized mice. These observations define the first virally encoded differentiation factor with significant implications not only for HCMV reactivation but also for alteration of the hematopoietic compartment in transplant patients.