Influences of 23 different equations used to calculate gene copies of SARS-CoV-2 during wastewater-based epidemiology.

Following the emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in late 2019, the use of wastewater-based surveillance (WBS) has increased dramatically along with associated infrastructure globally. However, due to the global nature of its application, and various workflow adaptations (e.g., sample collection, water concentration, RNA extraction kits), numerous methods for back-calculation of gene copies per volume (gc/L) of sewage have also emerged. Many studies have considered the comparability of processing methods (e.g., water concentration, RNA extraction); however, for equations used to calculate gene copies in a wastewater sample and subsequent influences on monitoring viral trends in a community and its association with epidemiological data, less is known. Due to limited information on how many formulas exist for the calculation of SARS-CoV-2 gene copies in wastewater, we initially attempted to quantify how many equations existed in the referred literature. We identified 23 unique equations, which were subsequently applied to an existing wastewater dataset. We observed a range of gene copies based on use of different equations, along with variability of AUC curve values, and results from correlation and regression analyses. Though a number of individual laboratories appear to have independently converged on a similar formula for back-calculation of viral load in wastewater, and share similar relationships with epidemiological data, differential influences of various equations were observed for variation in PCR volumes, RNA extraction volumes, or PCR assay parameters. Such observations highlight challenges when performing comparisons among WBS studies when numerous methodologies and back-calculation methods exist. To facilitate reproducibility among studies, the different gc/L equations were packaged as an R Shiny app, which provides end users the ability to investigate variability within their datasets and support comparisons among studies.

The ASAS-OMERACT core domain set for axial spondyloarthritis.

BACKGROUND: The current core outcome set for ankylosing spondylitis (AS) has had only minor adaptations since its development 20 years ago. Considering the significant advances in this field during the preceding decades, an update of this core set is necessary. OBJECTIVE: To update the ASAS-OMERACT core outcome set for AS into the ASAS-OMERACT core outcome set for axial spondyloarthritis (axSpA). METHODS: Following OMERACT and COMET guidelines, an international working group representing key stakeholders (patients, rheumatologists, health professionals, pharmaceutical industry and drug regulatory agency representatives) defined the core domain set for axSpA. The development process consisted of: i) Identifying candidate domains using a systematic literature review and qualitative studies; ii) Selection of the most relevant domains for different stakeholders through a 3-round Delphi survey involving axSpA patients and axSpA experts; iii) Consensus and voting by ASAS; iv) Endorsement by OMERACT. Two scenarios are considered based on the type of therapy investigated in the trial: symptom modifying therapies and disease modifying therapies. RESULTS: The updated core outcome set for axSpA includes 7 mandatory domains for all trials (disease activity, pain, morning stiffness, fatigue, physical function, overall functioning and health, and adverse events including death). There are 3 additional domains (extra-musculoskeletal manifestations, peripheral manifestations and structural damage) that are mandatory for disease modifying therapies and important but optional for symptom modifying therapies. Finally, 3 other domains (spinal mobility, sleep, and work and employment) are defined as important but optional domains for all trials. CONCLUSION: The ASAS-OMERACT core domain set for AS has been updated into the ASAS-OMERACT core domain set for axSpA. The next step is the selection of instruments for each domain.

Investigating diagnosis, treatment, and burden of disease in patients with ankylosing spondylitis in Central Eastern Europe and the United States: a real-world study.

INTRODUCTION/OBJECTIVES: Ankylosing spondylitis (AS) is a chronic inflammatory immune-mediated condition. We compared AS diagnosis, treatment, and burden in Central Eastern European countries (CEE), where this has been less researched, and the United States (US) from a real-world perspective. METHODS: Point-in-time survey of rheumatologists and their AS patients was conducted in the US (Apr-Oct 2018) and CEE (Aug-Nov 2019) via physician- and patient-completed record forms, including clinical and patient-reported outcomes. Statistical analysis included descriptive statistics, t-tests, Fisher's exact tests, and generalized linear models. RESULTS: In total, 487 patients were recruited from 88 rheumatologists in the US and 922 patients from 126 rheumatologists in CEE. Time from onset of symptoms to final AS diagnosis was longer in CEE than the US (4.2 vs 2.7 years, p < 0.05). At diagnosis, a greater use of conventional synthetic disease-modifying antirheumatic drugs (DMARDs) and injected steroids was reported in CEE vs the US (43.7% vs 27.6%, p < 0.05; 19.3% vs 8.7%, p < 0.05). 22.9% of US patients received a biologic DMARD at diagnosis vs 10% of CEE patients (p < 0.05). At current consultation, biologic DMARD use in CEE was lower vs the US (27.9% vs 71.0%, p < 0.05). CEE vs US patients had greater disease activity (mean Bath Ankylosing Spondylitis Disease Activity Index 4.2 vs 3.1, p < 0.05) and worse quality of life (QoL; mean Ankylosing Spondylitis Quality of Life Questionnaire score 6.2 vs 8.4, p < 0.05). CONCLUSIONS: AS patients in CEE vs the US faced slower diagnosis and worse access to biologics, disease activity, and QoL. Whether early access to biologics can improve symptoms, QoL, and daily activities in AS patients in CEE remains to be seen. Key Points • The study provided evidence on the real-world approach to the diagnosis, treatment, and burden of axSpA (axial spondyloarthritis) in CEE compared with the US. • The study reported patients in CEE experienced longer delays in diagnosis and poorer access to biologics than in the US. • This may have resulted in higher disease activity, greater levels of pain, and poorer outcomes, as reported by patients with axSpA in CEE.

The flexibility of ACE2 in the context of SARS-CoV-2 infection.

The COVID-19 pandemic has swept over the world in the past months, causing significant loss of life and consequences to human health. Although numerous drug and vaccine developments efforts are underway, many questions remain outstanding on the mechanism of SARS-CoV-2 viral association to angiotensin-converting enzyme 2 (ACE2), its main host receptor, and entry in the cell. Structural and biophysical studies indicate some degree of flexibility in the viral extracellular Spike glycoprotein and at the receptor binding domain-receptor interface, suggesting a role in infection. Here, we perform all-atom molecular dynamics simulations of the glycosylated, full-length membrane-bound ACE2 receptor, in both an apo and spike receptor binding domain (RBD) bound state, in order to probe the intrinsic dynamics of the ACE2 receptor in the context of the cell surface. A large degree of fluctuation in the full length structure is observed, indicating hinge bending motions at the linker region connecting the head to the transmembrane helix, while still not disrupting the ACE2 homodimer or ACE2-RBD interfaces. This flexibility translates into an ensemble of ACE2 homodimer conformations that could sterically accommodate binding of the spike trimer to more than one ACE2 homodimer, and suggests a mechanical contribution of the host receptor towards the large spike conformational changes required for cell fusion. This work presents further structural and functional insights into the role of ACE2 in viral infection that can be exploited for the rational design of effective SARS-CoV-2 therapeutics. STATEMENT OF SIGNIFICANCE: As the host receptor of SARS-CoV-2, ACE2 has been the subject of extensive structural and antibody design efforts in aims to curtail COVID-19 spread. Here, we perform molecular dynamics simulations of the homodimer ACE2 full-length structure to study the dynamics of this protein in the context of the cellular membrane. The simulations evidence exceptional plasticity in the protein structure due to flexible hinge motions in the head-transmembrane domain linker region and helix mobility in the membrane, resulting in a varied ensemble of conformations distinct from the experimental structures. Our findings suggest a dynamical contribution of ACE2 to the spike glycoprotein shedding required for infection, and contribute to the question of stoichiometry of the Spike-ACE2 complex.

Results of a global, patient-based survey assessing the impact of psoriatic arthritis discussed in the context of the Psoriatic Arthritis Impact of Disease (PsAID) questionnaire.

BACKGROUND: Psoriatic arthritis (PsA) is a chronic immune-mediated inflammatory musculoskeletal disease, manifesting as peripheral arthritis, enthesitis, dactylitis, spondylitis, and skin and nail psoriasis. A core set of domains for measuring the impact of PsA has been developed, including pain, patient global assessment, physical function, health-related quality of life (HRQoL), and fatigue. To understand the impact of PsA on health domains from a patient's perspective, a global survey was developed and results reported in the context of the 12-item Psoriatic Arthritis Impact of Disease (PsAID-12) questionnaire. METHODS: An online patient-based global survey was conducted by The Harris Poll in Australia, Brazil, Canada, France, Spain, Taiwan, the UK, and the US between November 2, 2017 and March 12, 2018. Eligible patients were ≥ 18 years old with a diagnosis of PsA for > 1 year, had visited a rheumatologist/dermatologist in the past 12 months and reported using ≥ 1 synthetic/biologic disease-modifying antirheumatic drug for PsA. Patients reported on PsA severity and symptoms, and the impact of PsA on HRQoL. After survey completion, responses were aligned with PsAID health domains. Descriptive statistics and chi-square tests were conducted. RESULTS: This analysis included 1286 patients from eight countries. Most patients (97%) reported musculoskeletal symptoms relating to PsA in the past year. Common moderate/major impacts of PsA were on physical activity (78%), ability to perform certain activities (76%), work productivity (62%), and career path (57%). Skin/nail symptoms occurred in 80% of patients. Overall, 69% of patients reported that PsA had a moderate/major impact on emotional/mental wellbeing, 56% on romantic relationships/intimacy, and 44% on relationships with family and friends. Social impacts included emotional distress (58%), social shame or disapproval (32%), and ceased participation in social activities (45%). Over half of all patients experienced unusual fatigue over the past 12 months (52%). The health domains that patients reported as being impacted by PsA aligned with life impact domains of the patient-derived PsAID health domains. CONCLUSION: These results highlight the impact of PsA on multiple health domains from a patient perspective that should be considered during shared decision-making processes between healthcare providers and patients.

Blue light from individual light masks directed at a single eye advances the breeding season in mares.

REASONS FOR PERFORMING STUDY: Artificial lighting is commonly used to advance the breeding season in horses. Light masks have been developed that direct light at a single eye to inhibit the production of melatonin, the decoder of photoperiod for seasonally breeding animals. OBJECTIVES: To investigate whether low-intensity blue light from light masks was effective at advancing the breeding season in mares. STUDY DESIGN: Controlled experiment. METHODS: Data on reproductive activity was collected from 3 groups of mares maintained on Kentucky horse farms under various lighting conditions between 20 November 2011 and 10 February 2012: 59 nonpregnant, healthy Thoroughbred mares were used. On 1 December 2011, Group 1 (n = 16) was housed indoors under barn lighting (250 Lux) until 23.00 h daily. Group 2 (n = 25) wore light masks programmed to turn on from 16.30 h until 23.00 h daily and was maintained outdoors. Group 3 (n = 19) was maintained outdoors under the natural photoperiod as control. At 2-week intervals, rectal ultrasound examinations were performed and blood was collected for progesterone analysis. Oestrous cyclicity was defined as the presence of follicles >20 mm diameter detected in conjunction with serum progesterone >1 ng/ml and confirmation of ovulation by transrectal ultrasound examination. RESULTS: On 10 February, the number of mares exhibiting oestrous cyclicity was 14/16 (87.5%) in Group 1; 20/25 (80%) in Group 2; and 4/19 (21%), in Group 3. Pairwise comparison of groups revealed no difference in the number of cycling mares between Groups 1 and 2 (χ(2) test, P = 0.3348) whereas differences were observed between Groups 1 and 3 (χ(2) test, P<0.0001) and Groups 2 and 3 (χ(2) test, P<0.0003). CONCLUSIONS: Low-intensity blue light to a single eye from a light mask is an effective alternative to maintenance of mares indoors under lights for advancing the breeding season. Mobile light therapy for horses could have economic benefits for the breeder by reducing the costs of maintaining mares indoors, and welfare benefits for horses by permitting outdoor maintenance.

Can voter-approved tax levies provide fiscal advantages and stability for local public health agencies?

CONTEXT: Major funding cuts have occurred throughout the United States public health system during the past several years. Funding for local public health agency (LPHA) services and programs is obtained through a patchwork of sources that vary both within and among states. Even though local city and county sources provide a significant proportion of funding for LPHAs, information available in the literature about these revenues is sparse and is not clearly described. OBJECTIVE: This study focused on a single specific revenue stream included in the local sources (local city and county) category: funds voted on directly by the public. The primary purpose of this study was to examine whether this type of funding source provided fiscal advantages for LPHAs. Specifically, we wanted to see how sensitive levy votes were to changing general economic conditions. METHODS: A questionnaire to collect LPHA levy data was developed, approved, and mailed to county boards of elections in Ohio (n = 88). Elections officials were asked to provide voting results for all LPHA levy ballot attempts since 1994 regardless of outcome. RESULTS: In the study period (1994 through 2011), 250 LPHA property tax levies were placed on election ballots in Ohio. LPHAs were successful in 155 (62.0%) and unsuccessful in 95 (38.0%) attempts. Over the 18-year period, the most noteworthy outcome was a 94.6% pass rate for renewal levies. CONCLUSION: Our study demonstrated that voter-approved tax levies provide some fiscal advantages for LPHAs: higher per capita revenues than those who have to rely on other sources of income and predictable revenue streams. This translates into more funds being available for public health programs and services. Property tax levies allow citizens to make direct investments in their local health departments.

Search for photon-linelike signatures from dark matter annihilations with H.E.S.S.

Gamma-ray line signatures can be expected in the very-high-energy (E(γ)>100 GeV) domain due to self-annihilation or decay of dark matter (DM) particles in space. Such a signal would be readily distinguishable from astrophysical γ-ray sources that in most cases produce continuous spectra that span over several orders of magnitude in energy. Using data collected with the H.E.S.S. γ-ray instrument, upper limits on linelike emission are obtained in the energy range between ∼ 500 GeV and ∼ 25 TeV for the central part of the Milky Way halo and for extragalactic observations, complementing recent limits obtained with the Fermi-LAT instrument at lower energies. No statistically significant signal could be found. For monochromatic γ-ray line emission, flux limits of (2 × 10(-7) -2 × 10(-5)) m(-2) s(-1) sr(-1) and (1 × 10(-8) -2 × 10(-6)) m(-2) s(-1)sr(-1) are obtained for the central part of the Milky Way halo and extragalactic observations, respectively. For a DM particle mass of 1 TeV, limits on the velocity-averaged DM annihilation cross section ⟨σv⟩(χχ → γγ) reach ∼ 10(-27) cm(3)s(-1), based on the Einasto parametrization of the Galactic DM halo density profile.

Immunisation of mares with binding domains of toxins A and B of Clostridium difficile elicits serum and colostral antibodies that block toxin binding.

REASONS FOR PERFORMING STUDY: Enterocolitis caused by Clostridium difficile (C. difficile) is a serious, sometimes fatal, disease of neonatal foals and older horses. Toxins A and B (TcdA and B) produced by C. difficile are important virulence factors. Immunisation of mares with receptor binding domains of toxins may prevent or reduce the severity of C. difficile colitis in foals. OBJECTIVES: To determine whether antibodies generated in the pregnant mare to the binding regions of TcdA and B will neutralise TcdA and B toxicity. METHODS: Sequences encoding the binding domains of each toxin were isolated by PCR amplification from C. difficile JF09, a foal isolate, and cloned and expressed into pET15b. Thirteen mares were immunised twice 2 weeks apart with 200 µg of each recombinant protein with Quil A 2 months prior to foaling. Antibodies were assayed in the sera and colostrum by ELISA and for ability to block the cytopathic activity of each of toxin for equine endothelial cells. RESULTS: All mares produced strong serum antibody responses to the binding domain of each toxin. A high level of toxin-specific antibodies was also detected in colostrum and in most foal sera 2 days after suckling. Diluted sera and colostrum premixed with either TcdA or B had no effect on the morphology of equine endothelial cells. Application of the same concentration of toxins alone or premixed with nonimmune mare/foal serum or colostrum led to an unambiguous cytopathic effect that ranged from complete degradation to varying degrees of cell rounding. CONCLUSIONS: Immunisation of pregnant mares with recombinant binding domains of TcdA and B of C. difficile resulted in the production of specific antibodies in serum and colostrum that blocked the cytopathic activity of toxins. POTENTIAL RELEVANCE: Results of studies support the feasibility of a prepartum vaccine against C. difficile enterocolitis in foals.

Reducing barriers for migratory agricultural workers to receive oral health care services.

Between one and two million migratory agricultural workers leave their homes each year to plant, cultivate, harvest and pack fruits, vegetables and nuts in the United States. While in the United States, most lack dental insurance and have no regular dentist. The primary purpose of this study was to describe issues associated with barriers to access and receiving oral health care from the perspective of migratory agricultural workers rather than the perspective of providers. Views and experiences regarding their use of oral health care services were collected from focus groups. Information from focus groups was used to design a questionnaire. Among the 157 respondents the most significant barriers hampering access to oral health care services were crop demands, travel distance, and transportation. Cost and the lack of an interpreter were ranked as the top two barriers to receiving oral health care. The most convenient times for respondents to visit a dentist were between 1 and 6 p.m. The most convenient day was Monday, followed by Sunday and Saturday.

Reducing barriers associated with delivering health care services to migratory agricultural workers.

INTRODUCTION: Between one and two million migratory agricultural workers (MAWs), primarily from Mexico and Central America, leave their homes each year to plant, cultivate, harvest and pack fruits, vegetables, and nuts in the USA. While in the USA, most lack health insurance, a permanent residence, and a regular healthcare provider. Publications over the past two decades in the USA have reported that a majority of MAWs encounter barriers to receiving medical services. Migratory agricultural workers experience high rates of occupational illness and injury. Poor access to medical care continues to exacerbate health problems among members of this population related to their working environments. In most studies concerning healthcare access issues for this population, researchers collected their information from healthcare service providers; rarely have they included input from migratory agricultural workers. This study was different in that opinions about healthcare access issues were collected directly from MAWs. The primary purpose of this study was to describe issues related to barriers associated with the delivery of healthcare services to migratory agricultural workers. A secondary purpose was to suggest strategies for reducing these barriers. METHODS: In this study, data from focus group sessions were used to develop a survey questionnaire. Four certified bilingual interpreters were trained to administer the questionnaire. A total of 157 usable questionnaires were returned from MAWs living in employer-provided camps in Northwest Ohio. The statistical analyses were primarily descriptive. RESULTS: The most significant barriers hampering access to medical services among the 157 respondents were cost (n=113; 72.0%), crop demands (n=102; 65.0%), the lack of an interpreter (n=98; 62.4%), travel distance (n=88; 56.1%) and transportation (n=82; 52.2%). Approximately half (n=82; 52.2%) said that they had access to transportation for traveling to a medical clinic. As a group, respondents were willing to travel an average of 29.1 km (18.1 miles) (range 0-129 km [0-80 miles]) to obtain medical services. Female heads of households had significantly less access to transportation compared with male heads of households (t=2.35; df=74; p<0.05). CONCLUSIONS: Three general categories of barriers to health care for MAWs surfaced in this study: (1) work environment; (2) migratory agricultural worker resources; and (3) healthcare clinic practices. Work environment issues relate mostly to the employers. Resources are barriers for MAWs because they are poor and have limited funds for the cost of transportation to clinics and the fees associated with accessing health care. Most of the barriers identified related to healthcare clinic practices. Some strategies to address healthcare clinic practice barriers were developed by the group conducting the study. By listening to what MAWs described as barriers to health care, providers can help improve access which can reduce the use of high cost hospital emergency room care.

Search for a dark matter annihilation signal from the galactic center halo with H.E.S.S.

A search for a very-high-energy (VHE; ≥100 GeV) γ-ray signal from self-annihilating particle dark matter (DM) is performed towards a region of projected distance r∼45-150 pc from the Galactic center. The background-subtracted γ-ray spectrum measured with the High Energy Stereoscopic System (H.E.S.S.) γ-ray instrument in the energy range between 300 GeV and 30 TeV shows no hint of a residual γ-ray flux. Assuming conventional Navarro-Frenk-White and Einasto density profiles, limits are derived on the velocity-weighted annihilation cross section (σv) as a function of the DM particle mass. These are among the best reported so far for this energy range and in particular differ only little between the chosen density profile parametrizations. In particular, for the DM particle mass of ∼1 TeV, values for (σv) above 3×10(-25) cm(3) s(-1) are excluded for the Einasto density profile.

Communications between local health departments and the public during emergencies: the importance of standardized web sites.

Of the 2790 local health departments (LHDs) in the United States, Internet homepages were located for 1986. We reviewed each homepage to document the presence of 9 elements deemed to be critical for effective communications during emergency or disaster situations. LHD Web site homepages had a mean of 4.1 (±1.4) elements. Among the findings, this review revealed that 4 of 5 (80.5%) of the LHDs included the agency phone number, half (49.4%) provided links to emergency information, and about 1 in 5 (19.6%) listed an agency e-mail address. Fewer than 1 in 20 (4.3%) of the LHD homepages reviewed allowed visitors to sign up for automatic alerts or notifications. We suggest that these results be used as a starting point in developing a standardized template containing the 9 homepage elements. Such a template complements National Incident Management System protocols and can provide a recognizable source of consistent and reliable information for people during a public health emergency or disaster.

Creating a regional dental center serving six rural county health districts.

People with Medicaid or no dental insurance have a difficult time accessing dentists in private practice. The problem of access is more profound in rural than urban areas. Safety net dental clinics operated by small rural local health districts are difficult to start up, operate, and maintain. The number of these facilities in the United States is small and not evenly distributed to meet needs. This article describes how a full-service dental clinic was established to serve six rural county health districts in Northwest Ohio. Retired volunteer dentists were instrumental in the success of creating the clinic, starting with a field-type operation in 2001 serving 316 persons and building into a full-time regional dental center that served 1,306 individuals in 2007.