Virus replication in the honey bee parasite, Varroa destructor.

IMPORTANCE: The parasitic mite Varroa destructor is a significant driver of worldwide colony losses of our most important commercial pollinator, the Western honey bee Apis mellifera. Declines in honey bee health are frequently attributed to the viruses that mites vector to honey bees, yet whether mites passively transmit viruses as a mechanical vector or actively participate in viral amplification and facilitate replication of honey bee viruses is debated. Our work investigating the antiviral RNA interference response in V. destructor demonstrates that key viruses associated with honey bee declines actively replicate in mites, indicating that they are biological vectors, and the host range of bee-associated viruses extends to their parasites, which could impact virus evolution, pathogenicity, and spread.

Adaptation to vector-based transmission in a honeybee virus.

Global pollinator declines as a result of emerging infectious diseases are of major concern. Managed honeybees Apis mellifera are susceptible to numerous parasites and pathogens, many of which appear to be transmissible to sympatric non-Apis taxa. The ectoparasitic mite Varroa destructor is considered to be the most significant threat to honeybees due to its role in vectoring RNA viruses, particularly Deformed wing virus (DWV). Vector transmission of DWV has resulted in the accumulation of high viral loads in honeybees and is often associated with colony death. DWV has two main genotypes, A and B. DWV-A was more prevalent during the initial phase of V. destructor establishment. In recent years, the global prevalence of DWV-B has increased, suggesting that DWV-B is better adapted to vector transmission than DWV-A. We aimed to determine the role vector transmission plays in DWV genotype prevalence at a colony level. We experimentally increased or decreased the number of V. destructor mites in honeybee colonies, and tracked DWV-A and DWV-B loads over a period of 10 months. Our results show that the two DWV genotypes differ in their response to mite numbers. DWV-A accumulation in honeybees was positively correlated with mite numbers yet DWV-A was largely undetected in the absence of the mite. In contrast, colonies had high loads of DWV-B even when mite numbers were low. DWV-B loads persisted in miticide-treated colonies, indicating that this genotype has a competitive advantage over DWV-A irrespective of mite numbers. Our findings suggest that the global increase in DWV-B prevalence is not driven by selective pressure by the vector. Rather, DWV-B is able to persist in colonies at higher viral loads relative to DWV-A in the presence and absence of V. destructor. The interplay between V. destructor and DWV genotypes within honeybee colonies may have broad consequences upon viral diversity in sympatric taxa as a result of spillover.

Systematic generation of high-resolution deletion coverage of the Drosophila melanogaster genome.

In fruit fly research, chromosomal deletions are indispensable tools for mapping mutations, characterizing alleles and identifying interacting loci. Most widely used deletions were generated by irradiation or chemical mutagenesis. These methods are labor-intensive, generate random breakpoints and result in unwanted secondary mutations that can confound phenotypic analyses. Most of the existing deletions are large, have molecularly undefined endpoints and are maintained in genetically complex stocks. Furthermore, the existence of haplolethal or haplosterile loci makes the recovery of deletions of certain regions exceedingly difficult by traditional methods, resulting in gaps in coverage. Here we describe two methods that address these problems by providing for the systematic isolation of targeted deletions in the D. melanogaster genome. The first strategy used a P element-based technique to generate deletions that closely flank haploinsufficient genes and minimize undeleted regions. This deletion set has increased overall genomic coverage by 5-7%. The second strategy used FLP recombinase and the large array of FRT-bearing insertions described in the accompanying paper to generate 519 isogenic deletions with molecularly defined endpoints. This second deletion collection provides 56% genome coverage so far. The latter methodology enables the generation of small custom deletions with predictable endpoints throughout the genome and should make their isolation a simple and routine task.

Are at-risk sociodemographic attributes stable across COVID-19 transmission waves?

COVID-19 health impacts and risks have been disproportionate across social, economic, and racial gradients (Chen et al., 2021; Thompson et al., 2021; Mamuji et al., 2021; COVID-19 and Ethnicity, 2020). By examining the first five waves of the pandemic in Ontario, we identify if Forward Sortation Area (FSAs)based measures of sociodemographic status and their relationship to COVID-19 cases are stable or vary by time. COVID-19 waves were defined using a time-series graph of COVID-19 case counts by epi-week. Percent Black visible minority, percent Southeast Asian visible minority and percent Chinese visible minority at the FSA level were then integrated into spatial error models with other established vulnerability characteristics. The models indicate that area-based sociodemographic patterns associated with COVID-19 infection change over time. If sociodemographic characteristics are identified as high risk (increased COVID-19 case rates) increased testing, public health messaging, and other preventative care may be implemented to protect populations from the inequitable burden of disease.

Going home: identifying and overcoming barriers to nursing home discharge.

This article describes barriers to nursing home discharge encountered in an intervention designed to transition nursing home residents to the community. Staff in the intervention ("Project Home") provided intensive case management and discharge planning services to nursing home residents who expressed a desire to return to community-based living arrangements. Sixty program participants took part in the program evaluation that informs this article. With the exception of Medicaid status, no differences were found between the social, demographic, and health characteristics of individuals who remained in the nursing home and those who were discharged. A qualitative analysis was conducted to describe barriers to discharge and strategies intervention staff used to leverage each client's strengths and work around obstacles. Three main barriers to discharge were found: having an unstable or complex medical condition, lacking family or social support, and being unable to obtain suitable housing. Intervention staff advocated on the behalf of clients, encouraged clients to build skills toward independent living. and contributed extensive knowledge of local resources to advance client goals. Cases of successful transition suggest that a person-centered approach from intervention staff combined with a flexible organizational structure is a promising model for future interventions.

Brain-computer interface-based assessment of color vision.

Objective.Present methods for assessing color vision require the person's active participation. Here we describe a brain-computer interface-based method for assessing color vision that does not require the person's participation.Approach.This method uses steady-state visual evoked potentials to identify metamers-two light sources that have different spectral distributions but appear to the person to be the same color.Main results.We demonstrate that: minimization of the visual evoked potential elicited by two flickering light sources identifies the metamer; this approach can distinguish people with color-vision deficits from those with normal color vision; and this metamer-identification process can be automated.Significance.This new method has numerous potential clinical, scientific, and industrial applications.

Improving Cancer Screening Rates in Primary Care via Practice Facilitation and Academic Detailing: A Multi-PBRN Quality Improvement Project.

PURPOSE: In the United States, cancer screening rates are often below national targets. This project implemented practice facilitation and academic detailing aimed at increasing breast, cervical, and colorectal cancer screening rates in safety-net primary care practices. METHODS: Three practice-based research networks across western and central New York State partnered to provide quality improvement strategies on breast, cervical, and colorectal cancer screening. Pre/postintervention screening rates for all participating practices were collected annually, as were means across all practices over 7 years. Simple ordinary least squares linear regression was used to calculate the trend for each cancer type and test for statistical significance (ie, P≤0.05), using the ordinal time point as a fixed effect. RESULTS: An overall increase in mean screening rates was seen over the duration of this project for colorectal (24.6% preintervention to 48.0% in year 7 of intervention; P<0.001) and breast cancer (37.0% preintervention to 48.6% in year 7; P=0.460). Mean cervical cancer screening rates decreased (35.5% preintervention to 31.4% in year 7; P=0.209). Success in increasing screening rates varied across regions of New York State. CONCLUSIONS: Practice facilitation and academic detailing were successful in significantly increasing, on average, colorectal cancer screening rate. Cervical cancer screening showed an overall decrease, likely due to difficulties for primary care practices in tracking and implementation, as many patients seek this service at outside gynecology facilities. Regional differences, guideline changes, and practice reorganization each may have played a part in observed trends. A standardization of queries being used to pull screening rates is an important step in increasing the reliability of these data.

Increasing Breast, Cervical, and Colorectal Cancer Screenings: A Qualitative Assessment of Barriers and Promoters in Safety-Net Practices.

PURPOSE: Breast, cervical, and colorectal cancer screening rates are suboptimal in underserved populations. A 7-year quality improvement (QI) project implemented academic detailing and practice facilitation in safety-net primary care practices to increase cancer screening rates. This manuscript assesses barriers and promoters. METHODS: Primary care practices providing care to underserved patients were recruited in New York cities Buffalo, Rochester, and Syracuse. Enrollment totaled 31 practices, with 12 practices participating throughout. Annually, each practice received 6 months of practice facilitation support for development and implementation of evidence-based interventions to increase screening rates for the three cancer types. At the end of each practice facilitation period, focus groups and key informant interviews were conducted with participating personnel. Content analysis was performed annually to identify barriers and promoters. A comprehensive final analysis was performed at project end. RESULTS: Barriers included system-level (inconsistent communication with specialists, electronic health record system transitions, ownership changes) and practice-level challenges (staff turnover, inconsistent data entry, QI fatigue) that compound patient-level challenges of transportation, cost, and health literacy. Cyclical barriers like staff turnover returned despite attempts to resolve them, while successful implementation was promoted by reducing patients' structural barriers, adapting interventions to existing practice priorities, and enacting officewide policies. During the QI project, practices became aware of the impact of social determinants of health on patients' screening decisions. CONCLUSIONS: The project's longitudinal design enabled identification of key barriers that reduced accuracy of practices' screening rates and increased risk of patients falling through the cracks. Identified promoters can help sustain interventions to increase screenings.

Impact of COVID-19 on Screening Rates for Colorectal, Breast, and Cervical Cancer: Practice Feedback From a Quality Improvement Project in Primary Care.

PURPOSE: Three New York State practice-based research networks provided quality improvement strategies to improve screening rates for breast, cervical, and colorectal (BCC) cancers in safety-net primary care, over 7 years. In the final year (Y7), the United States experienced the COVID-19 pandemic. The impact of the COVID-19 pandemic on BCC cancer screening rates was assessed qualitatively. METHODS: A total of 12 primary care practices participated in Y7 of the quality improvement project. BCC cancer screening rates at year beginning and end were assessed. Practice staff were asked about how COVID-19 impacted screening. Average pre/postintervention screening rates and qualitative thematic analysis regarding how COVID-19 impacted cancer screening were ascertained. RESULTS: In Y7, there was an increase in breast cancer and a decrease in colorectal and cervical cancer screening rates compared to the previous project year. Many practices were able to continue pre-COVID-19 cancer screening processes. Overall, practices reported loss of staff, changes in data entry, and a shift from preventive screening to care of sick patients. Telehealth was vital for practices to continue serving patients but had a less positive impact on patients with financial/technological disadvantages. BCC cancer screenings were impacted at various levels. CONCLUSIONS: The COVID-19 pandemic negatively impacted primary care practice cancer screening; however, some practices were able to mitigate effects by shifting focus to processes supporting screening outside of in-person office visits.

Accumulation and Competition Amongst Deformed Wing Virus Genotypes in Naïve Australian Honeybees Provides Insight Into the Increasing Global Prevalence of Genotype B.

Honeybee colony deaths are often attributed to the ectoparasitic mite Varroa destructor and deformed wing virus (DWV), vectored by the mite. In the presence of V. destructor both main genotypes (DWV-A and DWV-B) have been correlated with colony loss. Studies show that DWV-B is the most prevalent genotype in the United Kingdom and Europe. More recently DWV-B has increased in prevalence in the United States. The increasing prevalence of DWV-B at the expense of DWV-A suggests that competition exists between the genotypes. Competition may be due to disparities in virulence between genotypes, differences in fitness, such as rate of replication, or a combination of factors. In this study we investigated if DWV genotypes differ in their rate of accumulation in Australian honeybees naïve to both V. destructor and DWV, and if viral load was associated with mortality in honeybee pupae. We singly and co-infected pupae with DWV-A, DWV-B, and a recombinant strain isolated from a V. destructor tolerant bee population. We monitored viral accumulation throughout pupation, up to 192 h post-injection. We found significant differences in accumulation, where DWV-A accumulated to significantly lower loads than DWV-B and the DWV-recombinant. We also found evidence of competition, where DWV-B loads were significantly reduced in the presence of DWV-A, but still accumulated to the highest loads overall. In contrast to previous studies, we found significant differences in virulence between pupae injected with DWV-A and DWV-B. The average mortality associated with DWV-B (0.4% ± 0.33 SE) and DWV-recombinant (2.2% ± 0.83 SE) injection were significantly less than observed for DWV-A (11% ± 1.2 SE). Our results suggest that a higher proportion of DWV-B infected pupae will emerge into adults, compared to DWV-A. Overall, our data suggest that low mortality in pupae and the ability of DWV-B to accumulate to higher loads relative to DWV-A even during co-infection may favor vector transmission by V. destructor, and may thus be contributing factors to the increasing prevalence of DWV-B globally.

Project RedDE: Cluster Randomized Trial to Reduce Missed or Delayed Abnormal Laboratory Value Actions.

Failure of timely abnormal laboratory result follow-up is relatively common and may lead to harm. This study hypothesized that a quality improvement collaborative (QIC) could reduce the frequency of missed or delayed action on abnormal laboratory values. METHODS: A national cohort of pediatric practices was cluster-randomized to sequentially receive a QIC intervention: video conferences, transparent data sharing, a "focus on failures," QI coaching, and tools to help reduce missed or delayed action on abnormal laboratory values. Practices recorded the percentage of patients with 5 specific abnormal laboratory values who received an appropriate provider action (control), and then, during an 8-month intervention phase, implemented QI strategies to reduce errors (intervention). Subsequently, practices collected data on laboratory errors while working to reduce unrelated second (sustain phase), and third (maintenance phase) errors. Generalized mixed-effects regression models compared the mean percentage of patients with appropriate actions. RESULTS: We randomized 43 practices, of which 31 were included in analyses. Control and intervention phases included 1,357 and 1,426 patients with abnormal laboratory values, respectively. The mean percentage of patients who received appropriate actions did not change comparing control and intervention phases [risk difference (RD) 1%; 95% CI -1%, 3%]. In post-hoc analyses, practices significantly improved comparing control to sustain (RD 3%; 95% CI 0.3%, 6%) and maintenance phases (RD 6%; 95% CI 3%, 9%). CONCLUSION: Implementation of a QIC did not reduce the frequency of abnormal laboratory errors in the initial 8-month intervention phase. A significant reduction was appreciated comparing sustain and maintenance phases (months 9-24) to the control phase.

Diagnostic Errors in Primary Care Pediatrics: Project RedDE.

OBJECTIVE: Diagnostic errors (DEs), which encompass failures of accuracy, timeliness, or patient communication, cause appreciable morbidity but are understudied in pediatrics. Pediatricians have expressed interest in reducing high-frequency/subacute DEs, but their epidemiology remains unknown. The objective of this study was to investigate the frequency of two high-frequency/subacute DEs and one missed opportunity for diagnosis (MOD) in primary care pediatrics. METHODS: As part of a national quality improvement collaborative, 25 primary care pediatric practices were randomized to collect 5 months of retrospective data on one DE or MOD: elevated blood pressure (BP) and abnormal laboratory values (DEs), or adolescent depression evaluation (MOD). Relationships between DE or MOD proportions and patient age, gender, and insurance status were explored with mixed-effects logistic regression models. RESULTS: DE or MOD rates in pediatric primary care were found to be 54% for patients with elevated BP (n = 389), 11% for patients with abnormal laboratory values (n = 381), and 62% for adolescents with an opportunity to evaluate for depression (n = 400). When examining the number of times a pediatrician may have recognized an abnormal condition but either knowingly or unknowingly did not act according to recommended guidelines, providers did not document recognition of an elevated BP in 51% of patients with elevated BP, and they did not document recognition of an abnormal laboratory value without a delay in 9% of patients with abnormal laboratory values. CONCLUSIONS: DEs and MODs occur at an appreciable frequency in pediatric primary care. These errors may contribute to care delays and patient harm.

Synthetic lethality of retinoblastoma mutant cells in the Drosophila eye by mutation of a novel peptidyl prolyl isomerase gene.

Mutations that inactivate the retinoblastoma (Rb) pathway are common in human tumors. Such mutations promote tumor growth by deregulating the G1 cell cycle checkpoint. However, uncontrolled cell cycle progression can also produce new liabilities for cell survival. To uncover such liabilities in Rb mutant cells, we performed a clonal screen in the Drosophila eye to identify second-site mutations that eliminate Rbf(-) cells, but allow Rbf(+) cells to survive. Here we report the identification of a mutation in a novel highly conserved peptidyl prolyl isomerase (PPIase) that selectively eliminates Rbf(-) cells from the Drosophila eye.

A Practice Facilitation and Academic Detailing Intervention Can Improve Cancer Screening Rates in Primary Care Safety Net Clinics.

BACKGROUND: Despite the current evidence of preventive screening effectiveness, rates of breast, cervical, and colorectal cancer in the United States fall below national targets. We evaluated the efficacy and feasibility of combining practice facilitation and academic detailing quality improvement (QI) strategies to help primary care practices increase breast, cervical, and colorectal cancer screening among patients. METHODS: Practices received a 1-hour academic detailing session addressing current cancer screening guidelines and best practices, followed by 6 months of practice facilitation to implement evidence-based interventions aimed at increasing patient screening. One-way repeated measures analysis of variance compared screening rates before and after the intervention, provider surveys, and TRANSLATE model scores. Qualitative data were gathered via participant focus groups and interviews. RESULTS: Twenty-three practices enrolled in the project: 4 federally qualified health centers, 10 practices affiliated with larger health systems, 4 physician-owned practices, 4 university hospital clinics, and 1 nonprofit clinic. Average screening rates for breast cancer increased by 13% (P = .001), and rates for colorectal cancer increased by 5.6% (P = .001). Practices implemented a mix of electronic health record data cleaning workflows, provider audits and feedback, reminder systems streamlining, and patient education and outreach interventions. Practice facilitators assisted practices in tailoring interventions to practice-specific priorities and constraints and in connecting with community resources. Practices with resource constraints benefited from the engagement of all levels of staff in the quality improvement processes and from team-based adaptations to office workflows and policies. Many practices aligned quality improvement interventions in this project with patient-centered medical home and other regulatory reporting targets. CONCLUSIONS: Combining practice facilitation and academic detailing is 1 method through which primary care practices can achieve systems-level changes to better manage patient population health.

Quantitation of Dihydroxyacetone in Australian Leptospermum Nectar via High-Performance Liquid Chromatography.

The nonperoxide antibacterial activity of New Zealand ma̅nuka honey originates from dihydroxyacetone (DHA) within Leptospermum scoparium nectar. This study determined if DHA was present within the nectar of four Australian Leptospermum species: L. laevigatum, L. polygalifolium, L. trinervium, and L. whitei. A rapid and convenient new method was developed, which quantitated DHA/sugar ratios (ppm). The DHA and sugars were derivatized with o-(2,3,4,5,6-pentafluorobenzyl) hydroxylamine hydrochloride and analyzed via RP-HPLC with diode array detection at two wavelengths (200 and 243 nm). DHA was detected in all L. whitei and L. polygalifolium samples, where DHA/sugar ratios ranged from 10169 to 24199 ppm and from 9321 to 20174 ppm, respectively. DHA was undetected in any of the L. laevigatum and L. trinervium samples, and nectar activity was <100 ppm. The results of this study have implications for the Australian beekeeping industry, as the findings indicated that not all species of Leptospermum will produce active honey.