Strengthening Bordetella pertussis genomic surveillance by direct sequencing of residual positive specimens.

Whole-genome sequencing (WGS) of microbial pathogens recovered from patients with infectious disease facilitates high-resolution strain characterization and molecular epidemiology. However, increasing reliance on culture-independent methods to diagnose infectious diseases has resulted in few isolates available for WGS. Here, we report a novel culture-independent approach to genome characterization of Bordetella pertussis, the causative agent of pertussis and a paradigm for insufficient genomic surveillance due to limited culture of clinical isolates. Sequencing libraries constructed directly from residual pertussis-positive diagnostic nasopharyngeal specimens were hybridized with biotinylated RNA "baits" targeting B. pertussis fragments within complex mixtures that contained high concentrations of host and microbial background DNA. Recovery of B. pertussis genome sequence data was evaluated with mock and pooled negative clinical specimens spiked with reducing concentrations of either purified DNA or inactivated cells. Targeted enrichment increased the yield of B. pertussis sequencing reads up to 90% while simultaneously decreasing host reads to less than 10%. Filtered sequencing reads provided sufficient genome coverage to perform characterization via whole-genome single nucleotide polymorphisms and whole-genome multilocus sequencing typing. Moreover, these data were concordant with sequenced isolates recovered from the same specimens such that phylogenetic reconstructions from either consistently clustered the same putatively linked cases. The optimized protocol is suitable for nasopharyngeal specimens with diagnostic IS481 Ct < 35 and >10 ng DNA. Routine implementation of these methods could strengthen surveillance and study of pertussis resurgence by capturing additional cases with genomic characterization.

Interventions for Instrumental Activities of Daily Living Among Adults With Multiple Sclerosis: A Systematic Review.

IMPORTANCE: Occupational therapy practitioners need updated information about interventions that may improve or maintain functional changes in instrumental activity of daily living (IADL) engagement caused by multiple sclerosis (MS). OBJECTIVE: To conduct a narrative synthesis of updated evidence on interventions within the scope of occupational therapy to improve or maintain performance of and participation in IADLs among adults with MS. DATA SOURCES: CINAHL, MEDLINE in PubMed, Cochrane, OTseeker, and PsycINFO. Study Selection and Data Collection: This systematic review followed the Cochrane Collaboration methodology and is reported according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines for conducting a systematic review. Inclusion criteria were Level 1 or 2 evidence, published in English, published from January 2011 to December 2018, intervention within the occupational therapy scope of practice, and at least one IADL outcome measure. FINDINGS: Nineteen randomized controlled trials (including pilot and feasibility trials) and 1 preintervention-postintervention study met inclusion criteria. Results of this review show strong strength of evidence for coaching interventions in addressing physical activity (PA) routines and moderate support in addressing medication routines. Moderate strength of evidence was found with mixed results for interventions involving coaching plus prescribed PA in improving PA participation. CONCLUSIONS AND RELEVANCE: This systematic review supports occupational therapy practitioners addressing PA and medication health management and maintenance IADLs through the use of coaching interventions when treating people with MS. Other IADLs were addressed by the articles in this review but require more evidence to make clinical recommendations. What This Article Adds: Occupational therapy practitioners' skills in promoting habits and routines paired with utilization of evidence-supported coaching interventions can support independence with health management and reduce the negative impact of MS on daily activity participation.

Occupational Therapy Practice Guidelines for Adults With Multiple Sclerosis.

IMPORTANCE: Occupational therapy practitioners play an important role in addressing the occupational performance and participation needs of adults with multiple sclerosis (MS) and their caregivers. OBJECTIVE: This Practice Guideline aims to help occupational therapy practitioners, as well as the people who manage, reimburse, or set policy regarding occupational therapy services, understand occupational therapy's role in providing services to adults with MS and their caregivers. This guideline can also serve as a reference for health care professionals, health care managers, educators, regulators, third-party payers, managed care organizations, and researchers. METHOD: We examined, synthesized, and integrated the results of four systematic reviews into clinical recommendations. Three systematic reviews specified occupational domains as outcomes of interest, and one focused on interventions for caregivers of people with MS. RESULTS: Twenty-one articles from the systematic reviews with strong or moderate strength of evidence served as the basis for the clinical recommendations. CONCLUSIONS AND RECOMMENDATIONS: Four interventions benefited participation in the domains of activities of daily living or sleep. Two interventions improved participation in the domain of instrumental activities of daily living. One intervention improved work participation. Two interventions benefited caregivers and the quality of the relationship between the caregiver and person with MS. Other potentially appropriate interventions or areas to address on the basis of existing or emerging evidence are discussed in the Limitations: Gaps in the Evidence and Conclusions sections. What This Article Adds: This Practice Guideline provides a summary and application of the current evidence supporting occupational therapy intervention for adults with MS. It includes case examples and decision-making algorithms to support occupational therapy practitioners in addressing client goals.

Detection and Characterization of Diphtheria Toxin Gene-Bearing Corynebacterium Species through a New Real-Time PCR Assay.

Respiratory diphtheria, characterized by a firmly adherent pseudomembrane, is caused by toxin-producing strains of Corynebacterium diphtheriae, with similar illness produced occasionally by toxigenic Corynebacterium ulcerans or, rarely, Corynebacterium pseudotuberculosis While diphtheria laboratory confirmation requires culture methods to determine toxigenicity, real-time PCR (RT-PCR) provides a faster method to detect the toxin gene (tox). Nontoxigenic tox-bearing (NTTB) Corynebacterium isolates have been described, but impact of these isolates on the accuracy of molecular diagnostics is not well characterized. Here, we describe a new triplex RT-PCR assay to detect tox and distinguish C. diphtheriae from the closely related species C. ulcerans and C. pseudotuberculosis Analytical sensitivity and specificity of the assay were assessed in comparison to culture using 690 previously characterized microbial isolates. The new triplex assay characterized Corynebacterium isolates accurately, with 100% analytical sensitivity for all targets. Analytical specificity with isolates was 94.1%, 100%, and 99.5% for tox, Diph_rpoB, and CUP_rpoB targets, respectively. Twenty-nine NTTB Corynebacterium isolates, representing 5.9% of 494 nontoxigenic isolates tested, were detected by RT-PCR. Whole-genome sequencing of NTTB isolates revealed varied mutations putatively underlying their lack of toxin production, as well as eight isolates with no mutation in tox or the promoter region. This new Corynebacterium RT-PCR method provides a rapid tool to screen isolates and identify probable diphtheria cases directly from specimens. However, the sporadic occurrence of NTTB isolates reinforces the viewpoint that diphtheria culture diagnostics continue to provide the most accurate case confirmation.

Comparative Cost of Virtual Reality Training and Live Exercises for Training Hospital Workers for Evacuation.

Adoption of virtual reality technology may be delayed due to high up-front costs with unknown returns on that investment. In this article, we present a cost analysis of using virtual reality as a training tool. Virtual reality was used to train neonatal intensive care workers in hospital evacuation. A live disaster exercise with mannequins was also conducted that approximated the virtual experience. Comparative costs are presented for the planning, development, and implementation of both interventions. Initially, virtual reality is more expensive, with a cost of $229.79 per participant (total cost $18 617.54 per exercise) for the live drill versus $327.78 (total cost $106 951.14) for virtual reality. When development costs are extrapolated to repeated training over 3 years, however, the virtual exercise becomes less expensive with a cost of $115.43 per participant, while the cost of live exercises remains fixed. The larger initial investment in virtual reality can be spread across a large number of trainees and a longer time period with little additional cost, while each live drill requires additional costs that scale with the number of participants.

The training and practice of crew resource management: recommendations from an inductive in vivo study of the flight deck.

Crew resource management (CRM) is credited with saving 185 lives at Sioux City. While the theory behind CRM is well documented, there are few studies of how CRM manifests on the line. This inductive in vivo study had three objectives. First, to describe how CRM manifests. Secondly, to evaluate the efficacy of CRM vis-à-vis flight safety. Thirdly, to suggest improvements to the CRM training syllabus. The study produced five conclusions: First, CRM is durable under conditions of moderate strain. Secondly, crews embed and refine CRM through reflection and action. Thirdly, CRM facilitates and shapes social relations. Fourthly, mindlessness (Langer 1989 ) undermines CRM. Finally, the interruption of flight-deck routines by third-parties poses a threat to flight-safety. The paper recommends multi-profession CRM training as a means of improving communication and co-ordination in and around aircraft. The study's limitations include a monocultural flight-deck: flights were operated by pilots with European backgrounds. Mindful of Hofstede's ( 1980 ), Engle's ( 2000 ) and Helmreich and Merritt's ( 2001 ) examination of the relationship between culture and performance, the author suggests the study be repeated with carriers that employ pilots from a variety of cultures. Practitioner Summary: This in vivo study evaluates the efficacy of CRM vis-à-vis flight safety and supports a critique of the CRM syllabus. The author observed twenty sectors and attended a CRM training day. To improve safety and efficiency, it is recommended that airlines run multi-profession (inclusive) CRM training courses.

Multidisciplinary Team Treatment Approaches to Chronic Daily Headaches.

OBJECTIVE: In this review, we focus on nonmedication treatment approaches to chronic daily headaches and chronic migraine. We review the current scientific data on studies using multimodal treatments, especially physical therapy and occupational therapy, and provide recommendations on the formation of interdisciplinary headache teams. BACKGROUND: Chronic daily headache, which includes chronic migraine, is a particularly challenging clinical entity which often involves multiple headache types and comorbidities. A team approach in treating these patients may be particularly useful. DESIGN/METHODS: We review all current studies performed with at least one or more other modality in addition to usual medical treatment, with a focus on physical and occupational therapy. Emphasis on physical and occupational therapy with an explanation of their methods and role in multidisciplinary treatment is a pivotal part of this review. We also suggest approaches to setting up a multimodality clinic for the busy headache clinician. CONCLUSION: Setting up a collaborative, multidisciplinary team of specialists in headache practices with the goal of modifying physical, environmental, and psychological triggers for chronic daily headaches may facilitate treatment of these refractory patients.

Lifestyle Redesign® for Chronic Pain Management: A Retrospective Clinical Efficacy Study.

OBJECTIVE: Our objective was to determine the efficacy of a Lifestyle Redesign® intervention for people living with chronic pain on quality of life (QOL), function, self-efficacy, and pain levels. METHOD: Clinical outcomes were collected from 45 patients who completed an individual outpatient Lifestyle Redesign occupational therapy program for chronic pain as part of their usual plan of medical care. Outcome measures included the Canadian Occupational Performance Measure, the 36-Item Short-Form Survey, the Brief Pain Inventory, and the Pain Self-Efficacy Questionnaire. We analyzed scores using paired-samples t tests. RESULTS: Significant changes were observed in occupational performance and satisfaction scores, physical and social functioning, role limitations due to physical and emotional problems, energy and fatigue, general health, and pain self-efficacy. CONCLUSION: Lifestyle Redesign interventions, when integrated into a patient's medical plan of care, can improve patient functioning, self-efficacy, and QOL.

Effectiveness of positive pressure ventilation during newborn care unit evacuation.

OBJECTIVE: Assess the utility of high fidelity simulation in understanding effectiveness of bag-valve ventilation in a simulated newborn intensive care unit vertical evacuation. PARTICIPANTS: A total of 70 participants, (13 teams of 4-6 staff) including physicians, nurses, respiratory therapists and other support personnel participated in a 90-min evacuation sessions. METHODS: Two wireless high-fidelity newborn mannequins (Gaumand ScientificR) provided real-time data of ventilation support during a NICU evacuation exercise. Trained evaluators also recorded data related to performance. Following the exercises, the simulator data were downloaded and analyzed for rate and consistency of respirations. RESULTS: Using the data from the simulators and evaluator comments, it was found the infants received proper airway management during the evacuation only 58% of the time. This study highlights the need for ongoing training for NICU staff around safe, effective, coordinated, and timely care of these fragile newborns in the event of an evacuation.

shinyMBA: a novel R shiny application for quality control of the multiplex bead assay for serosurveillance studies.

The multiplex bead assay (MBA) based on Luminex xMAP technology can be used as a tool to measure seroprevalence as part of population immunity evaluations to multiple antigens in large-scale serosurveys. However, multiplexing several antigens presents challenges for quality control (QC) assessments of the data because multiple parameters must be evaluated for each antigen. MBA QC parameters include monitoring bead counts and median fluorescence intensity (MFI) for each antigen in plate wells, and performance of assay controls included on each plate. Analyzing these large datasets to identify plates failing QC standards presents challenges for many laboratories. We developed a novel R Shiny application, shinyMBA, to expedite the MBA QC processes and reduce the risk of user error. The app allows users to rapidly merge multi-plate assay outputs to evaluate bead count, MFI, and performance of assay controls using statistical process control charts for all antigen targets simultaneously. The utility of the shinyMBA application and its various outputs are demonstrated using data from 32 synthetic xPONENT files with 3 multiplex antigens and two population serosurveillance studies that evaluated 1200 and 3871 samples, respectively, for 20 multiplexed antigens. The shinyMBA open-source code is available for download and modification at https://github.com/CDCgov/shinyMBA . Incorporation of shinyMBA into Luminex serosurveillance workflows can vastly improve the speed and accuracy of QC processes.

Evaluation of Asymptomatic Bordetella Carriage in a Convenience Sample of Children and Adolescents in Atlanta, Georgia, United States.

Few data exist on asymptomatic carriage of Bordetella species among populations receiving acellular pertussis vaccine. We conducted a cross-sectional study among acellular-vaccinated children presenting to an emergency department (ED). Bordetella pertussis carriage prevalence was <1% in this population, a lower prevalence than that found in recent studies among whole-cell pertussis-vaccinated participants.

Comparison of Bordetella species identification among differing rt-PCR assays in the United States.

In the United States, the general laboratory method for diagnosing pertussis, caused by Bordetella pertussis, is real-time PCR (rt-PCR) targeting insertion sequence 481 (IS481). Other Bordetella species (parapertussis, holmesii, and bronchiseptica) can also cause a pertussis-like syndrome, and some commercial laboratory assays include the insertion sequence 1001 (pIS1001) that can detect B. parapertussis/B. bronchiseptica (BppBb). Because IS481 exists in B. pertussis and B. holmesii, current commercial assays cannot differentiate these two species. We used a multiplex rt-PCR assay containing species-specific targets to Bordetella to evaluate clinical specimens detected as B. pertussis/B. holmesii (BpBh) or BppBb by commercial laboratories. A sample of 3,984 clinical specimens positive for IS481 or pIS1001 from two commercial laboratories during 2012-2019 were re-tested at CDC. Agreement of Bordetella species between the CDC and commercial laboratory assays, and the proportion of commercial laboratory specimens that were non-B. pertussis by CDC's assay was assessed. Overall agreement in Bordetella species detection and identification between the CDC and commercial lab assays was 85%. Agreement for identifying B. pertussis was 87% for 3,663 BpBh specimens and 98% for identifying B. parapertussis in 310 BppBb specimens. CDC's assay detected B. holmesii in 55/3,984 (1.4%) specimens. Most discrepant results (410/490, 82%) were BpBh specimens interpreted as indeterminate B. pertussis at CDC. We found a small portion of B. holmesii in a sample of IS481-positive clinical specimens originally identified by commercial laboratory rt-PCR assays, suggesting that commercial PCR assays are a reliable diagnostic tool for correctly identifying Bordetella species in most patients with suspected pertussis. IMPORTANCE: When testing specimens collected from patients with suspected pertussis, large-scale commercial laboratories in the United States employ an IS481-based assay that cannot differentiate between Bordetella pertussis and Bordetella holmseii. The level of B. holmesii causing pertussis-like illness in the United States is not well-understood given that only B. pertussis is nationally notifiable. After re-testing with a multiplex, species-specific rt-PCR assay, our data show low levels of B. holmesii identified in a sample of IS481-positive clinical specimens originally identified by commercial laboratory rt-PCR assays. These results reinforce the validity of large-scale commercial rt-PCR testing as a reliable diagnostic tool for pertussis in the United States.

Effects of Virtual Reality Simulation on Worker Emergency Evacuation of Neonates.

OBJECTIVE: This study examined differences in learning outcomes among newborn intensive care unit (NICU) workers who underwent virtual reality simulation (VRS) emergency evacuation training versus those who received web-based clinical updates (CU). Learning outcomes included a) knowledge gained, b) confidence with evacuation, and c) performance in a live evacuation exercise. METHODS: A longitudinal, mixed-method, quasi-experimental design was implemented utilizing a sample of NICU workers randomly assigned to VRS training or CUs. Four VRS scenarios were created that augmented neonate evacuation training materials. Learning was measured using cognitive assessments, self-efficacy questionnaire (baseline, 0, 4, 8, 12 months), and performance in a live drill (baseline, 12 months). Data were collected following training and analyzed using mixed model analysis. Focus groups captured VRS participant experiences. RESULTS: The VRS and CU groups did not statistically differ based upon the scores on the Cognitive Assessment or perceived self-efficacy. The virtual reality group performance in the live exercise was statistically (P<.0001) and clinically (effect size of 1.71) better than that of the CU group. CONCLUSIONS: Training using VRS is effective in promoting positive performance outcomes and should be included as a method for disaster training. VRS can allow an organization to train, test, and identify gaps in current emergency operation plans. In the unique case of disasters, which are low-volume and high-risk events, the participant can have access to an environment without endangering themselves or clients. (Disaster Med Public Health Preparedness. 2019;13:301-308).

KDM1A/LSD1 regulates the differentiation and maintenance of spermatogonia in mice.

The proper regulation of spermatogenesis is crucial to ensure the continued production of sperm and fertility. Here, we investigated the function of the H3K4me2 demethylase KDM1A/LSD1 during spermatogenesis in developing and adult mice. Conditional deletion of Kdm1a in the testis just prior to birth leads to fewer spermatogonia and germ cell loss before 3 weeks of age. These results demonstrate that KDM1A is required for spermatogonial differentiation, as well as germ cell survival, in the developing testis. In addition, inducible deletion of Kdm1a in the adult testis results in the abnormal accumulation of meiotic spermatocytes, as well as apoptosis and progressive germ cell loss. These results demonstrate that KDM1A is also required during adult spermatogenesis. Furthermore, without KDM1A, the stem cell factor OCT4 is ectopically maintained in differentiating germ cells. This requirement for KDM1A is similar to what has been observed in other stem cell populations, suggesting a common function. Taken together, we propose that KDM1A is a key regulator of spermatogenesis and germ cell maintenance in the mouse.

Evacuation performance evaluation tool.

OBJECTIVE: Hospitals conduct evacuation exercises to improve performance during emergency events. An essential aspect in this process is the creation of reliable and valid evaluation tools. The objective of this article is to describe the development and implications of a disaster evacuation performance tool that measures one portion of the very complex process of evacuation. DESIGN: Through the application of the Delphi technique and DeVellis's framework, disaster and neonatal experts provided input in developing this performance evaluation tool. Following development, content validity and reliability of this tool were assessed. SETTING: Large pediatric hospital and medical center in the Midwest. PARTICIPANTS: The tool was pilot tested with an administrative, medical, and nursing leadership group and then implemented with a group of 68 healthcare workers during a disaster exercise of a neonatal intensive care unit (NICU). RESULTS: The tool has demonstrated high content validity with a scale validity index of 0.979 and inter-rater reliability G coefficient (0.984, 95% CI: 0.948-0.9952). CONCLUSIONS: The Delphi process based on the conceptual framework of DeVellis yielded a psychometrically sound evacuation performance evaluation tool for a NICU.

Maternally provided LSD1/KDM1A enables the maternal-to-zygotic transition and prevents defects that manifest postnatally.

Somatic cell nuclear transfer has established that the oocyte contains maternal factors with epigenetic reprogramming capacity. Yet the identity and function of these maternal factors during the gamete to embryo transition remains poorly understood. In C. elegans, LSD1/KDM1A enables this transition by removing H3K4me2 and preventing the transgenerational inheritance of transcription patterns. Here we show that loss of maternal LSD1/KDM1A in mice results in embryonic arrest at the 1-2 cell stage, with arrested embryos failing to undergo the maternal-to-zygotic transition. This suggests that LSD1/KDM1A maternal reprogramming is conserved. Moreover, partial loss of maternal LSD1/KDM1A results in striking phenotypes weeks after fertilization; including perinatal lethality and abnormal behavior in surviving adults. These maternal effect hypomorphic phenotypes are associated with alterations in DNA methylation and expression at imprinted genes. These results establish a novel mammalian paradigm where defects in early epigenetic reprogramming can lead to defects that manifest later in development.

A collaborative outcome resource network (ACORN): Tools for increasing the value of psychotherapy.

The authors describe a collaborative outcomes resource network (ACORN) and the suite of measurement and decision support tools (ACORN Toolkit) that have emerged from this collaboration for the purpose of providing clinical feedback to therapists. The ACORN Toolkit is most accurately described as a comprehensive clinical information system designed to increase the value of mental health services across large systems of care. It was built to integrate large datasets from multiple sources including outcome data, client demographics and diagnostic data, therapist credentialing information, pharmacy data, and service claims data. For the limited purposes of this article, the authors focus on the ACORN Toolkit for measuring and how it has contributed to improving outcomes in psychotherapy. Implications to current practice and future training are provided.