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Frequent and widespread testing of members of the population who are asymptomatic for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is essential for the mitigation of the transmission of the virus. Despite the recent increases in testing capacity, tests based on quantitative polymerase chain reaction (qPCR) assays cannot be easily deployed at the scale required for population-wide screening. Here, we show that next-generation sequencing of pooled samples tagged with sample-specific molecular barcodes enables the testing of thousands of nasal or saliva samples for SARS-CoV-2 RNA in a single run without the need for RNA extraction. The assay, which we named SwabSeq, incorporates a synthetic RNA standard that facilitates end-point quantification and the calling of true negatives, and that reduces the requirements for automation, purification and sample-to-sample normalization. We used SwabSeq to perform 80,000 tests, with an analytical sensitivity and specificity comparable to or better than traditional qPCR tests, in less than two months with turnaround times of less than 24 h. SwabSeq could be rapidly adapted for the detection of other pathogens.

ABSTRACT The rapid spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is due to the high rates of transmission by individuals who are asymptomatic at the time of transmission 1, 2 . Frequent, widespread testing of the asymptomatic population for SARS-CoV-2 is essential to suppress viral transmission. Despite increases in testing capacity, multiple challenges remain in deploying traditional reverse transcription and quantitative PCR (RT-qPCR) tests at the scale required for population screening of asymptomatic individuals. We have developed SwabSeq, a high-throughput testing platform for SARS-CoV-2 that uses next-generation sequencing as a readout. SwabSeq employs sample-specific molecular barcodes to enable thousands of samples to be combined and simultaneously analyzed for the presence or absence of SARS-CoV-2 in a single run. Importantly, SwabSeq incorporates an in vitro RNA standard that mimics the viral amplicon, but can be distinguished by sequencing. This standard allows for end-point rather than quantitative PCR, improves quantitation, reduces requirements for automation and sample-to-sample normalization, enables purification-free detection, and gives better ability to call true negatives. After setting up SwabSeq in a high-complexity CLIA laboratory, we performed more than 80,000 tests for COVID-19 in less than two months, confirming in a real world setting that SwabSeq inexpensively delivers highly sensitive and specific results at scale, with a turn-around of less than 24 hours. Our clinical laboratory uses SwabSeq to test both nasal and saliva samples without RNA extraction, while maintaining analytical sensitivity comparable to or better than traditional RT-qPCR tests. Moving forward, SwabSeq can rapidly scale up testing to mitigate devastating spread of novel pathogens.

"Helping General Physical Educators and Adapted Physical Educators Address the Office of Civil Rights Dear Colleague Guidance Letter: Part IV--Sport Groups" provides the the following articles: (1) "Sport Programming Offered by Camp Abilities and the United States Association for Blind Athletes" (Lauren Lieberman and Mark Lucas) describes Camp Abilities, and the United States Association for Blind Athletes (USABA) Sports Camps, These two organizations often combine to provide sport opportunities that should be considered by local school districts for addressing extracurricular athletics and the Office of Civil Rights (OCR) Dear Colleague Letter. (2) "BlazeSports America: Addressing OCR's Guidance for Sport Opportunities for Students with Disabilities" (Jeffery Jones, Dan Humphreys, and Ann Cody) describes the legacy program of the 1996 Atlanta Paralympic Games. BlazeSports America accomplishes its mission to change the lives of children and adults with physical disabilities through sport through two distinctive pathways (a) community-based sport and recreation programs for children and adults with physical disabilities; and (b) professional development. Professional development is accomplished through training front -line professionals working in sport, physical education and physical activity,and recreation and fitness settings. (3) "Building an Adapted Sport Infrastructure Guided by the American Association of Adapted Sports Programs" (Bev Vaughn and Tommie Storms) alerts the reader to the resources available through the American Association of Adapted Sports Programs (AAASP). The AAASP seeks to assure that students with physical disabilities can have the same or similar benefits of athletic participation as their non-disabled peers. All of the articles presented in Part IV of this feature share information from leading adapted sport groups to serve as resources for inservices or for the creation of infrastructures and events in local school districts.

The purpose of this study was to investigate the factors that influence adolescent males to enroll in school choir as an elective class and to assess their attitudes about singing in general, self-concept of their own voices, and perception of others' view of adolescent males' participation in choir. Data were obtained from 101 adolescent males who were enrolled in choir at one of six participating schools, and were in Grade 7 or Grade 8. Data analysis showed that most participants enrolled in choir because they thought it was fun and/or they were good singers. Results also indicated that peer pressure factored less than the enjoyment of singing in influencing their decision to enroll in choir. Other results suggested that the participants perceived support from their families, principals, and nonmusic teachers in their decision to take choir and that they perceived slightly less support from coaches at their school. The study indicates a need for future research of adolescent males who have chosen not to participate in choir and their attitudes about choir participation. Future research studying the attitudes of adolescent males from other regions and their attitudes about choir participation is also recommended. (Contains 5 tables.)

Although the bundle-forming pilus (BFP) of enteropathogenic Escherichia coli (EPEC) mediates microcolony formation on epithelial cells, the adherence of BFP-deficient mutants is significantly abrogated, but the mutants are still adherent due to the presence of intimin and possibly other adhesins. In this study we investigated the contribution of the recently described E. coli common pilus (ECP) to the overall adherence properties of EPEC. We found that ECP and BFP structures can be simultaneously observed in the course (between zero time and 7 h during infection) of formation of localized adherence on cultured epithelial cells. These two pilus types colocalized at different levels of the microcolony topology, tethering the adhering bacteria. No evidence of BFP disappearance was found after prolonged infection. When expressed from a plasmid present in nonadherent E. coli HB101, ECP rendered this organism highly adherent at levels comparable to those of HB101 expressing the BFP. Purified ECP bound in a dose-dependent manner to epithelial cells, and the binding was blocked with anti-ECP antibodies, confirming that the pili possess adhesin properties. An ECP mutant showed only a modest reduction in adherence to cultured cells due to background expression levels of BFP and intimin. However, isogenic mutants not expressing EspA or BFP were significantly less adherent when the ecpA gene was also deleted. Furthermore, a [DELTA]espA [DELTA]ecpA double mutant (unable to translocate Tir and to establish intimate adhesion) was at least 10-fold less adherent than the [DELTA]espA and [DELTA]ecpA single mutants, even in the presence of BFP. A [DELTA]bfp [DELTA]espA [DELTA]ecpA triple mutant showed the least adherence compared to the wild type and all the isogenic mutant strains tested, suggesting that ECP plays a synergistic role in adherence. Our data indicate that ECP is an accessory factor that, in association with BFP and other adhesins, contributes to the multifactorial complex interaction of EPEC with host epithelial cells.

Socket grafting or immediate implant placement are established treatment pathways for management of tooth extraction sites. Immediate implant placement reduces treatment time, patient discomfort, and cost for implant-supported restorations. Clinicians utilize various material, technology, and technique approaches to maximize functional, esthetic, and longevity outcomes. This article, in which two case reports are illustrated, describes the use of minimally invasive immediate implant treatment using dynamic navigation, bone grafting, and dehydrated deepithelialized human amnion/chorion membrane (dHACM). dHACM is an important adjunct when performing minimally invasive grafting for implant procedures, as it possesses signaling proteins that can facilitate wound healing and regulate inflammation and pain. Considerations and implications for this treatment approach and material selection are presented. [ABSTRACT FROM AUTHOR]