Structural variants of the synthetic opioid fentanyl are a major threat to public health. Following an investigation showing that many derivatives are poorly detected by commercial lateral flow and related assays, we created hapten conjugate vaccines using an immunogenic virus-like particle carrier and eight synthetic fentanyl derivatives designed to mimic the structural features of several of the more dangerous analogues. Immunization of mice elicited strong antihapten humoral responses, allowing the screening of hundreds of hapten-specific hybridomas for binding strength and specificity. A panel of 13 monoclonal IgG antibodies were selected, each showing a different pattern of recognition of fentanyl structural variations, and all proving to be highly efficient at capturing parent fentanyl compounds in competition ELISA experiments. These results provide antibody reagents for assay development as well as a demonstration of the power of the immune system to create binding agents capable of both broad and specific recognition of small-molecule targets.

The need for high-affinity, SARS-CoV-2-specific monoclonal antibodies (mAbs) is critical in the face of the global COVID-19 pandemic, as such reagents can have important diagnostic, research, and therapeutic applications. Of greatest interest is the ~300 amino acid receptor binding domain (RBD) within the S1 subunit of the spike protein because of its key interaction with the human angiotensin converting enzyme 2 (hACE2) receptor present on many cell types, especially lung epithelial cells. We report here the development and functional characterization of 29 nanomolar-affinity mouse SARS-CoV-2 mAbs created by an accelerated immunization and hybridoma screening process. Differing functions, including binding of diverse protein epitopes, viral neutralization, impact on RBD-hACE2 binding, and immunohistochemical staining of infected lung tissue, were correlated with variable gene usage and sequence.Competing Interest StatementThe authors have declared no competing interest.

BACKGROUND: The cross-contamination of cell lines in culture is a persistent problem. Genetically modified L20B (Mouse) and RD (Human Rhabdomyosarcoma) cell lines are commonly used in poliovirus research, surveillance, and diagnostics. Cross-contamination between these cell lines leads to unreproducible results and unreliable surveillance data, negatively affecting public health. The gold standard method for cell authentication is Short Tandem Repeats analysis, which is time-consuming and expensive. The disadvantage of STR is limited detection of interspecies contamination. METHODS: This assay targets the mitochondrial cytochrome c oxidase subunit I (MTCO1) gene, a highly conserved and emergent DNA barcode region for detection of cross-contamination in RD and L20B cell lines. The MagNA Pure Compact instrument and ABI 7500 Fast Dx Real-time PCR systems were used for DNA extraction and to perform real-time PCR respectively. RESULTS: The newly developed assay is very sensitive with a limit of detection of 100 RD cells/1 million L20B/mL. The amplification efficiency and R(2)-value were 102.26% and 0.9969 respectively. We evaluated specificity of the assay with five human and four mouse cell lines, as well as monkey and rat cell lines. The assay showed no cross-reactivity with genomic DNA from human, mouse, rat, or monkey cell lines. The analytical sensitivity was also evaluated by spiking varying amounts of RD cells (0.001% - 10%) into L20B cells. There was no difference in C(T) values when running single-plex or duplex PCR reactions with similar experimental conditions. CONCLUSIONS: We have developed and validated a TaqMan real-time PCR assay, a sensitive method for the detection of cross-contamination of RD and L20B cell lines.

The need for high-affinity, SARS-CoV-2-specific monoclonal antibodies (mAbs) is critical in the face of the global COVID-19 pandemic, as such reagents can have important diagnostic, research, and therapeutic applications. Of greatest interest is the ~ 300 amino acid receptor binding domain (RBD) within the S1 subunit of the spike protein because of its key interaction with the human angiotensin converting enzyme 2 (hACE2) receptor present on many cell types, especially lung epithelial cells. We report here the development and functional characterization of 29 nM-affinity mouse SARS-CoV-2 mAbs created by an accelerated immunization and hybridoma screening process. Differing functions, including binding of diverse protein epitopes, viral neutralization, impact on RBD-hACE2 binding, and immunohistochemical staining of infected lung tissue, were correlated with variable gene usage and sequence.

A cluster of Achromobacter xylosoxidans, an emerging multidrug-resistant aquaphilic bacterium, was identified in three long-term-care facility residents. As Pseudomonas aeruginosa and Serratia marcescens were also present in clinical specimens, we conducted an investigation of all three water-associated species and identified P. aerguniosa and S. marcesens contamination at the facility. Sequencing analysis linked P. aeruginosa to a clinical isolate. Findings highlight the need for precautionary measures to prevent transmission of water-associated multidrug-resistant bacteria in long-term-care facilities.

Objectives: Limited research has characterized nonfatal injury/illness in Alaska's hazardous fishing industry. This study aimed to determine (a) the utility of linking datasets to conduct surveillance, and (b) injury/illness patterns during 2012-2016. Methods: Data were obtained from the Alaska Trauma Registry (ATR), Fishermen's Fund (FF), and US Coast Guard (USCG). Datasets were coded to identify patterns in injury/illness characteristics and circumstances. Probabilistic linkage methods were utilized to identify unique incidents that appeared in more than one dataset. Results: After linking datasets, 3,014 unique injury/illness cases were identified. By dataset, 2,365 cases appeared only in FF, 486 only in USCG, 110 only in ATR, 25 in ATR and FF, 15 in ATR and USCG, 10 in USCG and FF, and 3 in all datasets. FF mainly captured claims submitted by small, independently-owned vessels in Southcentral and Southeastern Alaska. In contrast, USCG mainly captured reports from large, company-owned vessels in Western Alaska. By nature, cases were most frequently sprains, strain, and tears (27%), cuts (15%), and fractures (11%). Across fleets, injuries/illnesses most frequently resulted from contact with objects and equipment (41%), overexertion and bodily reaction (27%), and slips, trips, and falls (20%). Work processes associated with traumatic injuries were most frequently hauling gear (18%) and walking, climbing, and descending (18%). Half of all injuries were of moderate severity (53%). Conclusion: Linking datasets, which capture different segments of Alaska's fishing industry, provides the most comprehensive understanding of nonfatal injury/illness to date. These results, stratified by fleet and severity, will inform prevention strategies.

Aviation operations in Alaska often occur in remote locations and during inclement weather. Limited infrastructure and staff in some locations often requires aviation workers to perform tasks outside of their specific job descriptions. Researchers identified workers' compensation claims as a valuable data source to characterise nonfatal injuries among Alaskan aviation workers. Keyword searches of injury claim narrative fields and industry codes were used to identify potentially aviation-related workers' compensation claims during 2014-2015. These claims were manually reviewed to verify whether aviation related and manually coded according to the US Bureau of Labour Statistics' Occupational Injury and Illness Classification System. There were 875 aviation-related injury claims accepted during 2014-2015. Ramp/baggage/cargo agents incurred the most injuries (35%), followed by mechanics/maintenance workers (15%). Among all workers, Overexertion and Bodily Reaction (40%) was most often cited as the injury event, followed by Contact with Objects and Equipment (28%), and Falls, Slips, Trips (22%). Sprains, strains, tears were the most frequent nature of injury (55%). Cargo/freight/luggage was the most frequent source of injury (24%). The 3 most frequently identified injury event types were responsible for over 90% of all injuries, which indicates that preventive interventions should be directed towards tasks rather than occupational groups.

Background: To gain a better understanding of nonfatal injuries in Alaska, underutilized data sources such as workers’ compensation claims must be analyzed. The purpose of the current study was to utilize workers’ compensation claims data to estimate the risk of nonfatal, work-related injuries among occupations in Alaska, characterize injury patterns, and prioritize future research. Methods: A dataset with information on all submitted claims during 2014–2015 was provided for analysis. Claims were manually reviewed and coded. For inclusion in this study, claims had to represent incidents that resulted in a nonfatal acute traumatic injury, occurred in Alaska during 2014–2015, and were approved for compensation. Results: Construction workers had the highest number of injuries (2,220), but a rate lower than the overall rate (34 per 1,000 construction workers, compared to 40 per 1,000 workers overall). Fire fighters had the highest rate of injuries on the job, with 162 injuries per 1,000 workers, followed by law enforcement officers with 121 injuries per 1,000 workers. The most common types of injuries across all occupations were sprains/strains/tears, contusions, and lacerations. Conclusion: The successful use of Alaska workers’ compensation data demonstrates that the information provided in the claims dataset is meaningful for epidemiologic research. The predominance of sprains, strains, and tears among all occupations in Alaska indicates that ergonomic interventions to prevent overexertion are needed. These findings will be used to promote and guide future injury prevention research and interventions.