[{"command":"settings","settings":{"pluralDelimiter":"\u0003","suppressDeprecationErrors":true,"entitySetting":{"type":"bibcite_reference","bundle":"journal_article","mapping":{"node":{"blog":"blog","class":"classes","events":"calendar","faq":"faq","link":"links","news":"news","page":"","person":"people","presentation":"presentations","software_project":"software","software_release":"software"},"bibcite_reference":{"*":"publications"},"paragraph":{"class_material":"classes"}},"viewmode":"teaser"},"user":{"uid":0,"permissionsHash":"d2bc148dcc39b09733b00df53ed69c94d6c4af2d41c2517e0f84d66722ecd3c8"}},"merge":true},{"command":"add_js","selector":"body","data":[{"src":"\/files\/js\/js_tyCxUflIoByHJZq7vNaKY7VRk1zut-9zIJBV9tLHNrg.js?scope=footer\u0026delta=0\u0026language=en\u0026theme=eligendi_lab_bilh\u0026include=eJzLL44vKE3KyUxOLMnMzyvWTykqLUjM0ctHFdbLzSxO1ikrzixJ1U_OzytJrSgpTcxxK83JCctMLQcAsjEbVw"}]},{"command":"insert","method":"replaceWith","selector":"#","data":"\n\u003Cul  id=\u0022list-of-posts\u0022 more_link_id=\u0022node-readmore\u0022 class=\u0022publications view-teaser list-view\u0022\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EKirby, James E, and Ramy Arnaout. (2026) 2026. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/why-are-we-doing-alone-collaborative-framework-ldt-development-and-validation\u0022 hreflang=\u0022en\u0022\u003EWhy Are We Doing This Alone? A Collaborative Framework for LDT Development and Validation.\u003C\/a\u003E\u201d. \u003Ci\u003EJournal of Clinical Microbiology\u003C\/i\u003E, e0186725. https:\/\/doi.org\/10.1128\/jcm.01867-25.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/41854436\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003ELaboratory-developed tests (LDTs) play a critical role in meeting unmet diagnostic needs, particularly for rare infections and high-acuity or immunocompromised patient populations. However, current US regulatory and reimbursement frameworks have constrained innovation and delayed implementation of many essential laboratory diagnostic tests. Here, we propose a pragmatic, collaborative model focusing on infectious disease molecular diagnostics that maintains analytical rigor while allowing clinical validity to be supported by evidence from the medical literature and clinical judgment. To address the resource constraints faced by hospital laboratories, we envision a voluntary, national repository where laboratories and manufacturers contribute standardized analytical and, where appropriate, clinical validation data for both newly developed and modified Food and Drug Administration (FDA)-cleared assays. Data elements such as accuracy, limit of detection, analytical measurement range, analytical specificity, and inclusivity compared with reference determinations would be aggregated within groups representing technically identical assays, enabling cumulative validation and shared use of high-quality evidence. By leveraging these shared data sets, hospital laboratories could implement LDTs in-house with minimal redundant validation, enabling broader access to testing near the point of patient care and faster turnaround times. Manufacturers could use the same data to support applications for expanded intended use of existing FDA-cleared and -approved tests. This framework would strengthen hospital laboratory diagnostic capacity, accelerate test implementation, and improve patient outcomes and healthcare system efficiency.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EUy, Matthew T J, Andrea Kirmaier, Lindsey M Rudtner, Aidan Pine, and James E Kirby. (2026) 2026. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/poloxamer-dilution-demand-alternative-agar-dilution-based-antimicrobial-susceptibility\u0022 hreflang=\u0022en\u0022\u003EPoloxamer Dilution As an On-Demand Alternative to Agar Dilution-Based Antimicrobial Susceptibility Testing.\u003C\/a\u003E\u201d. \u003Ci\u003EJournal of Clinical Microbiology\u003C\/i\u003E, e0182225. https:\/\/doi.org\/10.1128\/jcm.01822-25.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/41784364\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003E\u003Cb\u003EUNLABELLED: \u003C\/b\u003EAgar dilution is a reference susceptibility testing method uniquely or preferentially recommended for certain antimicrobials. However, the effort required to pour individual agar plates spanning a doubling dilution range precludes its practical implementation in hospital clinical laboratories. Here, we describe an on-demand replacement for agar dilution, specifically substituting Poloxamer 407 (also known as Pluronic F-127) for Bacto agar as the solidifying agent. Notably, 20% Poloxamer 407 solutions (e.g., with Mueller-Hinton broth) remain liquid at refrigerated temperatures but solidify upon warming, enabling facile setup of poloxamer dilution testing in Petri dish or microwell format. For fosfomycin susceptibility testing, poloxamer dilution and reference agar dilution showed excellent categorical agreement (CA) and essential agreement (EA) for \u003Ci\u003EEscherichia coli\u003C\/i\u003E (100% and 87%, respectively, \u003Ci\u003En\u003C\/i\u003E = 31). For other \u003Ci\u003EEnterobacterales\u003C\/i\u003E, excluding \u003Ci\u003EKlebsiella\u003C\/i\u003E spp., CA and EA were both 82% (\u003Ci\u003En\u003C\/i\u003E = 17, respectively). For \u003Ci\u003EPseudomonas aeruginosa\u003C\/i\u003E, CA and EA were 60% and 100% (\u003Ci\u003En\u003C\/i\u003E = 10), respectively, with the lower CA reflecting the large number of strains tested with minimal inhibitory concentrations near categorical breakpoints. There were no very major errors, while major errors were only observed for \u003Ci\u003EKlebsiella\u003C\/i\u003E spp. Additionally, poloxamer dilution substantially reduced the number of skipped dilutions sixfold for \u003Ci\u003EE. coli\u003C\/i\u003E (\u003Ci\u003EP\u003C\/i\u003E \u0026lt; 0.0001) and inhibited swarming of \u003Ci\u003EProteus\u003C\/i\u003E spp. We conclude that poloxamer dilution and agar dilution, an imperfect gold standard, have essentially equivalent practical performance and that poloxamer dilution can therefore serve as an on-demand alternative testing methodology in clinical laboratories for fosfomycin testing of gram-negative pathogens. A broader exploration of poloxamer dilution\u0027s utility is thus warranted.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003EIMPORTANCE: \u003C\/b\u003EAccurate antibiotic susceptibility testing is essential for guiding treatment of bacterial infections. For the antibiotic fosfomycin, used to treat \u003Ci\u003EEscherichia coli\u003C\/i\u003E urinary tract infections, the most reliable testing method requires solid media prepared by hand for each antibiotic concentration, which is too time-consuming for most clinical laboratories to perform. Our study shows that replacing agar with an alternative temperature-sensitive gelling agent called poloxamer enables laboratories to prepare solid test plates rapidly without special equipment. This approach, which is essentially identical to traditional agar dilution, provides a practical means for performing reference-quality minimal inhibitory concentration (MIC) testing near the point of patient care, as demonstrated for fosfomycin, for which current FDA-cleared methods do not provide MIC data. This strategy may also be applicable to other drugs for which agar dilution is the preferred testing method, supporting expedited testing to inform treatment decisions for bacterial infections.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EKang, Yoon-Suk, and James E Kirby. (2026) 2026. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/host-gpcr-camp-signaling-balances-gas-and-gai-activity-control-intracellular-brucella\u0022 hreflang=\u0022en\u0022\u003EHost GPCR-CAMP Signaling Balances G\u03b1s and G\u03b1i Activity to Control Intracellular Brucella Infection.\u003C\/a\u003E\u201d. \u003Ci\u003EBioRxiv : The Preprint Server for Biology\u003C\/i\u003E. https:\/\/doi.org\/10.64898\/2026.01.06.697936.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/41542404\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003EIn this study, we investigated the impact of G protein-coupled receptor (GPCR) signaling on the intracellular replication of the model pathogen \u003Ci\u003EBrucella neotomae\u003C\/i\u003E. Building on a prior chemical genetics screen, we identified agonists of the G\u03b1i-coupled adenosine A1 and dopamine D4 receptors as potent inhibitors of intracellular Brucella replication. In contrast, agonists of G\u03b1s-coupled adenosine A2A or dopamine D1 receptors, as well as antagonists of A1 or D4 receptors, either failed to inhibit or enhanced intracellular replication. Wild-type \u003Ci\u003EB. neotomae\u003C\/i\u003E induced a rapid, type IV secretion system-dependent increase in host-cell cAMP during early infection. ENBA and cilostamide prevented this infection-associated cAMP increase and completely inhibited intracellular growth; this effect was partially reversed by cell-permeable cAMP analogs Using a real-time NanoBRET biosensor, we detected rapid G\u03b1s activation within minutes of infection that was sustained during wild-type but not \u0394virB4 infection and was abrogated by ENBA or cilostamide. Disruption of early G\u03b1s-cAMP signaling redirected BCVs to replication-incompatible phagolysosomal and autophagy-associated compartments. Collectively, these data support a model in which early GPCR signaling dynamics, balancing G\u03b1s and G\u03b1i pathways, are critical for establishment of productive intracellular Brucella infection.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EKassu, Mintesinot, Katelyn E Zulauf, Jessica N Ross, James E Kirby, and Roman Manetsch. (2025) 2025. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/development-cgs-15943-adjunctives-disruption-plasmid-maintenance-multidrug-resistant-e\u0022 hreflang=\u0022en\u0022\u003EDevelopment of CGS-15943 Adjunctives for the Disruption of Plasmid Maintenance in Multidrug Resistant E. Coli.\u003C\/a\u003E\u201d. \u003Ci\u003EACS Infectious Diseases\u003C\/i\u003E 11 (1): 80-87. https:\/\/doi.org\/10.1021\/acsinfecdis.4c00587.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/39741382\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003ECarbapenemase producing \u003Ci\u003EEnterobacterales\u003C\/i\u003E (CPEs) represent a group of multidrug resistant pathogens for which few, if any, therapeutics options remain available. CPEs generally harbor plasmids that encode resistance to last resort carbapenems and many other antibiotics. We previously performed a high throughput screen to identify compounds that can disrupt the maintenance and replication of resistance conferring plasmids through use of a synthetic screening plasmid introduced into \u003Ci\u003EEscherichia coli\u003C\/i\u003E K-12 \u003Ci\u003EtolC\u003C\/i\u003E cells. Despite being identified as a potent and selective antiplasmid agent through this screening effort, CGS-15943 was inactive in wild-type \u003Ci\u003EE. coli\u003C\/i\u003E, suggesting that it is susceptible to TolC-mediated efflux. Herein, a series of analogues were developed to confirm the activity of the triazoloquinazoline chemotype and overcome efflux observed in wild-type \u003Ci\u003EE. coli\u003C\/i\u003E K-12. Two analogues demonstrated superior antiplasmid activity to CGS-15943 in \u003Ci\u003EE. coli tolC\u003C\/i\u003E mutants, while one compound displayed moderate activity in wild-type \u003Ci\u003EE. coli\u003C\/i\u003E at low concentrations.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EKang, Yoon-Suk, Simone C Silva, Kenneth Smith, Krissty Sumida, Yuhan Wang, Lucius Chiaraviglio, Ramachandra Reddy Donthiri, Alhanouf Z Aljahdali, James E Kirby, and George A O\u2019Doherty. (2025) 2025. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/exploration-fusidic-acid-structure-activity-space-antibiotic-activity\u0022 hreflang=\u0022en\u0022\u003EExploration of the Fusidic Acid Structure Activity Space for Antibiotic Activity.\u003C\/a\u003E\u201d. \u003Ci\u003EMolecules (Basel, Switzerland)\u003C\/i\u003E 30 (3). https:\/\/doi.org\/10.3390\/molecules30030465.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/39942570\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003EFusidic acid is a translation inhibitor with activity against major Gram-positive bacterial pathogens such as \u003Ci\u003ES. aureus\u003C\/i\u003E. However, its activity against Gram-negatives is poor based on an inability to access its cytoplasmic target in these organisms. Opportunities for functionalization of the fusidic acid scaffold to enhance activity against Gram-negative pathogens have not been explored. Using an activity-guided synthetic strategy, the tolerance of the tetracyclic natural product to derivatization at the A- and C-rings and its carboxylic acid side chain was explored with the goal of enhancing its activity spectrum and pharmacological properties. All side-chain carboxylic acid esters were inactive. Oxidation of the C-ring alcohol and oxime were not tolerated either. A number of esters of the A-ring alcohol retained modest activity against Gram-positive bacteria and were informative for future activity-guided studies. For the A-ring esters, differences in antibacterial activity relative to inhibitory activity in a ribosome in vitro translation assay suggested the possibility of a pro-druglike effect for the fusidic acid pyrazine-2-carboxylate. This study furthers the understanding of the activity of the fusidic acid scaffold against Gram-positive bacteria. These results suggest promise for future modification of the A-ring alcohol of fusidic acid in the advancement of its antibiotic properties.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EGregor, William D, Rakesh Maharjan, Zhemin Zhang, Lucius Chiaraviglio, Nithya Sastry, Meng Cui, James E Kirby, and Edward W Yu. (2025) 2025. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/cryo-em-structure-pseudomonas-aeruginosa-mexy-multidrug-efflux-pump\u0022 hreflang=\u0022en\u0022\u003ECryo-EM Structure of the Pseudomonas Aeruginosa MexY Multidrug Efflux Pump.\u003C\/a\u003E\u201d. \u003Ci\u003EMBio\u003C\/i\u003E 16 (4): e0382624. https:\/\/doi.org\/10.1128\/mbio.03826-24.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/40042268\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003E\u003Cb\u003EUNLABELLED: \u003C\/b\u003E\u003Ci\u003EPseudomonas aeruginosa\u003C\/i\u003E, a Gram-negative pathogen, has emerged as one of the most highly antibiotic-resistant bacteria worldwide and subsequently has become a leading cause of healthcare-associated, life-threatening infections. \u003Ci\u003EP. aeruginosa\u003C\/i\u003E multidrug efflux Y (MexY) is an efflux pump that belongs to the resistance-nodulation-cell division (RND) superfamily. It is a major determinant for resistance to aminoglycosides in this opportunistic pathogen. However, the detailed molecular mechanisms involved in aminoglycoside recognition and extrusion by MexY have not been elucidated. Here, we report the cryo-electron microscopy structure of MexY to a resolution of 3.63 \u00c5. The structure directly indicates two plausible pathways for drug export. It also suggests that MexY is capable of picking up antibiotics via the ceiling of the central cavity formed by the MexY trimer. Molecular dynamics simulations depict that MexY is able to use a tunnel connecting the central cavity to the funnel of the trimer to export its substrates.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003EIMPORTANCE: \u003C\/b\u003EHere, we report the cryo-electron microscopy structure of the MexY multidrug efflux pump, posing the possibility that this pump is capable of capturing antibiotics from both the central cavity and the periplasmic cleft of the pump. The results indicate that MexY may utilize charged residues to bind and export drugs, mediating resistance to these antibiotics.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EHuang, Yanqin, Katherine A Truelson, Isabella A Stewart, George A O\u2019Doherty, and James E Kirby. (2025) 2025. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/enhanced-activity-apramycin-and-apramycin-based-combinations-against-mycobacteroides-0\u0022 hreflang=\u0022en\u0022\u003EEnhanced Activity of Apramycin and Apramycin-Based Combinations Against Mycobacteroides Abscessus.\u003C\/a\u003E\u201d. \u003Ci\u003EBioRxiv : The Preprint Server for Biology\u003C\/i\u003E. https:\/\/doi.org\/10.1101\/2025.04.09.648020.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/40391321\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003E\u003Cb\u003EBACKGROUND: \u003C\/b\u003E\u003Ci\u003EMycobacteroides abscessus\u003C\/i\u003E are rapidly growing non-tuberculous mycobacteria that cause chronic lung and soft tissue infections. Treatment options are often severely limited due to intrinsic resistance to most antimicrobials. Amikacin has historically been a mainstay of combination treatment regimens. However, irreversible hearing loss and vestibular toxicity have led to a search for alternative agents. Apramycin is a novel aminoglycoside currently in phase I clinical trials that may offer lower potential for ototoxic and renal toxic side effects.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003EOBJECTIVES: \u003C\/b\u003EThe goal of this study was to compare apramycin\u0027s \u003Ci\u003Ein vitro\u003C\/i\u003E activity with amikacin and other aminoglycosides against a large collection of \u003Ci\u003EM. abscessus\u003C\/i\u003E clinical isolates, both alone and in combination with clofazimine or linezolid. We also tested the activity of apramycin against a more limited collection of other species of rapidly growing mycobacteria.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003EMETHODS: \u003C\/b\u003EAnalysis was performed using reference broth microdilution minimal inhibitory concentration testing, inkjet printer-assisted checkerboard assays, and time-kill assays.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003ERESULTS: \u003C\/b\u003EAgainst \u003Ci\u003EM. abscessus\u003C\/i\u003E, the MIC50\/90 for apramycin (2 \u03bcg\/mL) was 8-fold lower than for amikacin (16 \u03bcg\/mL). Plazomicin was inactive, and organisms were rarely susceptible to tobramycin. Synergy was not detected by checkerboard assay. In time-kill studies, clofazimine modestly potentiated activity of apramycin and. to a lesser extent, amikacin. Apramycin and amikacin showed delayed bacterial killing that either achieved or approached a bactericidal threshold. Apramycin was similarly potent against other rapidly growing mycobacteria tested.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003ECONCLUSIONS: \u003C\/b\u003EApramycin exhibits more potent \u003Ci\u003Ein vitro\u003C\/i\u003E activity against a diverse set of \u003Ci\u003EM. abscessus\u003C\/i\u003E and other rapidly growing mycobacteria than approved aminoglycosides.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EHuang, Yanqin, Lucius Chiaraviglio, Ibidunni Bode-Sojobi, and James E Kirby. (2025) 2025. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/triple-antimicrobial-combinations-potent-synergistic-activity-against-m-abscessus-0\u0022 hreflang=\u0022en\u0022\u003ETriple Antimicrobial Combinations With Potent Synergistic Activity Against M. Abscessus.\u003C\/a\u003E\u201d. \u003Ci\u003EBioRxiv : The Preprint Server for Biology\u003C\/i\u003E. https:\/\/doi.org\/10.1101\/2025.02.13.638181.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/39990351\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003ESynergy of antimicrobial combinations was tested against contemporary \u003Ci\u003EMycobacteroides\u003C\/i\u003E abscessus isolates using 2-D and 3-D checkerboard assays. Triple combinations of omadacycline-azithromycin-clofazimine, tigecycline-azithromycin-clofazimine, omadacycline-azithromycin-linezolid and omadacycline-azithromycin-contezolid demonstrated synergy (FIC \u2264 0.5) against 33%, 31%, 62%, and 66% of isolates, respectively. Notably, in all triple combinations, macrolide resistant \u003Ci\u003EM. abscessus\u003C\/i\u003E subsp. \u003Ci\u003Eabscessus\u003C\/i\u003E and \u003Ci\u003EM. abscessus\u003C\/i\u003E subsp. \u003Ci\u003Ebolletii\u003C\/i\u003E isolates were fully sensitized to azithromycin at the FIC index, as were isolates with elevated clofazimine MICs.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EHuang, Yanqin, Lucius Chiaraviglio, Ibidunni Bode-Sojobi, and James E Kirby. (2025) 2025. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/triple-antimicrobial-combinations-potent-synergistic-activity-against-m-abscessus\u0022 hreflang=\u0022en\u0022\u003ETriple Antimicrobial Combinations With Potent Synergistic Activity Against M. Abscessus.\u003C\/a\u003E\u201d. \u003Ci\u003EAntimicrobial Agents and Chemotherapy\u003C\/i\u003E 69 (4): e0182824. https:\/\/doi.org\/10.1128\/aac.01828-24.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/40084880\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003ESynergy of antimicrobial combinations was tested against contemporary \u003Ci\u003EMycobacteroides abscessus\u003C\/i\u003E isolates using 2D and 3D checkerboard assays. Triple combinations of omadacycline-azithromycin-clofazimine, tigecycline-azithromycin-clofazimine, omadacycline-azithromycin-linezolid, and omadacycline-azithromycin-contezolid demonstrated synergy (fractional inhibitory concentration index \u2264 0.5) against 33%, 31%, 62%, and 66% of isolates, respectively. Notably, in all triple combinations, macrolide-resistant \u003Ci\u003EM. abscessus\u003C\/i\u003E subsp. \u003Ci\u003Eabscessus\u003C\/i\u003E and \u003Ci\u003EM. abscessus\u003C\/i\u003E subsp. \u003Ci\u003Ebolletii\u003C\/i\u003E isolates were fully sensitized to azithromycin at the FIC index, as were isolates with elevated clofazimine MICs.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EHicks, Ian, Ryan Moradei, Lucius Chiaraviglio, Alhanouf Z Aljahdali, Ramachandra Reddy Donthiri, James E Kirby, and George A O\u2019Doherty. (2025) 2025. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/de-novo-design-and-asymmetric-synthesis-c-12-benzodioxin-fused-glycoside-analogue\u0022 hreflang=\u0022en\u0022\u003EDe Novo Design and Asymmetric Synthesis of a C-1\/2 Benzodioxin Fused Glycoside Analogue of Lincomycin.\u003C\/a\u003E\u201d. \u003Ci\u003EOrganic Letters\u003C\/i\u003E 27 (37): 10532-36. https:\/\/doi.org\/10.1021\/acs.orglett.5c03402.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/40911788\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003EAn \u003Ci\u003Ein silico\u003C\/i\u003E designed benzodioxin fused analogue of a des-(1-hydroxyethyl)-lincomycin analogue was synthesized in a \u003Ci\u003Ede novo\u003C\/i\u003E asymmetric fashion from an achiral acylfuran, a 4-(\u003Ci\u003En\u003C\/i\u003E-Pr)-\u003Ci\u003EN\u003C\/i\u003E-methyl-proline, and catechol. The synthesis of the 6-amino-galactose portion of the lincomycin analogue necessitated the development of a novel stereospecific tandem Pd-glycosylation\/1,4-addition reaction between catechol and an \u003Ci\u003EN\u003C\/i\u003E-Cbz-protected 6-amino-pyranone with a Pd-\u03c0-allyl leaving group at the \u003Ci\u003EC\u003C\/i\u003E-1 position. The desired \u003Ci\u003Egalacto\u003C\/i\u003E-stereochemistry was installed by a subsequent stereoselective ketone reduction, alcohol elimination, and diastereoselective dihydroxylation of the \u003Ci\u003EC\u003C\/i\u003E-3\/4 alkene.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EHuang, Yanqin, Katherine A Truelson, Isabella A Stewart, George A O\u2019Doherty, and James E Kirby. (2025) 2025. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/enhanced-activity-apramycin-and-apramycin-based-combinations-against-mycobacteroides\u0022 hreflang=\u0022en\u0022\u003EEnhanced Activity of Apramycin and Apramycin-Based Combinations Against Mycobacteroides Abscessus.\u003C\/a\u003E\u201d. \u003Ci\u003EThe Journal of Antimicrobial Chemotherapy\u003C\/i\u003E. https:\/\/doi.org\/10.1093\/jac\/dkaf433.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/41313250\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003E\u003Cb\u003EBACKGROUND: \u003C\/b\u003EMycobacteroides abscessus is a rapidly growing non-tuberculous mycobacterium that causes chronic lung and soft tissue infections. Treatment options are severely limited. Amikacin has historically been a mainstay of combination treatment regimens. However, irreversible hearing loss and vestibular toxicity have led to a search for alternative agents. Apramycin is a novel aminoglycoside currently in Phase I clinical trials that may offer lower potential for ototoxic and renal toxic side effects.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003EOBJECTIVES: \u003C\/b\u003EThe goal of this study was to compare apramycin\u0027s in vitro activity with amikacin and other aminoglycosides against a large collection of M. abscessus clinical isolates, alone, and for apramycin and amikacin, in combination with clofazimine or linezolid. We also examined activity against a more limited collection of other rapidly growing mycobacteria.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003EMETHODS: \u003C\/b\u003EAnalysis was performed using reference broth microdilution MIC testing, inkjet printer-assisted checkerboard assays, and time-kill assays.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003ERESULTS: \u003C\/b\u003EAgainst M. abscessus, the MIC50\/90 for apramycin (2\u2005mg\/L) was one-eighth that of amikacin (16\u2005mg\/L). Plazomicin was inactive, and organisms were rarely susceptible to tobramycin. Synergy of either apramycin or amikacin with clofazimine or linezolid was not detected by checkerboard assay. In time-kill studies, clofazimine modestly increased activity of apramycin and, to a lesser extent, amikacin. Apramycin and amikacin showed delayed bacterial killing that either achieved or approached a bactericidal threshold. Apramycin was similarly potent against other rapidly growing mycobacteria tested.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003ECONCLUSIONS: \u003C\/b\u003EApramycin exhibits more potent in vitro activity against a diverse set of M. abscessus and other rapidly growing mycobacteria than currently recommended aminoglycosides.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003ELee, Yong Woo, Roy Blum, Tyler Mrozowich, Bojan Bujisic, James E Kirby, and Jeannie T Lee. (2025) 2025. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/pharmacologically-stabilizing-rna-g-quadruplexes-coronavirus-genome-reduces\u0022 hreflang=\u0022en\u0022\u003EPharmacologically Stabilizing RNA G-Quadruplexes in Coronavirus Genome Reduces Infectivity.\u003C\/a\u003E\u201d. \u003Ci\u003ERNA (New York, N.Y.)\u003C\/i\u003E 31 (11): 1632-51. https:\/\/doi.org\/10.1261\/rna.080321.124.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/40877136\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003ERNA G-quadruplexes (rG4s) are unusual RNA secondary structures formed by stacking arrays of guanine tetrads. Although thousands of potential rG4-forming motifs occur throughout the mammalian transcriptome, many single-stranded RNA (ssRNA) viruses are thought to be depleted of rG4-forming sequences. Using in silico methods, we examine rG4-forming potential in single-stranded RNA (ssRNA) viruses and observe that, while canonical rG4 motifs are depleted, noncanonical rG4 motifs occur at comparable or higher frequencies relative to the mammalian transcriptome. We ask if the noncanonical rG4\u0027s can be leveraged to block viral replication and control infection using OC43, the coronavirus believed to be responsible for the 1889 \u0022Russian flu\u0022 pandemic. Profiling with \u0022d-rG4-seq\u0022 confirms a dearth of folded rG4 in the OC43 RNA genome during natural infection. Intriguingly, rG4 ligands induce synthetic rG4 structures of a noncanonical nature. Significantly, induced rG4 inhibits viral replication and reduces infectivity. We show that the rG4 ligands act by disrupting the unique pattern of OC43 \u0022discontinuous transcription.\u0022 Thus, rG4-targeting compounds present a potential therapeutic approach for targeting ssRNA viruses.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EKim, Hyo In, Anupamaa J Seshadri, James Harbison, Eva Csizmadia, Jinbong Park, David Gallo, Vanessa A Voltarelli, et al. (2025) 2025. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/traumatic-liver-injury-increases-susceptibility-bacterial-pneumonia-swine\u0022 hreflang=\u0022en\u0022\u003ETraumatic Liver Injury Increases Susceptibility to Bacterial Pneumonia in Swine.\u003C\/a\u003E\u201d. \u003Ci\u003EThe Journal of Trauma and Acute Care Surgery\u003C\/i\u003E. https:\/\/doi.org\/10.1097\/TA.0000000000004768.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/40875191\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003E\u003Cb\u003EBACKGROUND: \u003C\/b\u003EIn this study, we develop a standardized porcine model of distant injury plus lung bacterial inoculation to allow translational investigations of the effects of tissue injury on susceptibility to infection. This generalizable model will allow testing of immune interventions on the evolution of infection.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003EMETHODS: \u003C\/b\u003EA standardized liver crush (5 cm \u00d7 2.5 cm\/3 kg) plus hemoperitoneum (6 mL\/kg) or sham procedure was performed in 30-kg Yorkshire pigs, followed by intratracheal inoculation of bacteria (Actinobacillus pleuropneumoniae). We then compared gross pathology, histology, lung bacterial counts, danger-associated molecular pattern molecules, and serum cytokines between the two groups.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003ERESULTS: \u003C\/b\u003EThe lungs of injured pigs demonstrated significantly enhanced responses to infection compared with sham injured pigs, both on the macroscopic and microscopic levels. Lung bacterial clearance was significantly impaired after trauma, with increased infiltration of neutrophils and differential location of myeloid cells on immunostaining. In lung parenchyma expression of the stress response genes, Hmox1 and Nrf2 were increased in both trauma alone and trauma plus infection. Plasma from pigs subjected to trauma showed increased levels of the danger-associated molecular patters heme and mitochondrial DNA and promoted bacterial growth in vitro compared with plasma from uninjured pigs.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003ECONCLUSION: \u003C\/b\u003EWe have developed a novel, clinically relevant, reproducible porcine model of abdominal injury with subsequent A. pleuropneumoniae pneumonia for the study and development of therapeutics against immune dysregulation induced by trauma. Additionally, a novel finding is that plasma from traumatized pigs provides a permissive environment for bacterial growth.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EKirby, James E, and Ramy Arnaout. (2025) 2025. \u201cWh\u003Ca href=\u0022\/james-kirby\/publications\/whats-not-learn-ai-meets-parasitology\u0022 hreflang=\u0022en\u0022\u003Eat\u2019s Not to Learn? AI Meets Parasitology.\u003C\/a\u003E\u201d. \u003Ci\u003EJournal of Clinical Microbiology\u003C\/i\u003E, e0145125. https:\/\/doi.org\/10.1128\/jcm.01451-25.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/41358779\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003EAlthough artificial intelligence-particularly large-language models-receives daily attention, the application of AI to image-recognition challenges in clinical microbiology has been under development for several years. In the accompanying article, B. A. Mathison, K. Knight, J. Potts, B. Black, et al. (J Clin Microbiol 63:e01062-25, 2025, \u003Ca href=\u0022https:\/\/doi.org\/10.1128\/jcm.01062-25\u0022\u003Ehttps:\/\/doi.org\/10.1128\/jcm.01062-25\u003C\/a\u003E) (in collaboration with ARUP Laboratories and TechCyte) describe a trained convolutional neural network (CNN) that reviews wet-mount parasitology smears with accuracy and analytical sensitivity exceeding that of a cohort of highly trained medical technologists. The impressive results were enabled by an extensive, globally sourced training set. These findings constitute Part\u202fII of the authors\u0027 earlier Journal of Clinical Microbiology publication on CNN-based diagnosis of trichrome-stained smears and provide a robust proof-of-concept for integrating AI into clinical microbiology workflows. We comment on the translatability of this technology to routine clinical laboratories.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EKirmaier, Andrea, Leslie Blackshear, Matthew Shou Lun Lee, and James E Kirby. (2024) 2024. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/cellulitis-and-bacteremia-caused-fish-pathogenstreptococcus-iniae-immunocompromised\u0022 hreflang=\u0022en\u0022\u003ECellulitis and Bacteremia Caused by the Fish Pathogen,Streptococcus Iniae, in an Immunocompromised Patient: Case Report and Mini-Review of Zoonotic Disease, Lab Identification, and Antimicrobial Susceptibility.\u003C\/a\u003E\u201d. \u003Ci\u003EDiagnostic Microbiology and Infectious Disease\u003C\/i\u003E 108 (4): 116189. https:\/\/doi.org\/10.1016\/j.diagmicrobio.2024.116189.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/38278004\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003EStreptococcus iniae is a fish pathogen that can also infect mammals including dolphins and humans. Its prevalence in farmed fish, particularly tilapia, provides potential for zoonotic infections, as documented by multiple case reports. Systematic clinical data beyond cellulitis for S. iniae infection in humans, including antimicrobial susceptibility data, are unfortunately rare. Here, we present a case of cellulitis progressing to bacteremia caused by Streptococcus iniae in a functionally immunocompromised patient based on CDK4\/CDK6 inhibitor and endocrine therapy, and we discuss risk factors, identification, and antimicrobial susceptibility of this rare pathogen.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003ECunha, Vitor L S, George A O\u2019Doherty, and Todd L Lowary. (2024) 2024. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/exploring-de-novo-route-bradyrhizose-synthesis-and-isomeric-equilibrium-bradyrhizose\u0022 hreflang=\u0022en\u0022\u003EExploring a De Novo Route to Bradyrhizose: Synthesis and Isomeric Equilibrium of Bradyrhizose Diastereomers\u2260.\u003C\/a\u003E\u201d. \u003Ci\u003EChemistry (Weinheim an Der Bergstrasse, Germany)\u003C\/i\u003E 30 (33): e202400886. https:\/\/doi.org\/10.1002\/chem.202400886.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/38590211\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003EA de\u2005novo asymmetric strategy for the synthesis of d-bradyrhizose diastereomers from an achiral ketoenolester precursor is described. Key transformations used in the stereodivergent approach include two Noyori asymmetric reductions, an Achmatowicz rearrangement, diastereoselective alkene oxidations, and the first example of a palladium(0)-catalyzed glycosylation of a vinylogous pyranone. The isomeric composition of the bicyclic reducing sugars obtained was analyzed and their behaviour was compared to the natural product, revealing key stereocentres that impact the overall distribution.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EReimche, Jennifer L, Cau D Pham, Sandeep J Joseph, Shelby Hutton, John C Cartee, Yuheng Ruan, Mandy Breaux, et al. (2024) 2024. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/novel-strain-multidrug-non-susceptible-neisseria-gonorrhoeae-usa\u0022 hreflang=\u0022en\u0022\u003ENovel Strain of Multidrug Non-Susceptible Neisseria Gonorrhoeae in the USA.\u003C\/a\u003E\u201d. \u003Ci\u003EThe Lancet. Infectious Diseases\u003C\/i\u003E 24 (3): e149-e151. https:\/\/doi.org\/10.1016\/S1473-3099(23)00785-5.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/38215769\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n\n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003ETheel, Elitza S, James E Kirby, and Nira R Pollock. (2024) 2024. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/testing-sars-cov-2-lessons-learned-and-current-use-cases\u0022 hreflang=\u0022en\u0022\u003ETesting for SARS-CoV-2: Lessons Learned and Current Use Cases.\u003C\/a\u003E\u201d. \u003Ci\u003EClinical Microbiology Reviews\u003C\/i\u003E 37 (2): e0007223. https:\/\/doi.org\/10.1128\/cmr.00072-23.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/38488364\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003ESUMMARYThe emergence and worldwide dissemination of SARS-CoV-2 required both urgent development of new diagnostic tests and expansion of diagnostic testing capacity on an unprecedented scale. The rapid evolution of technologies that allowed testing to move out of traditional laboratories and into point-of-care testing centers and the home transformed the diagnostic landscape. Four years later, with the end of the formal public health emergency but continued global circulation of the virus, it is important to take a fresh look at available SARS-CoV-2 testing technologies and consider how they should be used going forward. This review considers current use case scenarios for SARS-CoV-2 antigen, nucleic acid amplification, and immunologic tests, incorporating the latest evidence for analytical\/clinical performance characteristics and advantages\/limitations for each test type to inform current debates about how tests should or should not be used.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EKirby, James E, Stefan Riedel, Sanjucta Dutta, Ramy Arnaout, Annie Cheng, Sarah Ditelberg, Donald J Hamel, Charlotte A Chang, and Phyllis J Kanki. (2023) 2023. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/sars-cov-2-antigen-tests-predict-infectivity-based-viral-culture-comparison-antigen\u0022 hreflang=\u0022en\u0022\u003ESars-Cov-2 Antigen Tests Predict Infectivity Based on Viral Culture: Comparison of Antigen, PCR Viral Load, and Viral Culture Testing on a Large Sample Cohort.\u003C\/a\u003E\u201d. \u003Ci\u003EClinical Microbiology and Infection : The Official Publication of the European Society of Clinical Microbiology and Infectious Diseases\u003C\/i\u003E 29 (1): 94-100. https:\/\/doi.org\/10.1016\/j.cmi.2022.07.010.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/35863629\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003E\u003Cb\u003EOBJECTIVE: \u003C\/b\u003ETo define the relationship of SARS-CoV-2 antigen, viral load determined by RT-qPCR, and viral culture detection. Presumptively, viral culture can provide a surrogate measure for infectivity of sampled individuals and thereby inform how and where to most appropriately deploy antigen and nucleic acid amplification-based diagnostic testing modalities.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003EMETHODS: \u003C\/b\u003EWe compared the antigen testing results from three lateral flow and one microfluidics assay to viral culture detection and viral load determination performed in parallel in up to 189 nasopharyngeal swab samples positive for SARS-CoV-2. Sample viral loads, determined by RT-qPCR, were distributed across the range of viral load values observed in our testing population.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003ERESULTS: \u003C\/b\u003EAntigen tests were predictive of viral culture positivity, with the LumiraDx microfluidics method showing enhanced sensitivity (90%; 95% CI 83-94%) compared with the BD Veritor (74%, 95% CI 65-81%), CareStart (74%, 95% CI 65-81%) and Oscar Corona (74%, 95% CI 65-82%) lateral flow antigen tests. Antigen and viral culture positivity were also highly correlated with sample viral load, with areas under the receiver operator characteristic curves of 0.94 to 0.97 and 0.92, respectively. A viral load threshold of 100\u00a0000 copies\/mL was 95% sensitive (95% CI, 90-98%) and 72% specific (95% CI, 60-81%) for predicting viral culture positivity. Adjusting for sample dilution inherent in our study design, sensitivities of antigen tests were \u226595% for detection of viral culture positive samples with viral loads \u0026gt;106 genome copies\/mL, although specificity of antigen testing was imperfect.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003EDISCUSSION: \u003C\/b\u003EAntigen testing results and viral culture were correlated. For culture positive samples, the sensitivity of antigen tests was high at high viral loads that are likely associated with significant infectivity. Therefore, our data provides support for use of antigen testing in ruling out infectivity at the time of sampling.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EKang, Yoon-Suk, and James E Kirby. (2023) 2023. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/versatile-nanoluciferase-reporter-reveals-structural-properties-associated-highly\u0022 hreflang=\u0022en\u0022\u003EA Versatile Nanoluciferase Reporter Reveals Structural Properties Associated With a Highly Efficient, N-Terminal Legionella Pneumophila Type IV Secretion Translocation Signal.\u003C\/a\u003E\u201d. \u003Ci\u003EMicrobiology Spectrum\u003C\/i\u003E 11 (2): e0233822. https:\/\/doi.org\/10.1128\/spectrum.02338-22.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/36815834\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003EMany Gram-negative pathogens rely on type IV secretion systems (T4SS) for infection. One limitation has been the lack of ideal reporters to identify T4SS translocated effectors and study T4SS function. Most reporter systems make use of fusions to reporter proteins, in particular, \u03b2-lactamase (TEM) and calmodulin-dependent adenylate cyclase (CYA), that allow detection of translocated enzymatic activity inside host cells. However, both systems require costly reagents and use complex, multistep procedures for loading host cells with substrate (TEM) or for analysis (CYA). Therefore, we have developed and characterized a novel reporter system using nanoluciferase (NLuc) fusions to address these limitations. Serendipitously, we discovered that Nluc itself is efficiently translocated by Legionella pneumophila T4SS in an IcmSW chaperone-dependent manner via an N-terminal translocation signal. Extensive mutagenesis in the NLuc N terminus suggested the importance of an \u03b1-helical domain spanning D5 to V9, as mutations predicted to disrupt this structure, with one exception, were translocation defective. Notably, NLuc was capable of translocating several proteins examined when fused to the N or C terminus, while maintaining robust luciferase activity. In particular, it delivered the split GFP11 fragment into J774 macrophages transfected with GFPopt, thereby resulting in \u003Ci\u003Ein vivo\u003C\/i\u003E assembly of superfolder green fluorescent protein (GFP). This provided a bifunctional assay in which translocation could be assayed by fluorescence microplate, confocal microscopy, and\/or luciferase assays. We further identified an optimal NLuc substrate which allowed a robust, inexpensive, one-step, high-throughput screening assay to identify T4SS translocation substrates and inhibitors. Taken together, these results indicate that NLuc provides both new insight into and also tools for studying T4SS biology. IMPORTANCE Type IV secretion systems (T4SS) are used by Gram-negative pathogens to coopt host cell function. However, the translocation signals recognized by T4SS are not fully explained by primary amino acid sequence, suggesting yet-to-be-defined contributions of secondary and tertiary structure. Here, we unexpectedly identified nanoluciferase (NLuc) as an efficient IcmSW-dependent translocated T4SS substrate, and we provide extensive mutagenesis data suggesting that the first N-terminal, alpha-helix domain is a critical translocation recognition motif. Notably, most existing reporter systems for studying translocated proteins make use of fusions to reporters to permit detection of translocated enzymatic activity inside the host cell. However, existing systems require extremely costly substrates, complex technical procedures to isolate eukaryotic cytoplasm for analysis, and\/or are insensitive. Importantly, we found that NLuc provides a powerful, cost-effective new tool to address these limitations and facilitate high-throughput exploration of secretion system biology.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EFrescas, Brian E, Christopher M McCoy, James Kirby, Robert Bowden, and Nicholas J Mercuro. (2023) 2023. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/outcomes-associated-empiric-cefepime-bloodstream-infections-caused-ceftriaxone\u0022 hreflang=\u0022en\u0022\u003EOutcomes Associated With Empiric Cefepime for Bloodstream Infections Caused by Ceftriaxone-Resistant, Cefepime-Susceptible\u00a0Escherichia Coli\u00a0and\u00a0Klebsiella Pneumoniae.\u003C\/a\u003E\u201d. \u003Ci\u003EInternational Journal of Antimicrobial Agents\u003C\/i\u003E 61 (5): 106762. https:\/\/doi.org\/10.1016\/j.ijantimicag.2023.106762.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/36804369\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003E\u003Cb\u003EBACKGROUND: \u003C\/b\u003ECefepime is a first-line agent for empiric sepsis therapy; however, cefepime use may be associated with increased mortality for extended-spectrum beta-lactamase-producing Enterobacterales (ESBL-E) in an MIC-dependent manner. This study aimed to compare the efficacy of empiric cefepime versus meropenem for bloodstream infections (BSI) caused by ceftriaxone-resistant Escherichia coli and Klebsiella pneumoniae with cefepime MICs \u2264 2 mg\/L.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003EMETHODS: \u003C\/b\u003EThis single-center retrospective cohort study included patients admitted from October 2010 to August 2020 who received cefepime or meropenem empirically for sepsis with a blood culture growing ceftriaxone-resistant Escherichia coli or Klebsiella pneumoniae. The primary outcome was 30-day mortality; secondary endpoints included 14-day mortality, recurrent BSI, readmission and recurrent infection within 90 days, time to clinical resolution of infection, time to clinical stability, and clinical stability at 48 hours.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003ERESULTS: \u003C\/b\u003EFifty-four patients met inclusion criteria: 36 received meropenem and 18 received cefepime. The median (IQR) treatment durations of cefepime and meropenem were 3 (2-6) days and 7 (5-10) days, respectively. Thirty-day and 14-day mortality were similar between cefepime and meropenem (11.1% vs. 2.8%; P\u00a0=\u00a00.255 and 5.6% vs. 2.8%; P\u00a0=\u00a01.00, respectively). Cefepime was associated with longer time to clinical stability compared with meropenem (median 38.48 hours vs. 21.26; P\u00a0=\u00a00.016).\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003ECONCLUSION: \u003C\/b\u003EMortality was similar between groups, although most patients who received cefepime empirically were ultimately transitioned to a carbapenem to complete the full treatment course. Empiric cefepime was associated with a delay in achieving clinical stability when compared with meropenem to treat BSI caused by ceftriaxone-resistant Enterobacterales, even when cefepime-susceptible.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EGreen, Alex B, Lucius Chiaraviglio, Katherine A Truelson, Katelyn E Zulauf, Meng Cui, Zhemin Zhang, Matthew P Ware, et al. (2023) 2023. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/rnd-pump-mediated-efflux-amotosalen-compound-used-pathogen-inactivation-technology\u0022 hreflang=\u0022en\u0022\u003ERND Pump-Mediated Efflux of Amotosalen, a Compound Used in Pathogen Inactivation Technology to Enhance Safety of Blood Transfusion Products, May Compromise Its Gram-Negative Anti-Bacterial Activity.\u003C\/a\u003E\u201d. \u003Ci\u003EMSphere\u003C\/i\u003E 8 (2): e0067322. https:\/\/doi.org\/10.1128\/msphere.00673-22.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/36853056\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003EPathogen inactivation is a strategy to improve the safety of transfusion products. The only pathogen reduction technology for blood products currently approved in the US utilizes a psoralen compound, called amotosalen, in combination with UVA light to inactivate bacteria, viruses, and protozoa. Psoralens have structural similarity to bacterial multidrug efflux pump substrates. As these efflux pumps are often overexpressed in multidrug-resistant pathogens, we tested whether contemporary drug-resistant pathogens might show resistance to amotosalen and other psoralens based on multidrug efflux mechanisms through genetic, biophysical, and molecular modeling analysis. The main efflux systems in \u003Ci\u003EEnterobacterales\u003C\/i\u003E, Acinetobacter baumannii, and Pseudomonas aeruginosa are tripartite resistance-nodulation-cell division (RND) systems, which span the inner and outer membranes of Gram-negative pathogens, and expel antibiotics from the bacterial cytoplasm into the extracellular space. We provide evidence that amotosalen is an efflux substrate for the E. coli AcrAB, Acinetobacter baumannii AdeABC, and P. aeruginosa MexXY RND efflux pumps. Furthermore, we show that the MICs for contemporary Gram-negative bacterial isolates for these species and others \u003Ci\u003Ein vitro\u003C\/i\u003E approached and exceeded the concentration of amotosalen used in the approved platelet and plasma inactivation procedures. These findings suggest that otherwise safe and effective inactivation methods should be further studied to identify possible gaps in their ability to inactivate contemporary, multidrug-resistant bacterial pathogens. IMPORTANCE Pathogen inactivation is a strategy to enhance the safety of transfused blood products. We identify the compound, amotosalen, widely used for pathogen inactivation, as a bacterial multidrug efflux substrate. Specifically, experiments suggest that amotosalen is pumped out of bacteria by major efflux pumps in E. coli, Acinetobacter baumannii, and Pseudomonas aeruginosa. Such efflux pumps are often overexpressed in multidrug-resistant pathogens. Importantly, the MICs for contemporary multidrug-resistant \u003Ci\u003EEnterobacterales\u003C\/i\u003E, Acinetobacter baumannii, Pseudomonas aeruginosa, \u003Ci\u003EBurkholderia\u003C\/i\u003E spp., and Stenotrophomonas maltophilia isolates approached or exceeded the amotosalen concentration used in approved platelet and plasma inactivation procedures, potentially as a result of efflux pump activity. Although there are important differences in methodology between our experiments and blood product pathogen inactivation, these findings suggest that otherwise safe and effective inactivation methods should be further studied to identify possible gaps in their ability to inactivate contemporary, multidrug-resistant bacterial pathogens.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EMorgan, Christopher E, Yoon-Suk Kang, Alex B Green, Kenneth P Smith, Matthew G Dowgiallo, Brandon C Miller, Lucius Chiaraviglio, et al. (2023) 2023. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/streptothricin-f-bactericidal-antibiotic-effective-against-highly-drug-resistant-gram\u0022 hreflang=\u0022en\u0022\u003EStreptothricin F Is a Bactericidal Antibiotic Effective Against Highly Drug-Resistant Gram-Negative Bacteria That Interacts With the 30S Subunit of the 70S Ribosome.\u003C\/a\u003E\u201d. \u003Ci\u003EPLoS Biology\u003C\/i\u003E 21 (5): e3002091. https:\/\/doi.org\/10.1371\/journal.pbio.3002091.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/37192172\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003EThe streptothricin natural product mixture (also known as nourseothricin) was discovered in the early 1940s, generating intense initial interest because of excellent gram-negative activity. Here, we establish the activity spectrum of nourseothricin and its main components, streptothricin F (S-F, 1 lysine) and streptothricin D (S-D, 3 lysines), purified to homogeneity, against highly drug-resistant, carbapenem-resistant Enterobacterales (CRE) and Acinetobacter baumannii. For CRE, the MIC50 and MIC90 for S-F and S-D were 2 and 4 \u03bcM, and 0.25 and 0.5 \u03bcM, respectively. S-F and nourseothricin showed rapid, bactericidal activity. S-F and S-D both showed approximately 40-fold greater selectivity for prokaryotic than eukaryotic ribosomes in in vitro translation assays. In vivo, delayed renal toxicity occurred at \u0026gt;10-fold higher doses of S-F compared with S-D. Substantial treatment effect of S-F in the murine thigh model was observed against the otherwise pandrug-resistant, NDM-1-expressing Klebsiella pneumoniae Nevada strain with minimal or no toxicity. Cryo-EM characterization of S-F bound to the A. baumannii 70S ribosome defines extensive hydrogen bonding of the S-F steptolidine moiety, as a guanine mimetic, to the 16S rRNA C1054 nucleobase (Escherichia coli numbering) in helix 34, and the carbamoylated gulosamine moiety of S-F with A1196, explaining the high-level resistance conferred by corresponding mutations at the residues identified in single rrn operon E. coli. Structural analysis suggests that S-F probes the A-decoding site, which potentially may account for its miscoding activity. Based on unique and promising activity, we suggest that the streptothricin scaffold deserves further preclinical exploration as a potential therapeutic for drug-resistant, gram-negative pathogens.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EMatysiak, Colette, Annie Cheng, and James E Kirby. (2023) 2023. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/evaluation-abbott-alinity-m-sti-assay-diagnosis-primary-causes-sexually-transmitted\u0022 hreflang=\u0022en\u0022\u003EEvaluation of the Abbott Alinity M STI Assay for Diagnosis of the Primary Causes of Sexually Transmitted Infections in the United States.\u003C\/a\u003E\u201d. \u003Ci\u003EPractical Laboratory Medicine\u003C\/i\u003E 36: e00332. https:\/\/doi.org\/10.1016\/j.plabm.2023.e00332.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/37705589\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003E\u003Cb\u003EOBJECTIVES: \u003C\/b\u003EThe sexually transmitted infections, \u003Ci\u003EChlamydia trachomatis\u003C\/i\u003E (CT), \u003Ci\u003ENeisseria gonorrhoeae\u003C\/i\u003E (NG), \u003Ci\u003ETrichomonas vaginalis\u003C\/i\u003E (TV), and \u003Ci\u003EMycoplasma genitalium\u003C\/i\u003E (MG), have similar risk factors and symptoms, supporting use of a quadruplex test as an efficient diagnostic modality.We assessed the clinical and analytical performance of the Abbott Alinity m STI assay to detect these pathogens.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003EDESIGN AND METHODS: \u003C\/b\u003EUrine and genital swabs from 142 patient samples were tested from an adult outpatient population in the Northeast United States. The positive and negative percent agreement for CT, NG, and TV were determined by comparison with the Hologic Panther Aptima assay. The analytical sensitivity was determined through serial dilution of standards for CT, NG, TV, and MG in negative urine and swab matrix.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003ERESULTS: \u003C\/b\u003EThe positive and negative percent agreement of the Alinity m assay in comparison with the Hologic Panther Aptima assay were, respectively: CT [100.0% (90.6-100.0%) and 99.1% (94.8-100.0%)], NG [100.0% (89.6-100.0%) and 99.1% (94.9-100.0%)]; and TV [96.3% (81.7-99.8%) and 99.1% (95.2-100.0%)]. The limits of detection in urine and swab matrix were, respectively: CT\u00a0\u2264\u00a05, \u22641; NG\u00a0\u2264\u00a05, \u22645; TV\u00a0\u2264\u00a00.5, \u22640.5; and MG\u00a0\u2264\u00a0500, \u2264250 genome copies\/mL.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003ECONCLUSIONS: \u003C\/b\u003EThe Alinity m assay demonstrated excellent performance characteristics and identifies CT, NG, and TV accurately compared with a well-established comparator.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EPawar, Rahul Dnyaneshwar, Lakshman Balaji, Shivani Mehta, Andrew Cole, Xiaowen Liu, Natia Peradze, Anne Victoria Grossestreuer, et al. (2022) 2022. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/viral-load-and-disease-severity-covid-19\u0022 hreflang=\u0022en\u0022\u003EViral Load and Disease Severity in COVID-19.\u003C\/a\u003E\u201d. \u003Ci\u003EInternal and Emergency Medicine\u003C\/i\u003E 17 (2): 359-67. https:\/\/doi.org\/10.1007\/s11739-021-02786-w.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/34133005\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003EThe relationship between COVID-19 severity and viral load is unknown. Our objective was to assess the association between viral load and disease severity in COVID-19. In this single center observational study of adults with laboratory confirmed SARS-CoV-2, the first positive in-hospital nasopharyngeal swab was used to calculate the log10 copies\/ml [log10 copy number (CN)] of SARS-CoV-2. Four categories based on level of care and modified sequential organ failure assessment score (mSOFA) at time of swab were determined. Median log10CN was compared between different levels of care and mSOFA quartiles. Median log10CN was compared in patients who did and did not receive influenza vaccine, and the correlation between log10CN and D-dimer was examined. We found that\u00a0of 396 patients, 54.3% were male, and 25% had no major comorbidity. Hospital mortality was 15.7%. Median mSOFA was 2 (IQR 0-3). Median log10CN was 5.5 (IQR 3.3-8.0). Median log10CN was highest in non-intubated ICU patients [6.4 (IQR 4.4-8.1)] and lowest in intubated ICU patients [3.6 (IQR 2.6-6.9)] (p value\u2009\u0026lt;\u20090.01). In adjusted analyses, this difference remained significant [mean difference 1.16 (95% CI 0.18-2.14)]. There was no significant difference in log10CN between other groups in the remaining pairwise comparisons. There was no association between median log10CN and mSOFA in either unadjusted or adjusted analyses or between median log10CN in patients with and without influenza immunization. There was no correlation between log10CN and D-dimer. We conclude, in our cohort, we did not find a clear association between viral load and disease severity in COVID-19 patients. Though viral load was higher in non-intubated ICU patients than in intubated ICU patients there were no other significant differences in viral load by disease severity.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EDowgiallo, Matthew G, Brandon C Miller, Mintesinot Kassu, Kenneth P Smith, Andrew D Fetigan, Jason J Guo, James E Kirby, and Roman Manetsch. (2022) 2022. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/convergent-total-synthesis-and-antibacterial-profile-natural-product-streptothricin-f\u0022 hreflang=\u0022en\u0022\u003EThe Convergent Total Synthesis and Antibacterial Profile of the Natural Product Streptothricin F.\u003C\/a\u003E\u201d. \u003Ci\u003EChemical Science\u003C\/i\u003E 13 (12): 3447-53. https:\/\/doi.org\/10.1039\/d1sc06445b.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/35432870\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003EA convergent, diversity-enabling total synthesis of the natural product streptothricin F has been achieved. Herein, we describe the potent antimicrobial activity of streptothricin F and highlight the importance of a total synthesis that allows for the installation of practical divergent steps for medicinal chemistry exploits. Key features of our synthesis include a Burgess reagent-mediated 1,2-\u003Ci\u003Eanti\u003C\/i\u003E-diamine installation, diastereoselective azidation of a lactam enolate, and a mercury(ii) chloride-mediated desulfurization-guanidination. The development of this chemistry enables the synthesis and structure-activity studies of streptothricin F analogs.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EBrennan-Krohn, Thea, Alexandra Grote, Shade Rodriguez, James E Kirby, and Ashlee M Earl. (2022) 2022. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/transcriptomics-reveals-how-minocycline-colistin-synergy-overcomes-antibiotic\u0022 hreflang=\u0022en\u0022\u003ETranscriptomics Reveals How Minocycline-Colistin Synergy Overcomes Antibiotic Resistance in Multidrug-Resistant Klebsiella Pneumoniae.\u003C\/a\u003E\u201d. \u003Ci\u003EAntimicrobial Agents and Chemotherapy\u003C\/i\u003E 66 (3): e0196921. https:\/\/doi.org\/10.1128\/aac.01969-21.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/35041511\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003EMultidrug-resistant Gram-negative bacteria are a rapidly growing public health threat, and the development of novel antimicrobials has failed to keep pace with their emergence. Synergistic combinations of individually ineffective drugs present a potential solution, yet little is understood about the mechanisms of most such combinations. Here, we show that the combination of colistin (polymyxin E) and minocycline has a high rate of synergy against colistin-resistant and minocycline-intermediate or -resistant strains of Klebsiella pneumoniae. Furthermore, using transcriptome sequencing (RNA-Seq), we characterized the transcriptional profiles of these strains when treated with the drugs individually and in combination. We found a striking similarity between the transcriptional profiles of bacteria treated with the combination of colistin and minocycline at individually subinhibitory concentrations and those of the same isolates treated with minocycline alone. We observed a similar pattern with the combination of polymyxin B nonapeptide (a polymyxin B analogue that lacks intrinsic antimicrobial activity) and minocycline. We also found that genes involved in polymyxin resistance and peptidoglycan biosynthesis showed significant differential gene expression in the different treatment conditions, suggesting possible mechanisms for the antibacterial activity observed in the combination. These findings suggest that the synergistic activity of this combination against bacteria resistant to each drug alone involves sublethal outer membrane disruption by colistin, which permits increased intracellular accumulation of minocycline.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003ECheng, Annie, Stefan Riedel, Ramy Arnaout, and James E Kirby. (2022) 2022. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/verification-abbott-alinity-m-resp-4-plex-assay-detection-sars-cov-2-influenza-ab-and\u0022 hreflang=\u0022en\u0022\u003EVerification of the Abbott Alinity M Resp-4-Plex Assay for Detection of SARS-CoV-2, Influenza A\/B, and Respiratory Syncytial Virus.\u003C\/a\u003E\u201d. \u003Ci\u003EDiagnostic Microbiology and Infectious Disease\u003C\/i\u003E 102 (2): 115575. https:\/\/doi.org\/10.1016\/j.diagmicrobio.2021.115575.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/34839127\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003ECOVID-19 symptomology may overlap with other circulating respiratory viruses that may also cause severe disease and for which there are specific and potentially life-saving treatments. The Abbott Alinity m Resp-4-Plex assay is a multiplex PCR assay that simultaneously detects and differentiates infection with SARS-CoV-2, influenza A, influenza B, and respiratory syncytial virus (RSV). We characterized its accuracy, precision, and analytical sensitivity. All were found to be robust for measures examined. In the context of sample-to-answer, near random access automation on the Alinity m platform, we believe that the Resp-4-Plex assay offers significant utility in addressing the current needs of the SARS-CoV-2 pandemic and future needs during anticipated endemic circulation of SARS-CoV-2 with other respiratory viruses.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003ESalamzade, Rauf, Abigail L Manson, Bruce J Walker, Thea Brennan-Krohn, Colin J Worby, Peijun Ma, Lorrie L He, et al. (2022) 2022. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/inter-species-geographic-signatures-tracing-horizontal-gene-transfer-and-long-term\u0022 hreflang=\u0022en\u0022\u003EInter-Species Geographic Signatures for Tracing Horizontal Gene Transfer and Long-Term Persistence of Carbapenem Resistance.\u003C\/a\u003E\u201d. \u003Ci\u003EGenome Medicine\u003C\/i\u003E 14 (1): 37. https:\/\/doi.org\/10.1186\/s13073-022-01040-y.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/35379360\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n                \u003Cdiv class=\u0022field--label field--abstract\u0022\u003E\n      \u003Cbutton class=\u0022btn-abstract collapsed\u0022 data-toggle=\u0022collapse\u0022 data-target=\u0022#collapseAbstract\u0022 aria-expanded=\u0022false\u0022 aria-controls=\u0022collapseAbstract\u0022\u003EAbstract \u003C\/button\u003E\n    \u003C\/div\u003E\n                  \u003Cdiv class=\u0022field--item abstract--content collapse\u0022 id=\u0022collapseAbstract\u0022 aria-expanded=\u0026quot;false\u0026quot;\u003E\u003Cp\u003E\u003Cb\u003EBACKGROUND: \u003C\/b\u003ECarbapenem-resistant Enterobacterales (CRE) are an urgent global health threat. Inferring the dynamics of local CRE dissemination is currently limited by our inability to confidently trace the spread of resistance determinants to unrelated bacterial hosts. Whole-genome sequence comparison is useful for identifying CRE clonal transmission and outbreaks, but high-frequency horizontal gene transfer (HGT) of carbapenem resistance genes and subsequent genome rearrangement complicate tracing the local persistence and mobilization of these genes across organisms.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003EMETHODS: \u003C\/b\u003ETo overcome this limitation, we developed a new approach to identify recent HGT of large, near-identical plasmid segments across species boundaries, which also allowed us to overcome technical challenges with genome assembly. We applied this to complete and near-complete genome assemblies to examine the local spread of CRE in a systematic, prospective collection of all CRE, as well as time- and species-matched carbapenem-susceptible Enterobacterales, isolated from patients from four US hospitals over nearly 5 years.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003ERESULTS: \u003C\/b\u003EOur CRE collection comprised a diverse range of species, lineages, and carbapenem resistance mechanisms, many of which were encoded on a variety of promiscuous plasmid types. We found and quantified rearrangement, persistence, and repeated transfer of plasmid segments, including those harboring carbapenemases, between organisms over multiple years. Some plasmid segments were found to be strongly associated with specific locales, thus representing geographic signatures that make it possible to trace recent and localized HGT events. Functional analysis of these signatures revealed genes commonly found in plasmids of nosocomial pathogens, such as functions required for plasmid retention and spread, as well survival against a variety of antibiotic and antiseptics common to the hospital environment.\u003C\/p\u003E\u003Cp\u003E\u003Cb\u003ECONCLUSIONS: \u003C\/b\u003ECollectively, the framework we developed provides a clearer, high-resolution picture of the epidemiology of antibiotic resistance importation, spread, and persistence in patients and healthcare networks.\u003C\/p\u003E\u003C\/div\u003E\n        \n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n \u003Cli\u003E\n\u003Carticle class=\u0022bibcite-reference bibcite bibcite--teaser\u0022\u003E\n  \n  \n  \n\n  \u003Cdiv class=\u0022bibcite__content\u0022\u003E\n    \u003Cdiv class=\u0022bibcite-citation\u0022\u003E\n      \u003Cdiv class=\u0022csl-bib-body\u0022\u003E\u003Cdiv class=\u0022csl-entry\u0022\u003EStanley, Sydney, Donald J Hamel, Ian D Wolf, Stefan Riedel, Sanjucta Dutta, Elisa Contreras, Cody J Callahan, et al. (2022) 2022. \u201c\u003Ca href=\u0022\/james-kirby\/publications\/limit-detection-rapid-antigen-testing-sars-cov-2-omicron-and-delta-variants-concern\u0022 hreflang=\u0022en\u0022\u003ELimit of Detection for Rapid Antigen Testing of the SARS-CoV-2 Omicron and Delta Variants of Concern Using Live-Virus Culture.\u003C\/a\u003E\u201d. \u003Ci\u003EJournal of Clinical Microbiology\u003C\/i\u003E 60 (5): e0014022. https:\/\/doi.org\/10.1128\/jcm.00140-22.\u003C\/div\u003E\u003C\/div\u003E\n  \u003C\/div\u003E\n\n  \u003Cdiv class=\u0022field field--name-publishers-version field--type-link field--label-visually_hidden field--mode-teaser\u0022\u003E\n    \u003Cdiv class=\u0022field--label sr-only\u0022\u003EPublisher\u0027s Version\u003C\/div\u003E\n              \u003Cdiv class=\u0022field--item\u0022\u003E\u003Ca href=\u0022https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/35440165\u0022\u003EPublisher\u0026#039;s Version\u003C\/a\u003E\u003C\/div\u003E\n          \u003C\/div\u003E\n\n  \u003C\/div\u003E\n\u003C\/article\u003E\n\u003C\/li\u003E\n\u003C\/ul\u003E\n  \u003Cnav role=\u0022navigation\u0022 aria-labelledby=\u0022pagination-for-lop-publications\u0022 id=pager-heading\u003E\n    \u003Ch3 id=\u0022pagination-for-lop-publications\u0022 class=\u0022visually-hidden\u0022\u003Epagination for lop publications\u003C\/h3\u003E\n    \u003Cul class=\u0022js-pager__items pager-mini\u0022\u003E\n            \u003Cli class=\u0022current\u0022\u003E\n        \u003Cspan aria-live=\u0022polite\u0022\u003E\n            \u003Cspan class=\u0022visually-hidden\u0022\u003ELOP - Publications\u003C\/span\u003E\n            1 of 6\n          \u003C\/span\u003E      \u003C\/li\u003E\n              \u003Cli\u003E\n          \u003Ca href=\u0022\/james-kirby\/refresh-widget-content\/29557?page=1\u0026amp;selector=list-of-posts\u0026amp;pagerid=pager-heading\u0026amp;moreid=node-readmore\u0022 class=\u0022use-ajax next\u0022 rel=\u0022next\u0022\u003E\u003Cspan aria-hidden=\u0022true\u0022\u003E\u203a\u203a\u003C\/span\u003E\u003Cspan class=\u0022visually-hidden\u0022\u003ENext page\u003C\/span\u003E\u003C\/a\u003E\n        \u003C\/li\u003E\n          \u003C\/ul\u003E\n  \u003C\/nav\u003E\n\n\u003Cdiv class=\u0022node-readmore\u0022 id=node-readmore\u003E\u003C\/div\u003E\n","settings":null},{"command":"insert","method":"replaceWith","selector":"#","data":"","settings":null},{"command":"insert","method":"replaceWith","selector":"#","data":"","settings":null},{"command":"insert","method":"replaceWith","selector":".field--name-field-widget-title","data":"","settings":null}]