Fluoroquinolones and isoniazid-resistant tuberculosis: implications for the 2018 WHO guidance.

INTRODUCTION: 2018 World Health Organization (WHO) guidelines for the treatment of isoniazid (H)-resistant (Hr) tuberculosis recommend a four-drug regimen: rifampicin (R), ethambutol (E), pyrazinamide (Z) and levofloxacin (Lfx), with or without H ([H]RZE-Lfx). This is used once Hr is known, such that patients complete 6 months of Lfx (≥6[H]RZE-6Lfx). This cohort study assessed the impact of fluoroquinolones (Fq) on treatment effectiveness, accounting for Hr mutations and degree of phenotypic resistance. METHODS: This was a retrospective cohort study of 626 Hr tuberculosis patients notified in London, 2009-2013. Regimens were described and logistic regression undertaken of the association between regimen and negative regimen-specific outcomes (broadly, death due to tuberculosis, treatment failure or disease recurrence). RESULTS: Of 594 individuals with regimen information, 330 (55.6%) were treated with (H)RfZE (Rf=rifamycins) and 211 (35.5%) with (H)RfZE-Fq. The median overall treatment period was 11.9 months and median Z duration 2.1 months. In a univariable logistic regression model comparing (H)RfZE with and without Fqs, there was no difference in the odds of a negative regimen-specific outcome (baseline (H)RfZE, cluster-specific odds ratio 1.05 (95% CI 0.60-1.82), p=0.87; cluster NHS trust). Results varied minimally in a multivariable model. This odds ratio dropped (0.57, 95% CI 0.14-2.28) when Hr genotype was included, but this analysis lacked power (p=0.42). CONCLUSIONS: In a high-income setting, we found a 12-month (H)RfZE regimen with a short Z duration to be similarly effective for Hr tuberculosis with or without a Fq. This regimen may result in fewer adverse events than the WHO recommendations

AB0846 CONCEPTION AND FEASIBILITY OF A DIGITAL TELE-GUIDED ABDOMEN, THORAX AND THYROID GLAND ULTRASOUND COURSE FOR MEDICAL STUDENTS

Background:Over the past few decades, technological advances in both ultrasound (US) and the application of telemedicine have been made [1]. Medical education has shifted to online classes during the COVID-19 pandemic, creating challenges in adequate training of US.Objectives:In the context of the current COVID-19 pandemic and the discontinuation of traditional classroom teaching, a tele-didactic US course for undergraduate medical students was developed. This study is examining the educational outcome of a new digital US course of the abdomen, thorax and the thyroid gland with the implementation of a modern portable US system.Methods:A tele-guided US course was established between April 1st and June 20th, 2020, at the University Hospital in Bonn, Germany. Students completed pre-and post-course surveys and underwent six US organ modules. Each module took place in a flipped-classroom concept including a digital learning platform. An objective structured assessment of US skills (OSAUS) [2] was implemented as final exam. Further, US images of the course and exam were rated by the Brightness Mode Quality Ultrasound Imaging Examination Technique (B-QUIET) [3]. Achieved points in image rating were compared to the OSAUS exam and survey results.Results:A total of 15 medical students were enrolled. There was a significant increase in self-assessed theoretical and practical ability (p &lt; 0.001). Students achieved an average score of 154.5 (SD ± 11.72) out of 175 points (88.29%) in OSAUS, which corresponded to the image rating using B-QUIET and self-assessment data obtained in surveys. Interrater analysis of US images showed a favorable agreement with an ICC (2,1) of 0.895 (95% confidence interval 0.858 &lt; ICC &lt; 0.924).Conclusion:US training via teleguidance should be considered in medical education. Our pilot study demonstrates the feasibility of this concept that can be used in the future to improve US training of medical students even during a pandemic. The digital implementation with an affordable, portable point-of-care-US device could be an incredible opportunity to expedite US training worldwide.References:[1]Law J, Macbeth PB. Ultrasound: from Earth to space. Mcgill J Med 2011; 13(2): 59 [PMID: 22399873][2]Tolsgaard MG, Todsen T, Sorensen JL, et al. International multispecialty consensus on how to evaluate ultrasound competence: a Delphi consensus survey. PLoS One 2013; 8(2): e57687[https://doi.org/10.1371/journal.pone.0057687][PMID: 23469051][3]Bahner DP, Adkins EJ, Nagel R, Way D, Werman HA, Royall NA. Brightness mode quality ultrasound imaging examination technique (B-QUIET): quantifying quality in ultrasound imaging. J Ultrasound Med 2011; 30(12): 1649–55[4][https://doi.org/10.7863/jum.2011.30.12.1649][PMID: 22124000]Figure 1.Implementation of the digital ultrasound course conceptClassroom setting is displayed, demonstrating simultaneous screen transmission of ultrasound (US) and camera images. A: The peer tutor’s camera image is shared with the students while demonstrating the US examination, B: At the same time the tutor’s mobile phone screen is shared with the students in order to visualize the US image acquisition, C: Students performing the US examination on their own, D: Example of a worksheet, which had to be completed within the app and was stored along with the US images in the cloudDisclosure of Interests:None declared</jats:sec

THU0650-HPR THE USE OF GAMIFICATION TO MOTIVATE HEALTH PROFESSIONALS IN RHEUMATOLOGY TO PARTICIPATE IN BLENDED LEARNING.

Background:Blended learning is an increasingly popular learning supplement for traditional classroom-based courses in medical education. Once implemented, many factors influence its success. This was demonstrated by Shivetts et al., who concluded that student motivation plays a major role. In particular, if a learner is not self-motivated, e-learning may not represent the best learning environment.1However, gamification methods are known to enhance motivation in medical education and, if used correctly, can overcome this deficit.2For this purpose, a quiz duel was created and used as a blended learning approach for health professional training in rheumatology. We hypothesize that the use of the quiz duel gamification technique improves learners’ motivation to successfully complete their blended learning course.Objectives:To investigate the potential of gamification methods in motivating health professionals to answer multiple choice questions (MCQs) in a pilot blended learning scenario.Methods:Four hundred and sixty MCQs were developed in accordance with the learning objectives of a certified training course and integrated into a learning management system (LMS). As a gamification technique, a duel mode was created. Course participants had access via an individual user account and used personal smartphones. After each answer was provided, the learners received corrective and explanatory feedback, as well as information on how the duel opponent answered. Incorrectly answered MCQs were repeated in further duels.Information on the number of MCQs answered (1), days learned (2), and learning time spent (3) was collected and analyzed. Each day on which at least one MCQ was answered counted as a learning day per user. The learning time was calculated with 1.5 min per MCQ answered. Analysis was performed over a 15-week period (08/19–12/19).The training event (“RFAplus”) was organized by the Rheumatologische Fortbildungsakademie GmbH and took place on three weekends in intervals of four weeks in Germany. The LMS used was Humeo (Humeo GmbH). All users agreed to the terms and conditions of use and data protection before participating in the blended learning intervention.Results:Nineteen female health professionals in rheumatology participated in the study. The mean age of participants was 43.5 years (range, 21–60 years). The 460 MCQs were answered 20,397 times, with 1039 MCQs per user (range, 247-1839 MCQs) during the 15-week period (105 days). Each MCQ was answered 2.33 times. In total, there were 1167 learning days, with 60.8 days per user (range, 15-95 days). The users spent 30,596 min (509.9 h) answering the MCQs, resulting in 1,610 min (or 26.8 h) per user. Furthermore, each user answered 17.5 MCQs and spent an average of 26 min per learning day.Conclusion:Blended learning is an interactive method to potentially extend learning time over several weeks. However, the success of this technique lies in motivating the participants to continue learning after the event. A quiz duel as a gamification technique proved to be effective in motivating participants to learn daily. In our study, learners spent an average of 27 h, i.e., almost half of the total attendance time of 60 h, learning. Correspondingly, this technique could also replace parts of lengthy face-to-face courses in an attempt to save costs in the future. Information drawn from the MCQs could potentially serve as promising learning analytics.References:[1]Shivetts, C. E-Learning and Blended Learning: The Importance of the Learner A Research Literature Review.Int. J. E-Learn.10, 331–337 (2011).[2]Pesare, E., Roselli, T., Corriero, N. &amp; Rossano, V. Game-based learning and Gamification to promote engagement and motivation in medical learning contexts.Smart Learn. Environ.3, 5 (2016).Disclosure of Interests:Ruben Sengewein: None declared, Patricia Steffens-Korbanka Consultant of: Abbvie, Chugai, Novartis, Sanofi, Mylan, Lilly, Speakers bureau: Abbvie, Chugai, Novartis, Sanofi, Lilly, Joerg Wendler Consultant of: Janssen, AbbVie, Sanofi, Speakers bureau: Roche, Chugai, Janssen, AbbVie, Novartis, Max Kieslich: None declared, Erik Schmok: None declared, Georg Gauler Consultant of: Abbvie, Lilly, MSD, Speakers bureau: Abbvie, Celgene, Novartis, Sanofi,</jats:sec

Large-scale tethered function assays identify factors that regulate mRNA stability and translation

The molecular functions of the majority of RNA-binding proteins (RBPs) remain unclear, highlighting a major bottleneck to a full understanding of gene expression regulation. Here, we develop a plasmid resource of 690 human RBPs that we subject to luciferase-based 3'-untranslated-region tethered function assays to pinpoint RBPs that regulate RNA stability or translation. Enhanced UV-cross-linking and immunoprecipitation of these RBPs identifies thousands of endogenous mRNA targets that respond to changes in RBP level, recapitulating effects observed in tethered function assays. Among these RBPs, the ubiquitin-associated protein 2-like (UBAP2L) protein interacts with RNA via its RGG domain and cross-links to mRNA and rRNA. Fusion of UBAP2L to RNA-targeting CRISPR-Cas9 demonstrates programmable translational enhancement. Polysome profiling indicates that UBAP2L promotes translation of target mRNAs, particularly global regulators of translation. Our tethering survey allows rapid assignment of the molecular activity of proteins, such as UBAP2L, to specific steps of mRNA metabolism

Fluoroquinolones and isoniazid resistant TB: implications for the 2018 WHO guidance

Introduction: 2018 World Health Organization (WHO) guidelines for the treatment of isoniazid (H)-resistant (Hr) tuberculosis recommend a four-drug regimen: rifampicin (R), ethambutol (E), pyrazinamide (Z) and levofloxacin (Lfx), with or without H ([H]RZE-Lfx). This is used once Hr is known, such that patients complete 6 months of Lfx (≥6[H]RZE-6Lfx). This cohort study assessed the impact of fluoroquinolones (Fq) on treatment effectiveness, accounting for Hr mutations and degree of phenotypic resistance. / Methods: This was a retrospective cohort study of 626 Hr tuberculosis patients notified in London, 2009–2013. Regimens were described and logistic regression undertaken of the association between regimen and negative regimen-specific outcomes (broadly, death due to tuberculosis, treatment failure or disease recurrence). / Results: Of 594 individuals with regimen information, 330 (55.6%) were treated with (H)RfZE (Rf=rifamycins) and 211 (35.5%) with (H)RfZE-Fq. The median overall treatment period was 11.9 months and median Z duration 2.1 months. In a univariable logistic regression model comparing (H)RfZE with and without Fqs, there was no difference in the odds of a negative regimen-specific outcome (baseline (H)RfZE, cluster-specific odds ratio 1.05 (95% CI 0.60–1.82), p=0.87; cluster NHS trust). Results varied minimally in a multivariable model. This odds ratio dropped (0.57, 95% CI 0.14–2.28) when Hr genotype was included, but this analysis lacked power (p=0.42). / Conclusions: In a high-income setting, we found a 12-month (H)RfZE regimen with a short Z duration to be similarly effective for Hr tuberculosis with or without a Fq. This regimen may result in fewer adverse events than the WHO recommendations

Discovery and functional interrogation of SARS-CoV-2 protein-RNA interactions

AbstractThe COVID-19 pandemic is caused by severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2). The betacoronvirus has a positive sense RNA genome which encodes for several RNA binding proteins. Here, we use enhanced crosslinking and immunoprecipitation to investigate SARS-CoV-2 protein interactions with viral and host RNAs in authentic virus-infected cells. SARS-CoV-2 proteins, NSP8, NSP12, and nucleocapsid display distinct preferences to specific regions in the RNA viral genome, providing evidence for their shared and separate roles in replication, transcription, and viral packaging. SARS-CoV-2 proteins expressed in human lung epithelial cells bind to 4773 unique host coding RNAs. Nine SARS-CoV-2 proteins upregulate target gene expression, including NSP12 and ORF9c, whose RNA substrates are associated with pathways in protein N-linked glycosylation ER processing and mitochondrial processes. Furthermore, siRNA knockdown of host genes targeted by viral proteins in human lung organoid cells identify potential antiviral host targets across different SARS-CoV-2 variants. Conversely, NSP9 inhibits host gene expression by blocking mRNA export and dampens cytokine productions, including interleukin-1α/β. Our viral protein-RNA interactome provides a catalog of potential therapeutic targets and offers insight into the etiology of COVID-19 as a safeguard against future pandemics.</jats:p