The Type III Effectors NleE and NleB from Enteropathogenic E. coli and OspZ from Shigella Block Nuclear Translocation of NF-κB p65

Many bacterial pathogens utilize a type III secretion system to deliver multiple effector proteins into host cells. Here we found that the type III effectors, NleE from enteropathogenic E. coli (EPEC) and OspZ from Shigella, blocked translocation of the p65 subunit of the transcription factor, NF-κB, to the host cell nucleus. NF-κB inhibition by NleE was associated with decreased IL-8 expression in EPEC-infected intestinal epithelial cells. Ectopically expressed NleE also blocked nuclear translocation of p65 and c-Rel, but not p50 or STAT1/2. NleE homologues from other attaching and effacing pathogens as well OspZ from Shigella flexneri 6 and Shigella boydii, also inhibited NF-κB activation and p65 nuclear import; however, a truncated form of OspZ from S. flexneri 2a that carries a 36 amino acid deletion at the C-terminus had no inhibitory activity. We determined that the C-termini of NleE and full length OspZ were functionally interchangeable and identified a six amino acid motif, IDSY(M/I)K, that was important for both NleE- and OspZ-mediated inhibition of NF-κB activity. We also established that NleB, encoded directly upstream from NleE, suppressed NF-κB activation. Whereas NleE inhibited both TNFα and IL-1β stimulated p65 nuclear translocation and IκB degradation, NleB inhibited the TNFα pathway only. Neither NleE nor NleB inhibited AP-1 activation, suggesting that the modulatory activity of the effectors was specific for NF-κB signaling. Overall our data show that EPEC and Shigella have evolved similar T3SS-dependent means to manipulate host inflammatory pathways by interfering with the activation of selected host transcriptional regulators

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Vancomycin Flight Simulator: A Team-Based Learning Exercise

BACKGROUND: Team-based learning (TBL) encourages learners to think critically to solve problems they will face in practice. Pharmacokinetic dosing and monitoring are complex skills requiring the application of learned knowledge. The study sought to assess the impact of a TBL, vancomycin dosing activity in a Pharmaceutical Skills IV course measured with exam question performance during the second professional year. METHODS: This retrospective, descriptive study relates a TBL activity, assigned to 85 students, which included an individual student pre-preparation quiz, assigned readings, in-class individual and team-based readiness assessments, small group application of a vancomycin patient case, and group discussion/feedback on clinical decisions with supportive reasoning. The class year before and class year of the TBL implementation were compared using the total percentage of points possible earned by the class years, by topic. To minimize potential confounding, the primary outcome was the change in topic performance by the rank difficulty (e.g., the largest possible benefit being the hardest topic becoming the easiest with no other variation in topic rank difficulty). RESULTS: In the year of implementation, the mean individual readiness assurance test (IRAT) performance was 5.5 ± 1.88 (10 points possible, 55%). The mean team readiness assurance test (TRAT) performance was 10 of 10 points possible (100%). The class exam item performance in the year before (n = 101) and year of (n = 84) TBL implementation showed a general decline in exam scores. However, the vancomycin topic difficultly went from fifth easiest, to second easiest, with less than 1% change in raw score. CONCLUSIONS: Implementation of a pharmacokinetic TBL activity appeared to moderately support the students’ vancomycin learning. Additional studies are warranted on APPE readiness and performance

Self-Directed, Higher-Level Learning Through Journal Club Debates

Purpose: Journal clubs are frequently used by healthcare educators to support learner’s critical thinking and clinical application. However, there is a lack of evidence to support the effectiveness of this learning tool for both the learner utilizing this tool and those in attendance. Debates have been proven to support critical thinking and communication skills. Method: A journal club debate format and rubric were created to promote an engaging format for effective interpretation and application of drug information among fourth year pharmacy students and residents. Sections from relevant articles identified in a literature search were used to create a journal club debate format and rubric. This updated tool was used with pairs of pharmacy students and pharmacy residents. Feedback was obtained from the learners and the pharmacy preceptors in attendance. Results: A journal club debate is generally well accepted by both learners and preceptors. Benefits include promotion of critical thinking, interpretation of evidence in the scope of supporting literature, and deeper understanding of the relevant disease state and treatment. Opportunities to improve may be the inclusion of limited visual aids for supporting evidence and the expansion of time for discussion and rebuttals. Conclusion: Journal club debates are an engaging learning tool to promote and develop drug information identification, evaluation, and application to clinical practice among pharmacy students and residents

Longitudinal Assessment of Empathy and Burnout Across a Single Pharmacy Class Cohort.

OBJECTIVE: This is a final analysis of longitudinal evaluation of burnout and empathy among a cohort of Doctor of Pharmacy students throughout their four-year enrollment. METHODS: The Class of 2021 received sequential Qualtrics RESULTS: Matched survey responses were included for 91 students (85.8% response rate). Across all years, a decrease in empathy and professional efficacy and an increase in exhaustion and cynicism was seen. High categorical levels of exhaustion and cynicism indicated evidence of burnout throughout the program. Year-to-year analysis indicated statistically significant increases in exhaustion and cynicism between PY1 CONCLUSION: Students reported trends of decreasing empathy and professional efficacy, with a simultaneous increase in exhaustion and cynicism. Further evaluation of the impact of COVID-19 on these results, as well as additional methods to support overall student wellness, is needed

Measures of Burnout and Empathy in United States Doctor of Pharmacy Students: Time for a Change?

PURPOSE: To review interim data regarding longitudinal burnout and empathy levels in a single Doctor of Pharmacy class cohort. METHODS: Students were emailed an electronic survey during their first semester and annually at the end of each academic year for a total of 3 years (2017-2020). Validated survey tools included the Jefferson Scale of Empathy (JSE) and the Maslach Burnout Inventory (MBI) student version. The JSE survey consists of 20 questions, with higher scores denoting more empathy. The MBI student version contains 3 subscales: exhaustion (higher scores are worse), cynicism (higher scores are worse) and professional efficacy (higher scores are better). RESULTS: The median JSE score at the end of the third academic year (PY3) was 110, with females scoring significantly higher (114.5 vs. 103.5; p CONCLUSION: This interim data suggests high degrees of pharmacy student burnout. Empathy levels remained stable throughout the duration of the study. Pharmacy schools may need to focus on reform regarding well-being and prevention of burnout

The Genetic Architecture of Skeletal Convergence and Sex Determination in Ninespine Sticklebacks

SummaryThe history of life offers plentiful examples of convergent evolution, the independent derivation of similar phenotypes in distinct lineages [1]. The emergence of convergent phenotypes among closely related lineages (frequently termed “parallel” evolution) is often assumed to result from changes in similar genes or developmental pathways [2], but the genetic origins of convergence remains poorly understood. Ninespine (Pungitius pungitius) and threespine (Gasterosteus aculeatus) stickleback fish provide many examples of convergent evolution of adaptive phenotypes, both within and between genera. The genetic architecture of several important traits is now known for threespine sticklebacks [3–10]; thus, ninespine sticklebacks provide a unique opportunity to critically test whether similar or different chromosome regions control similar phenotypes in these lineages. We have generated the first genome-wide linkage map for ninespine sticklebacks and used quantitative trait locus mapping to identify chromosome regions controlling several skeletal traits and sex determination. In ninespine sticklebacks, these traits mapped to chromosome regions not previously known to control the corresponding traits in threespine sticklebacks. Therefore, convergent morphological evolution in these related, but independent, vertebrate lineages might have different genetic origins. Comparative genetics in sticklebacks provides an exciting opportunity to study the mechanisms controlling similar phenotypic changes in different animal groups