We present a proximity ligation-based multiplexed protein detection procedure in which several selected proteins can be detected via unique nucleic-acid identifiers and subsequently quantified by real-time PCR. The assay requires a 1-microl sample, has low-femtomolar sensitivity as well as five-log linear range and allows for modular multiplexing without cross-reactivity. The procedure can use a single polyclonal antibody batch for each target protein, simplifying affinity-reagent creation for new biomarker candidates.
Summary Each Pseudomonas aeruginosa cell localizes two types of motility structures, a single flagellum and one or two clusters of type IV pili, to the cell poles. Previous studies suggested that these motility structures arrive at the pole through distinct mechanisms. Here we performed a swimming motility screen to identify polar flagellum localization factors and discovered three genes homologous to the TonB/ExbB/ExbD complex that have defects in both flagella-mediated swimming and pilus-mediated twitching motility. We found that deletion of tonB3, PA2983 or PA2982 led to non-polar localization of the flagellum and FlhF, which was thought to sit at the top of the flagellar localization hierarchy. Surprisingly, these mutants also exhibited pronounced changes in pilus formation or localization, indicating that these proteins may co-ordinate both the pilus and flagellum motility systems. Thus, we have renamed PA2983 and PA2982, pocA and pocB, respectively, for polar organelle co-ordinator to reflect this function. Our results suggest that TonB3, PocA and PocB may form a membrane-associated complex, which we term the Poc complex. These proteins do not exhibit polar localization themselves, but are required for increased expression of pilus genes upon surface association, indicating that they regulate motility structures through either localization or transcriptional mechanisms.
Introduction: Distal radius fractures are common orthopedic injuries managed in emergency departments. Simulation-based mastery learning is widely recognized to improve provider competence for bedside procedures but has not been studied to teach fracture management. This study evaluated the effectiveness of a simulation-based mastery learning curriculum to teach distal radius fracture reduction to novice orthopedic surgery and emergency medicine residents. Methods: We created a novel mastery learning checklist using the Mastery Angoff method of standard setting, paired with a new simulation model designed for this project, to teach orthopedic surgery and emergency medicine interns (N = 22) at the study site. Orthopedic surgery and emergency medicine faculty members participated in checklist development, curriculum design, and implementation. Training included just-in-time asynchronous education with a readiness assessment test, in-classroom expert demonstration, and deliberate practice with feedback. Residents completed a pretest/posttest skills examination and a presurvey/postsurvey assessing procedural confidence. Results: Standard setting resulted in a 41-item checklist with minimum passing score of 37/41 items. All participants met or surpassed the minimum passing score on postexamination. Postsurvey confidence levels were significantly higher than presurvey in all aspects of the distal radius fracture procedure (P < 0.05). Conclusions: This study demonstrated that a simulation-based mastery learning curriculum improved skills and confidence performing distal radius fracture reductions for orthopedic surgery and emergency medicine interns. Future planned studies include curriculum testing across additional institutions, examination of clinical impact, and application of mastery learning for other orthopedic procedures.
Introduction: The Covid-19 pandemic limited educational and career development opportunities for medical students, requiring innovative programs to accelerate professional identity formation and clinical skills acquisition. Methods: We developed a brief coaching intervention that took place over the advanced (sub-internship) emergency medicine rotation at our institution. We trained coaches using a newly developed workshop, who met with students for an average of 4.5 hours over 3 weeks. Impact/effectiveness: We showed that this coaching program was both feasible and impactful for faculty coaches and medical students.
AbstracrThis paper describes the rationale and the experimental program undertaken to develop an industry-standard surface-charge monitor wafer employing EEPROM-based test structures to monitor the driving forces behind charging damage associated with ion-and plasma-based IC processes. B c t i o sThe spectacular increases in performance and density of VLSI integrated circuits during the past decade are largely due to the scaling of device structures, and the use of advanced ion-beam and plasma-based processes. Regrettably, scaling of device structures makes them more fragile, and thus more susceptible to damage by such processes [1,2]. The trend to higher ion-beam currents and higher RF powers to achieve greater equipment throughput only aggravates the problem. Having, at one time or another, suffered yield losses or degradation of device performance or reliability on their products, IC manufacturers now employ extensive monitoring procedures to ensure product quality. The growing concern in this area was evident at the SEMATECH Plasma Damage Workshop [3], where upwards of forty different device parameters were identified as possible plasma-etch related damage monitors. Although efforts to standardize the monitor structures and measurement procedures are being made under the auspices of SEMATECH, virtually all of them revolve around the measurement of changes in electrical characteristics of different device structures to monitor device damage. Although essential to ensuring product reliability, these types of monitors frequently provide little insight into the causeeffect relationships between device degradation and process recipes. Moreover, the fabrication of these structures invariably involves several processing steps, making it more difficult to assess the integrity of individual process steps. A Case for Fundamentals-Based Wafer Surface Charge MonitorsIn exploring other options, it is worthwhile to consider what attributes other charge monitors would have to offer to compete with, or supplement, the device-structure oriented monitors presently in use. Certainly, the ability to accurately assess the integrity of individual ion-or plasma-based process steps independently of other process steps is extremely desirable: it would promote more focused process improvement efforts, and provide equipment manufacturers with the ability to verify the capabilities of their equipment without the likely complications of other process steps. This could enable equipment manufacturers to develop more benign wafer-processing equipment. Moreover, the monitors would have to be incorporated onto silicon wafers to pass through the wafer transport systems, and not contaminate the process equipment. And, most importantly, they would have to measure those attributes of a process which are responsible for the damage observed on IC device structures.These combined requirements, especially the last one, require a change in approach which becomes apparent when we recognize that most of the observed damage effects are due to the transpo...
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