Search

Background The integration of entrustable professional activities (EPAs) within objective structured clinical examinations (OSCEs) has yielded a valuable avenue for delivering timely feedback to residents. However, concerns about feedback quality persist. Objective This study aimed to assess the quality and content alignment of verbal feedback provided by examiners during an entrustment-based OSCE. Methods We conducted a progress test OSCE for internal medicine residents in 2022, assessing 7 EPAs. The immediate 2-minute feedback provided by examiners was recorded and analyzed using the Quality of Assessment of Learning (QuAL) score. We also analyzed the degree of alignment with EPA learning objectives: competency milestones and task-specific abilities. In a randomized crossover experiment, we compared the impact of 2 scoring methods used to assess residents' clinical performance (3-point entrustability scales vs task-specific checklists) on feedback quality and alignment. Results Twenty-one examiners provided feedback to 67 residents. The feedback demonstrated high quality (mean QuAL score 4.3 of 5) and significant alignment with the learning objectives of the EPAs. On average, examiners addressed in their feedback 2.5 milestones (61%) and 1.2 task-specific abilities (46%). The scoring methods used had no significant impact on QuAL scores (95% CI -0.3, 0.1, P =.28), alignment with competency milestones (95% CI -0.4, 0.1, P =.13), or alignment with task-specific abilities (95% CI -0.3, 0.1, P =.29). Conclusions In our entrustment-based OSCE, examiners consistently offered valuable feedback aligned with intended learning outcomes. Notably, we explored high-quality feedback and alignment as separate dimensions, finding no significant impact from our 2 scoring methods on either aspect.

Background: As the scope of pharmacy practice is expanding, a growing number of pharmacists perform physical examination (PE) to gather additional information to monitor the effectiveness and safety of their patients' therapy. This professional activity calls for the development of comprehensive and valuable PE training. We sought to determine by consensus which PE tests should be given teaching priority in pharmacy education., Methods: Using existing PE literature in pharmacy, we conducted an online Delphi survey from December 2021 to April 2022 with 16 pharmacists who practise in a variety of settings and/or who are considered experts in PE., Results: After 2 Delphi rounds, consensus was reached to either include or exclude 27 PE tests in entry-to-practice programs. One last round allowed prioritizing the agreed-upon PE tests in terms of educational needs. Clinicians agreed that measuring blood pressure is indispensable and should be given teaching priority, followed by pulse rate, weight and blood glucose measurements. Endocrine system and head and neck examinations should be included in pharmacy programs, but their clinical usefulness was considered less important., Discussion: We compared our results with PE literature in other health care disciplines. We found that only a few PE tests truly influence drug therapy management, that some examinations can be quite difficult to perform accurately and that without proper training and opportunities to retrain, skill decay can lead to dangerous misinterpretations. Pharmacy programs should consider focusing on teaching PE tests supported by evidence as having an impact on drug therapy management. Can Pharm J (Ott) 2024;157:xx-xx., Competing Interests: The authors declared no potential conflicts of interest with respect to the research, authorship and/or publication of this article., (© The Author(s) 2024.)

Introduction: In simulation, students often observe their peers perform a task. It is still unclear how different types of instructional guidance can turn the observational phase into an active learning experience for novices. This mixed-method study aims to understand similarities and differences between use of collaboration scripts and checklists by observers in terms of cognitive load and perception of learning., Methods: Second-year pharmacy students ( N = 162) were randomly assigned to 1 of 4 conditions when observing a simulation: collaboration scripts (heuristic to analyze in dyads while observing), checklists, both, or no guidance. We measured observers' intrinsic and extraneous cognitive load and self-perceived learning and conducted focus group interviews., Results: Intrinsic cognitive load was significantly lower for checklists (M = 3.6/10) than for scripts (M = 4.7/10) or scripts and checklists combined (M = 4.7/10). Extraneous cognitive load was significantly lower for checklists (M = 1.5/10) than for scripts combined with checklists (M = 2.6/10) or no guidance (M = 1.8/10). There was no statistical difference between conditions for self-perceived learning. Coding of focus group interviews revealed 6 themes on observers' perception of learning under different conditions of instructional guidance. Students explained that collaboration scripts felt more complex, whereas checklists were perceived as a simple fact-checking exercise. Observing the simulation, regardless of guidance, was a meaningful learning experience., Conclusions: With or without guidance, observers are actively engaged with the simulation, but their effort differed depending on instructions. When choosing between checklists or collaboration scripts, educators should be guided by the type of simulation task., Competing Interests: The authors declare no conflict of interest., (Copyright © 2023 Society for Simulation in Healthcare.)

Purpose: In vivo immune cell tracking using MRI can be a valuable tool for studying the mechanisms underlying successful cancer therapies. Current cell labeling methods using superparamagnetic iron oxide (SPIO) lack the persistence to track the fate and location of transplanted cells long-term. Magnetospirillum magneticum is a commercially available, iron-producing bacterium that can be taken up by and live harmoniously within mammalian cells as magneto-endosymbionts (MEs). MEs have shown promise as labeling agents for in vivo stem and cancer cell tracking but have yet to be evaluated in immune cells. This pilot study examined ME labeling in myeloid-derived suppressor cells (MDSCs), cytotoxic T lymphocytes (CTLs), and dendritic cells (DCs) and its effects on cell purity, function, and MRI contrast., Procedures: MDSCs, CTLs, and DCs were incubated with MEs at various ME labeling ratios (MLR), and various biological metrics and iron uptake were assessed. For in vivo imaging, MDSCs were labeled overnight with either MEs or SPIO (Molday ION Rhodamine B) and injected into C3 tumor-bearing mice via tail vein injection 24 days post-implant and scanned daily with MRI for 1 week to assess cellular quantification., Results: Following incubations, MDSCs contained > 0.6 pg Fe/cell. CTLs achieved Fe loading of < 0.5 pg/cell, and DCs achieved Fe loading of ~ 1.4 pg/cell. The suppressive functionality of MDSCs at 1000 MLR was not affected by ME labeling but was affected at 2000 MLR. Markers of CTL dysfunction were not markedly affected by ME labeling nor were DC markers. In vivo data demonstrated that the MDSCs labeled with MEs generated sufficient contrast to be detectable using TurboSPI, similar to SPIO-labeled cells., Conclusions: Cells can be labeled with sufficient numbers of MEs to be detectable with MRI without compromising cell viability. Care must be taken at higher concentrations of MEs, which may affect some cell types' functional activity and/or morphology. Immune cells with minimal phagocytic behavior have much lower iron content per cell after incubation with MEs vs SPIO; however, MEs can successfully be used as a contrast agent for phagocytic immune cells., (© 2023. The Author(s), under exclusive licence to World Molecular Imaging Society.)

Introduction: When designing simulation for novices, educators aim to design tasks and environments that are complex enough to promote learning but not too complex to compromise task performance and cause cognitive overload. This study aimed to determine the impact of modulating task and environment complexity on novices' performance and cognitive load during simulation., Methods: Second-year pharmacy students (N = 162) were randomly assigned to one of four conditions (2 × 2 factorial design) in simulation: simple task in simple environment, complex task in simple environment, simple task in complex environment and complex task in complex environment. Using video recordings, two raters assessed students' performance during the simulation. We measured intrinsic cognitive load (ICL) and extraneous cognitive load (ECL) with questionnaires after the task and tested knowledge after task and debriefing., Results: Mean performance scores in simple environment were 28.2/32 (SD = 3.8) for simple task and 25.8/32 (SD = 4.2) for complex task. In complex environment, mean performance scores were 24.6/32 (SD = 5.2) for simple task and 25.6/32 (SD = 5.3) for complex task. We found significant interaction effects between task and environment complexity for performance. In simple environment, mean ICL scores were 4.2/10 (SD = 2.2) for simple task and 5.7/10 (SD = 1.5) for complex task. In complex environment, mean ICL scores were 4.9/10 (SD = 1.8) for simple task and 5.1/10 (SD = 1.9) for complex task. There was a main effect of task complexity on ICL. For ECL, we found neither an interaction effect nor main effects of task and environment complexity. There was a main effect of task complexity on knowledge test after task and main effects of both task and environment complexity on knowledge after debriefing., Conclusions: Performance was good, and cognitive load remained reasonable in all conditions, which suggests that, despite increased complexity, students seemed to strategically manage their own cognitive load and learn from the simulations. Our findings also indicate that environmental complexity contributes to ICL., (© 2022 Association for the Study of Medical Education and John Wiley & Sons Ltd.)

DPX is a unique T cell activating formulation that generates robust immune responses (both clinically and preclinically) which can be tailored to various cancers via the use of tumor-specific antigens and adjuvants. While DPX-based immunotherapies may act complementary with checkpoint inhibitors, combination therapy is not always easily predictable based on individual therapeutic responses. Optimizing these combinations can be improved by understanding the mechanism of action underlying the individual therapies. Magnetic Resonance Imaging (MRI) allows tracking of cells labeled with superparamagnetic iron oxide (SPIO), which can yield valuable information about the localization of crucial immune cell subsets. In this work, we evaluated the use of a multi-echo, single point MRI pulse sequence, TurboSPI, for tracking and quantifying cytotoxic T lymphocytes (CTLs) and myeloid lineage cells (MLCs). In a subcutaneous cervical cancer model (C3) we compared untreated mice to mice treated with either a single therapy (anti-PD-1 or DPX-R9F) or a combination of both therapies. We were able to detect, using TurboSPI, significant increases in CTL recruitment dynamics in response to combination therapy. We also observed differences in MLC recruitment to therapy-draining (DPX-R9F) lymph nodes in response to treatment with DPX-R9F (alone or in combination with anti-PD-1). We demonstrated that the therapies presented herein induced time-varying changes in cell recruitment. This work establishes that these quantitative molecular MRI techniques can be expanded to study a number of cancer and immunotherapy combinations to improve our understanding of longitudinal immunological changes and mechanisms of action., Competing Interests: At the time of writing this manuscript, MLT, GW, and MS were employees of IMV Inc., who manufactures DPX-based immunotherapies. KDB also had a research contract with IMV for an unrelated project., (© 2020 The Author(s). Published with license by Taylor & Francis Group, LLC.)

Background: In self-regulated procedural simulation, learners practise on many simulators (e.g. paracentesis), self-regulating their choice of simulators, time and goals. Current needs assessments cannot predict the number of simulators needed to plan cost-effective self-regulated simulation. Knowing the ratios of simulators and participants would allow for better-informed purchase decisions to be made., Methods: We designed 90-minute sessions of self-regulated procedural simulation for internal medicine residents. In Phase 1, 51 participants (8.5 per group) could use 22 simulators (US$69 925): ultrasound-guided central (n = 6) and peripheral (n = 2) venous catheterisation; thoracocentesis (n = 2); paracentesis (n = 2); lumbar puncture (n = 6); and arthrocentesis (n = 4). We calculated minimal numbers of simulators based on the time that participants used each simulator in order to design a resource-effective Phase 2, with 24 participants (with 12 per group) using 14 simulators (US$48 720) to meet their needs., Results: Calculated from time of use (83 minutes in total), the optimal ratios of simulators expressed for 10 participants were 9.2: 3.7 for jugular and subclavian venous catheterisation (33 minutes); 1.5 for thoracocentesis (13 minutes), 1.0 for femoral venous catheterisation (9 minutes), 1.0 for lumbar puncture (9 minutes), 0.8 for peripheral venous catheterisation (8 minutes), 0.7 for paracentesis (6 minutes) and 0.5 for arthrocentesis (5 minutes). In Phase 2, the usage rate of simulators increased from 35.5% to 76.6%, maintaining the total time of use at 80.4 minutes., Conclusions: We present a replicable method for the cost-effective planning of self-regulated simulation by measuring the use of simulators. Expressed as ratios of simulators per participant, this information can support purchase decisions and be shared with similar programmes., (© 2019 John Wiley & Sons Ltd and The Association for the Study of Medical Education.)

Context: Simulated clinical immersion (SCI), in which clinical situations are simulated in a realistic environment, safely and gradually exposes novices to complex problems. Given their limited experience, undergraduate students can potentially be quite overwhelmed by SCI learning tasks, which may result in misleading learning outcomes. Although task complexity should be adapted to the learner's level of expertise, many factors, both intrinsic and extraneous to the learning task, can influence perceived task complexity and its impact on cognitive processes., Objectives: The purpose of this mixed-methods study was to understand the effects of task complexity on undergraduate pharmacy students' cognitive load, task performance and perception of learning in SCI., Methods: A total of 167 second-year pharmacy students were randomly assigned to undertake one simple and one complex learning task in SCI consecutively. Participants' cognitive load was measured after each task and debriefing. Task performance and time on task were also assessed. As part of a sequential explanatory design, semi-structured interviews were conducted with students showing maximal variations in intrinsic cognitive load to elucidate their perceptions of learning when dealing with complexity., Results: Although the complex task generated significantly higher cognitive load and time on task than the simple task, performance was high for both tasks. Qualitative results revealed that a lack of clinical experience, an unfamiliar resource in the environment and the constraints inherent to SCI, such as time limitations, hindered the clinical reasoning process and led to poorer self-evaluation of performance. Simple tasks helped students gain more self-confidence, whereas complex tasks further encouraged reflective practice during debriefings., Conclusions: Although complex tasks in SCI were more cognitively demanding and took longer to execute, students indicated that they learned more from them than they did from simple tasks. Complex tasks constitute an additional challenge in terms of clinical reasoning and thus provide a more valuable learning experience from the student's perspective., (© 2018 John Wiley & Sons Ltd and The Association for the Study of Medical Education.)

Objective. To describe strategies for implementation of simulation-based crisis resource management (CRM) in pharmacy education and present students' appreciation of an interdisciplinary CRM training at a university in Canada. Methods. In fall 2016, third-year undergraduate pharmacy students at Laval University and pharmacy technician students from Fierbourg school participated in a CRM activity and completed a five-item survey to assess the quality of the CRM activity they had just experienced. Paired t-tests were computed to detect differences of appreciation between pharmacy technician students and pharmacy students. Results. Students rated each item as very good or excellent varying from 81% to 97%. The only difference found between the two types of students was on their overall appreciation of the experience. Pharmacy technician students rated their experience as very good while pharmacy students rated it as excellent. Conclusion. CRM training can easily be adapted to the context of pharmacy education because its key concepts of team management, resource allocation, awareness of environment and dynamic decision-making directly apply to pharmacy practice. Based on the results of this study, students greatly value their CRM training experience. Future research is needed to measure the transfer into practice of CRM principles.

Purpose: MRI cell tracking can be used to monitor immune cells involved in the immunotherapy response, providing insight into the mechanism of action, temporal progression of tumor growth, and individual potency of therapies. To evaluate whether MRI could be used to track immune cell populations in response to immunotherapy, CD8 + cytotoxic T cells, CD4 + CD25 + FoxP3 + regulatory T cells, and myeloid-derived suppressor cells were labeled with superparamagnetic iron oxide particles., Methods: Superparamagnetic iron oxide-labeled cells were injected into mice (one cell type/mouse) implanted with a human papillomavirus-based cervical cancer model. Half of these mice were also vaccinated with DepoVax TM (ImmunoVaccine, Inc., Halifax, Nova Scotia, Canada), a lipid-based vaccine platform that was developed to enhance the potency of peptide-based vaccines., Results: MRI visualization of CD8 + cytotoxic T cells, regulatory T cells, and myeloid-derived suppressor cells was apparent 24 h post-injection, with hypointensities due to iron-labeled cells clearing approximately 72 h post-injection. Vaccination resulted in increased recruitment of CD8 + cytotoxic T cells, and decreased recruitment of myeloid-derived suppressor cells and regulatory T cells to the tumor. We also found that myeloid-derived suppressor cell and regulatory T cell recruitment were positively correlated with final tumor volume., Conclusion: This type of analysis can be used to noninvasively study changes in immune cell recruitment in individual mice over time, potentially allowing improved application and combination of immunotherapies. Magn Reson Med 80:304-316, 2018. © 2017 International Society for Magnetic Resonance in Medicine., (© 2017 International Society for Magnetic Resonance in Medicine.)

Context: Simulated clinical immersion (SCI) is used in undergraduate healthcare programs to expose the learner to real-life situations in authentic simulated clinical environments. For novices, the environment in which the simulation occurs can be distracting and stressful, hence potentially compromising learning., Objectives: This study aims to determine whether SCI (with environment) imposes greater extraneous cognitive load and stress on undergraduate pharmacy students than simulated patients (SP) (without environment). It also aims to explore how features of the simulated environment influence students' perception of learning., Methods: In this mixed-methods study, 143 undergraduate pharmacy students experienced both SCI and SP in a crossover design. After the simulations, participants rated their cognitive load and emotions. Thirty-five students met in focus groups to explore their perception of learning in simulation., Results: Intrinsic and extraneous cognitive load and stress scores in SCI were significantly but modestly higher compared to SP. Qualitative findings reveal that the physical environment in SCI generated more stress and affected students? focus. In SP, students concentrated on clinical reasoning. SCI stimulated a focus on data collection but impeded in-depth problem solving processes., Conclusion: The physical environment in simulation influences what and how students learn. SCI was reported as more cognitively demanding than SP. Our findings emphasize the need for the development of adapted instructional design guidelines in simulation for novices.

Spider aciniform (wrapping) silk is a remarkable fibrillar biomaterial with outstanding mechanical properties. It is a modular protein consisting, in Argiope trifasciata, of a core repetitive domain of 200 amino acid units (W units). In solution, the W units comprise a globular folded core, with five α-helices, and disordered tails that are linked to form a ~63-residue intrinsically disordered linker in concatemers. Herein, we present nuclear magnetic resonance (NMR) spectroscopy-based (15)N spin relaxation analysis, allowing characterization of backbone dynamics as a function of residue on the ps-ns timescale in the context of the single W unit (W₁) and the two unit concatemer (W₂). Unambiguous mapping of backbone dynamics throughout W₂ was made possible by segmental NMR active isotope-enrichment through split intein-mediated trans-splicing. Spectral density mapping for W₁ and W₂ reveals a striking disparity in dynamics between the folded core and the disordered linker and tail regions. These data are also consistent with rotational diffusion behaviour where each globular domain tumbles almost independently of its neighbour. At a localized level, helix 5 exhibits elevated high frequency dynamics relative to the proximal helix 4, supporting a model of fibrillogenesis where this helix unfolds as part of the transition to a mixed α-helix/β-sheet fibre.

Aciniform silk protein (AcSp1) is the primary component of wrapping silk, the toughest of the spider silks because of a combination of high tensile strength and extensibility. Argiope trifasciata AcSp1 contains a core repetitive domain with at least 14 homogeneous 200-amino acid units ("W" units). Upon fibrillogenesis, AcSp1 converts from an α-helix-rich soluble state to a mixed α-helical/β-sheet conformation. Solution-state nuclear magnetic resonance (NMR) spectroscopy allowed demonstration of variable local stability within the W unit, but comprehensive characterization was confounded by spectral overlap, which was exacerbated by decreased chemical shift dispersion upon denaturation. Here, (19)F NMR spectroscopy, in the context of a single W unit (W1), is applied to track changes in structure and dynamics. Four strategic positions in the W unit were mutated to tryptophan and biosynthetically labeled with 5-fluorotryptophan (5F-Trp). Simulated annealing-based structure calculations implied that these substitutions should be tolerated, while circular dichroism (CD) spectroscopy and (1)H-(15)N chemical shift displacements indicated minimal structural perturbation in W1 mutants. Fiber formation by W2 concatemers containing 5F-Trp substitutions in both W units demonstrated retention of functionality, a somewhat surprising finding in light of sequence conservation between species. Each 5F-Trp-labeled W1 exhibited a unique (19)F chemical shift, line width, longitudinal relaxation time constant (T1), and solvent isotope shift. Perturbation to (19)F chemical shift and nuclear spin relaxation parameters reflected changes in the conformation and dynamics at each 5F-Trp site upon addition of urea and dodecylphosphocholine (DPC). (19)F NMR spectroscopy allowed unambiguous localized tracking throughout titration with each perturbant, demonstrating distinct behavior for each perturbant not previously revealed by heteronuclear NMR experiments.

Spiders store spidroins in their silk glands as high concentration aqueous solutions, spinning these dopes into fibres with outstanding mechanical properties. Aciniform (or wrapping) silk is the toughest spider silk and is devoid of the short amino acid sequence motifs characteristic of the other spidroins. Using solution-state NMR spectroscopy, we demonstrate that the 200 amino acid Argiope trifasciata AcSp1 repeat unit contrasts with previously characterized spidroins, adopting a globular 5-helix bundle flanked by intrinsically disordered N- and C-terminal tails. Split-intein-mediated segmental NMR-active isotope-enrichment allowed unambiguous demonstration of modular and malleable "beads-on-a-string" concatemeric behaviour. Concatemers form fibres upon manual drawing with silk-like morphology and mechanical properties, alongside secondary structuring and orientation consistent with native AcSp1 fibres. AcSp1 structural stability varies locally, with the fifth helix denaturing most readily. The structural transition of aciniform spidroin from a mostly α-helical dope to a mixed α-helix/β-sheet-containing fibre can be directly related to spidroin architecture and stability.

Artificial spider silk proteins may form fibers with exceptional strength and elasticity. Wrapping silk, or aciniform silk, is the toughest of the spider silks, and has a very different protein composition than other spider silks. Here, we present the characterization of an aciniform protein (AcSp1) subunit named W1, consisting of one AcSp1 199 residue repeat unit from Argiope trifasciata. The structural integrity of recombinant W1 is demonstrated in a variety of buffer conditions and time points. Furthermore, we show that W1 has a high thermal stability with reversible denaturation at ∼71°C and forms self-assembled nanoparticle in near-physiological conditions. W1 therefore represents a highly stable and structurally robust module for protein-based nanoparticle formation., (Copyright © 2013 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)

Experimental embolic particles based on a novel zinc-silicate glass system have been biologically evaluated for potential consideration in transcatheter arterial embolization procedures. In addition to controlling the cytotoxicity and haemocompatibility for such embolic particles, its glass structure may mediate specific responses via dissolution in the physiological environment. In a 120 h in-vitro dissolution study, ion release levels for silicon (Si4+), sodium (Na+), calcium (Ca2+), zinc (Zn2+), titanium (Ti4+), lanthanum (La3+), strontium (Sr2+), and magnesium (Mg2+), were found to range from 0.04 to 5.41 ppm, 0.27-2.28 ppm, 2.32-8.47 ppm, 0.16-0.20 ppm, 0.12-2.15 ppm, 0.16-0.49 ppm and 0.01-0.12 ppm, respectively for the series of glass compositions evaluated. Initial release of Zn2+ (1.93-10.40 ppm) was only evident after 120 h. All compositions showed levels of cell viabilities ranging from 61.31 ± 4.33% to 153.7 ± 1.25% at 25%-100% serial extract dilutions. The conformational state of fibrinogen, known to induce thrombi, indicated that no changes were induced with respect of the materials dissolution by-products. Furthermore, the best-in-class experimental composition showed equivalency to contour PVA in terms of inducing platelet adhesion. The data generated here provides requisite evidence to continue to in-vivo pre-clinical evaluation using the best-in-class experimental composition evaluated.

Spider silk is one of nature's most remarkable biomaterials due to extraordinary strength and toughness not found in today's synthetic materials. Of the seven types of silk, wrapping silk (AcSp1) is the most extensible of the types of silks and has no sequence similarity to the other types. Here we report the chemical shifts for the AcSp1 199 amino acid protein repeat unit and its anticipated secondary structure based on secondary chemical shifts.

Biomolecular NMR spectroscopy frequently employs estimates of protein secondary structure using secondary chemical shift (Deltadelta) values, measured as the difference between experimental and random coil chemical shifts (RCCS). Most published random coil data have been determined in aqueous conditions, reasonable for non-membrane proteins, but potentially less relevant for membrane proteins. Two new RCCS sets are presented here, determined in dimethyl sulfoxide (DMSO) and chloroform:methanol:water (4:4:1 by volume) at 298 K. A web-based program, CS-CHEMeleon, has been implemented to determine the accuracy of secondary structure assessment by calculating and comparing Deltadelta values for various RCCS datasets. Using CS-CHEMeleon, Deltadelta predicted versus experimentally determined secondary structures were compared for large datasets of membrane and non-membrane proteins as a function of RCCS dataset, Deltadelta threshold, nucleus, localized parameter averaging and secondary structure type. Optimized Deltadelta thresholds are presented both for published and for the DMSO and chloroform:methanol:water derived RCCS tables. Despite obvious RCCS variations between datasets, prediction of secondary structure was consistently similar. Strikingly, predictive accuracy seems to be most dependent upon the type of secondary structure, with helices being the most accurately predicted by Deltadelta values using five different RCCS tables. We suggest caution when using Deltadelta-based restraints in structure calculations as the underlying dataset may be biased. Comparative assessment of multiple RCCS datasets should be performed, and resulting Deltadelta-based restraints weighted appropriately relative to other experimental restraints.