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Single-molecule imaging provides a powerful way to study biochemical processes in live cells, yet it remains challenging to track single molecules while simultaneously detecting their interactions. Here, we describe a novel property of rhodamine dyes, proximity-assisted photoactivation (PAPA), in which one fluorophore (the ‘sender’) can reactivate a second fluorophore (the ‘receiver’) from a dark state. PAPA requires proximity between the two fluorophores, yet it operates at a longer average intermolecular distance than Förster resonance energy transfer (FRET). We show that PAPA can be used in live cells both to detect protein–protein interactions and to highlight a subpopulation of labeled protein complexes in which two different labels are in proximity. In proof-of-concept experiments, PAPA detected the expected correlation between androgen receptor self-association and chromatin binding at the single-cell level. These results establish a new way in which a photophysical property of fluorophores can be harnessed to study molecular interactions in single-molecule imaging of live cells.

How molecules interact governs how they move. Single-molecule tracking (SMT) thus provides a unique window into the dynamic interactions of biomolecules within live cells. Using transcription regulation as a case study, we describe how SMT works, what it can tell us about molecular biology, and how it has changed our perspective on the inner workings of the nucleus. We also describe what SMT cannot yet tell us and how new technical advances seek to overcome its limitations. This ongoing progress will be imperative to address outstanding questions about how dynamic molecular machines function in live cells.

ObjectivesTo identify incident SARS-CoV-2 infections and inform effective mitigation strategies in university settings, we piloted an integrated symptom and exposure monitoring and testing system among a cohort of university students and employees.DesignProspective cohort study.SettingA public university in California from June to August 2020.Participants2180 university students and 738 university employees.Primary outcome measuresAt baseline and endline, we tested participants for active SARS-CoV-2 infection via quantitative PCR (qPCR) test and collected blood samples for antibody testing. Participants received notifications to complete additional qPCR tests throughout the study if they reported symptoms or exposures in daily surveys or were selected for surveillance testing. Viral whole genome sequencing was performed on positive qPCR samples, and phylogenetic trees were constructed with these genomes and external genomes.ResultsOver the study period, 57 students (2.6%) and 3 employees (0.4%) were diagnosed with SARS-CoV-2 infection via qPCR test. Phylogenetic analyses revealed that a super-spreader event among undergraduates in congregate housing accounted for at least 48% of cases among study participants but did not spread beyond campus. Test positivity was higher among participants who self-reported symptoms (incidence rate ratio (IRR) 12.7; 95% CI 7.4 to 21.8) or had household exposures (IRR 10.3; 95% CI 4.8 to 22.0) that triggered notifications to test. Most (91%) participants with newly identified antibodies at endline had been diagnosed with incident infection via qPCR test during the study.ConclusionsOur findings suggest that integrated monitoring systems can successfully identify and link at-risk students to SARS-CoV-2 testing. As the study took place before the evolution of highly transmissible variants and widespread availability of vaccines and rapid antigen tests, further research is necessary to adapt and evaluate similar systems in the present context.

SARS coronavirus 2 (SARS-CoV-2) has caused an ongoing global pandemic with significant mortality and morbidity. At this time, the only FDA-approved therapeutic for COVID-19 is remdesivir, a broad-spectrum antiviral nucleoside analog. Efficacy is only moderate, and improved treatment strategies are urgently needed. To accomplish this goal, we devised a strategy to identify compounds that act synergistically with remdesivir in preventing SARS-CoV-2 replication. We conducted combinatorial high-throughput screening in the presence of submaximal remdesivir concentrations, using a human lung epithelial cell line infected with a clinical isolate of SARS-CoV-2. This identified 20 approved drugs that act synergistically with remdesivir, many with favorable pharmacokinetic and safety profiles. Strongest effects were observed with established antivirals, Hepatitis C virus nonstructural protein 5A (HCV NS5A) inhibitors velpatasvir and elbasvir. Combination with their partner drugs sofosbuvir and grazoprevir further increased efficacy, increasing remdesivir's apparent potency > 25-fold. We report that HCV NS5A inhibitors act on the SARS-CoV-2 exonuclease proofreader, providing a possible explanation for the synergy observed with nucleoside analog remdesivir. FDA-approved Hepatitis C therapeutics Epclusa® (velpatasvir/sofosbuvir) and Zepatier® (elbasvir/grazoprevir) could be further optimized to achieve potency and pharmacokinetic properties that support clinical evaluation in combination with remdesivir.

Single-molecule imaging provides a powerful way to study biochemical processes in live cells, yet it remains challenging to track single molecules while simultaneously detecting their interactions. Here, we describe a novel property of rhodamine dyes, proximity-assisted photoactivation (PAPA), in which one fluorophore (the 'sender') can reactivate a second fluorophore (the 'receiver') from a dark state. PAPA requires proximity between the two fluorophores, yet it operates at a longer average intermolecular distance than Förster resonance energy transfer (FRET). We show that PAPA can be used in live cells both to detect protein-protein interactions and to highlight a subpopulation of labeled protein complexes in which two different labels are in proximity. In proof-of-concept experiments, PAPA detected the expected correlation between androgen receptor self-association and chromatin binding at the single-cell level. These results establish a new way in which a photophysical property of fluorophores can be harnessed to study molecular interactions in single-molecule imaging of live cells.

Abstract This contribution summarizes the current state of research regarding so‐called sensor‐integrating machine elements as an enabler of digitalization in mechanical engineering and——if available—their application in industry. The focus is on the methodical aspects of the development of these machine elements in general as well as specific sensor‐integrating machine elements that are either already in use or currently under development. Developmental aspects include the robust design of initially evaluated concepts for sensor‐integrating machine elements as well as their modularization. Smart materials with sensory functions are included in the analysis as well as the differentiation with regard to add‐on sensors. The aim of the authors interlinked by a special research program funded by the German Research Foundation (DFG) is to facilitate the exchange with other researchers with the help of the comprehensive overview given in this contribution. The contribution concludes with a brief discussion of open challenges, such as the energy supply and data transfer in rotating systems and also data security.

Abstract This retrospective observational study evaluated racial disparities among Black and White patients with multiple myeloma (MM). We included patients from a longitudinal de-identified EHR-derived database who had ≥2 visits recorded on or after 1/1/2011, documented treatment, and race listed as White or Black. Black patients (n = 1172) were more likely female (54.8%/42.9%) and younger (

The design of reliable systems is a key challenge in product engineering [...]

Using hydrogen fuel cells for power systems, temperature conditions are important for efficient and reliable operations, especially in low-temperature environments. A heating system with an electrical energy buffer is therefore required for reliable operation. There is a research gap in finding an appropriate control strategy regarding energy efficiency and reliable operations for different environmental conditions. This paper investigates heating strategies for the subfreezing start of a fuel cell for portable applications at an early development stage to enable frontloading in product engineering. The strategies were investigated by simulation and experiment. A prototype for such a system was built and tested for subfreezing start-ups and non-subfreezing start-ups. This was done by heating the fuel cell system with different control strategies to test their efficiency. It was found that operating strategies to heat up the fuel cell system can ensure a more reliable and energy-efficient operation. The heating strategy needs to be adjusted according to the ambient conditions, as this influences the required heating energy, efficiency, and reliable operation of the system. A differentiation in the control strategy between subfreezing and non-subfreezing temperatures is recommended due to reliability reasons.

For the control of dynamic systems such as an Electro-Hydraulic Actuator (EHA), there is a need to optimize the control based on simulations, since a prototype or a physical system is usually not available during system design. In consequence, no system identification can be performed. Therefore, it is unclear how well a simulation model of an EHA can be used for multicriteria optimization of the position control due to the uncertain model quality. To evaluate the suitability for control optimization, the EHA is modeled and parameterized as a grey-box model using existing parameters independent of test bench experiments. A method for multi-objective optimization of a controller is used to optimize the position control of the EHA. Finally, the step responses are compared with the test bench. The evaluation of the step responses for different loads and control parameters shows similar behavior between the simulation model and the physical system on the test bench, although the essential phenomena could not be reproduced. This means that the model quality achieved by modeling is suitable as an indication for the optimization of the control by simulation without a physical system.

Repetitive overhead work with a heavy load increases the risk for work-related shoulder disorders. Occupational exoskeletons supporting arm elevation are potential solutions to reduce that risk by lowering the physical strains on the shoulder. Many studies have reported a reduction in shoulder stress in various overhead tasks by using such exoskeletons. However, the support demand can vary in each phase of motion as well as in each individual task. This paper presents a laboratory study with five participants to evaluate the influence of the support level of an active shoulder exoskeleton in different motion phases (e.g., arm lifting, screw-in, and arm lowering of two overhead tasks) to identify the potential optimization of support at each phase. Results show that the support level of the exoskeleton should be adapted to the motion phase of the two chosen tasks. A higher support force is desired for the screw phase compared to the arm lifting and lowering phases, and the support level needs to be reduced immediately for arm lowering after the screw phase. The time for switching the support levels can be recognized by the electric current of the screwdriver.

Validation is important for a high product quality of drive components. An X-in-the-Loop test bench enables the integration of scaled prototypes through coupling systems and scaling models even before serial parts are available. In the context of X-in-the-loop investigations, it is still unclear whether a scaling model enables the early investigation of geometry variants in powertrain subsystems. In this paper, scaled geometry experiments taking into account the interacting system are considered to evaluate the scaling model in terms of early investigation of geometry variants. The aim of this paper is the functional investigation of geometrically scaled drive components by integrating scaled prototypes in an X-in-the-Loop test bench. Using an overload clutch with detents, component variants of different size levels are investigated in scaled experiments with a scaling model. The results confirm possibilities of X-in-the-Loop integration of scaled prototypes and their investigation on geometrically scaled drive components. The investigations show, therefore, the opportunities of integrating scaled drive components through the scaling model to support the investigation of geometry variants before serial parts are available. Scaled geometry investigations considering the interacting system can, thus, support product development.

Type 2 Nuclear Receptors (T2NRs) require heterodimerization with a common partner, the Retinoid X Receptor (RXR), to bind cognate DNA recognition sites in chromatin. Based on previous biochemical and over-expression studies, binding of T2NRs to chromatin is proposed to be regulated by competition for a limiting pool of the core RXR subunit. However, this mechanism has not yet been tested for endogenous proteins in live cells. Using single molecule tracking (SMT) and proximity-assisted photoactivation (PAPA), we monitored interactions between endogenously tagged retinoid X receptor (RXR) and retinoic acid receptor (RAR) in live cells. Unexpectedly, we find that higher expression of RAR, but not RXR increases heterodimerization and chromatin binding in U2OS cells. This surprising finding indicates the limiting factor is not RXR but likely its cadre of obligate dimer binding partners. SMT and PAPA thus provide a direct way to probe which components are functionally limiting within a complex TF interaction network providing new insights into mechanisms of gene regulation in vivo with implications for drug development targeting nuclear receptors.

Psychomotor skill and decision-making efficiency in surgical wire navigation can be objectively evaluated by analysis of intraoperative fluoroscopic image sequences. Prior work suggests that such image-based behavior analysis of operating room (OR) performance can predict performer experience level (R 2  = 0.62) and agree with expert opinion (the current standard) on the quality of a final implant construct (R 2  = 0.59). However, it is unclear how objective image-based evaluation compares with expert assessments for entire technical OR performances. This study examines the relationships between three key variables: (1) objective image-based criteria, (2) expert opinions, and (3) performing surgeon experience level. A paired-comparison survey of seven experts, based upon eight OR fluoroscopic wire navigation image sequences, shows that the experts' preferences are best explained by objective metrics that reflect psychomotor and decision-making behaviors which are counter-productive to successful implant placement, like image count (R 2  = 0.83) and behavior tally (R 2  = 0.74). One such behavior, adjustments away from goal, uniquely correlated well with all three key variables: a fluoroscopic image-based analysis composite score (R 2  = 0.40), expert consensus (R 2  = 0.76), and performer experience (R 2  = 0.41). These results confirm that experts view less efficient technical behavior as indicative of lesser technical proficiency. While expert assessments of technical skill were reliable and consistent, neither individual nor consensus expert opinion appears to correlate with performer experience (R 2  = 0.11)., (© 2023 The Authors. Journal of Orthopaedic Research® published by Wiley Periodicals LLC on behalf of Orthopaedic Research Society.)

Background: Antegrade femoral intramedullary nailing (IMN) is a common orthopedic procedure that residents are exposed to early in their training. A key component to this procedure is placing the initial guide wire with fluoroscopic guidance. A simulator was developed to train residents on this key skill, building off an existing simulation platform originally developed for wire navigation during a compression hip screw placement. The objective of this study was to assess the construct validity of the IMN simulator., Methods: Thirty orthopedic surgeons participated in the study: 12 had participated in fewer than 10 hip fracture or IMN related procedures and were categorized as novices; 18 were faculty, categorized as experts. Both cohorts were instructed on the goal of the task, placing a guide wire for an IM nail, and the ideal wire position reference that their wire placement would be graded against. Participants completed 2 assessments with the simulator. Performance was graded on the distance from the ideal starting point, distance from the ideal end point, wire trajectory, duration, fluoroscopy image count, and other elements of surgical decision making. A two-way ANOVA analysis was used to analyze the data looking at experience level and trial number., Results: The expert cohort performed significantly better than the novice cohort on all metrics but one (overuse of fluoroscopy). The expert cohort had a more accurate starting point and completed the task while using fewer images and less overall time., Conclusion: This initial study shows that the IMN application of a wire navigation simulator demonstrates good construct validity. With such a large cohort of expert participants, we can be confident that this study captures the performance of active surgeons today. Implementing a training curriculum on this simulator has the potential to increase the performance of the novice level residents prior to their operating on a vulnerable patient. Level of Evidence: III ., (Copyright © The Iowa Orthopaedic Journal 2023.)

Introduction: Arthroscopy simulation is increasingly used in orthopaedic residency training. The implementation of a curriculum to accommodate these new training tools is a point of interest. We assessed the use of a high-fidelity arthroscopy simulator in a strictly voluntary curriculum to gauge resident interest and educational return., Methods: Fifty-eight months of simulator use data were collected from a single institution to analyze trends in resident use. Comparable data from two additional residency programs were analyzed as well, for comparison. Orthopaedic residents were surveyed to gauge interest in continued simulation training., Results: Average annual simulator use at the study institution was 27.7 hours (standard deviation = 26.8 hours). Orthopaedic residents spent an average of 1.7 hours practicing on the simulation trainer during the observation period. A total of 21% of residents met or exceeded a minimum of 3 hours of simulation time required for skill improvement defined by literature. Most (86%) of the residents agreed that the simulator in use should become a mandated component of a junior resident training., Conclusion: Although surgical simulation has a role in orthopaedic training, voluntary simulator use is sporadic, resulting in many residents not receiving the full educational benefits of such training. Implementation of a mandated simulation training curriculum is desired by residents and could improve the educational return of surgical simulators in residency training., (Copyright © 2021 by the American Academy of Orthopaedic Surgeons.)

Skill assessment in orthopedics has traditionally relied on subjective impressions from a supervising surgeon. The feedback derived from these tools may be limited by bias and other practical issues. Objective analysis of intraoperative fluoroscopic images offers an inexpensive, repeatable, and precise assessment strategy without bias. Assessors generally refrain from using the scores of images obtained throughout the operation to evaluate skill for practical reasons. A new system was designed to facilitate rapid analysis of this fluoroscopy via minimally trained analysts. Four expert and four novice analysts independently measured one objective metric for skill using both a custom analysis software and a commercial alternative. Analysts were able to measure the objective metric three times faster when using the custom software, and without a practical difference in accuracy in comparison to the expert analysts using the commercial software. These results suggest that a well-designed fluoroscopy analysis system can facilitate inexpensive, reliable, and objective assessment of surgical skills., Competing Interests: Disclosure of Interest The authors report that there are no conflict of interests for this work.