Distal weight bearing in transtibial prosthesis users wearing pin suspension.

Introduction: Low-level distal weight bearing in transtibial prosthesis users may help maintain perfusion and improve both proprioception and residual limb tissue health. Methods: The primary objectives of this research were to develop a sensor to continuously measure distal weight bearing, evaluate how prosthesis design variables affected weight bearing levels, and assess fluctuations in distal weight bearing during at-home and community use. Results: In-lab testing on a small group of participants wearing adjustable sockets demonstrated that if distal contact was present, when socket size was increased distal weight bearing increased and when socket size was reduced distal weight bearing decreased. During take-home use, participants accepted the distal weight bearing level set by the research team. It ranged between 1.1% and 6.4% BW for all days tested. The coefficient of variation (standard deviation/mean) ranged from 25% to 43% and was expected due in part to differences in walking style, speed, terrain, direction of ambulation, and bout duration. Two participants commented that they preferred presence of distal weight bearing to non-presence. Discussion: Next steps in this research are to develop clinical practices to determine target distal weight bearing levels and ranges, and to simplify the design of the sensor and weight bearing adjustment mechanism for clinical use.

Mechanically and physiologically optimizing prosthetic elevated vacuum systems in people with transtibial amputation: a pilot study.

Introduction: The most suitable elevated vacuum (EV) pressure may differ for each individual prosthesis user depending on suspension needs, socket fit, prosthetic components, and health. Mechanical and physiological effects of EV were evaluated in an effort to determine the optimal vacuum pressure for three individuals. Methods: Instrumented EV sockets were created based on the participants' regular EV sockets. Inductive distance sensors were embedded into the wall of the socket at select locations to measure limb movement relative to the socket. Each participant conducted an activity protocol while limb movement, limb fluid volume, and user-reported comfort were measured at various socket vacuum pressure settings. Results: Increased socket vacuum pressure resulted in reduced limb-socket displacement for each participant; however, 81-93% of limb movement was eliminated by a vacuum pressure setting of 12 (approximately -9 inHg). Relative limb-socket displacement by sensor location varied for each participant, suggesting distinct differences related to socket fit or residual limb tissue content. The rate of limb fluid volume change and the change in socket comfort did not consistently differ with socket vacuum pressure, suggesting a more complex relationship unique to each individual. Conclusions: Practitioners may use individual responses to optimize socket vacuum pressure settings, balancing mechanical and physiological effects of EV for improved clinical outcomes.

Using magnetic panels to enlarge a transtibial prosthetic socket.

A novel method is described to connect a prosthetic liner to the panels of an adjustable socket to facilitate limb fluid volume stabilization in prosthesis users. Magnets are placed in the socket panels, and iron powder is embedded in the user's prosthetic liner. When the magnet is in close proximity to the liner, a firm connection is formed. The system's capability to execute panel pull on transtibial prosthesis users was tested. The backs of the panels were supported by a bracket mounted to the external surface of the socket that allowed the radial position of the panels to be adjusted. Bench testing demonstrated an optimized strength-to-weight ratio using 1.27-cm thick annular-shaped magnets supported by 0.32-cm thick backplates. Testing on four people with transtibial amputation showed that the maximum socket increase achieved using magnetic panel pull ranged from 5.3% to 13.8% of the initial (panels flush) socket volume. The results indicate that magnetic panel pull induces a meaningful increase in socket volume during sitting. The clinical relevance is a novel strategy that may help stabilize prosthesis users' limb fluid volume over the day.

Science of the Van Allen Probes Science Operations Centers.

The Van Allen Probes mission operations materialized through a distributed model in which operational responsibility was divided between the Mission Operations Center (MOC) and separate instrument specific SOCs. The sole MOC handled all aspects of telemetering and receiving tasks as well as certain scientifically relevant ancillary tasks. Each instrument science team developed individual instrument specific SOCs proficient in unique capabilities in support of science data acquisition, data processing, instrument performance, and tools for the instrument team scientists. In parallel activities, project scientists took on the task of providing a significant modeling tool base usable by the instrument science teams and the larger scientific community. With a mission as complex as Van Allen Probes, scientific inquiry occurred due to constant and significant collaboration between the SOCs and in concert with the project science team. Planned cross-instrument coordinated observations resulted in critical discoveries during the seven-year mission. Instrument cross-calibration activities elucidated a more seamless set of data products. Specific topics include post-launch changes and enhancements to the SOCs, discussion of coordination activities between the SOCs, SOC specific analysis software, modeling software provided by the Van Allen Probes project, and a section on lessons learned. One of the most significant lessons learned was the importance of the original decision to implement individual team SOCs providing timely and well-documented instrument data for the NASA Van Allen Probes Mission scientists and the larger magnetospheric and radiation belt scientific community.

A novel portable sensor to monitor bodily positions and activities in transtibial prosthesis users.

BACKGROUND: Step activity monitors provide insight into the amount of physical activity prosthesis users conduct but not how they use their prosthesis. The purpose of this research was to help fill this void by developing and testing a technology to monitor bodily position and type of activity. METHODS: Thin inductive distance sensors were adhered to the insides of sockets of a small group of transtibial prosthesis users, two at proximal locations and two at distal locations. An in-lab structured protocol and a semi-structured out-of-lab protocol were video recorded, and then participants wore the sensing system for up to 7 days. A data processing algorithm was developed to identify sit, seated shift, stand, standing weight-shift, walk, partial doff, and non-use. Sensed distance data from the structured and semi-structured protocols were compared against the video data to characterize accuracy. Bodily positions and activities during take-home testing were tabulated to characterize participants' use of the prosthesis. FINDINGS: Sit and walk detection accuracies were above 95% for all four participants tested. Stand detection accuracy was above 90% for three participants and 62.5% for one participant. The reduced accuracy may have been due to limited stand data from that participant. Step count was not proportional to active use time (sum of stand, walk, and standing weight-shift times). INTERPRETATION: Step count may provide an incomplete picture of prosthesis use. Larger studies should be pursued to investigate how bodily position and type of activity may facilitate clinical decision-making and improve the lives of people with lower limb amputation.

Performance of an auto-adjusting prosthetic socket during walking with intermittent socket release.

Introduction: A challenge in the engineering of auto-adjusting prosthetic sockets is to maintain stable operation of the control system while users change their bodily position and activity. The purpose of this study was to test the stability of a socket that automatically adjusted socket size to maintain fit. Socket release during sitting was conducted between bouts of walking. Methods: Adjustable sockets with sensors that monitored distance between the liner and socket were fabricated. Motor-driven panels and a microprocessor-based control system adjusted socket size during walking to maintain a target sensed distance. Limb fluid volume was recorded continuously. During eight sit/walk cycles, the socket panels were released upon sitting and then returned to position for walking, either the size at the end of the prior bout or a size 1.0% larger in volume. Results: In six transtibial prosthesis users, the control system maintained stable operation and did not saturate (move to and remain at the end of the actuator's range) during 98% of the walking bouts. Limb fluid volume changes generally matched the panel position changes executed by the control system. Conclusions: Stable operation of the control system suggests that the auto-adjusting socket is ready for testing in users' at-home settings.

Protocol for drug screening of patient-derived tumor organoids using high-content fluorescent imaging.

High-content imaging of tumor organoids (TOs) treated with therapeutic agents provides detailed cell viability readouts at the organoid level. In contrast, most used protocols provide one number per well. While requiring the use of inverted microscopy with an automated stage, this protocol can provide critical information about heterogeneous responses of TOs to various treatments. This protocol describes a technique for culturing and drug testing TOs using fluorescent indicators of cell viability with high reproducibility. For complete details on the use and execution of this protocol, please refer to Larsen et al. (2021).

Cyclic socket enlargement and reduction during walking to minimize limb fluid volume loss in transtibial prosthesis users.

The purpose of this research was to pursue an innovative cyclic panel-pull strategy during ambulation to minimize limb fluid volume loss in transtibial prosthesis users. Participants' traditional socket shapes were duplicated, and test sockets prepared with three adjustable motor-driven panels that were controlled by a microprocessor. After donning the prosthesis, participants' liners were fastened to the panels. During a 40 min test session, participants conducted three cycles of sitting (5 min) and walking (8 min). During the 5th and 6th min of each cycle of walking, the panels were cyclically pulled outward in late stance phase, decreasing pressure on the residual limb. Panels were returned to their original position in swing phase. Eight of twelve participants gained more fluid volume while walking when panel-pull was added than when it was removed. When the liner was uncoupled from the panels and panel-pull was executed, eight of twelve participants gained less fluid volume compared to when the liner was fastened to the panels. Panel-pull may facilitate limb fluid volume retention in transtibial prosthesis users. Efforts to simplify the design so that it can be implemented in long-term testing during at-home use should be considered.

Cancer cells use self-inflicted DNA breaks to evade growth limits imposed by genotoxic stress.

Genotoxic therapy such as radiation serves as a frontline cancer treatment, yet acquired resistance that leads to tumor reoccurrence is frequent. We found that cancer cells maintain viability during irradiation by reversibly increasing genome-wide DNA breaks, thereby limiting premature mitotic progression. We identify caspase-activated DNase (CAD) as the nuclease inflicting these de novo DNA lesions at defined loci, which are in proximity to chromatin-modifying CCCTC-binding factor (CTCF) sites. CAD nuclease activity is governed through phosphorylation by DNA damage response kinases, independent of caspase activity. In turn, loss of CAD activity impairs cell fate decisions, rendering cancer cells vulnerable to radiation-induced DNA double-strand breaks. Our observations highlight a cancer-selective survival adaptation, whereby tumor cells deploy regulated DNA breaks to delimit the detrimental effects of therapy-evoked DNA damage.

A pan-cancer organoid platform for precision medicine.

Patient-derived tumor organoids (TOs) are emerging as high-fidelity models to study cancer biology and develop novel precision medicine therapeutics. However, utilizing TOs for systems-biology-based approaches has been limited by a lack of scalable and reproducible methods to develop and profile these models. We describe a robust pan-cancer TO platform with chemically defined media optimized on cultures acquired from over 1,000 patients. Crucially, we demonstrate tumor genetic and transcriptomic concordance utilizing this approach and further optimize defined minimal media for organoid initiation and propagation. Additionally, we demonstrate a neural-network-based high-throughput approach for label-free, light-microscopy-based drug assays capable of predicting patient-specific heterogeneity in drug responses with applicability across solid cancers. The pan-cancer platform, molecular data, and neural-network-based drug assay serve as resources to accelerate the broad implementation of organoid models in precision medicine research and personalized therapeutic profiling programs.

A sensor to monitor limb depth in transtibial sockets with locking pin suspension: a technical note.

BACKGROUND: Monitoring of limb depth in transtibial sockets may provide useful information toward patient education and care. OBJECTIVE: The objective was to develop a sensor to detect the depth of a locking pin in the shuttle lock of a transtibial socket and to monitor the small motions between ratchet clicks during ambulation. STUDY DESIGN: Controlled bench testing and single-participant study. METHODS: A copper wire coil positioned beneath the socket shuttle lock was used with an inductive sensing chip to monitor locking pin depth. A custom jig was used to calibrate the sensor and bench test the system. Repeatability, drift, and the effects of pin length, carbon fiber presence, temperature change, and pin angulation on sensor performance were tested. Testing was conducted on a participant wearing an adjustable socket, walking with the panels at four different radial positions. RESULTS: The sensor demonstrated a root mean square error of 0.21% of the full-scale output. Different pins, different pin lengths, and the presence of carbon fiber affected calibration, indicating that the sensor must be calibrated to the individual user's socket and pin. Ratchet clicks and cyclic motion between clicks during walking were evident in the data. During participant testing, enlarging the socket at 1.00 mm radial increments caused significant changes in pin peak-to-peak distance (up and down motion) within a step. CONCLUSIONS: The sensor is sufficiently accurate to pursue studies investigating utility of the data toward clinical monitoring of socket fit.

Socket release/relock: An innovative mechanism to maintain residual limb volume.

Management of socket fit is challenging for people using lower-limb prostheses because of residual limb volume fluctuation throughout the day. Releasing socket pressures during sitting (partial doffing) may help users increase their limb volume after they have undergone volume loss earlier in the day. The purpose of this research was to develop and evaluate a system to allow for quick and easy locking pin and socket panel release during sitting and relock upon standing. The system was to allow the partial doff tether length to be custom set for each user, accomplish release and relock in less than 2.0 s each, require only one hand, and require a finger push force comparable to a push button on a phone. A motor-driven release/relock system (<240 g build weight) housed within the socket adjusts locking pin tether length, and an instrumented ratcheting dial adjusts socket panel position. Three participants with a trans-tibial amputation operated the system properly using one hand. For a partial doff, users preferred a tether length between 5 and 6 cm. All users executed release within 1.5 s and relock within 1.5 s.

Clearance of activity-evoked K+ transients and associated glia cell swelling occur independently of AQP4: A study with an isoform-selective AQP4 inhibitor.

The mammalian brain consists of 80% water, which is continuously shifted between different compartments and cellular structures by mechanisms that are, to a large extent, unresolved. Aquaporin 4 (AQP4) is abundantly expressed in glia and ependymal cells of the mammalian brain and has been proposed to act as a gatekeeper for brain water dynamics, predominantly based on studies utilizing AQP4-deficient mice. However, these mice have a range of secondary effects due to the gene deletion. An efficient and selective AQP4 inhibitor has thus been sorely needed to validate the results obtained in the AQP4-/- mice to quantify the contribution of AQP4 to brain fluid dynamics. In AQP4-expressing Xenopus laevis oocytes monitored by a high-resolution volume recording system, we here demonstrate that the compound TGN-020 is such a selective AQP4 inhibitor. TGN-020 targets the tested species of AQP4 with an IC50 of ~3.5 µM, but displays no inhibitory effect on the other AQPs (AQP1-AQP9). With this tool, we employed rat hippocampal slices and ion-sensitive microelectrodes to determine the role of AQP4 in glia cell swelling following neuronal activity. TGN-020-mediated inhibition of AQP4 did not prevent stimulus-induced extracellular space shrinkage, nor did it slow clearance of the activity-evoked K+ transient. These data, obtained with a verified isoform-selective AQP4 inhibitor, indicate that AQP4 is not required for the astrocytic contribution to the K+ clearance or the associated extracellular space shrinkage.

Automatic Control of Prosthetic Socket Size for People WithTranstibial Amputation: Implementation and Evaluation.

OBJECTIVE: The purpose was to design, implement, and test a control system for a motor-actuated, cable-panel prosthetic socket that automatically maintains socket fit by continuous adjustment of the socket size. METHODS: Sockets with motor-driven adjustable panels were fabricated for participants with transtibial amputation. A proportional-integral control system was implemented to adjust socket size based on Socket Fit Metric (SFM) data collected by an inductive sensor embedded within the socket wall. The sensed distance was representative of limb-to-socket distance. Testing was conducted with participants walking on a treadmill to characterize the system's capability to maintain a set point and to respond to a change in the set point. RESULTS: Test results from 10 participants with transtibial amputation showed that the Integral of Absolute Error (IAE) to maintain a set point ranged from 0.001 to 0.046 mm with a median of 0.003 mm. When the set point was changed, IAE errors ranged from 0.001 to 0.005 mm, with a median of 0.003 mm. An IAE of 0.003 mm corresponded to approximately a 0.08% socket volume error, which was considered clinically acceptable. CONCLUSION: The capability of the control system to maintain and respond to a change in set point indicates that it is ready for evaluation outside of the laboratory. SIGNIFICANCE: Integration of the developed control system into everyday prostheses may improve quality of life of prosthesis users by relieving them of the burden of continually adjusting socket size to maintain fit.

Photochemotherapy Induces Interferon Type III Expression via STING Pathway.

DNA-damaging cancer therapies induce interferon expression and stimulate the immune system, promoting therapy responses. The immune-activating STING (Stimulator of Interferon Genes) pathway is induced when DNA or double-stranded RNA (dsRNA) is detected in the cell cytoplasm, which can be caused by viral infection or by DNA damage following chemo- or radiotherapy. Here, we investigated the responses of cutaneous T-cell lymphoma (CTCL) cells to the clinically applied DNA crosslinking photochemotherapy (combination of 8-methoxypsoralen and UVA light; 8-MOP + UVA). We showed that this treatment evokes interferon expression and that the type III interferon IFNL1 is the major cytokine induced. IFNL1 upregulation is dependent on STING and on the cytoplasmic DNA sensor cyclic GMP-AMP synthase (cGAS). Furthermore, 8-MOP + UVA treatment induced the expression of genes in pathways involved in response to the tumor necrosis factor, innate immune system and acute inflammatory response. Notably, a subset of these genes was under control of the STING-IFNL1 pathway. In conclusion, our data connected DNA damage with immune system activation via the STING pathway and contributed to a better understanding of the effectiveness of photochemotherapy.

Incorporating a Ferrous Polymer Target into Elastomeric Liners for Socket Fit Sensing in Prosthesis Users.

Liner-to-socket distance measurement using inductive sensing may be an effective means to continuously monitor socket fit in people using trans-tibial prostheses. A practical limitation, however, is a means to incorporate a thin uniform-thickness layer of conductive or magnetically permeable target material into the wide range of prosthetic liner products that people with limb amputation commonly use. In this paper, a method is presented whereby a 0.50-mm thickness ferrous polymer made from a SEEPS polymer and iron powder that is formed adjacent to a 0.25-mm thick non-ferrous layer of SEEPS polymer is assembled between two sheets of elastic fabric material. Bench testing showed that the fabrication procedure achieved a root-mean-square error in the thickness of this construct of 58 µm, helping to create a consistent calibration result over the entire surface. The original fabric backing of an off-the-shelf prosthetic liner was removed and replaced with the developed construct. When worn in the shoe of an able-bodied participant for 7.5 h per day for 28 days, the sensor well maintained the shape of its calibration curve at the start of wear, but a distance offset (shifting of the y-intercept) was introduced that increased during the initial approximately 12 days of wear. When the distance offset was corrected, for the primary distance range of clinical interest for this application (0.00-5.00 mm), the sensor maintained its calibration within 4.4%. Before being used in clinical application for liner-to-socket distance monitoring, new ferrous liners may need to be pre-worn so as to achieve a consistent distance reference.

Does actively enlarging socket volume during resting facilitate residual limb fluid volume recovery in trans-tibial prosthesis users?

BACKGROUND: Residual limb volume loss is a source of prosthetic socket fit problems in people with lower-limb amputation. The aim of this study was to investigate a novel volume recovery strategy for people with trans-tibial amputation. METHODS: Test sockets for people with trans-tibial amputation were created that allowed panels of an adjustable socket and the underlying elastomeric liner to be pulled radially outward, using small motors mounted to the socket. One Control and one Intervention session were conducted with each participant. During Intervention sessions, panel-pull was executed during the sits of a multi-cycle sit/walk protocol. No panel-pull was executed during the Control sessions. Residual limb fluid volume was monitored in anterior and posterior regions using bioimpedance analysis. FINDINGS: Results from 12 participants demonstrated that short-term (12 min after the intervention was applied) median posterior residual limb fluid volume change for Intervention (0.44%) was higher than that for Control (-0.02%) (P = .015). Long-term (40 min after the intervention was applied) median posterior residual limb fluid volume change for Intervention (0.95%) was higher than that for Control (-0.26%) (P = .002). INTERPRETATION: If a panel-pull mechanism that was easy to assemble and operate could be created, then panel-pull may be an effective accommodation strategy to reduce daily limb volume loss in trans-tibial prosthesis users.

Fluid Volume Management in Prosthesis Users: Augmenting Panel Release with Pin Release.

BACKGROUND: Management of fluid in the limbs is a challenge faced by people with disabilities. In prosthetics, a means for transtibial prosthesis users to stabilize their residual limb fluid volume during the day may improve socket fit. OBJECTIVE: To determine if releasing the panels and locking pin of a cabled-panel adjustable socket during socket release significantly improved limb fluid volume recovery and retention over releasing the panels alone. DESIGN: Repeated-measures experiment to assess the effects on limb fluid volume retention. SETTING: Participants were tested in a laboratory setting while walking on a treadmill. INTERVENTION: Release of a locking pin tether during sitting as a limb volume accommodation strategy. MAIN OUTCOME MEASURE: Percent limb fluid volume retention for panel and pin release compared with panel release alone at 2 minutes (short term) and 50 minutes (long term) after subsequent activity. Limb fluid volume was monitored using bioimpedance analysis. RESULTS: Median percent limb fluid volume retention for the panel and pin release was significantly greater than panel release alone for both anterior and posterior regions for the long term (P = .0499 and .0096, respectively) but not the short term (P = .0712 and .1580, respectively). CONCLUSION: Augmenting panel release with pin release may be an effective accommodation strategy for prosthesis users with transtibial amputation to better retain limb fluid volume.