Visuospatial Skills Explain Differences in the Ability to Use Propulsion Biofeedback Post-stroke.

BACKGROUND AND PURPOSE: Visual biofeedback can be used to help people post-stroke reduce biomechanical gait impairments. Using visual biofeedback engages an explicit, cognitively demanding motor learning process. Participants with better overall cognitive function are better able to use visual biofeedback to promote locomotor learning; however, which specific cognitive domains are responsible for this effect are unknown. We aimed to understand which cognitive domains were associated with performance during acquisition and immediate retention when using visual biofeedback to increase paretic propulsion in individuals post-stroke. METHODS: Participants post-stroke completed cognitive testing, which provided scores for different cognitive domains, including executive function, immediate memory, visuospatial/constructional skills, language, attention, and delayed memory. Next, participants completed a single session of paretic propulsion biofeedback training, where we collected treadmill-walking data for 20 min with biofeedback and 2 min without biofeedback. We fit separate regression models to determine if cognitive domain scores, motor impairment (measured with the lower-extremity Fugl-Meyer), and gait speed could explain propulsion error and variability during biofeedback use and recall error during immediate retention. RESULTS: Visuospatial/constructional skills and motor impairment best-explained propulsion error during biofeedback use (adjusted R 2  = 0.56, P = 0.0008), and attention best-explained performance variability (adjusted R 2  = 0.17, P = 0.048). Language skills best-explained recall error during immediate retention (adjusted R 2  = 0.37, P = 0.02). DISCUSSION AND CONCLUSIONS: These results demonstrate that specific cognitive domain impairments explain variability in locomotor learning outcomes in individuals with chronic stroke. This suggests that with further investigation, specific cognitive impairment information may be useful to predict responsiveness to interventions and personalize training parameters to facilitate locomotor learning.

NSF DARE-Transforming modeling in neurorehabilitation: Four threads for catalyzing progress.

We present an overview of the Conference on Transformative Opportunities for Modeling in Neurorehabilitation held in March 2023. It was supported by the Disability and Rehabilitation Engineering (DARE) program from the National Science Foundation's Engineering Biology and Health Cluster. The conference brought together experts and trainees from around the world to discuss critical questions, challenges, and opportunities at the intersection of computational modeling and neurorehabilitation to understand, optimize, and improve clinical translation of neurorehabilitation. We organized the conference around four key, relevant, and promising Focus Areas for modeling: Adaptation & Plasticity, Personalization, Human-Device Interactions, and Modeling 'In-the-Wild'. We identified four common threads across the Focus Areas that, if addressed, can catalyze progress in the short, medium, and long terms. These were: (i) the need to capture and curate appropriate and useful data necessary to develop, validate, and deploy useful computational models (ii) the need to create multi-scale models that span the personalization spectrum from individuals to populations, and from cellular to behavioral levels (iii) the need for algorithms that extract as much information from available data, while requiring as little data as possible from each client (iv) the insistence on leveraging readily available sensors and data systems to push model-driven treatments from the lab, and into the clinic, home, workplace, and community. The conference archive can be found at (dare2023.usc.edu). These topics are also extended by three perspective papers prepared by trainees and junior faculty, clinician researchers, and federal funding agency representatives who attended the conference.

Patterns of asymmetry and energy cost generated from predictive simulations of hemiparetic gait.

Hemiparesis, defined as unilateral muscle weakness, often occurs in people post-stroke or people with cerebral palsy, however it is difficult to understand how this hemiparesis affects movement patterns as it often presents alongside a variety of other neuromuscular impairments. Predictive musculoskeletal modeling presents an opportunity to investigate how impairments affect gait performance assuming a particular cost function. Here, we use predictive simulation to quantify the spatiotemporal asymmetries and changes to metabolic cost that emerge when muscle strength is unilaterally reduced and how reducing spatiotemporal symmetry affects metabolic cost. We modified a 2-D musculoskeletal model by uniformly reducing the peak isometric muscle force unilaterally. We then solved optimal control simulations of walking across a range of speeds by minimizing the sum of the cubed muscle excitations. Lastly, we ran additional optimizations to test if reducing spatiotemporal asymmetry would result in an increase in metabolic cost. Our results showed that the magnitude and direction of effort-optimal spatiotemporal asymmetries depends on both the gait speed and level of weakness. Also, the optimal speed was 1.25 m/s for the symmetrical and 20% weakness models but slower (1.00 m/s) for the 40% and 60% weakness models, suggesting that hemiparesis can account for a portion of the slower gait speed seen in people with hemiparesis. Modifying the cost function to minimize spatiotemporal asymmetry resulted in small increases (~4%) in metabolic cost. Overall, our results indicate that spatiotemporal asymmetry may be optimal for people with hemiparesis. Additionally, the effect of speed and the level of weakness on spatiotemporal asymmetry may help explain the well-known heterogenous distribution of spatiotemporal asymmetries observed in the clinic. Future work could extend our results by testing the effects of other neuromuscular impairments on optimal gait strategies, and therefore build a more comprehensive understanding of the gait patterns observed in clinical populations.

Speed-dependent biomechanical changes vary across individual gait metrics post-stroke relative to neurotypical adults.

BACKGROUND: Gait training at fast speeds is recommended to reduce walking activity limitations post-stroke. Fast walking may also reduce gait kinematic impairments post-stroke. However, it is unknown if differences in gait kinematics between people post-stroke and neurotypical adults decrease when walking at faster speeds. OBJECTIVE: To determine the effect of faster walking speeds on gait kinematics post-stroke relative to neurotypical adults walking at similar speeds. METHODS: We performed a secondary analysis with data from 28 people post-stroke and 50 neurotypical adults treadmill walking at multiple speeds. We evaluated the effects of speed and group on individual spatiotemporal and kinematic metrics and performed k-means clustering with all metrics at self-selected and fast speeds. RESULTS: People post-stroke decreased step length asymmetry and trailing limb angle impairment, reducing between-group differences at fast speeds. Speed-dependent changes in peak swing knee flexion, hip hiking, and temporal asymmetries exaggerated between-group differences. Our clustering analyses revealed two clusters. One represented neurotypical gait behavior, composed of neurotypical and post-stroke participants. The other characterized stroke gait behavior-comprised entirely of participants post-stroke with smaller lower extremity Fugl-Meyer scores than the post-stroke participants in the neurotypical gait behavior cluster. Cluster composition was largely consistent at both speeds, and the distance between clusters increased at fast speeds. CONCLUSIONS: The biomechanical effect of fast walking post-stroke varied across individual gait metrics. For participants within the stroke gait behavior cluster, walking faster led to an overall gait pattern more different than neurotypical adults compared to the self-selected speed. This suggests that to potentiate the biomechanical benefits of walking at faster speeds and improve the overall gait pattern post-stroke, gait metrics with smaller speed-dependent changes may need to be specifically targeted within the context of fast walking.

Manual stabilization reveals a transient role for balance control during locomotor adaptation.

A fundamental feature of human locomotor control is the need to adapt walking patterns in response to changes in the environment. For example, when people walk on a split-belt treadmill, which has belts that move at different speeds, they adapt to the asymmetric speed constraints by reducing spatiotemporal asymmetry. Here, we aim to understand the role of balance control as a potential factor driving this adaptation process. We recruited 24 healthy, young adults to adapt to walking on a split-belt treadmill while either holding on to a handrail or walking with free arm swing. We measured whole body angular momentum and step length asymmetry as measures of dynamic balance and spatiotemporal asymmetry, respectively. To understand how changes in intersegmental coordination influenced whole body angular momentum, we also measured segmental angular momenta and the coefficient of cancellation. When participants were initially exposed to the asymmetry in belt speeds, we observed an increase in whole body angular momentum that was due to both an increase in the momentum of individual segments and a reduction in the coefficient of cancellation. Holding on to a handrail reduced the perturbation to asymmetry during the early phase of adaptation and resulted in a smaller aftereffect during early postadaptation. In addition, the stabilization provided by holding on to a handrail led to reductions in the coupling between angular momentum and asymmetry. These results suggest that regulation of dynamic balance is most important during the initial, transient phase of adaptation to walking on a split-belt treadmill.NEW & NOTEWORTHY We investigated the role of dynamic balance during adaptation to a split-belt treadmill by measuring whole body angular momentum with or without holding on to a handrail. The initial step length asymmetry and associations between balance and asymmetry reduced when holding on to a handrail during early adaptation. These findings indicate that dynamic balance mostly contributes to the initial phase of adaptation when people are exposed to an asymmetric walking constraint.

Using Bayesian inference to estimate plausible muscle forces in musculoskeletal models.

BACKGROUND: Musculoskeletal modeling is currently a preferred method for estimating the muscle forces that underlie observed movements. However, these estimates are sensitive to a variety of assumptions and uncertainties, which creates difficulty when trying to interpret the muscle forces from musculoskeletal simulations. Here, we describe an approach that uses Bayesian inference to identify plausible ranges of muscle forces for a simple motion while representing uncertainty in the measurement of the motion and the objective function used to solve the muscle redundancy problem. METHODS: We generated a reference elbow flexion-extension motion and computed a set of reference forces that would produce the motion while minimizing muscle excitations cubed via OpenSim Moco. We then used a Markov Chain Monte Carlo (MCMC) algorithm to sample from a posterior probability distribution of muscle excitations that would result in the reference elbow motion. We constructed a prior over the excitation parameters which down-weighted regions of the parameter space with greater muscle excitations. We used muscle excitations to find the corresponding kinematics using OpenSim, where the error in position and velocity trajectories (likelihood function) was combined with the sum of the cubed muscle excitations integrated over time (prior function) to compute the posterior probability density. RESULTS: We evaluated the muscle forces that resulted from the set of excitations that were visited in the MCMC chain (seven parallel chains, 500,000 iterations per chain). The estimated muscle forces compared favorably with the reference forces generated with OpenSim Moco, while the elbow angle and velocity from MCMC matched closely with the reference (average RMSE for elbow angle = 2°; and angular velocity = 32°/s). However, our rank plot analyses and potential scale reduction statistics, which we used to evaluate convergence of the algorithm, indicated that the chains did not fully mix. CONCLUSIONS: While the results from this process are a promising step towards characterizing uncertainty in muscle force estimation, the computational time required to search the solution space with, and the lack of MCMC convergence indicates that further developments in MCMC algorithms are necessary for this process to become feasible for larger-scale models.

Using asymmetry to your advantage: learning to acquire and accept external assistance during prolonged split-belt walking.

People can learn to exploit external assistance during walking to reduce energetic cost. For example, walking on a split-belt treadmill affords the opportunity for people to redistribute the mechanical work performed by the legs to gain assistance from the difference in belts' speed and reduce energetic cost. Though we know what people should do to acquire this assistance, this strategy is not observed during typical adaptation studies. We hypothesized that extending the time allotted for adaptation would result in participants adopting asymmetric step lengths to increase the assistance they can acquire from the treadmill. Here, participants walked on a split-belt treadmill for 45 min while we measured spatiotemporal gait variables, metabolic cost, and mechanical work. We show that when people are given sufficient time to adapt, they naturally learn to step further forward on the fast belt, acquire positive mechanical work from the treadmill, and reduce the positive work performed by the legs. We also show that spatiotemporal adaptation and energy optimization operate over different timescales: people continue to reduce energetic cost even after spatiotemporal changes have plateaued. Our findings support the idea that walking with symmetric step lengths, which is traditionally thought of as the endpoint of adaptation, is only a point in the process by which people learn to take advantage of the assistance provided by the treadmill. These results provide further evidence that reducing energetic cost is central in shaping adaptive locomotion, but this process occurs over more extended timescales than those used in typical studies.NEW & NOTEWORTHY Split-belt treadmill adaptation can be seen as a process where people learn to acquire positive work from the treadmill to reduce energetic cost. Though we know what people should do to reduce energetic cost, this strategy is not observed during adaptation studies. We extended the duration of adaptation and show that people continuously adapt their gait to acquire positive work from the treadmill to reduce energetic cost. This process requires longer exposure than traditionally allotted.

General Practice and Digital Methods to Recruit Stroke Survivors to a Clinical Mobility Study: Comparative Analysis.

BACKGROUND: Participant recruitment remains a barrier to conducting clinical research. The disabling nature of a stroke, which often includes functional and cognitive impairments, and the acute stage of illness at which patients are appropriate for many trials make recruiting patients particularly complex and challenging. In addition, people aged 65 years and older, which includes most stroke survivors, have been identified as a group that is difficult to reach and is commonly underrepresented in health research, particularly clinical trials. Digital media may provide effective tools to support enrollment efforts of stroke survivors in clinical trials. OBJECTIVE: The objective of this study was to compare the effectiveness of general practice (traditional) and digital (online) methods of recruiting stroke survivors to a clinical mobility study. METHODS: Recruitment for a clinical mobility study began in July 2018. Eligible study participants included individuals 18 years and older who had a single stroke and were currently ambulatory in the community. General recruiting practice included calling individuals listed in a stroke registry, contacting local physical therapists, and placing study flyers throughout a university campus. Between May 21, 2019, and June 26, 2019, the study was also promoted digitally using the social network Facebook and the search engine marketing tool Google AdWords. The recruitment advertisements (ads) included a link to the study page to which users who clicked were referred. Primary outcomes of interest for both general practice and digital methods included recruitment speed (enrollment rate) and sample characteristics. The data were analyzed using the Lilliefors test, the Welch two-sample t test, and the Mann-Whitney test. Significance was set at P=.05. All statistical analyses were performed in MATLAB 2019b. RESULTS: Our results indicate that digital recruitment methods can address recruitment challenges regarding stroke survivors. Digital recruitment methods allowed us to enroll study participants at a faster rate (1.8 participants/week) compared to using general practice methods (0.57 participants/week). Our findings also demonstrate that digital and general recruitment practices can achieve an equivalent level of sample representativeness. The characteristics of the enrolled stroke survivors did not differ significantly by age (P=.95) or clinical scores (P=.22; P=.82). Comparing the cost-effectiveness of Facebook and Google, we found that the use of Facebook resulted in a lower cost per click and cost per enrollee per ad. CONCLUSIONS: Digital recruitment can be used to expedite participant recruitment of stroke survivors compared to more traditional recruitment practices, while also achieving equivalent sample representativeness. Both general practice and digital recruitment methods will be important to the successful recruitment of stroke survivors. Future studies could focus on testing the effectiveness of additional general practice and digital media approaches and include robust cost-effectiveness analyses. Examining the effectiveness of different messaging and visual approaches tailored to culturally diverse and underrepresented target subgroups could provide further data to move toward evidence-based recruitment strategies.

A Virtual Reality Muscle-Computer Interface for Neurorehabilitation in Chronic Stroke: A Pilot Study.

Severe impairment of limb movement after stroke can be challenging to address in the chronic stage of stroke (e.g., greater than 6 months post stroke). Recent evidence suggests that physical therapy can still promote meaningful recovery after this stage, but the required high amount of therapy is difficult to deliver within the scope of standard clinical practice. Digital gaming technologies are now being combined with brain-computer interfaces to motivate engaging and frequent exercise and promote neural recovery. However, the complexity and expense of acquiring brain signals has held back widespread utilization of these rehabilitation systems. Furthermore, for people that have residual muscle activity, electromyography (EMG) might be a simpler and equally effective alternative. In this pilot study, we evaluate the feasibility and efficacy of an EMG-based variant of our REINVENT virtual reality (VR) neurofeedback rehabilitation system to increase volitional muscle activity while reducing unintended co-contractions. We recruited four participants in the chronic stage of stroke recovery, all with severely restricted active wrist movement. They completed seven 1-hour training sessions during which our head-mounted VR system reinforced activation of the wrist extensor muscles without flexor activation. Before and after training, participants underwent a battery of clinical and neuromuscular assessments. We found that training improved scores on standardized clinical assessments, equivalent to those previously reported for brain-computer interfaces. Additionally, training may have induced changes in corticospinal communication, as indexed by an increase in 12-30 Hz corticomuscular coherence and by an improved ability to maintain a constant level of wrist muscle activity. Our data support the feasibility of using muscle-computer interfaces in severe chronic stroke, as well as their potential to promote functional recovery and trigger neural plasticity.

Taking advantage of external mechanical work to reduce metabolic cost: the mechanics and energetics of split-belt treadmill walking.

KEY POINTS: The neuromotor system generates flexible motor patterns that can adapt to changes in our bodies or environment and also take advantage of assistance provided by the environment. We ask how energy minimization influences adaptive learning during human locomotion to improve economy when walking on a split-belt treadmill. We use a model-based approach to predict how people should adjust their walking pattern to take advantage of the assistance provided by the treadmill, and we validate these predictions empirically. We show that adaptation to a split-belt treadmill can be explained as a process by which people reduce step length asymmetry to take advantage of the work performed by the treadmill to reduce metabolic cost. Our results also have implications for the evaluation of devices designed to reduce effort during walking, as locomotor adaptation may serve as a model approach to understand how people learn to take advantage of external assistance. ABSTRACT: In everyday tasks such as walking and running, we often exploit the work performed by external sources to reduce effort. Recent research has focused on designing assistive devices capable of performing mechanical work to reduce the work performed by muscles and improve walking function. The success of these devices relies on the user learning to take advantage of this external assistance. Although adaptation is central to this process, the study of adaptation is often done using approaches that seem to have little in common with the use of external assistance. We show in 16 young, healthy participants that a common approach for studying adaptation, split-belt treadmill walking, can be understood from a perspective in which people learn to take advantage of mechanical work performed by the treadmill. Initially, during split-belt walking, people step further forward on the slow belt than the fast belt which we measure as a negative step length asymmetry, but this asymmetry is reduced with practice. We demonstrate that reductions in asymmetry allow people to extract positive work from the treadmill, reduce the positive work performed by the legs, and reduce metabolic cost. We also show that walking with positive step length asymmetries, defined by longer steps on the fast belt, minimizes metabolic cost, and people choose this pattern after guided experience of a wide range of asymmetries. Our results suggest that split-belt adaptation can be interpreted as a process by which people learn to take advantage of mechanical work performed by an external device to improve economy.

Microtargeting for conservation.

Widespread human action and behavior change is needed to achieve many conservation goals. Doing so at the requisite scale and pace will require the efficient delivery of outreach campaigns. Conservation gains will be greatest when efforts are directed toward places of high conservation value (or need) and tailored to critical actors. Recent strategic conservation planning has relied primarily on spatial assessments of biophysical attributes, largely ignoring the human dimensions. Elsewhere, marketers, political campaigns, and others use microtargeting-predictive analytics of big data-to identify people most likely to respond positively to particular messages or interventions. Conservationists have not yet widely capitalized on these techniques. To investigate the effectiveness of microtargeting to improve conservation, we developed a propensity model to predict restoration behavior among 203,645 private landowners in a 5,200,000 ha study area in the Chesapeake Bay Watershed (U.S.A.). To isolate the additional value microtargeting may offer beyond geospatial prioritization, we analyzed a new high-resolution land-cover data set and cadastral data to identify private owners of riparian areas needing restoration. Subsequently, we developed and evaluated a restoration propensity model based on a database of landowners who had conducted restoration in the past and those who had not (n = 4978). Model validation in a parallel database (n = 4989) showed owners with the highest scorers for propensity to conduct restoration (i.e., top decile) were over twice as likely as average landowners to have conducted restoration (135%). These results demonstrate that microtargeting techniques can dramatically increase the efficiency and efficacy of conservation programs, above and beyond the advances offered by biophysical prioritizations alone, as well as facilitate more robust research of many social-ecological systems.

Micro-focalización para la Conservación Resumen Se necesitan acciones humanas diseminadas y un cambio en el comportamiento para lograr muchos objetivos de conservación. Que se logre esto a la escala y al ritmo requerido requerirá de la entrega eficiente de las campañas de participación. Las ganancias de la conservación serán mayores cuando los esfuerzos estén dirigidos hacia sitios con un alto valor (o necesidad) de conservación y estén personalizados para los actores más importantes. La reciente planeación estratégica de la conservación ha dependido principalmente de las evaluaciones espaciales de los atributos biofísicos, ignorando generalmente las dimensiones humanas. En otros ámbitos, los mercadólogos, las campañas políticas, así como otros, usan la micro-focalización - el análisis predictivo de datos masivos - para identificar a las personas con mayor probabilidad de responder positivamente a mensajes o intervenciones particulares. Los conservacionistas todavía no han capitalizado extensamente estas técnicas. Desarrollamos un modelo de tendencia para predecir el comportamiento de restauración entre 203,645 terratenientes privados en un área de estudio de 5,200,000 ha en la cuenca de la Bahía de Chesapeake (E.U.A.) y así investigar la eficiencia de la micro-focalización en el aumento de la conservación. Para aislar el valor adicional que puede ofrecer la micro-focalización más allá de la priorización geoespacial, analizamos un nuevo conjunto de alta resolución de datos sobre la cobertura del suelo y datos catastrales para identificar a los terratenientes privados de áreas ribereñas que necesitan restauración. Después de esto, desarrollamos y evaluamos el modelo de tendencia a la restauración basado en una base de datos de terratenientes que han realizado restauraciones en el pasado y aquellos que no (n = 4,978). La validación del modelo en una base de datos paralela (n = 4,989) mostró que los terratenientes con los puntajes más altos para la tendencia a la restauración (es decir, el decil superior) tenían el doble de probabilidad de haber realizado acciones de conservación que el terrateniente promedio (135%). Estos resultados muestran que las técnicas de micro-focalización pueden incrementar dramáticamente la eficiencia y la eficacia de los programas de conservación, más allá de los avances ofrecidos sólo por las priorizaciones biofísicas, así como facilitar la investigación más sólida sobre muchos sistemas socio-ecológicos.

Locomotor skill acquisition in virtual reality shows sustained transfer to the real world.

BACKGROUND: Virtual reality (VR) is a potentially promising tool for enhancing real-world locomotion in individuals with mobility impairment through its ability to provide personalized performance feedback and simulate real-world challenges. However, it is unknown whether novel locomotor skills learned in VR show sustained transfer to the real world. Here, as an initial step towards developing a VR-based clinical intervention, we study how young adults learn and transfer a treadmill-based virtual obstacle negotiation skill to the real world. METHODS: On Day 1, participants crossed virtual obstacles while walking on a treadmill, with the instruction to minimize foot clearance during obstacle crossing. Gradual changes in performance during training were fit via non-linear mixed effect models. Immediate transfer was measured by foot clearance during physical obstacle crossing while walking over-ground. Retention of the obstacle negotiation skill in VR and retention of over-ground transfer were assessed after 24 h. RESULTS: On Day 1, participants systematically reduced foot clearance throughout practice by an average of 5 cm (SD 4 cm) and transferred 3 cm (SD 1 cm) of this reduction to over-ground walking. The acquired reduction in foot clearance was also retained after 24 h in VR and over-ground. There was only a small, but significant 0.8 cm increase in foot clearance in VR and no significant increase in clearance over-ground on Day 2. Moreover, individual differences in final performance at the end of practice on Day 1 predicted retention both in VR and in the real environment. CONCLUSIONS: Overall, our results support the use of VR for locomotor training as skills learned in a virtual environment readily transfer to real-world locomotion. Future work is needed to determine if VR-based locomotor training leads to sustained transfer in clinical populations with mobility impairments, such as individuals with Parkinson's disease and stroke survivors.

Receptor-Mediated Delivery of CRISPR-Cas9 Endonuclease for Cell-Type-Specific Gene Editing.

CRISPR-Cas RNA-guided endonucleases hold great promise for disrupting or correcting genomic sequences through site-specific DNA cleavage and repair. However, the lack of methods for cell- and tissue-selective delivery currently limits both research and clinical uses of these enzymes. We report the design and in vitro evaluation of S. pyogenes Cas9 proteins harboring asialoglycoprotein receptor ligands (ASGPrL). In particular, we demonstrate that the resulting ribonucleoproteins (Cas9-ASGPrL RNP) can be engineered to be preferentially internalized into cells expressing the corresponding receptor on their surface. Uptake of such fluorescently labeled proteins in liver-derived cell lines HEPG2 (ASGPr+) and SKHEP (control; diminished ASGPr) was studied by live cell imaging and demonstrates increased accumulation of Cas9-ASGPrL RNP in HEPG2 cells as a result of effective ASGPr-mediated endocytosis. When uptake occurred in the presence of a peptide with endosomolytic properties, we observed receptor-facilitated and cell-type specific gene editing that did not rely on electroporation or the use of transfection reagents. Overall, these in vitro results validate the receptor-mediated delivery of genome-editing enzymes as an approach for cell-selective gene editing and provide a framework for future potential applications to hepatoselective gene editing in vivo.

The quality of visual information about the lower extremities influences visuomotor coordination during virtual obstacle negotiation.

Successful negotiation of obstacles during walking relies on the integration of visual information about the environment with ongoing locomotor commands. When information about the body and the environment is removed through occlusion of the lower visual field, individuals increase downward head pitch angle, reduce foot placement precision, and increase safety margins during crossing. However, whether these effects are mediated by loss of visual information about the lower extremities, the obstacle, or both remains to be seen. Here we used a fully immersive, virtual obstacle negotiation task to investigate how visual information about the lower extremities is integrated with information about the environment to facilitate skillful obstacle negotiation. Participants stepped over virtual obstacles while walking on a treadmill with one of three types of visual feedback about the lower extremities: no feedback, end-point feedback, and a link-segment model. We found that absence of visual information about the lower extremities led to an increase in the variability of leading foot placement after crossing. The presence of a visual representation of the lower extremities promoted greater downward head pitch angle during the approach to and subsequent crossing of an obstacle. In addition, having greater downward head pitch was associated with closer placement of the trailing foot to the obstacle, further placement of the leading foot after the obstacle, and higher trailing foot clearance. These results demonstrate that the fidelity of visual information about the lower extremities influences both feedforward and feedback aspects of visuomotor coordination during obstacle negotiation. NEW & NOTEWORTHY Here we demonstrate that visual information about the lower extremities is utilized for precise foot placement and control of safety margins during obstacle negotiation. We also found that when a visual representation of the lower extremities is present, this information is used in the online control of foot trajectory. Together, our results highlight how visual information about the body and the environment is integrated with motor commands for planning and online control of obstacle negotiation.

Efficient Liver Targeting by Polyvalent Display of a Compact Ligand for the Asialoglycoprotein Receptor.

A compact and stable bicyclic bridged ketal was developed as a ligand for the asialoglycoprotein receptor (ASGPR). This compound showed excellent ligand efficiency, and the molecular details of binding were revealed by the first X-ray crystal structures of ligand-bound ASGPR. This analogue was used to make potent di- and trivalent binders of ASGPR. Extensive characterization of the function of these compounds showed rapid ASGPR-dependent cellular uptake in vitro and high levels of liver/plasma selectivity in vivo. Assessment of the biodistribution in rodents of a prototypical Alexa647-labeled trivalent conjugate showed selective hepatocyte targeting with no detectable distribution in nonparenchymal cells. This molecule also exhibited increased ASGPR-directed hepatocellular uptake and prolonged retention compared to a similar GalNAc derived trimer conjugate. Selective release in the liver of a passively permeable small-molecule cargo was achieved by retro-Diels-Alder cleavage of an oxanorbornadiene linkage, presumably upon encountering intracellular thiol. Therefore, the multicomponent construct described here represents a highly efficient delivery vehicle to hepatocytes.

Development of Fluorophore-Labeled Thailanstatin Antibody-Drug Conjugates for Cellular Trafficking Studies.

As the antibody-drug conjugate (ADC) field grows increasingly important for cancer treatment, it is vital for researchers to establish a firm understanding of how ADCs function at the molecular level. To gain insight into ADC uptake, trafficking, and catabolism-processes that are critical to ADC efficacy and toxicity-imaging studies have been performed with fluorophore-labeled conjugates. However, such labels may alter the properties and behavior of the ADC under investigation. As an alternative approach, we present here the development of a "clickable" ADC bearing an azide-functionalized linker-payload (LP) poised for "click" reaction with alkyne fluorophores; the azide group represents a significantly smaller structural perturbation to the LP than most fluorophores. Notably, the clickable ADC shows excellent potency in target-expressing cells, whereas the fluorophore-labeled product ADC suffers from a significant loss of activity, underscoring the impact of the label itself on the payload. Live-cell confocal microscopy reveals robust uptake of the clickable ADC, which reacts selectively in situ with a derivatized fluorescent label. Time-course trafficking studies show greater and more rapid net internalization of the ADCs than the parent antibody. More generally, the application of chemical biology tools to the study of ADCs should improve our understanding of how ADCs are processed in biological systems.

Walking in fully immersive virtual environments: an evaluation of potential adverse effects in older adults and individuals with Parkinson's disease.

BACKGROUND: Virtual reality (VR) has recently been explored as a tool for neurorehabilitation to enable individuals with Parkinson's disease (PD) to practice challenging skills in a safe environment. Current technological advances have enabled the use of affordable, fully immersive head-mounted displays (HMDs) for potential therapeutic applications. However, while previous studies have used HMDs in individuals with PD, these were only used for short bouts of walking. Clinical applications of VR for gait training would likely involve an extended exposure to the virtual environment, which has the potential to cause individuals with PD to experience simulator-related adverse effects due to their age or pathology. Thus, our objective was to evaluate the safety of using an HMD for longer bouts of walking in fully immersive VR for older adults and individuals with PD. METHODS: Thirty-three participants (11 healthy young, 11 healthy older adults, and 11 individuals with PD) were recruited for this study. Participants walked for 20 min while viewing a virtual city scene through an HMD (Oculus Rift DK2). Safety was evaluated using the mini-BESTest, measures of center of pressure (CoP) excursion, and questionnaires addressing symptoms of simulator sickness (SSQ) and measures of stress and arousal. RESULTS: Most participants successfully completed all trials without any discomfort. There were no significant changes for any of our groups in symptoms of simulator sickness or measures of static and dynamic balance after exposure to the virtual environment. Surprisingly, measures of stress decreased in all groups while the PD group also increased the level of arousal after exposure. CONCLUSIONS: Older adults and individuals with PD were able to successfully use immersive VR during walking without adverse effects. This provides systematic evidence supporting the safety of immersive VR for gait training in these populations.

A marching-walking hybrid induces step length adaptation and transfers to natural walking.

Walking is highly adaptable to new demands and environments. We have previously studied adaptation of locomotor patterns via a split-belt treadmill, where subjects learn to walk with one foot moving faster than the other. Subjects learn to adapt their walking pattern by changing the location (spatial) and time (temporal) of foot placement. Here we asked whether we can induce adaptation of a specific walking pattern when one limb does not "walk" but instead marches in place (i.e., marching-walking hybrid). The marching leg's movement is limited during the stance phase, and thus certain sensory signals important for walking may be reduced. We hypothesized that this would produce a spatial-temporal strategy different from that of normal split-belt adaptation. Healthy subjects performed two experiments to determine whether they could adapt their spatial-temporal pattern of step lengths during the marching-walking hybrid and whether the learning transfers to over ground walking. Results showed that the hybrid group did adapt their step lengths, but the time course of adaptation and deadaption was slower than that for the split-belt group. We also observed that the hybrid group utilized a mostly spatial strategy whereas the split-belt group utilized both spatial and temporal strategies. Surprisingly, we found no significant difference between the hybrid and split-belt groups in over ground transfer. Moreover, the hybrid group retained more of the learned pattern when they returned to the treadmill. These findings suggest that physical rehabilitation with this marching-walking paradigm on conventional treadmills may produce changes in symmetry comparable to what is observed during split-belt training.

Drift during overground locomotion in newly hatched chicks varies with light exposure during embryogenesis.

In an earlier study of newly hatched chicks we reported that continuous bright light exposure throughout incubation accelerated locomotor development and continuous dark exposure delayed it, compared to less intense, intermittent light exposure. Commonly studied gait parameters indicated locomotor skill was similar across groups. However, dark incubated chicks walked with a greater step width, raising the possibility of differences in dynamic balance and control of forward progression. In this study, we established methods to retrospectively examine the previously published locomotor data for differences in lateral drift. We hypothesized that chicks incubated in darkness would exhibit more drift than chicks incubated in light. Analyses identified differences in forward progression between chicks incubated in the two extreme light conditions, supporting the study's hypothesis. We discuss the significance of our findings and potential design considerations for future studies of light-accelerated motor development in precocial and nonprecocial animals.

Daily Quantity of Infant Leg Movement: Wearable Sensor Algorithm and Relationship to Walking Onset.

BACKGROUND: Normative values are lacking for daily quantity of infant leg movements. This is critical for understanding the relationship between the quantity of leg movements and onset of independent walking, and will begin to inform early therapy intervention for infants at risk for developmental delay. METHODS: We used wearable inertial movement sensors to record full-day leg movement activity from 12 infants with typical development, ages 1-12 months. Each infant was tested three times across 5 months, and followed until the onset of independent walking. We developed and validated an algorithm to identify infant-produced leg movements. RESULTS: Infants moved their legs tens of thousands of times per day. There was a significant effect of leg movement quantity on walking onset. Infants who moved their legs more walked later than infants who moved their legs less, even when adjusting for age, developmental level or percentile length. We will need a much larger sample to adequately capture and describe the effect of movement experience on developmental rate. Our algorithm defines a leg movement in a specific way (each pause or change in direction is counted as a new movement), and further assessment of movement characteristics are necessary before we can fully understand and interpret our finding that infants who moved their legs more walked later than infants who moved their legs less. CONCLUSIONS: We have shown that typically-developing infants produce thousands of leg movements in a typical day, and that this can be accurately captured in the home environment using wearable sensors. In our small sample we can identify there is an effect of leg movement quantity on walking onset, however we cannot fully explain it.