Electroencephalographic Study on Sensory Integration in Visually Induced Postural Sway.

A periodically reversing optic flow animation, experienced while standing, induces an involuntary sway termed visually induced postural sway (VIPS). Interestingly, VIPS is suppressed during light finger touch to a stationary object. Here, we explored whether VIPS is mediated by parietal field activity in the dorsal visual stream as well as by activity in early visual areas, as has been suggested. We performed a mobile brain/body imaging study using high-density electroencephalographic recording from human participants (11 men and 3 women) standing during exposure to periodically reversing optic flow with and without light finger touch to a stable surface. We also performed recording their visuo-postural tracking movements as a typical visually guided movement to explore differences of cortical process of VIPS from the voluntary visuomotor process involving the dorsal stream. In the visuo-postural tracking condition, the participants moved their center of pressure in time with a slowly oscillating (expanding, shrinking) target rectangle. Source-resolved results showed that alpha band (8-13 Hz) activity in the medial and right occipital cortex during VIPS was modulated by the direction and velocity of optic flow and increased significantly during light finger touch. However, source-resolved potentials from the parietal association cortex showed no such modulation. During voluntary postural sway with feedback (but no visual flow) in which the dorsal stream is involved, sensorimotor areas produced more theta band (4-7 Hz) and less beta band (14-35 Hz) activity than during involuntary VIPS. These results suggest that VIPS involves cortical field dynamic changes in the early visual cortex rather than in the posterior parietal cortex of the visual dorsal stream.

Repetitive Control for Multi-Joint Arm Movements Based on Virtual Trajectories.

According to the neuromuscular model of virtual trajectory control, the postures and movements of limbs are performed by shifting the equilibrium positions determined by agonist and antagonist muscle activities. In this study, we develop virtual trajectory control for the reaching movements of a multi-joint arm, introducing a proportional-derivative feedback control scheme. In virtual trajectory control, it is crucial to design a suitable virtual trajectory such that the desired trajectory can be realized. To this end, we propose an algorithm for updating virtual trajectories in repetitive control, which can be regarded as a Newton-like method in a function space. In our repetitive control, the virtual trajectory is corrected without explicit calculation of the arm dynamics, and the actual trajectory converges to the desired trajectory. Using computer simulations, we assessed the proposed repetitive control for the trajectory tracking of a two-link arm. Our results confirmed that when the feedback gains were reasonably high and the sampling time was sufficiently small, the virtual trajectory was adequately updated, and the desired trajectory was almost achieved within approximately 10 iterative trials. We also propose a method for modifying the virtual trajectory to ensure that the formation of the actual trajectory is identical even when the feedback gains are changed. This modification method makes it possible to execute flexible control, in which the feedback gains are effectively altered according to motion tasks.

Study on AAV-mediated gene therapy for diabetes in humanized liver mouse to predict efficacy in humans.

Most in vivo studies on the conversion to insulin-producing cells with AAV carrying PDX1 gene are performed in rodents. However, there is little information regarding Adeno-associated virus (AAV) carrying PDX1 gene transduced to human liver in vivo because accidental death caused by unpredicted factors cannot be denied, such as the hypoglycemic agent troglitazone with hepatic failure. Here we aim to confirm insulin secretion from human liver transduced with AAV carrying PDX1 gene in vivo and any secondary effect using a humanized liver mouse. As the results, AAV2-PG succeeded to improve the hyperglycemia of STZ-induced diabetic humanized liver mice. Then, the analysis of humanized liver mice revealed that the AAV2-PG was more transducible to humanized liver area than to mouse liver area. In conclusion, the humanized liver mouse model could be used to examine AAV transduction of human hepatocytes in vivo and better predict clinical transduction efficiency than nonhumanized mice.

Whole-Body Reaching Movements Formulated by Minimum Muscle-Tension Change Criterion.

It is well known that planar reaching movements of the human shoulder and elbow joints have invariant features: roughly straight hand paths and bell-shaped velocity profiles. The optimal control models with the criteria of smoothness or precision, which determine a unique movement pattern, predict such features of hand trajectories. In this letter on expanding the research on simple arm reaching movements, we examine whether the smoothness criteria can be applied to whole-body reaching movements with many degrees of freedom. Determining a suitable joint trajectory in the whole-body reaching movement corresponds to the optimization problem with constraints, since body balance must be maintained during a motion task. First, we measured human joint trajectories and ground reaction forces during whole-body reaching movements, and confirmed that subjects formed similar movements with common characteristics in the trajectories of the hand position and body center of mass. Second, we calculated the optimal trajectories according to the criteria of torque and muscle-tension smoothness. While the minimum torque change trajectories were not consistent with the experimental data, the minimum muscle-tension change model was able to predict the stereotyped features of the measured trajectories. To explore the dominant effects of the extension from the torque change to the muscle-tension change, we introduced a weighted torque change cost function. Considering the maximum voluntary contraction (MVC) force of the muscle as the weighting factor of each joint torque, we formulated the weighted torque change cost as a simplified version of the minimum muscle-tension change cost. The trajectories owing to the minimum weighted torque change criterion also showed qualitative agreement with the common features of the measured data. Proper estimation of the MVC forces in the body joints is essential to reproduce human whole-body movements according to the minimum muscle-tension change criterion.

Change of a motor synergy for dampening hand vibration depending on a task difficulty.

The present study investigated the relationship between the number of usable degrees of freedom (DOFs) and joint coordination during a human-dampening hand vibration task. Participants stood on a platform generating an anterior-posterior directional oscillation and held a water-filled cup. Their usable DOFs were changed under the following conditions of limb constraint: (1) no constraint; (2) ankle constrained; and (3) ankle-knee constrained. Kinematic whole-body data were recorded using a three-dimensional position measurement system. The jerk of each body part was evaluated as an index of oscillation intensity. To quantify joint coordination, an uncontrolled manifold (UCM) analysis was applied and the variance of joints related to hand jerk divided into two components: a UCM component that did not affect hand jerk and an orthogonal (ORT) component that directly affected hand jerk. The results showed that hand jerk when the task used a cup filled with water was significantly smaller than when a cup containing stones was used, regardless of limb constraint condition. Thus, participants dampened their hand vibration utilizing usable joint DOFs. According to UCM analysis, increasing the oscillation velocity and the decrease in usable DOFs by the limb constraints led to an increase of total variance of the joints and the UCM component, indicating that a synergy-dampening hand vibration was enhanced. These results show that the variance of usable joint DOFs is more fitted to the UCM subspace when the joints are varied by increasing the velocity and limb constraints and suggest that humans adopt enhanced synergies to achieve more difficult tasks.

Motor synergies for dampening hand vibration during human walking.

This study investigated the motion required to carry a cup filled with water without spilling it, which is a common human dexterous task. This task requires the individual to dampen hand vibration while walking. We hypothesize that a reduction in hand jerk and a constant cup angle are required to achieve this task. We measured movements while human subjects carried a cup with water (WW task) and with stones (WS task) using a three-dimensional position measurement system and then analyzed joint coordination. We empirically confirmed that the value of hand jerk and the variance in cup angle in the WW task were smaller than those in the WS task. We used uncontrolled manifold (UCM) analysis to quantify joint coordination corresponding to the motor synergy required to reduce the hand jerk and variance of the cup angle. UCM components, which did not affect the hand jerk and cup angle, were larger than orthogonal components, which directly affected the hand jerk and cup angle in the WW task. These results suggest that there is a coordinated control mechanism that reduces hand jerk and maintains a constant cup angle when carrying a cup filled with water without spilling it. In addition, we suggest that humans adopt a flexible and coordinated control strategy of allowing variance independent of the variables that should be controlled to achieve this dexterous task.

Validation of Balance Map Analysis of Walking at Different Speeds.

Walking balance about falling in the forward direction is associated with the body's center of mass and placement of the swing foot during the swing phase. Balance map analysis evaluates walking balance based on the prediction of the reachability of an appropriate foot placement using a simple biomechanical model during the swing phase without active joint torque (ballistic walking model). The ballistic walking model can be justified in terms of the preferred walking speed because the metabolic energy consumption associated with muscle activity in faster and slower walking is higher than that in preferred speed walking. Therefore, the assumption that the active joint torque is sufficiently small during the swing phase may not hold in faster or slower walking, which can be a significant limitation of balance map analysis. In this study, it was hypothesized that steady-state walking at various walking speeds would be evaluated as stable for validation of the balance map analysis, and the gait patterns for three types of walking speeds (slow, normal, and fast) were examined. The results showed that the trajectories during the swing phase were within stable regions for all conditions, with a sufficient margin from the forward balance loss region. In addition, the margin from forward balance was reduced with an increase in walking velocity. The decrease in the margin during fast walking resulted from an increase in the forward velocity of the body's center of mass in relation to the velocity of the swing leg. These results suggest that balance map analysis effectively measures walking balance at various speeds.

Balance Map Analysis for Visualization and Quantification of Balance in Human Walking.

Evaluation of stability or loss of balance in walking persists as an open question. Although an inverted pendulum model is often adopted to evaluate stance leg balance, a stumbling-related balance loss should be associated with the swing leg. We propose a new framework based on a compass gait model that determines whether the current state (i.e., position and velocity) in the swing phase can maintain steady state walking or, instead, fall without active joint torque, which is termed as balance map analysis. The forward and backward balance loss regions are derived by a linear compass gait model. To test the balance map analysis, measurement experiments of steady state walking and stumbled walking are used to validate two hypotheses: 1) the state during steady-state walking is not located in the balance loss region; and 2) if stumbling occurs, the state moves toward the forward balance loss region. The results of the balance map analysis showed good agreement with our prediction in the hypotheses. The minimum Euclid distance from the balance loss region is defined as the margin, and the margin from forward balance loss rapidly decreased after the stumbling perturbation. The statistical results reveal that the margin from the forward balance loss region after perturbation is significantly smaller than the margin in steady state walking. These results suggest that balance map analysis provides a new aspect of walking balance, expanded for the stumbling and recovery behavior of human walking.The code for the balance map analysis is available at https://github.com/TakahiroKagawa/GaitAnalysis_BM.

Development of a novel humanized mouse model for improved evaluation of in vivo anti-cancer effects of anti-PD-1 antibody.

Immune checkpoint inhibitors (ICIs) have revolutionized the treatment of cancer in the clinic. Further discovery of novel drugs or therapeutic protocols that enhance efficacy requires reliable animal models that recapitulate human immune responses to ICI treatment in vivo. In this study, we utilized an immunodeficient NOG mouse substrain deficient for mouse FcγR genes, NOG-FcγR-/- mice, to evaluate the anti-cancer effects of nivolumab, an anti-programmed cell death-1 (PD-1) antibody. After reconstitution of human immune systems by human hematopoietic stem cell transplantation (huNOG-FcγR-/- mice), four different programmed death-ligand 1 (PD-L1)-positive human cancer cell lines were tested. Among them, the growth of three cell lines was strongly suppressed by nivolumab in huNOG-FcγR-/- mice, but not in conventional huNOG mice. Accordingly, immunohistochemistry demonstrated the enhanced infiltration of human T cells into tumor parenchyma in only nivolumab-treated huNOG-FcγR-/- mice. Consistently, the number of human T cells was increased in the spleen in huNOG-FcγR-/- mice by nivolumab but not in huNOG mice. Furthermore, human PD-L1 expression was strongly induced in the spleen of huNOG-FcγR-/- mice. Collectively, our results suggest that the anti-cancer effects of anti-PD-1 antibodies can be detected more clearly in NOG-FcγR-/- mice than in NOG mice.

Development of blastocyst complementation technology without contributions to gametes and the brain.

In Japan, it is possible to generate chimeric animals from specified embryos by combining animal blastocysts with human pluripotent stem (PS) cells (animal-human PS chimera). However, the production of animal-human PS chimeras has been restricted because of ethical concerns, such as the development of human-like intelligence and formation of humanized gametes in the animals, owing to the contributions of human PS cells to the brain and reproductive organs. To solve these problems, we established a novel blastocyst complementation technology that does not contribute to the gametes or the brain. First, we established GFP-expressing mouse embryonic stem cells (G-mESCs) in which the Prdm14 and Otx2 genes were knocked out and generated chimeric mice by injecting them into PDX-1-deficient blastocysts. The results showed that the G-mESCs did not contribute to the formation of gametes and the brain. Therefore, in the PDX-1-deficient mice complemented by G-mESCs without the Prdm14 and Otx2 genes, the germline was not transmitted to the next generations. This approach could address concerns regarding the development of both human gametes and a human-like brain upon mouse blastocyst complementation using human stem cells.

Adiponectin deficiency-induced diabetes increases TNFα and FFA via downregulation of PPARα.

Expression of peroxisome proliferator-activated receptor (PPAR) α was investigated in adiponectin knockout mice to elucidate the relationship between PPARα and adiponectin deficiency-induced diabetes. Adiponectin knockout (Adp-/-) mice were generated by gene targeting. Glucose tolerance test (GTT), insulin tolerance test (ITT), and organ sampling were performed in Adp-/- mice at the age of 10 weeks. PPARα, insulin, triglyceride, free fatty acid (FFA), and tumor necrosis factor α (TNFα) were analyzed from the sampled organs. Adp-/- mice showed impaired glucose tolerance and insulin resistance. Additionally, PPARα levels were decreased and plasma concentration of triglyceride, FFA and TNFα were increased. These data may indicate that insulin resistance in Adp-/- mice is likely caused by an increase in concentrations of TNFα and FFA via downregulation of PPARα.

Establishment of Immunodeficient Retinal Degeneration Model Mice and Functional Maturation of Human ESC-Derived Retinal Sheets after Transplantation.

Increasing demand for clinical retinal degeneration therapies featuring human ESC/iPSC-derived retinal tissue and cells warrants proof-of-concept studies. Here, we established two mouse models of end-stage retinal degeneration with immunodeficiency, NOG-rd1-2J and NOG-rd10, and characterized disease progress and immunodeficient status. We also transplanted human ESC-derived retinal sheets into NOG-rd1-2J and confirmed their long-term survival and maturation of the structured graft photoreceptor layer, without rejection or tumorigenesis. We recorded light responses from the host ganglion cells using a multi-electrode array system; this result was consistent with whole-mount immunostaining suggestive of host-graft synapse formation at the responding sites. This study demonstrates an application of our mouse models and provides a proof of concept for the clinical use of human ESC-derived retinal sheets.

Stability analysis of paraplegic standing while wearing an orthosis.

Paraplegics can maintain a standing posture, called the "C-posture", while wearing an orthosis. The significant feature of the C-posture is that the body's center of mass is located behind the hip joint. In this study, we investigate the C-posture mechanism and assess the relationship between posture and stability, the aim being to restore standing function. We first measured the standing postures of paraplegic subjects wearing an orthosis. The subjects maintained the standing posture for 30 s. Next, assuming the C-posture to be an equilibrium attractor in the musculoskeletal system, we used a dynamic model of the musculoskeletal system to analyze the relationship between posture and stability, and also to assess the influence of ankle stiffness. We calculated the standing posture on the basis of a return map. The calculated standing postures show some features of the C-posture. The stability analysis revealed that, despite a limitation in the range of stable postures, the C-posture is more stable than the postures of normal people. The results suggest that although the C-posture is an appropriate posture for paraplegic standing, sufficient ankle stiffness and an appropriate alignment of ankle angle are necessary and preventing hip flexion movements is desirable.

Uncontrolled Manifold Reference Feedback Control of Multi-Joint Robot Arms.

The brain must coordinate with redundant bodies to perform motion tasks. The aim of the present study is to propose a novel control model that predicts the characteristics of human joint coordination at a behavioral level. To evaluate the joint coordination, an uncontrolled manifold (UCM) analysis that focuses on the trial-to-trial variance of joints has been proposed. The UCM is a nonlinear manifold associated with redundant kinematics. In this study, we directly applied the notion of the UCM to our proposed control model called the "UCM reference feedback control." To simplify the problem, the present study considered how the redundant joints were controlled to regulate a given target hand position. We considered a conventional method that pre-determined a unique target joint trajectory by inverse kinematics or any other optimization method. In contrast, our proposed control method generates a UCM as a control target at each time step. The target UCM is a subspace of joint angles whose variability does not affect the hand position. The joint combination in the target UCM is then selected so as to minimize the cost function, which consisted of the joint torque and torque change. To examine whether the proposed method could reproduce human-like joint coordination, we conducted simulation and measurement experiments. In the simulation experiments, a three-link arm with a shoulder, elbow, and wrist regulates a one-dimensional target of a hand through proposed method. In the measurement experiments, subjects performed a one-dimensional target-tracking task. The kinematics, dynamics, and joint coordination were quantitatively compared with the simulation data of the proposed method. As a result, the UCM reference feedback control could quantitatively reproduce the difference of the mean value for the end hand position between the initial postures, the peaks of the bell-shape tangential hand velocity, the sum of the squared torque, the mean value for the torque change, the variance components, and the index of synergy as well as the human subjects. We concluded that UCM reference feedback control can reproduce human-like joint coordination. The inference for motor control of the human central nervous system based on the proposed method was discussed.

Changes in motor synergies for tracking movement and responses to perturbations depend on task-irrelevant dimension constraints.

We investigated the changes in the motor synergies of target-tracking movements of hands and the responses to perturbation when the dimensionalities of target positions were changed. We used uncontrolled manifold (UCM) analyses to quantify the motor synergies. The target was changed from one to two dimensions, and the direction orthogonal to the movement direction was switched from task-irrelevant directions to task-relevant directions. The movement direction was task-relevant in both task conditions. Hence, we evaluated the effects of constraints on the redundant dimensions on movement tracking. Moreover, we could compare the two types of responses to the same directional perturbations in one- and two-dimensional target tasks. In the one-dimensional target task, the perturbation along the movement direction and the orthogonal direction were task-relevant and -irrelevant perturbations, respectively. In the two-dimensional target task, the both perturbations were task-relevant perturbations. The results of the experiments showed that the variabilities of the hand positions in the two-dimensional target-tracking task decreased, but the variances of the joint angles did not significantly change. For the task-irrelevant perturbations, the variances of the joint angles within the UCM that did not affect hand position (UCM component) increased. For the task-relevant perturbations, the UCM component tended to increase when the available UCM was large. These results suggest that humans discriminate whether the perturbations were task-relevant or -irrelevant and then adjust the responses of the joints by utilizing the available UCM.

Control model for dampening hand vibrations using information of internal and external coordinates.

In the present study, we investigate a control mechanism that dampens hand vibrations. Here, we propose a control method with two components to suppress hand vibrations. The first is a passive suppression method that lowers the joint stiffness to passively dampen the hand vibrations. The second is an active suppression method that adjusts an equilibrium point based on skyhook control to actively dampen the hand vibrations. In a simulation experiment, we applied these two methods to dampen hand vibrations during the shoulder's horizontal oscillation. We also conducted a measurement experiment wherein a subject's shoulder was sinusoidally oscillated by a platform that generated horizontal oscillations. The results of the measurement experiments showed that the jerk of each part of the arm in a task using a cup filled with water was smaller than the shoulder jerk and that in a task with a cup filled with stones was larger than the shoulder jerk. Moreover, the amplitude of the hand trajectory in both horizontal and vertical directions was smaller in a task using a cup filled with water than in a task using a cup filled with stones. The results of the measurement experiments were accurately reproduced by the active suppression method based on skyhook control. These results suggest that humans dampen hand vibrations by controlling the equilibrium point through the information of the external workspace and the internal body state rather than by lowering joint stiffness only by using internal information.

Expression of pancreatic and duodenal homeobox1 (PDX1) protein in the interior and exterior regions of the intestine, revealed by development and analysis of Pdx1 knockout mice.

We developed pancreatic and duodenal homeobox1 (Pdx1) knockout mice to improve a compensatory hyperinsulinemia, which was induced by hyperplasia in the ß cells or Langerhans' islands, as the diabetic model mice. For targeting of Pdx1 gene by homologous recombination, ES cells derived from a 129(+Ter) /SvJcl×C57BL/6JJcl hybrid mouse were electroporated and subjected to positive-negative selection with hygromycin B and ganciclovir. As these results, one of the three chimeric mice succeeded to produce the next or F1 generation. Then, the mouse fetuses were extracted from the mother's uterus and analyzed immunohistologically for the existence of a pancreas. The fetuses were analyzed at embryonic day 14.5 (E14.5) because Pdx1 knockout could not alive after birth in this study. Immunohistochemical staining revealed that 10 fetuses out of 26 did not have any PDX1 positive primordium of the pancreas and that the PDX1 expresses in both the interior and exterior regions of intestine. In particular, one the exterior of the intestine PDX1 was expressed in glands that would be expected to form the pancreas. The result of PCR genotyping with extracted DNA from the paraffin sections showed existence of 10 Pdx1-knockout mice and corresponded to results of immunostaining. Thus, we succeeded to establish a Pdx1-knockout (Pdx1 (-/-)) mice.

State-dependent corrective reactions for backward balance losses during human walking.

We investigated corrective reactions for backward balance losses during walking. Several biomechanical studies have suggested that backward falling can be predicted from the horizontal position and velocity of the body center of mass (COM) related to the stance foot. Our hypothesis was that corrective reactions for backward balance losses depend on whether the body moves forward or backward after a perturbation. Using a split-belt treadmill, backward balance losses during walking were induced by rapid decreases of belt speed from 3.5 km/h to 2.5, 2.0, 1.5 and 1.0 km/h. We measured kinematic data and surface electromyography (EMG) during corrective reactions while walking on the treadmill. Phase portrait analysis of COM trajectories revealed that backward balance stability was decreased by the perturbations. When the perturbed belt speed was 1.0 km/h, the COM states at toe-off were significantly lower than the stability limit; a rapid touch-down of the swing foot posterior to the stance foot then occurred, and the gait rhythm was modulated so that the phase advanced. EMG recordings during perturbed steps revealed a bilateral response, including modulation of the swing leg during the recovery. For weaker perturbations, the swing foot placements were anterior to the stance foot and there was a phase delay. In contrast to the bilateral responses for stronger perturbations, unilateral EMG responses were observed for weaker perturbations. The differences in joint kinematics and EMG patterns in the unperturbed swing leg depended on the COM states at toe-off, suggesting the existence of different responses consisting of ongoing swing movements and rapid touch-down. Thus, we conclude that corrective reactions for backward balance losses are not only phase-dependent but also state-dependent. In addition, the control system for backward balance losses predicts the feasibility of forward progression and modulates swing movement and walking rhythm according to backward balance stability.

Necessary condition for forward progression in ballistic walking.

Ballistic walking requires an appropriate configuration of posture and velocity at toe-off to avoid backward falling. In this study, we investigated a determinant of the state of the body center of mass (COM) at the toe-off with regard to ballistic walking. We used an inverted pendulum model to represent ballistic trajectories and the necessary condition for forward progression by a simple relationship between the COM states (position and velocity) at toe-off. This condition was validated through a computer simulation of a 7-link musculoskeletal model and measurement experiments of human movements involving stepping and walking. The results of the model simulation were in good agreement with some of the results predicted by the inverted pendulum model. The measurement experiments of walking and stepping movements showed that most COM states at toe-off satisfied the condition for forward progression and the measured trajectories during single support phase were similar to the ballistic trajectories although humans are capable of walking in non-ballistic ways. These results suggested that the necessary condition for forward progression can predict the COM states at toe-off for efficient movement and for avoiding backward falling during single support phase.

Analysis of trunk movement in orthotic gait of paraplegics.

In an orthotic gait of paraplegics, a leg restriction and motor paralysis result in a significant loading. In this study, we address to quantify a relationship between the loading, leg restriction and motor paralysis, and analyze lumbar joint trajectories in the orthotic gait of paraplegic subjects and the ordinary and orthotic gaits of a normal subject using an inverted pendulum model. For the leg restriction, the trajectories are located anterior to an equilibrium point of the inverted pendulum, and the loading is higher due to the influence of gravity moment. Comparing the orthotic gait kinematics of paraplegic with that of normal in the horizontal plane, the lumbar joint trajectory in the paraplegic subjects was rectilinear shape, while that in the normal subject was curved in the direction to the equilibrium point. The loading is lower in the curved trajectory than in the straight trajectory because of the trade-off between gravity and inertia. These results suggest that the leg restriction and motor paralysis lead to the increase of the distance between the trunk movement and the equilibrium point of an inverted pendulum, which causes significant loading in the orthotic gate of paraplegics.