Club-mosses (Diphasiastrum, Lycopodiaceae) from the Far East - Introgression and possible cryptic speciation.

Hybridization occurs often in the genus Diphasiastrum (Lycopodiaceae), which corroborates reports for the two other recognized lycophyte families, Isoëtaceae and Selaginellaceae. Here we investigate the case of D. alpinum and D. sitchense from the Russian Far East (Kamchatka). Their hybrid, D. × takedae, was morphologically recognizable in 16 out of 22 accessions showing molecular signatures of hybridization; the remaining accessions displayed the morphology of either D. alpinum (3) or D. sitchense (3). We sequenced markers for chloroplast microsatellites (cp, 175 accessions from Kamchatka) and for the two nuclear markers RPB and LFY (175 and 152 accessions). A selection of 42 accessions, including all hybrid accessions, was analysed via genotyping by sequencing (GBS). We found multiple, but apparently uniparental hybridization, clearly characterized by a deviating group of haplotypes for D. sitchense and all hybrids. All accessions showing molecular signatures of hybridization in nuclear markers revealed the parental haplotype of D. sitchense, however only the LFY marker differentiated between the parent species. GBS, including 69,819 quality-filtered single nucleotid polymorphisms, unambiguously identified the hybrids and revealed introgression to occur. Most of the hybrids were F1, but three turned out to be backcrosses with D. alpinum (one) and with D. sitchense (two). These observations are in contrast to prior findings on three European species and their intermediates where all three hybrids turned out to be independent F1 crosses without evidence of recent backcrossing. In this study, backcrossing was detected, which indicates a limited fertility of the hybrid taxon D. × takedae. A comparison of accessions of Kamchatkian D. alpinum with plants from Europe indicated possible cryptic speciation. Accessions from the Far East had (i) a lower DNA content (7.0 vs. 7.5 pg/2C), (ii) different prevailing cp haplotypes, and (iii) RPB genotypes, and (iv) a clearly different SNP pattern in GBS. Diphasiastrum sitchense and the similar D. nikoënse, for the latter additional accessions from Japan were investigated, appeared as forms of one diverse species, sharing genotypes in both nuclear markers, although chloroplast haplotypes and DNA content show slight variations.

Left-Right Locomotor Coordination in Human Neonates.

Terrestrial locomotion requires coordinated bilateral activation of limb muscles, with left-right alternation in walking or running, and synchronous activation in hopping or skipping. The neural mechanisms involved in interlimb coordination at birth are well known in different mammalian species, but less so in humans. Here, 46 neonates (of either sex) performed bilateral and unilateral stepping with one leg blocked in different positions. By recording EMG activities of lower-limb muscles, we observed episodes of left-right alternating or synchronous coordination. In most cases, the frequency of EMG oscillations during sequences of consecutive steps was approximately similar between the two sides, but in some cases it was considerably different, with episodes of 2:1 interlimb coordination and episodes of activity deletions on the blocked side. Hip position of the blocked limb significantly affected ipsilateral, but not contralateral, muscle activities. Thus, hip extension backward engaged hip flexor muscle, and hip flexion engaged hip extensors. Moreover, the sudden release of the blocked limb in the posterior position elicited the immediate initiation of the swing phase of the limb, with hip flexion and a burst of an ankle flexor muscle. Extensor muscles showed load responses at midstance. The variable interlimb coordination and its incomplete sensory modulation suggest that the neonatal locomotor networks do not operate in the same manner as in mature locomotion, also because of the limited cortical control at birth. These neonatal mechanisms share many properties with spinal mammalian preparations (i.e., independent pattern generators for each limb, and for flexor and extensor muscles, load, and hip position feedback).SIGNIFICANCE STATEMENT Bilateral coupling and reciprocal activation of flexor and extensor burst generators represent the fundamental mechanisms used by mammalian limbed locomotion. Considerable progress has been made in deciphering the early development of the spinal networks and left-right coordination in different mammals, but less is known about human newborns. We compared bilateral and unilateral stepping in human neonates, where cortical control is still underdeveloped. We found neonatal mechanisms that share many properties with spinal mammalian preparations (i.e., independent pattern generators for each limb, the independent generators for flexor and extensor muscles, load, and hip-position feedback. The variable interlimb coordination and its incomplete sensory modulation suggest that the human neonatal locomotor networks do not operate in the same manner as in mature locomotion.

Influence of pregnancy related anthropometric changes on plantar pressure distribution during gait-A follow-up study.

BACKGROUND: As foot constitutes the base of support for the whole body, the pregnancy-related anthropometric changes can result in adaptive plantar pressure alterations. The present study aimed to investigate how pregnancy affects foot loading pattern in gait, and if it is related to body adjustments to growing foetus that occur in the course of pregnancy. METHODS: A prospective longitudinal study included 30 women. Three experimental sessions in accordance with the same procedure were carried out in the first, second and third trimesters of pregnancy. First, the anthropometric measures of the body mass and waist circumference were taken. Then walking trials at a self-selected speed along a ~6-m walkway were registered with the FreeMED force platform (Sensor Medica, Italy). Vertical foot pressure was recorded by the force plate located in the middle of the walkway. FINDINGS: The correlation of individual foot loading parameters across different trimesters was relatively high. Nevertheless, our results revealed a longitudinal foot arch flattening with the strongest effect in late pregnancy (P = 0.01). The anthropometric characteristics also influenced the foot loading pattern depending on the phase of pregnancy. In particular, arch flattening correlated with the body mass in all trimesters (r≥0.44, P≤0.006) while the medial-lateral loading index correlated only in the first (r = 0.45, P = 0.005) and second (r = 0.36, P = 0.03) trimesters. Waist circumference changes significantly influenced dynamic arch flattening but only in the late pregnancy (r≥0.46, P≤0.004). In the third trimester, a small though significant increase in the right foot angle was observed (P = 0.01). INTERPRETATION: The findings provided the characteristics of the relative foot areas loading throughout pregnancy. Growing abdominal size increases the risk of medial arch flattening, which can result in less stable gait. The observed increase in foot angle in late pregnancy may constitute a strategy to enhance gait stability.

Pendular energy transduction within the step during human walking on slopes at different speeds.

When ascending (descending) a slope, positive (negative) work must be performed to overcome changes in gravitational potential energy at the center of body mass (COM). This modifies the pendulum-like behavior of walking. The aim of this study is to analyze how energy exchange and mechanical work done vary within a step across slopes and speeds. Ten subjects walked on an instrumented treadmill at different slopes (from -9° to 9°), and speeds (between 0.56 and 2.22 m s-1). From the ground reaction forces, we evaluated energy of the COM, recovery (i.e. the potential-kinetic energy transduction) and pendular energy savings (i.e. the theoretical reduction in work due to this recovered energy) throughout the step. When walking uphill as compared to level, pendular energy savings increase during the first part of stance (when the COM is lifted) and decreases during the second part. Conversely in downhill walking, pendular energy savings decrease during the first part of stance and increase during the second part (when the COM is lowered). In uphill and downhill walking, the main phase of external work occurs around double support. Uphill, the positive work phase is extended during the beginning of single support to raise the body. Downhill, the negative work phase starts before double support, slowing the downward velocity of the body. Changes of the pendulum-like behavior as a function of slope can be illustrated by tilting the 'classical compass model' backwards (uphill) or forwards (downhill).

Drawing ellipses in water: evidence for dynamic constraints in the relation between velocity and path curvature.

Several types of continuous human movements comply with the so-called Two-Thirds Power Law (2/3-PL) stating that velocity (V) is a power function of the radius of curvature (R) of the endpoint trajectory. The origin of the 2/3-PL has been the object of much debate. An experiment investigated further this issue by comparing two-dimensional drawing movements performed in air and water. In both conditions, participants traced continuously quasi-elliptic trajectories (period T = 1.5 s). Other experimental factors were the movement plane (horizontal/vertical), and whether the movement was performed free-hand, or by following the edge of a template. In all cases a power function provided a good approximation to the V-R relation. The main result was that the exponent of the power function in water was significantly smaller than in air. This appears incompatible with the idea that the power relationship depends only on kinematic constraints and suggests a significant contribution of dynamic factors. We argue that a satisfactory explanation of the observed behavior must take into account the interplay between the properties of the central motor commands and the visco-elastic nature of the mechanical plant that implements the commands.

Planar Covariation of Hindlimb and Forelimb Elevation Angles during Terrestrial and Aquatic Locomotion of Dogs.

The rich repertoire of locomotor behaviors in quadrupedal animals requires flexible inter-limb and inter-segmental coordination. Here we studied the kinematic coordination of different gaits (walk, trot, gallop, and swim) of six dogs (Canis lupus familiaris) and, in particular, the planar covariation of limb segment elevation angles. The results showed significant variations in the relative duration of rearward limb movement, amplitude of angular motion, and inter-limb coordination, with gait patterns ranging from a lateral sequence of footfalls during walking to a diagonal sequence in swimming. Despite these differences, the planar law of inter-segmental coordination was maintained across different gaits in both forelimbs and hindlimbs. Notably, phase relationships and orientation of the covariation plane were highly limb specific, consistent with the functional differences in their neural control. Factor analysis of published muscle activity data also demonstrated differences in the characteristic timing of basic activation patterns of the forelimbs and hindlimbs. Overall, the results demonstrate that the planar covariation of inter-segmental coordination has emerged for both fore- and hindlimbs and all gaits, although in a limb-specific manner.

Tapping into rhythm generation circuitry in humans during simulated weightlessness conditions.

An ability to produce rhythmic activity is ubiquitous for locomotor pattern generation and modulation. The role that the rhythmogenesis capacity of the spinal cord plays in injured populations has become an area of interest and systematic investigation among researchers in recent years, despite its importance being long recognized by neurophysiologists and clinicians. Given that each individual interneuron, as a rule, receives a broad convergence of various supraspinal and sensory inputs and may contribute to a vast repertoire of motor actions, the importance of assessing the functional state of the spinal locomotor circuits becomes increasingly evident. Air-stepping can be used as a unique and important model for investigating human rhythmogenesis since its manifestation is largely facilitated by a reduction of external resistance. This article aims to provide a review on current issues related to the "locomotor" state and interactions between spinal and supraspinal influences on the central pattern generator (CPG) circuitry in humans, which may be important for developing gait rehabilitation strategies in individuals with spinal cord and brain injuries.

Visual gravity cues in the interpretation of biological movements: neural correlates in humans.

Our visual system takes into account the effects of Earth gravity to interpret biological motion (BM), but the neural substrates of this process remain unclear. Here we measured functional magnetic resonance (fMRI) signals while participants viewed intact or scrambled stick-figure animations of walking, running, hopping, and skipping recorded at normal or reduced gravity. We found that regions sensitive to BM configuration in the occipito-temporal cortex (OTC) were more active for reduced than normal gravity but with intact stimuli only. Effective connectivity analysis suggests that predictive coding of gravity effects underlies BM interpretation. This process might be implemented by a family of snapshot neurons involved in action monitoring.

Design and control of the MINDWALKER exoskeleton.

Powered exoskeletons can empower paraplegics to stand and walk. Actively controlled hip ab/adduction (HAA) is needed for weight shift and for lateral foot placement to support dynamic balance control and to counteract disturbances in the frontal plane. Here, we describe the design, control, and preliminary evaluation of a novel exoskeleton, MINDWALKER. Besides powered hip flexion/extension and knee flexion/extension, it also has powered HAA. Each of the powered joints has a series elastic actuator, which can deliver 100 Nm torque and 1 kW power. A finite-state machine based controller provides gait assistance in both the sagittal and frontal planes. State transitions, such as stepping, can be triggered by the displacement of the Center of Mass (CoM). A novel step-width adaptation algorithm was proposed to stabilize lateral balance. We tested this exoskeleton on both healthy subjects and paraplegics. Experimental results showed that all users could successfully trigger steps by CoM displacement. The step-width adaptation algorithm could actively counteract disturbances, such as pushes. With the current implementations, stable walking without crutches has been achieved for healthy subjects but not yet for SCI paraplegics. More research and development is needed to improve the gait stability.

Coordination of intrinsic and extrinsic foot muscles during walking.

PURPOSE: The human foot undergoes complex deformations during walking due to passive tissues and active muscles. However, based on prior recordings it is unclear if muscles that contribute to flexion/extension of the metatarsophalangeal (MTP) joints are activated synchronously to modulate joint impedance, or sequentially to perform distinct biomechanical functions. We investigated the coordination of MTP flexors and extensors with respect to each other, and to other ankle-foot muscles. METHODS: We analyzed surface electromyographic (EMG) recordings of intrinsic and extrinsic foot muscles for healthy individuals during level treadmill walking, and also during sideways and tiptoe gaits. We computed stride-averaged EMG envelopes and used the timing of peak muscle activity to assess synchronous vs. sequential coordination. RESULTS: We found that peak MTP flexor activity occurred significantly before peak MTP extensor activity during walking (P < 0.001). The period around stance-to-swing transition could be roughly characterized by sequential peak muscle activity from the ankle plantarflexors, MTP flexors, MTP extensors, and then ankle dorsiflexors. We found that foot muscles that activated synchronously during forward walking tended to dissociate during other locomotor tasks. For instance, extensor hallucis brevis and extensor digitorum brevis muscle activation peaks decoupled during sideways gait. CONCLUSIONS: The sequential peak activity of MTP flexors followed by MTP extensors suggests that their biomechanical contributions may be largely separable from each other and from other extrinsic foot muscles during walking. Meanwhile, the task-specific coordination of the foot muscles during other modes of locomotion indicates a high-level of specificity in their function and control.

Control of Leg Movements Driven by EMG Activity of Shoulder Muscles.

During human walking, there exists a functional neural coupling between arms and legs, and between cervical and lumbosacral pattern generators. Here, we present a novel approach for associating the electromyographic (EMG) activity from upper limb muscles with leg kinematics. Our methodology takes advantage of the high involvement of shoulder muscles in most locomotor-related movements and of the natural co-ordination between arms and legs. Nine healthy subjects were asked to walk at different constant and variable speeds (3-5 km/h), while EMG activity of shoulder (deltoid) muscles and the kinematics of walking were recorded. To ensure a high level of EMG activity in deltoid, the subjects performed slightly larger arm swinging than they usually do. The temporal structure of the burst-like EMG activity was used to predict the spatiotemporal kinematic pattern of the forthcoming step. A comparison of actual and predicted stride leg kinematics showed a high degree of correspondence (r > 0.9). This algorithm has been also implemented in pilot experiments for controlling avatar walking in a virtual reality setup and an exoskeleton during over-ground stepping. The proposed approach may have important implications for the design of human-machine interfaces and neuroprosthetic technologies such as those of assistive lower limb exoskeletons.

Human locomotion under reduced gravity conditions: biomechanical and neurophysiological considerations.

Reduced gravity offers unique opportunities to study motor behavior. This paper aims at providing a review on current issues of the known tools and techniques used for hypogravity simulation and their effects on human locomotion. Walking and running rely on the limb oscillatory mechanics, and one way to change its dynamic properties is to modify the level of gravity. Gravity has a strong effect on the optimal rate of limb oscillations, optimal walking speed, and muscle activity patterns, and gait transitions occur smoothly and at slower speeds at lower gravity levels. Altered center of mass movements and interplay between stance and swing leg dynamics may challenge new forms of locomotion in a heterogravity environment. Furthermore, observations in the lack of gravity effects help to reveal the intrinsic properties of locomotor pattern generators and make evident facilitation of nonvoluntary limb stepping. In view of that, space neurosciences research has participated in the development of new technologies that can be used as an effective tool for gait rehabilitation.

EMG patterns during assisted walking in the exoskeleton.

Neuroprosthetic technology and robotic exoskeletons are being developed to facilitate stepping, reduce muscle efforts, and promote motor recovery. Nevertheless, the guidance forces of an exoskeleton may influence the sensory inputs, sensorimotor interactions and resulting muscle activity patterns during stepping. The aim of this study was to report the muscle activation patterns in a sample of intact and injured subjects while walking with a robotic exoskeleton and, in particular, to quantify the level of muscle activity during assisted gait. We recorded electromyographic (EMG) activity of different leg and arm muscles during overground walking in an exoskeleton in six healthy individuals and four spinal cord injury (SCI) participants. In SCI patients, EMG activity of the upper limb muscles was augmented while activation of leg muscles was typically small. Contrary to our expectations, however, in neurologically intact subjects, EMG activity of leg muscles was similar or even larger during exoskeleton-assisted walking compared to normal overground walking. In addition, significant variations in the EMG waveforms were found across different walking conditions. The most variable pattern was observed in the hamstring muscles. Overall, the results are consistent with a non-linear reorganization of the locomotor output when using the robotic stepping devices. The findings may contribute to our understanding of human-machine interactions and adaptation of locomotor activity patterns.

Spinal motor outputs during step-to-step transitions of diverse human gaits.

Aspects of human motor control can be inferred from the coordination of muscles during movement. For instance, by combining multimuscle electromyographic (EMG) recordings with human neuroanatomy, it is possible to estimate alpha-motoneuron (MN) pool activations along the spinal cord. It has previously been shown that the spinal motor output fluctuates with the body's center-of-mass motion, with bursts of activity around foot-strike and foot lift-off during walking. However, it is not known whether these MN bursts are generalizable to other ambulation tasks, nor is it clear if the spatial locus of the activity (along the rostrocaudal axis of the spinal cord) is fixed or variable. Here we sought to address these questions by investigating the spatiotemporal characteristics of the spinal motor output during various tasks: walking forward, backward, tiptoe and uphill. We reconstructed spinal maps from 26 leg muscle EMGs, including some intrinsic foot muscles. We discovered that the various walking tasks shared qualitative similarities in their temporal spinal activation profiles, exhibiting peaks around foot-strike and foot-lift. However, we also observed differences in the segmental level and intensity of spinal activations, particularly following foot-strike. For example, forward level-ground walking exhibited a mean motor output roughly 2 times lower than the other gaits. Finally, we found that the reconstruction of the spinal motor output from multimuscle EMG recordings was relatively insensitive to the subset of muscles analyzed. In summary, our results suggested temporal similarities, but spatial differences in the segmental spinal motor outputs during the step-to-step transitions of disparate walking behaviors.

Function dictates the phase dependence of vision during human locomotion.

In human and animal locomotion, sensory input is thought to be processed in a phase-dependent manner. Here we use full-field transient visual scene motion toward or away from subjects walking on a treadmill. Perturbations were presented at three phases of walking to test 1) whether phase dependence is observed for visual input and 2) whether the nature of phase dependence differs across body segments. Results demonstrated that trunk responses to approaching perturbations were only weakly phase dependent and instead depended primarily on the delay from the perturbation. Recording of kinematic and muscle responses from both right and left lower limb allowed the analysis of six distinct phases of perturbation effects. In contrast to the trunk, leg responses were strongly phase dependent. Leg responses during the same gait cycle as the perturbation exhibited gating, occurring only when perturbations were applied in midstance. In contrast, during the postperturbation gait cycle, leg responses occurred at similar response phases of the gait cycle over a range of perturbation phases. These distinct responses reflect modulation of trunk orientation for upright equilibrium and modulation of leg segments for both hazard accommodation/avoidance and positional maintenance on the treadmill. Overall, these results support the idea that the phase dependence of responses to visual scene motion is determined by different functional tasks during walking.

Locomotor-like leg movements evoked by rhythmic arm movements in humans.

Motion of the upper limbs is often coupled to that of the lower limbs in human bipedal locomotion. It is unclear, however, whether the functional coupling between upper and lower limbs is bi-directional, i.e. whether arm movements can affect the lumbosacral locomotor circuitry. Here we tested the effects of voluntary rhythmic arm movements on the lower limbs. Participants lay horizontally on their side with each leg suspended in an unloading exoskeleton. They moved their arms on an overhead treadmill as if they walked on their hands. Hand-walking in the antero-posterior direction resulted in significant locomotor-like movements of the legs in 58% of the participants. We further investigated quantitatively the responses in a subset of the responsive subjects. We found that the electromyographic (EMG) activity of proximal leg muscles was modulated over each cycle with a timing similar to that of normal locomotion. The frequency of kinematic and EMG oscillations in the legs typically differed from that of arm oscillations. The effect of hand-walking was direction specific since medio-lateral arm movements did not evoke appreciably leg air-stepping. Using externally imposed trunk movements and biomechanical modelling, we ruled out that the leg movements associated with hand-walking were mainly due to the mechanical transmission of trunk oscillations. EMG activity in hamstring muscles associated with hand-walking often continued when the leg movements were transiently blocked by the experimenter or following the termination of arm movements. The present results reinforce the idea that there exists a functional neural coupling between arm and legs.

Can modular strategies simplify neural control of multidirectional human locomotion?

Each human lower limb contains over 50 muscles that are coordinated during locomotion. It has been hypothesized that the nervous system simplifies muscle control through modularity, using neural patterns to activate muscles in groups called synergies. Here we investigate how simple modular controllers based on invariant neural primitives (synergies or patterns) might generate muscle activity observed during multidirectional locomotion. We extracted neural primitives from unilateral electromyographic recordings of 25 lower limb muscles during five locomotor tasks: walking forward, backward, leftward and rightward, and stepping in place. A subset of subjects also performed five variations of forward (unidirectional) walking: self-selected cadence, fast cadence, slow cadence, tiptoe, and uphill (20% incline). We assessed the results in the context of dimensionality reduction, defined here as the number of neural signals needing to be controlled. For an individual task, we found that modular architectures could theoretically reduce dimensionality compared with independent muscle control, but we also found that modular strategies relying on neural primitives shared across different tasks were limited in their ability to account for muscle activations during multi- and unidirectional locomotion. The utility of shared primitives may thus depend on whether they can be adapted for specific task demands, for instance, by means of sensory feedback or by being embedded within a more complex sensorimotor controller. Our findings indicate the need for more sophisticated formulations of modular control or alternative motor control hypotheses in order to understand muscle coordination during locomotion.

Changes of gait kinematics in different simulators of reduced gravity.

Gravity reduction affects the energetics and natural speed of walking and running. But, it is less clear how segmental coordination is altered. Various devices have been developed in the past to study locomotion in simulated reduced gravity. However, most of these devices unload only the body center of mass. The authors reduced the effective gravity acting on the stance or swing leg to 0.16g using different simulators. Locomotion under these conditions was associated with a reduction in the foot velocity and significant changes in angular motion. Moreover, when simulated reduced gravity directly affected the swing limb, it resulted in significantly slower swing and longer foot excursions, suggesting an important role of the swing phase dynamics in shaping locomotor patterns.