M Bassolino

M Jacono

M Bove

L Fadiga

Thierry Pozzo

Sensorimotor

SENSORIMOTOR DEPRIVATION INDUCED BY IMMOBILIZATION MODIFIES THE KINEMATIC OF A REACHING-TO-GRASP

published or not. The documents may come

"SENSORIMOTOR DEPRIVATION INDUCED BY IMMOBILIZATION MODIFIES THE KINEMATIC OF A REACHING-TO-GRASP"

Michela Bassolino 1,2 , Marco Jacono 1 , Marco Bove 3 , Luciano Fadiga 1,4 , Thierry Pozzo 1,5 1.IIT, Istituto Italiano di Tecnologia, Genoa, Italy; 2. DIST, University of Genoa, Italy; 3. Departement of Experimental Medicine, Section of Human Physiology, University of Genoa, Italy; 4. DSBTA, Section of Human Physiology, University of Ferrara, Italy; 5. INSERM, U887, Motricité-plasticité, Dijon, France.

ACKNOWLEDGEMENTS INTRODUCTION

Reaching-to-grasp is a daily life common motor task, fundamental for interaction with the surrounding environment. It plays a central role in the early development of perception and action (see [START_REF] Piaget | The construction of reality in the child[END_REF][START_REF] Piaget | The construction of reality in the child[END_REF], probably because the hand provides an efficient tool to continuously verify motor and sensory predictions. Two main components contribute to organize these sensorimotor activities. A first progressive opening of the grip, with straightening of the fingers is followed by a gradual closure until it matches the object's dimension and orientation (the grasp component, [START_REF] Arbib | Perceptual structures and distributed motor control[END_REF][START_REF] Jeannerod | The timing of natural prehension movements[END_REF], for a review [START_REF] Castiello | The neuroscience of grasping[END_REF]. Contemporaneously, the hand, moving towards the object, follows a characteristic path and timing (the transport component, Jeannerod 1999.) A successful performance requires a sophisticated coordination between the two components [START_REF] Wing | Grasp size and accuracy of approach in reaching[END_REF]; [START_REF] Wallace | Temporal constraints in the control of prehensile movement[END_REF][START_REF] Jakobson | Factors affecting higher-order movement planning: a kinematic analysis of human prehension[END_REF][START_REF] Gentilucci | Temporal coupling between transport and grasp components during prehension movements: effects of visual perturbation[END_REF][START_REF] Paulignan | Selective perturbation of visual input during prehension movements. 2. The effects of changing object size[END_REF]. At last, a synergistic control of each finger's position is essential to correctly adapt the hand shape to the target object [START_REF] Santello | Postural hand synergies for tool use[END_REF]. All these phases involve a continuum of feedforward and feedback processes (Seidler et al., 2004). Predictions about the state of the motor system and environment in addition to sensory feedback can be used to update parameters of the next motion (e.g. [START_REF] Gentilucci | Tactile input of the hand and the control of reaching to grasp movements[END_REF]). Thus, a constant stream of sensorimotor information seems to be essential for action execution. The presence of theses mechanisms raises a fundamental question: what is the effect of a transient lack of sensory information and motor commands in a reaching-to-grasp? Differently stated, what is the behavioral consequence of a short period of immobilization that induces a temporary reduction in sensory and motor information?

Previous investigation showed that upper limb immobilization for 12 hours induced significant changes in somatosensory-evoked potentials (SEPs), motor-evoked potentials (MEPs) and slow wave activity during sleep over the sensorimotor areas corresponding to the immobilized limb [START_REF] Huber | Arm immobilization causes cortical plastic changes and locally decreases sleep slow wave activity[END_REF]. However, little is known on the functional effect of hand and arm immobilization [START_REF] Moisello | Short-term limb immobilization affects motor performance[END_REF]. The underlying assumption is that if action execution generates crucial input to the brain to update the next motor command, reaching-to-grasp movement would be significantly affected after upper limb immobilization. To this aim, the right hand and wrist joint of 7 healthy subjects was immobilized with a removable bandage for 10 hours. Two different effects of the short term immobilization on the kinematic of a reaching-to-grasp could occur: first, a short period of immobilization would not produce any behavioral effect, as it is sometimes the case during daily life situations in which the arm motion is limited, such as after a long travel or sleeping. Alternatively, immobilization may modify the kinematic of a task strongly dependent of sensorimotor information such as reaching-to-grasp. In this context, an adaptation process could be also expected after the immobilization period. If so, a detailed analysis of reaching and grasping, separately, might also reveal different recovery time courses in the two components.

METHODS

Participants.

Thirteen healthy volunteers (7 female, 6 male) participated in the study. All participants were right handed, as determined by the Edinburgh Handedness Inventory [START_REF] Oldfield | The assessment and analysis of handedness: the Edinburgh inventory[END_REF]. They reported normal or corrected-to-normal vision, no previous history of neurological disorders or recent orthopedic problems for the right upper limb. All subjects gave their informed consent to participate in the study, which was performed with approval of the local ethics committee and in accordance with the Declaration of Helsinki. All participants were naive to the purpose of the study and received an attendance fee at the end of experiment.

Experimental protocol.

If present, the effect of immobilization could be also dependent on the degree of precision required in the task. To this aim, subjects were instructed to grasp, lift and back down a little yellow wooden pencil (7 cm in length, 1 cm in diameter) with a precision grip and a yellow tennis ball (6 cm in diameter) with a power grip. A power grip is achieved by moving the whole hand, while a precision grip involves grasping an object between the tips of the thumb and index finger (see for example, [START_REF] Napier | The prehensile movements of the human hand[END_REF]) in such a way that there is precise control of the position of the object and the grasping forces (Jones &Lederman, 2006). Movement were shown by the experimenter at the beginning of each experimental section. Participants sat in front of a table where on each trial, the experimenter placed the objects to grasp on a marked point. They started and finished each trial in the same fixed position (called "pinch"): they kept the index in contact with the thumb, holding the two fingers on a marked point. This point was drawn on the table 30 centimeters far from the objects. After a brief familiarization phase, they repeated the movement 15 times for each object in a self-paced mode. Subjects were instructed to start the movement when they preferred, after a verbal go-signal given by the experimenter. When the subjects ended the movement, the object was removed and the next one was set. The order of objects presentation was randomized. We compared the kinematic of the reaching-to-grasp in a group of 6 subjects (Control Group, CG) who repeated the task in two following days (first day=Pre, second day=Post), with another group of 7 subjects (Immobilization Group, IG ) who did the same task after 10 hours of immobilization (Post) and one day before (Pre). All participants did the task at about 6.30 p.m. The two groups were matched for age and gender (IG: mean age 27.00 ± 1.07 years, range 25-29, 4 female; CG: mean age 27.17 ± 1.33 years, range 25-29, 3 female).

Immobilization procedure.

In the IG, subjects were instructed to not move the right hand and the wrist joint for 10 hours from the morning (at about 8.30 a.m.) to the evening (at about 6.30 p.m.). We wrapped subjects' hand and wrist with a soft painless bandage used in everyday clinical practice by physiotherapists.

Participants wore also a cotton support that limited the arm movement and hold the elbow joint in a comfortable way at 90 degree flexion. As soon as the experimenter removed the bandage, subjects performed the grasping task.

Data acquisition.

The 3d kinematic data were recorded with an optical motion capture system (Vicon), at a frequency of 100 Hz. Retro-reflective markers (4 and 6 mm in diameters) were placed on the hand (19 markers), in particular on the metacarpal-phalangeal and the proximal interphalangeal joints, on the nails of each fingers, on the radial and the ulnar styloid process and on the lateral epicondyle of the humerus. At the beginning of each trial, before assuming the "pinch position" (see above), to be sure that all markers were visible by the recording system, the subjects were requested to keep the hand palm-down on the table with the thumb on a marked point. This recorded position was also used as a reference value of participants' hand size and in particular, of the thumb-index distance.

Software made with Matlab® (Mathworks, Natick, MA) was employed to filter (Butterworth filter, second order fc = 5Hz) and analyze the data. Both reaching and grasping components were evaluated. Moreover, we measured also 16 hand angles to describe the hand posture at the time of the contact with the object and their co-variation during the whole motion.

Reach component.

To determine the onset and the end of the reaching movement, we considered the velocity profile of the wrist, applying on it a threshold of the 5% of its maximum (see [START_REF] Santello | Patterns of hand motion during grasping and the influence of sensory guidance[END_REF]. Thus, the onset and the end of the reaching movement were respectively set at the first and at the last frame in which subjects moved with a wrist velocity equal to this threshold. We measure the following reach parameters: reach duration: the total duration of reaching phase (ms); time to peak velocity: the ratio between the time at which the maximum velocity occurred and the total reach duration (%).

Grasp component.

The distance between the two markers located on the nails of thumb and index fingers was calculated during the whole grasping task. The velocity profile of the displacement of this distance was then considered to determine the onset and the end of the duration of the grasping movement.

Starting with the hand in "pinch position", the grasp time course is typically constituted by a phase of finger opening till a maximum (maximal finger aperture) followed by a phase of finger closing on the object [START_REF] Jeannerod | The timing of natural prehension movements[END_REF]. Thus, the velocity profile of the variation of the index-thumb distance started from an initial value of zero, when the fingers were in pinch position, was then characterized by a positive peak corresponding to the phase of finger opening, reached the zero at the time of maximal finger aperture and showed a negative peak in the phase of finger closing. The beginning of the grasp was set at the first frame in which the two fingers started to open from the pinch position (i.e., when the velocity of the variation of index-thumb distance increased respect to the initial zero value). On the contrary, the end of the grasp corresponded to the first frame after finger closing in which the time course of the index-thumb distance reached the zero and remained stable at this value at least for 10 following samples ( i.e., during the contact with the object). To avoid any effect due to different sizes of participants' hands, for each subjects we normalized the index-thumb distance measured during the motion with respect to the corresponding distance recorded when the participant kept the hand still on the table at the beginning of the task ("hand size", see Data Acquisition). We measured the following grasp parameters: maximum grip aperture: the maximum value of the index-thumb distance (aperture / "hand size", %); time to maximum grip aperture: the ratio between the time at which the maximum grip aperture occurred and the total duration of grasping phase (%); final grip aperture: the index-thumb distance at the end of the grasp movement (aperture / "hand size", %).

Angular displacements.

To describe the hand shape during the grasp movement, we considered 16 angles. In particular, we measured the angular excursions at the metacarpal-phalangeal of the second, third, fourth and fifth finger ( respectively I_MCP, M_MCP, R_MCP, L_MCP) and proximal interphalangeal joints (PIP: I_PIP, M_PIP, R_PIP, L_PIP) of the same fingers. For the thumb, we considered the metacarpalphalangeal (T_MCP) and interphalangeal joints (T_IP). Therefore, we evaluated the abduction angles (ABD) of all digits. During the movement, hand rotated with respect to the 3d space of the data acquisition system. To avoid any difference in the measure of the angles due to this 3d rotation, first we introduced a local Cartesian coordinate system X-Y-Z (F1) attached on the palm of hand defined by the plane passing through the index and little metacarpal-phalangeal joints and the radial styloid, as previously described by [START_REF] Carpinella | Experimental protocol for the kinematic analysis of the hand: definition and repeatability[END_REF] (see Fig. 1). The ABD angle of each finger was defined as the angle between the Y-axis and the projection on the XY plane of the MCP joint of each finger. Second, concerning the measure of MCP and PIP joint angles, to keep away from any difference due to the abduction, the angular excursions were evaluated in a new frame of reference (F2). This was defined by a new plane obtained by the rotation of F1 with respect to the X axis and attached on each finger at the MCP and PIP joints respectively. MCP and PIP angles were calculated according to Denavit-Hartenberg convention [START_REF] Denavit | A kinematic notation for lower-pair mechanisms based on matrices[END_REF]. The angular excursion at the PIP joints were evaluated as the angles between the markers placed on nails, proximal interphalangeal and metacarpal-phalangeal joints. Concerning the thumb, the metacarpal-phalangeal (T_MCP) and the interphalangeal joints (T_IP) angles were measured as the others fingers. Moreover, we also evaluated the thumb rotation (T_ROT) and abduction angle (T_ABD), respectively as elevation and azimuth angle (polar coordinates) of the metacarpalphalangeal joint referring to trapezio-metacarpal joint. Angles at the end of grasping movement were calculated to describe the posture of the hand at the time of the contact with the object.

INSERT FIGURE 1 AROUND HERE

Time series of finger angular displacement were further analyzed by means of Principal components analysis (PCA) to evaluate a potential effect of immobilization on fingers' coordination.

Principal component analysis.

PCA was applied to the angular displacements of all 16 angles in the Post condition both in CG and IG. All time series were time-normalized to 100 points by using Matlab. Each sample of time was considered as a single observation lying an ambient vector space whose dimension was given by the number of time series included in the analysis. For instance, consider a simple input dataset composed of thirty-two columns (16 angular displacements recorded during each trial for the pencil and 16 angular displacements for the ball) and 100 rows (normalized time). PCA could be thought as a generalization of a correlation analysis in a high dimensional space. In this case, PCA extracted the commonality between the angular displacements, which was sometimes referred as "waveforms" because it especially focused on the shape of the angular time series. To this aim, each eigenvector was defined as a one-dimensional vector subspace (i.e., a certain direction in the thirthy-two dimensional vector space). These eigenvectors represented a well adapted basis of the thirthy-two dimensional vector space, characterizing the most important directions (in the sense of the variance account for: VAF), and, in this setting, principal component was simply the projection of the data onto a subspace spanned by a certain eigenvector. The ratio between the first eigenvalue and the sum of all eigenvalues could be viewed as an index of the whole-hand coordination (this value is commonly called the VAF by the PC1 and is referred to as PC1%). A PC1% value equal to 100% mean that the trajectory in the space of angles was a straight line (i.e., all angles were linearly correlated together). However, a low PC1% value indicated that only one principal component could not describe precisely the whole-hand movement. Therefore, we also reported the second PC whose VAF was denoted by PC2%. Here, instead of using independent PCAs (a PCA for each trial, each condition, and each participant), we used two PCAs, one for the IG and one for CG, taking together the two objects (pencil and ball), whose input dataset consisted of respectively 3360 or 2880 columns (16 angles for each objects, that is 32 in total, 15 trials, 7 participants for IG or 6 for the CG) and 100 rows (normalized time), considering only the Post condition. In this manner, the PCA automatically extracted the commonality between the shapes of the angular displacements (as in the study by [START_REF] Berret | Modular control of pointing beyond arm's length[END_REF]. Therefore, to statistically compare the VAF by PC1 and PC2, we also computed PCAs subjects by subjects.

Statistical analysis.

Reach and grasp components. To evaluate the effect of 10 hours of immobilization, we compared the kinematic of the grasping task in the CG and the IG. To this aim, we performed separate analysis of variance (ANOVAs) with repeated measures on each parameter calculated in every trial for all subjects with CONDITION ("Pre" and "Post") and OBJECTS ("Pencil" and "Ball") as within-subjects factors and with GROUP (Immobilization vs Control) as between-subjects factor.

While no variation in the motor performance of the CG was expected in the task recorded during the two days, in the IG we hypothesized some differences concerning the kinematic of reaching-tograsp after immobilization .

Angular displacements. To directly compare the effect of immobilization on angular displacements, we considered each angle subtracting the value obtained in the Pre from the corresponding measure recorded in the Post. In this way, we can evaluate if the angular excursion remained constant or not in the two experimental conditions. In particular we compared the CG and the IG performing separate ANOVAs on the normalized data with OBJECTS ("Pencil" and "Ball") as within-subjects factor and GROUP (Immobilization vs Control) as between-subjects factor. While no difference was expected in the CG for all the angles, in the IG a higher positive value at MCP or PIP joints indicated a wider flexion after immobilization. On the contrary, a negative value revealed a more extended posture of fingers. For the ABD angles of middle, ring and little fingers a higher positive difference could be obtained if the fingers were more adducted after immobilization. On the contrary, for the index and thumb the adduction was indicated by negative values, while positive values meant abduction. Moreover, a wider negative value of the T_ROT corresponded to an increased of the internal rotation of this finger with respect to the hand space.

Principal Component Analysis. To understand if the covariation between the angular excursions of the digits were affected by immobilization, we compared the VAF for each subject by means of two separate ANOVAs, one for PC1 and the other for PC2, with GROUP (Immobilization vs Control) as between-subjects factor. Moreover, we evaluated the correlation coefficients of the most common and the second angular waveform in the dataset, respectively described by PC1 and PC2, in IG and CG.

Recovery. Finally, we check if, after immobilization, there was a recovery trial after trial (i.e. 15 repetitions for each object). Thus, we performed linear regression analysis on reach duration and maximum grip aperture as a function of the number of trials, for each subject of IG and CG in the Post condition. Then, we statistically compared the slopes of the linear regression lines by means of 2 separate ANOVAs (one for reach duration and one for maximum grip aperture) with "OBJECT" ("Pencil" and "Ball") as within-subjects factor and "GROUP" (Immobilization vs Control) as between-subjects factor. Moreover, in the IG and CG, we evaluated the coefficient of correlation between the reach duration and the maximum grip aperture as a function of the number of trials. To this aim, we considered the mean value among all participants in the IG and in the CG, at each trial.

Significance threshold was set at p< 0.05. If ANOVAs showed significant interaction effects, we performed post hoc tests using the Newman-Keuls procedure to directly compare the experimental factors.

RESULTS

As soon as we removed the bandage (10 hours of immobilization), subjects were equally able to correctly grasp the objects as before immobilization and comparable to the CG. However, while in the CG no difference was shown comparing the Pre and Post conditions, in the IG a different kinematic for reach and grasp components, both in precision and power grip, was described after immobilization (Fig. 2 and3). Therefore, the hand posture at the end of grasping task changed in the two groups (Fig. 4). Nevertheless, the patterns of co-variation between the angular excursion of the digits remained similar (Fig. 5). Finally, after immobilization reach duration decreased trial after trial in contrast to maximum grip aperture that increased during the repetitions (Fig. 6).

Reach and grasp components.

For all reach and grasp parameters, excepted for the time to maximum grip aperture (see below), the critical interaction GROUP X CONDITION is statistically significant (p<0.05). In particular, Newman-Keuls post-hoc comparisons revealed that in the IG Pre and Post conditions were different, while this was not the case in the CG (p>0.05). The results obtained in conditions without immobilization, that is in Pre for IG and in the Pre and Post for CG, were never statistically different (in both comparisons p>0.05). For all parameters, the interaction GROUP X CONDITION X OBJECT was not statistically significant, underlining that the effect of immobilization was similar both in the precision and the power grip.

IG participants spent more time in the reaching in the Post condition (802 ms) with respect to the Pre (776 ms, critical interaction GROUP X CONDITION: F(1, 193)=6.98, p<0.01 ). Moreover, after immobilization the time to peak velocity occurred significantly earlier (40%) than in the Pre (42%, GROUP X CONDITION: F(1, 193) = 4.46, p< 0.05). Thus, immobilization affected both duration and timing of the reaching movement.

INSERT FIGURE 2 AROUND HERE

After immobilization participant opened their hand wider compared to Pre values, or CG. Indeed, post hoc comparisons revealed a significant increase of maximum grip aperture after immobilization (70% of "hand size") compared with the Pre (66%, p<0.001) in the IG (critical interaction GROUP X CONDITION (F(1,193) =4, p< 0.05). However, even if, the maximum grip aperture increased after immobilization, the time in which the peak aperture occurred (time to maximum grip aperture) was similar in both groups (GROUP X CONDITION: F(1, 193)=0.11, p>0.05). Moreover, a significant increase of final grip aperture in the Post condition in the IG indicated that after immobilization participants kept their hand more open also at the end of the grasp (Pre=40%, Post=44%, GROUP X CONDITION: F(1,193) = 9.2, p<0.01 ).

INSERT FIGURE 3 AROUND HERE

Angular displacements.

In order to verify if after immobilization the increase in the final grip aperture was associated to a different hand posture, we compared the value of the 16 angles of the fingers at the time to contact with the object. The hand posture changed before and after immobilization. Indeed, in the IG the hand posture was characterized by wider flexed MCP joints of the middle, ring, little and thumb fingers associated with a higher extension of PIP joints of all fingers. This effect was comparable for the two objects, excepted for L_PIP and T_IP (see Table 1). Therefore, the index and ring fingers were closer each other, more adducted. A similar effect was present also for the little finger only during precision grip. Finally, after immobilization also the thumb was nearer the palm, being more adducted (for the Ball) or internal rotated (for the Pencil). Statistical analysis and values are reported in Table1.

INSERT TABLE 1 AROUND HERE INSERT FIGURE 4 AROUND HERE

Principal Component Analysis . contrary, there was no correlation in CG between maximum grip aperture and the reach duration (r= 0.3, p< 0.05) (Figure 6, panel D). In other words, while reach duration recover nominal value trial after trail in the Post, maximum grip aperture followed an opposite mechanism.

INSERT FIGURE 6 AROUND HERE

DISCUSSION

Findings demonstrated that 10 hours of immobilization were sufficient to modify the kinematic of reaching and grasping, anyhow preserving the capability of participants to perform the task. Further, a different recovery was observed in the two components. As far as we know, this is the first demonstration of modifications in the kinematics of a reaching-to-grasp induced by a short term period of immobilization in healthy subjects.

General behavioural effects on reach and grasp

Firstly, concerning the reach, after immobilization both duration and timing were affected. In particular, reach duration increased and the time to peak velocity occurred earlier, indicating a longer deceleration phase. Additionally, the movement remained under feedforward control with a bell shape velocity profile with any online adjustment. Secondly, immobilization increased the maximum grip aperture (the peak of index-thumb distance) that, as for the transport component, was produced without online adjustments. How can we explain these modifications? The concept of internal models suggests that specialized neural networks would relate motor commands to sensory signals of body motion (forward models) and desired movements to appropriate motor commands (inverse model) [START_REF] Desmurget | Forward modeling allows feedback control for fast reaching movements[END_REF][START_REF] Wolpert | Multiple paired forward and inverse models for motor control[END_REF]. As such, sensory information from the moving limbs strongly contributes to regulate the motor command and to define hand shaping.

Thus, successful reaching-to-grasp movement requires continuous adaptation of fingers coordinative patterns in order to accurately grasp the object. Immobilization induced a reduction of dynamic sensory information (mainly from proprioception and vision), usually generated by arm motion (i.e. dynamic proprioception, [START_REF] Burke | The responses of human muscle spindle endings to vibration of non-contracting muscles[END_REF]. Evidences from animal models and human studies showed that this input is thought to have direct access to contralateral sensory and motor cortical areas [START_REF] Heath | Inputs from low threshold muscle and cutaneous afferents of hand and forearm to areas 3a and 3b of baboon's cerebral cortex[END_REF][START_REF] Hore | Responses of cortical neurons (areas 3a and 4) to ramp stretch of hindlimb muscles in the baboon[END_REF]. For instance Huber and collegues (2006), using EEG recording in healthy subjects after upper limb immobilization, described a significant reduction of the amplitude of the P45 component which classically represents the proprioceptive information processing within the sensorimotor areas [START_REF] Allison | The relationship between human long-latency somatosensory evoked potentials recorded from the cortical surface and from the scalp[END_REF]). However, during immobilization motor planning and consecutive forward (prediction of sensory consequence of movement) and inverse (motor command) models remained available, but no more updated because of the lack of motor output. This could lead to an inaccurate prediction of sensory consequences of motor command. In support to this, variability of imagined walking [START_REF] Courtine | Gait-dependent motor memory facilitation in covert movement execution[END_REF] or writing [START_REF] Papaxanthis | Does order and timing in performance of imagined and actual movements affect the motor imagery process? The duration of walking and writing task[END_REF] movements steadily increased as time elapsed between overt (i.e. real execution) and covert (i.e. imagined action) performances. This resulted from the absence of sensory feedback, which appears essential to accurately execute the tasks. Consequently, the present increase in the duration of transport, in the deceleration phase and in the amplitude of the grip aperture may represent an adaptative process in order to maintain a successful motion without online regulation. A wide opening of the hand might be employed to maximize the likelihood to successfully grasp the object [START_REF] Grosskopf | Grasping with the left and right hand: a kinematic study[END_REF]) and it could be the consequence of a motor strategy design to increase the tolerance for programming errors [START_REF] Jakobson | Factors affecting higher-order movement planning: a kinematic analysis of human prehension[END_REF].

According to this, a higher maximum finger opening was previously described in conditions which impede the accuracy of reaching [START_REF] Grosskopf | Grasping with the left and right hand: a kinematic study[END_REF], such as fast movements [START_REF] Wing | Grasp size and accuracy of approach in reaching[END_REF][START_REF] Wallace | Temporal constraints in the control of prehensile movement[END_REF], movements in the dark [START_REF] Jakobson | Factors affecting higher-order movement planning: a kinematic analysis of human prehension[END_REF]Churchilll et al. 2000;Schettino et al.2003) or anaesthesia [START_REF] Gentilucci | Tactile input of the hand and the control of reaching to grasp movements[END_REF]. Extending the duration of the motion (and in particular, here, the deceleration phase) may contribute to improve the action accuracy (Fitts,1954,Tanaka et al.2006) after sensorimotor deprivation induced by immobilization.

We can also suppose that because of the decrease of dynamic proprioceptive information during immobilization, visual inputs could be more reliable when participants start to move again, as soon as the bandage was removed. Since the visual loop is slower respect to the proprioceptive one [START_REF] Jeannerod | The neural and behavioural organization of goal-directed arm movements[END_REF], it is likely that the increased reach duration reflects an attempt to perform the movement using mostly visual inputs. Comparably, in deafferented patient [START_REF] Gentilucci | The role of proprioception in the control of prehension movements: a kinematic study in a peripherally deafferented patient and in normal subjects[END_REF] described an abnormal lengthening of movement (and in particular of deceleration phase), reported as a tentative to compensate for the proprioceptive deficit through vision.

Final hand posture

Inspection of fingers' position after immobilization revealed that participants adopted a different hand posture characterized by wider flexed MCP joints (except for the index) accompanied by a higher extension of PIP joints, with all fingers (little only for the ball) closer each other and more adducted. A greater distance between the tips of the index and thumb fingers observed at the end of the grasping motion (see above "final grip aperture") is congruent with this hand posture. This effect could result from the hand position imposed by the bandage used to constraint the fingers extended and close each other during immobilization. Such possibility cannot be ruled out and could have been tested by immobilizing subjects in another position, i.e. with fingers flexed.

However, we choose a comfortable position avoiding an over-stretching of agonist/antagonist muscles. Moreover, this explanation seems not compatible with the specific thumb position (more internally rotated for the pencil and more adducted for the ball) that significantly changed with respect to the immobilization posture. One possibility to explain these postural effects is that the thumb would compensate for the extension of the fingers. Indeed, by rotating or moving more internally the thumb, subjects could increase the opposition between the digits and the thumb and better stabilize the grip. This is in agreement with the idea that thumb will drive the adaptations in contrast to the others fingers [START_REF] Frak | Orientation of the opposition axis in mentally simulated grasping[END_REF][START_REF] Galea | Thumb invariance during prehension movement: effects of object orientation[END_REF][START_REF] Smeets | A new view on grasping[END_REF][START_REF] Wing | The contribution of the thumb to reaching movements[END_REF]. Biomechanically, there is a structural separation of the extrinsic muscles of the thumb from the multi-tendoned muscles acting on the other digits (Landsmeer JMF et al., 1986), permitting more selective, independent movement of the thumb (Schieber MH. Et al., 1991).

Fingers coordination

We found that only two principal components could account for a large proportion of the variance, showing that the motion of the hand during the movement is characterized by consistent, jointspecific co-variation in angular excursion. The presence of these co-variation patterns indicates that not all the finger joints were controlled independently, resulting in a reduction in the number of mechanical degrees of freedom [START_REF] Santello | Postural hand synergies for tool use[END_REF][START_REF] Santello | Patterns of hand motion during grasping and the influence of sensory guidance[END_REF]. Interestingly, despite kinematic changes and modifications in hand posture after non-use, finger coupling was not affected. Such robustness of fingers coordination could be due to the well-documented presence of biomechanical couplings among digits [START_REF] Lang | Human finger independence: limitations due to passive mechanical coupling versus active neuromuscular control[END_REF]Schieber 2004, Schieber 1991). Zatiorsky et al. (2000), for instance, described the tendency of fingers to generate forces as a consequence of an "enslaving" phenomenon that activates simultaneously the other fingers. Moreover, other studies demonstrated the consistency of joint co-variation despite the modifications of hand postures and kinematics in grasping multiple objects under different conditions (i.e. modulating visual or proprioceptive input) [START_REF] Thakur | Multidigit movement synergies of the human hand in an unconstrained haptic exploration task[END_REF][START_REF] Mason | Hand synergies during reach-to-grasp[END_REF][START_REF] Santello | Postural hand synergies for tool use[END_REF], 2002) and in complicated hand gestures, such as finger spelling (Jerde et al. 2003;[START_REF] Weiss | Muscular and postural synergies of the human hand[END_REF] or typing [START_REF] Fish | Synergistic finger movements in a skilled motor task[END_REF][START_REF] Soechting | Flexibility and repeatability of finger movements during typing: analysis of multiple degrees of freedom[END_REF].

Reaching versus Grasping

When considering transport and grasp component trial by trial, CG participants always kept the same reach duration while maximum grip aperture decreased. Consistency of reach duration probably resulted from the lack of experimental constraint on movement velocity (i.e. participants were in a self-paced mode). In contrast, a reduced hand opening seems a consequence of a learning effect due to the repetitions of the task [START_REF] Lin | Effects of modified constraint-induced movement therapy on reach-to-grasp movements and functional performance after chronic stroke: a randomized controlled study[END_REF]. Accordingly, developmental studies in children have showed that, in parallel with the refinement of other hand motor skills, the grip aperture becomes progressively smaller during the first decade of life [START_REF] Kuhtz-Buschbeck | Development of prehension movements in children: a kinematic study[END_REF][START_REF] Smyth | The role of sight of the hand in the development of prehension in childhood[END_REF]). In IG participants increased reach duration after non-use. Thus, one may suppose a reduction of transport duration trial by trial. Surprisingly, we observed an opposite recovery processes for reach and grasp components: while reach duration returned to the baseline value at the lasts repetitions of the task, the maximum grip aperture increased trial after trial.

Similarly, when reaching is under time constraint, several authors noted an increase of maximum grip aperture [START_REF] Wing | Grasp size and accuracy of approach in reaching[END_REF][START_REF] Wallace | Temporal constraints in the control of prehensile movement[END_REF][START_REF] Bootsma | The speed-accuracy trade-off in manual prehension: effects of movement amplitude, object size and object width on kinematic characteristics[END_REF]Mason and Carnahan 1999), as a strategy to grasp with a greater "safety margin" [START_REF] Grosskopf | Grasping with the left and right hand: a kinematic study[END_REF]. Why reach and grasp recovered differently? One possibility concerns the effect of immobilization on distal (wrist and fingers for the grasp) and proximal (shoulder and elbow for the transport) parts.

Actually, the bandage worn by participants prevented any hand and wrist motions, but did not completely block shoulder movements. Further, reaching is organized bilaterally, so it can be controlled adequately by the ipsilateral hemisphere [START_REF] Brinkman | Cerebral control of contralateral and ipsilateral arm, hand and finger movements in the split-brain rhesus monkey[END_REF]. On the contrary, grasping depends more on controlateral primary motor cortex [START_REF] Jeannerod | The formation of finger grip during prehension. A cortically mediated visuomotor pattern[END_REF][START_REF] Porter | R Corticospinal Function and Vol-untary Movement[END_REF]. Thus, we can suppose that the activity of the ipsilateral "not immobilized" motor cortex could support the recovery for the reaching, but not for the grasp component. In support of thi hypothesis, in another study, we demonstrated that the cortical excitability is reduced in the motor cortex controlateral to the immobilized arm, while not in the ipsilateral one (Bassolino et al., 2009 XXX). This could confirm a potential role of the ipsilateral motor cortex in the fast improvement observed only for the transport component. Finally, the different recovery processes could also be related to an optimal strategy [START_REF] Todorov | Optimal feedback control as a theory of motor coordination[END_REF]Guignon et al., 2007). Indeed, reduction of reach duration would decrease the energetic cost mainly due to arm displacement, and become a priority compared to grip aperture.

Conclusion

The present findings demonstrated that ten hours of hand and forearm immobilization affect the reaching-to-grasp movement, lengthening the duration and modifying the timing of the transport phase, increasing the amplitude of grip aperture and altering the final hand posture. Thus, a transient lack of sensory information and motor commands modify the kinematic of a very well known motor task. These results could be due both to peripheral or cortical effects. Findings from previous studies support the latter hypothesis [START_REF] Huber | Arm immobilization causes cortical plastic changes and locally decreases sleep slow wave activity[END_REF]Facchini et al.2002;Bassolino et al., 2009XXX). However, to answer this question, direct evidence of cortical changes after ten hours of arm immobilization are required. FIGURE 1 METHODS. On the left, markers displacement and F1 with x and y axis, the system of reference built on the hand plane by means of A, B and C points. Upper on the right, the y axis of F1 and F2.

F2 was obtained by the rotation of F1 with respect to the X axis and attached on each finger at the MCP and PIP joints respectively. Lower on the right, angles at MCP (metacarpal-phalangeal) and PIP (proximal interphalangeal) joints of index finger. with the object before ("blue shadow color") and after immobilization ("skin natural color"). After non-use, the hand posture was characterized by more flexed fingers at the MCP joints and more extended PIP joints. Therefore, index, ring and little fingers were more adducted. Moreover, a stronger opposition of the thumb (more internal rotated for the pencil and more adducted for the ball) was evident. FIGURE 6 RECOVERY. The linear regression lines of reach duration (panel A) and maximum grip aperture (panel B) on the number of trials, for the IG (dark grey) and CG (light grey). Participants in the IG reduced the reach duration trial by trial, while this parameter did not change in the CG. Concerning maximum grip aperture, in the IG, participants increased the hand aperture during the repetitions, while in the CG, subjects progressively reduced that aperture. In panel C, the inverse correlation between the mean values of reach duration and the maximum grip aperture at each trial in the IG; on the contrary, no correlation between these two parameters is showed for the CG (panel D).

MCP

FIGURE 2 REACH

 2 FIGURE 2

FIGURE 4 STATIC

 4 FIGURE 4

FIGURE 5 PRINCIPAL

 5 FIGURE 5

  Post-Pre) in Immobilization (IG) and Control group (CG): post hoc comparisons of the Group or when significant, of the Group X Object interaction (P= pencil, B= ball) are shown. At MCP and PIP joints a positive and negative value point out respectively wider flexed or extended posture. For the middle, ring and little ABD angles a positive difference reveals that the fingers were more adducted. On the contrary, for the index and thumb the adduction is identified by negative values. A higher negative value of the T_ROT corresponds to an increased internal rotation of this finger toward the hand palm. Standard error of mean values are in parentheses. * indicates p<0.05, ** indicates p<0.005.

PCA was applied to the angular displacements of the sixteen angles to understand if different patterns of co-variation of the digitis were present after immobilization. PCA showed that the first two principal components could account for a large proportion of the variance, i.e., 90.6 % and 89.9% for the Post conditions in the IG and the CG, respectively, without any different for the two objects. This implied that before immobilization there was a high degree of co-variation between the fingers that remained unvaried after non-use. The present result is in line with previous studies in which the first two components could account for the majority of the variance, without distinguishing the type of grip (e.g., precision or power) or different target object (for example [START_REF] Santello | Patterns of hand motion during grasping and the influence of sensory guidance[END_REF]. Moreover, also comparing the VAF by PC1 and PC2 through separate PCAs for each subjects, no difference was showed in the IG and the CG. Furthermore, the waveforms of the first two principal components were remarkably similar (PC1: r 2= 0.99, p<0.00001; PC2: r 2= 0.99 , p<0.00001).

INSERT FIGURE 5 AROUND HERE

Recovery.

ANOVA on slopes of reach duration calculated on 15 successive trials revealed a statistical difference in IG and CG (F(1,11) =9.59, p<0.01, mean slope IG= -8.8, mean slope CG= -0.7).

Indeed, the reach duration in IG was inversely correlated with trials (r= -0.8 , p<0.001), while there was no correlation in the CG (r= -0.2, p= 0.53) (Figure 6, panel A). This showed that after immobilization, subjects took more time to perform the reach in the first trials, but during the task, this effect reduced until getting values comparable with those obtained in the CG at the last three repetitions. Similarly, concerning the maximum grip aperture, ANOVA on slopes demonstrated a significant difference in IG and CG (F(1,11) =6.85, p<0.02, slope IG= 0.004, slope CG= -0.004).

While the maximum grip aperture in CG was inversely correlated with trial number (r= -0.7, p<0.01), in the IG the correlation was positive (r= 0.7, p<0.01) (Figure 6, panel B). Thus, in the CG participants decreased the index-thumb distance during the repetitions; on the contrary this distance increased trial by trial in the IG. Consequently, in IG the maximum grip aperture and the reach duration were inversely correlated (r= -0.7, p<0.01): while participants reduced the reach duration (i.e. recovery of immobilization effect), they opened their hand wider (Figure 6, panel C). On the