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Regulation of the cardiac isoform of the ryanodine receptor by S-adenosyl-l-methionine

2011-11-08 , Gaboardi, Angela Kampfer

Activity of the Ryanodine Receptor (RyR2) (aka cardiac Ca2+ release channel) plays a pivotal role in contraction of the heart. S-adenosyl-l-methionine (SAM) is a biological methyl group donor that has close structural similarity to ATP, an important physiological regulator of RyR2. This work provides evidence that SAM can act as a RyR2 regulatory ligand in a manner independent from its recognized role as a biological methyl group donor. RyR2 activation appears to arise from the direct interaction of SAM, via its adenosyl moiety, with the RyR2 adenine nucleotide binding sites. Because uncertainty remains regarding the structural motifs involved in RyR2 modulation by ATP and its metabolites, this finding has important implications for clarifying the structural basis of ATP regulation of RyR2. During the course of this project, direct measurements of single RyR2 activity revealed that SAM has distinct effects on RyR2 conductance. From the cytosolic side of the channel, SAM produced a single clearly resolved subconductance state. The effects of SAM on channel conductance were dependent on SAM concentration and membrane holding potential. A second goal of this work was to distinguish between the two possible mechanisms by which SAM could reduce RyR2 conductance: i) SAM interfering directly with ion permeation via binding within the conduction pathway (pore block), or ii) SAM binding a regulatory (or allosteric) site thereby stabilizing or inducing a reduced conductance conformation of the channel. It was determined that SAM does not directly interact with the RyR2 conduction pathway. To account for these observations an allosteric model for the effect of SAM on RyR2 conductance is proposed. According to this model, SAM binding stabilizes an inherent RyR2 subconductance conformation. The voltage dependence of the SAM related subconductance state is accounted for by direct effects of voltage on channel conformation which indirectly alter the affinity of RyR2 for SAM. Patterns in the transitions between RyR2 conductance states in the presence of SAM may provide insight into the structure-activity relationship of RyR2 which can aid in the development of therapeutic strategies targeting this channel.

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Motor control in persons with a trans-tibial amputation during cycling

2011-07-06 , Childers, Walter Lee

Motor control of any movement task involves the integration of neural, muscular and skeletal systems. This integration must occur throughout the sensorimotor system and focus its efforts on controlling the system endpoint, e.g. the foot during locomotion. A person with a uni-lateral trans-tibial amputation has lost the foot, ankle joint, and muscles crossing those joints, hence the residuum becomes the new terminus of the motor system. The amputee must now adjust to the additional challenges of utilizing a compromised motor system as well as the challenges of controlling an external device, i.e. prosthesis, through the mechanical interface between the residuum and prosthetic socket. The obvious physical and physiologic asymmetries between the sound and amputated limbs are also involved in strategies for locomotion involving kinematic and kinetic asymmetries (Winter&Sienko, 1988). There are many questions as to why these asymmetric locomotor strategies are selected and what factors may be influencing that strategy. Factors influencing a change in locomotor strategy could be related to 1) the central nervous system accounting for the loss of sensorimotor feedback, 2) the altered mechanics of this new human/prosthetic system, or some combination of these factors. Understanding how the human motor system adjusts to the amputation and to the addition of an external mechanical device can provide useful insight into how robust the human control system may be and to adaptations in human motor control. This research uses a group of individuals with a uni-lateral trans-tibial amputation and a group of intact individuals using an Ankle Foot Orthosis (AFO) performing a cycling task to understand the "motor adjustments" necessary to utilize an external device for locomotion. Results of these experiments suggest 1) the motor system does account for the activation-contraction dynamics when coordinating muscle activity post amputation, 2) the motor system also changes joint kinetics and muscle activity, 3) these changes are related to control of the interface between the limb and the external device, and 4) the motor system does not alter kinetic asymmetries when kinematic asymmetries are minimized, contrary to a common practice in rehabilitation (Kapp, 2004). Results suggest that control of the external device, i.e. prosthesis or AFO, via the interface between the limb and the device reflect "motor adjustments" made by the nervous system and may be viewed in the context of tool use. Clinical goals in rehabilitation currently focus on minimizing gait deviations whereas the clinical application of these results suggest these deviations from normal locomotion are motor adjustments necessary to control a tool, i.e. prosthesis, by the motor system. Examining amputee locomotion in the context of tool use changes the clinical paradigm from one designed to minimize deviations to one intended to understand this behavior as related to interface control of the device thereby shifting the focus to improving function of the limb/prosthesis system. Kapp SL. (2004) Atlas amp limb def: surg pros rehab princ. 3rd ed: 385 - 394. Winter&Sienko. (1988) J Biomech, 21: 361 - 367.

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Motor learning and its transfer during bilateral arm reaching.

2011-06-09 , Harley, Linda Rosemary

Have you ever attempted to rub your abdomen with one hand while tapping your head with the other? Separately these movements are easy to perform but doing them together (bilateral task) requires motor adaptation. Motor adaptation is the process through which the Central Nervous System improves upon performance. Transfer of learning is the process through which learning a motor task in one condition improves performance in another condition. The purpose of this study was to determine whether transfer of learning occurs during bilateral goal-directed reaching tasks. It was hypothesized that transfer of learning would occur from the non-dominant to the dominant arm during bilateral tasks and that position and load feedback from the arms would affect the rate of adaptation and transfer of learning. During the experiments, subjects reached with one or both their index finger(s) to eight targets while a velocity dependent force perturbation (force environment) was applied to the arm(s). Three groups of bilateral tasks were examined: (1) unilateral reaching, where one arm learned to reach in a force environment, while the other arm remained stationary and therefore did not provide movement related position or load feedback; (2) bilateral reaching single load, where both arms performed reaching movements but only one arm learned a force environment and therefore the other arm provided movement related position feedback but not load feedback; (3) bilateral reaching two loads, where both arms performed reaching movements and both learned a force environment, while providing movement related position and load feedback. The rate of adaptation of the force environment was quantified as the speed at which the perturbed index finger trajectory became straight over the course of repeated task performance. The rate of adaptation was significantly slower for the dominant arm during the unilateral reaching tasks than during the bilateral reaching single load tasks (p<0.05). This indicates that the movement related position feedback from the non-dominant arm improved significantly the motor adaptation of the dominant arm; therefore transfer of learning occurred from the non-dominant to the dominant arm. The rate of adaptation for the non-dominant arm did not differ significantly (p>0.05) between the unilateral reaching and bilateral reaching single load tasks. Results also indicated that the rate of adaptation was significantly (p<0.05) faster for both the non-dominant and the dominant arms during the bilateral reaching two loads tasks than during the bilateral reaching single load tasks. The latter results indicate that transfer of learning occurred in both directions - from the dominant to the non-dominant arm and from the non-dominant to the dominant arm - when position and load feedback was available from both arms, but only when the force environment acted in the same joint direction. This study demonstrated that transfer of learning does occur during bilateral reaching tasks and that the direction and degree of transfer of learning may be modulated by the position and load feedback that is available to the central nervous system. This information may be used by physical therapists in order to improve rehabilitation strategies for the upper extremity.

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