Peptide aptamers in label-free protein detection: 2. Chemical optimization and detection of distinct protein isoforms.

The early detection and diagnosis of cancer lies central to successful treatment and improved patient outcome. Current techniques are limited by the nature of the biological receptor and the assays available. This paper reports the use of novel biological probes, peptide aptamers, in detecting cyclin-dependent protein kinases (CDKs) whose activity is important in proliferating and cancerous cells. We describe, specifically, the optimization of an orientated peptide aptamer surface and its utilization in establishing a highly specific, low-nanomolar sensitive, detection protocol for the active form of CDK2. In comparing target binding affinity of two different aptamers (pep6 and pep9), both constructed through the insertion of peptide sequences into the surface of a scaffold protein, one was observed to be consistently more effective. Significantly, the pep9 aptamers were able to detect subtle changes in the conformation of CDK2 associated with activation of its catalytic activity that may be caused by the phosphorylation of a single amino acid (threonine 160). A typical response toward the inactive form of CDK2 was in the range of 0.5-2% of the binding of the active form of CDK2 in the concentration range from 2 to 20 nM. Although antibodies are occasionally able to recognize conformations in their targets, this is the first time that a nonantibody protein probe has been used to detect an active protein isoform. Because peptide aptamers are usually raised against full-length proteins, this raises the possibility that peptide aptamers will be able to extend the repertoire of probes that recognize protein conformations, post-translational modifications (PTMs), or conformations stabilized by PTMs.

Interventions to promote more effective balance-recovery reactions in industrial settings: new perspectives on footwear and handrails.

"Change-in-support" balance-recovery reactions that involve rapid stepping or reaching movements play a critical role in preventing falls. Recent geriatrics studies have led to new interventions to improve ability to execute these reactions effectively. Some of these interventions have the potential to reduce fall risk for younger persons working in industrial settings. In this paper, we review research pertaining to two such interventions: 1) balance-enhancing footwear insoles designed to improve stepping reactions, and 2) proximity-triggered handrail cueing systems designed to improve reach-to-grasp reactions. The insole has a raised ridge around the perimeter that is intended to improve balance control by providing increased stimulation of sensory receptors on the footsole in situations where loss of balance may be imminent. The cueing system uses flashing lights and/or verbal prompts to attract attention to the handrail and ensure that the brain registers its location, thereby facilitating more rapid and accurate grasping of the rail if and when sudden loss of balance occurs. Results to date support the efficacy of both interventions in geriatric populations. There is also some evidence that these interventions may improve balance control in younger persons; however, further research is needed to confirm their efficacy in preventing falls in industrial settings.

Does the "eyes lead the hand" principle apply to reach-to-grasp movements evoked by unexpected balance perturbations?

A fundamental principle that has emerged from studies of natural gaze behavior is that goal-directed arm movements are typically guided by a saccade to the target. In this study, we evaluated a hypothesis that this principle does not apply to rapid reach-to-grasp movements evoked by sudden unexpected balance perturbations. These perturbations involved forward translation of a large (2 × 6 m) motion platform configured to simulate a "real-life" environment. Subjects performed a common "daily-life" visuo-cognitive task (find a telephone and make a call) that required walking to the end of the platform, which was triggered to move as they approached a handrail mounted alongside the travel path. A deception was used to ensure that the perturbation was truly unexpected. Eleven of 18 healthy young-adult subjects (age 22-30) reached to grasp or touch the rail in response to the balance perturbation. In support of the hypothesis, none of these arm reactions was guided by concurrent visual fixation of the handrail. Seven of the 11 looked at the rail upon first entering the environment, and hence may have used "stored" central-field information about the handrail location to guide the subsequent arm reaction. However, the other four subjects never looked directly at the rail, indicating a complete reliance on peripheral vision. These findings add to previous evidence of distinctions in the CNS control of volitional and perturbation-evoked arm movements. Future studies will determine whether similar visuo-motor behavior occurs when the available handhold is smaller or when subjects are not engaged in a concurrent visuo-cognitive task.

The moveable handhold: a new paradigm to study visual contributions to the control of balance-recovery reactions.

Balance-recovery reactions that involve rapid step or reach-to-grasp movements are prevalent and functionally important responses to instability. Successful use of these reactions to recover balance in daily life requires a capacity to modulate the reaction to deal with the continual variation in environmental constraints that occurs as the person moves, i.e. location of objects that can obstruct limb movements or serve as handholds to grasp. The most direct approach to study this involves applying balance perturbations as subjects move within a visually complex environment; however, this approach does not allow precise control over kinematic variables or visual inputs, and is susceptible to strong learning effects. We have therefore developed an alternate approach, wherein the subject is stationary and the relative motion between subject and constraints that normally occurs as a result of ambulation is instead introduced via movement of the surrounding obstacles or handholds. We previously developed a motor-driven "obstacle-mover" to manipulate constraints on step reactions, and now describe an analogous approach to study reach-to-grasp reactions, using a motor-driven "handhold-mover". We anticipate that this paradigm will provide new opportunities to probe CNS control of upright stance, by providing a sensitive indicator of limitations in the neuromusculoskeletal systems. It can also be used to test perturbation-evoked reactions in seated subjects, thereby allowing testing or training of persons who are unable to stand and use of techniques (e.g. TMS, EEG) that can be difficult to perform in free-standing subjects.