[Molecular and structural-biological analysis of Nicotiana plumbaginifolia mutants for identification of the site of beta-tubulins interaction with dinitroanilines and phosphorotioamidates].

The identification of point mutation locations on beta-tubulin molecules of amiprophosmethyl- and trifluralin-resistant Nicotiana plumbaginifolia lines have described in the work. It was shown that in the first case this mutation is connected with the substitution ofserine residue on proline in position 248; in the second case--with the substitution of phenilalanine on serine in position 317 of beta-tubulin amino acid sequence. Three-dimensional models of beta-tubulin molecule from Chlamydomonas with well-known location of mutations conferring dinitroaniline- and phosphorotioamidate resistance (substitution of lysine residue to methionine on position 350), and beta-tubulin from Nicotiana plumbaginifolia have been reconstructed. On the basis of analysis of site of interaction with dinitroanilines and phosphorotioamides on Chlamydomonas beta-tubulin molecule it was concluded that the revealed mutations on Nicotiana plumbaginifolia beta-tubulin affect amino acid residues participating in formation of this site.

Biomimetic sensory feedback through peripheral nerve stimulation improves dexterous use of a bionic hand.

We describe use of a bidirectional neuromyoelectric prosthetic hand that conveys biomimetic sensory feedback. Electromyographic recordings from residual arm muscles were decoded to provide independent and proportional control of a six-DOF prosthetic hand and wrist-the DEKA LUKE arm. Activation of contact sensors on the prosthesis resulted in intraneural microstimulation of residual sensory nerve fibers through chronically implanted Utah Slanted Electrode Arrays, thereby evoking tactile percepts on the phantom hand. With sensory feedback enabled, the participant exhibited greater precision in grip force and was better able to handle fragile objects. With active exploration, the participant was also able to distinguish between small and large objects and between soft and hard ones. When the sensory feedback was biomimetic-designed to mimic natural sensory signals-the participant was able to identify the objects significantly faster than with the use of traditional encoding algorithms that depended on only the present stimulus intensity. Thus, artificial touch can be sculpted by patterning the sensory feedback, and biologically inspired patterns elicit more interpretable and useful percepts.

Emotional learning. Fear and loathing in the amygdala.

The amygdala in the brain plays a critical role in learning emotional components of experience, such as conditioned fear; these processes in turn affect many other aspects of memory and cognition.

Restoring motor control and sensory feedback in people with upper extremity amputations using arrays of 96 microelectrodes implanted in the median and ulnar nerves.

OBJECTIVE: An important goal of neuroprosthetic research is to establish bidirectional communication between the user and new prosthetic limbs that are capable of controlling >20 different movements. One strategy for achieving this goal is to interface the prosthetic limb directly with efferent and afferent fibres in the peripheral nervous system using an array of intrafascicular microelectrodes. This approach would provide access to a large number of independent neural pathways for controlling high degree-of-freedom prosthetic limbs, as well as evoking multiple-complex sensory percepts. APPROACH: Utah Slanted Electrode Arrays (USEAs, 96 recording/stimulating electrodes) were implanted for 30 days into the median (Subject 1-M, 31 years post-amputation) or ulnar (Subject 2-U, 1.5 years post-amputation) nerves of two amputees. Neural activity was recorded during intended movements of the subject's phantom fingers and a linear Kalman filter was used to decode the neural data. Microelectrode stimulation of varying amplitudes and frequencies was delivered via single or multiple electrodes to investigate the number, size and quality of sensory percepts that could be evoked. Device performance over time was assessed by measuring: electrode impedances, signal-to-noise ratios (SNRs), stimulation thresholds, number and stability of evoked percepts. MAIN RESULTS: The subjects were able to proportionally, control individual fingers of a virtual robotic hand, with 13 different movements decoded offline (r = 0.48) and two movements decoded online. Electrical stimulation across one USEA evoked >80 sensory percepts. Varying the stimulation parameters modulated percept quality. Devices remained intrafascicularly implanted for the duration of the study with no significant changes in the SNRs or percept thresholds. SIGNIFICANCE: This study demonstrated that an array of 96 microelectrodes can be implanted into the human peripheral nervous system for up to 1 month durations. Such an array could provide intuitive control of a virtual prosthetic hand with broad sensory feedback.

In situ measurements of nitrate leaching implicate poor nitrogen and irrigation management on sandy soils.

Minimizing the risk of nitrate contamination along the waterways of the U.S. Great Plains is essential to continued irrigated corn production and quality water supplies. The objectives of this study were to quantify nitrate (NO(3)) leaching for irrigated sandy soils (Pratt loamy fine sand [sandy, mixed, mesic Lamellic Haplustalfs]) and to evaluate the effects of N fertilizer and irrigation management strategies on NO(3) leaching in irrigated corn. Two irrigation schedules (1.0x and 1.25x optimum) were combined with six N fertilizer treatments broadcast as NH(4)NO(3) (kg N ha(-1)): 300 and 250 applied pre-plant; 250 applied pre-plant and sidedress; 185 applied pre-plant and sidedress; 125 applied pre-plant and sidedress; and 0. Porous-cup tensiometers and solution samplers were installed in each of the four highest N treatments. Soil solution samples were collected during the 2001 and 2002 growing seasons. Maximum corn grain yield was achieved with 125 or 185 kg N ha(-1), regardless of the irrigation schedule (IS). The 1.25x IS exacerbated the amount of NO(3) leached below the 152-cm depth in the preplant N treatments, with a mean of 146 kg N ha(-1) for the 250 and 300 kg N preplant applications compared with 12 kg N ha(-1) for the same N treatments and 1.0x IS. With 185 kg N ha(-1), the 1.25x IS treatment resulted in 74 kg N ha(-1) leached compared with 10 kg N ha(-1) for the 1.0x IS. Appropriate irrigation scheduling and N fertilizer rates are essential to improving N management practices on these sandy soils.

Early undernutrition impairs hippocampal long-term potentiation in adult rats.

Following high-frequency stimulation of hippocampal dentate granule cells, potentiation was difficult to achieve in undernourished animals, showed a significant decline within 3 to 6 hr, and was completely absent at 24 hr. Further trains of stimulation resulted in only small benefits in undernourished animals. Coupled with previously reported morphological and behavioral deficits, these findings indicate a marked hippocampal dysfunction resulting from early undernutrition and provide a potentially valuable approach for relating nutritionally induced behavioral impairments to brain function.

Synaptic depression at type B to A photoreceptor connections in Hermissenda.

We quantified synaptic depression at the type B to A photoreceptor connections of Hermissenda. Type B cell action potentials were evoked at interstimulus intervals (ISIs) of 0.3, 3 or 30 s and the resulting inhibitory postsynaptic potentials (IPSPs) were recorded in a type A cell. A progressive decline in IPSP amplitude occurred at all three ISIs. Synaptic depression was greater at shorter ISIs, as was the level of recovery 2 min after the stimulus series. The profound level of synaptic depression observed (79.0+/-3.2% at the 0.3 s ISI) implies that synaptic depression is an important control process in the neuronal circuitry that drives phototactic behavior.

Increased responsivity of dentate granule cells during nictitating membrane response conditioning in rabbit.

We examined the responsivity of dentate gyrus granule cells to perforant path stimulation during classical conditioning of the rabbit nictitating membrane response. Dentate field potentials elicited by perforant path stimulation were recorded during training to test for changes in granule cell responsivity. Results showed above-baseline increases in dentate population spike amplitudes over the course of training in paired but not unpaired animals. In addition, population spike amplitudes were smaller when elicited during tone presentations in both paired and unpaired animals than between trials when no conditioning stimuli were present. While alternative interpretations remain, these results provide preliminary evidence that processes similar to long-term potentiation may occur in the hippocampus during behavioral conditioning.

Effects of lesions of cerebellar nuclei on conditioned behavioral and hippocampal neuronal responses.

Rabbits were overtrained using classical conditioning of the rabbit nictitating membrane (NM)/eyelid response. Unilateral electrolytic lesions were then made through electrodes previously implanted in dentate and interpositus cerebellar nuclei ipsilateral to the trained (left) eye. Lesions caused a complete or near-complete abolition of conditioned behavioral responses on the ipsilateral side, but had no effect on unconditioned responses to corneal airpuff. When training was switched to the contralateral (right) side, animals learned within the first few trials, but did not relearn when training was returned to the left (lesioned) side. Control animals in which lesions spared the deep nuclei showed no such learning deficits. Lesions of cerebellar nuclei also abolished conditioned increases in hippocampal CAl neural activity evoked by the tone conditioned stimulus in this paradigm. As with the behavior, training on the right (non-lesioned) side reinstated the conditioned neuronal response within the first few trials of training, even though behavioral responding on lesioned side showed little or no improvement. These results indicate that the cerebellum is an essential structure for the behavioral expression of learning, and plays an important role in the generation of conditioned hippocampal responses observed in this paradigm.

GABA-induced synaptic facilitation at type B to A photoreceptor connections in Hermissenda.

Gamma-aminobutyric acid (GABA) is a prevalent neurotransmitter in both vertebrate and invertebrate systems. Here we report that, in addition to its usual inhibitory actions, GABA induced synaptic facilitation at type B to A photoreceptor connections of the marine mollusk Hermissenda when applied transiently to the isolated nervous system. Synaptic facilitation also occurred in response to mechanical stimulation of the GABAergic hair cells, which are normally activated by rotational unconditioned stimuli during behavioral training of the intact animal. This synaptic facilitation represents a novel form of GABA-induced neuromodulation which may contribute to learning-dependent suppression of phototaxis in Hermissenda.

Can infant malnutrition cause adult vertebral stenosis?

Does infant malnutrition produce smaller adult spinal canals? Lumbar and thoracic vertebrae (n X 1073), from a prehistoric American Indian population (15-55 yrs of age), were measured for anteroposterior (AP) and transverse (TR) vertebral canal sizes, nerve root tunnel (intervertebral foramen) widths (NRT), vertebral heights (VH), vertebral osteophytosis (VO), and tibial lengths. They underwent a dietary change from hunting-gathering, protein rich (PR), to maize agriculture, protein deficient (PD), between 950 and 1300 A.D. Multivariate analyses controlled for age, sex, culture, NRT, VH, VO, and wedging. Canal size was significantly smaller in the PD. AP diameters were generally and highly correlated with NRT, and thus both spinal stenosis and sciatica may have a developmental basis. Canal size was independent of statural components. Consequently, canal size is a most powerful tool in assessing the presence infant malnutrition. Moreover, perhaps the association between canal size and low-back pain (LBP) found in living populations has been underestimated, and this component of LBP is preventable.

The foreign body response to the Utah Slant Electrode Array in the cat sciatic nerve.

As the field of neuroprosthetic research continues to grow, studies describing the foreign body reaction surrounding chronic indwelling electrodes or microelectrode arrays will be critical for assessing biocompatibility. Of particular importance is the reaction surrounding penetrating microelectrodes that are used to stimulate and record from peripheral nerves used for prosthetic control, where such studies on axially penetrating electrodes are limited. Using the Utah Slant Electrode Array and a variety of histological methods, we investigated the foreign body response to the implanted array and its surrounding silicone cuff over long indwelling periods in the cat sciatic nerve. We observed that implanted nerves were associated with increased numbers of activated macrophages at the implant site, as well as distal to the implant, at all time points examined, with the longest observation being 350 days after implantation. We found that implanted cat sciatic nerves undergo a compensatory regenerative response after the initial injury that is accompanied by shifts in nerve fiber composition toward nerve fibers of smaller diameter and evidence of axons growing around microelectrode shafts. Nerve fibers located in fascicles that were not penetrated by the array or were located more than a few hundred microns from the implant appeared normal when examined over the course of a year-long indwelling period.

Non-invasive method for selection of electrodes and stimulus parameters for FES applications with intrafascicular arrays.

High-channel-count intrafascicular electrode arrays provide comprehensive and selective access to the peripheral nervous system. One practical difficulty in using several electrode arrays to evoke coordinated movements in paralyzed limbs is the identification of the appropriate stimulation channels and stimulus parameters to evoke desired movements. Here we present the use of a six degree-of-freedom load cell placed under the foot of a feline to characterize the muscle activation produced by three 100-electrode Utah Slanted Electrode Arrays (USEAs) implanted into the femoral nerves, sciatic nerves, and muscular branches of the sciatic nerves of three cats. Intramuscular stimulation was used to identify the endpoint force directions produced by 15 muscles of the hind limb, and these directions were used to classify the forces produced by each intrafascicular USEA electrode as flexion or extension. For 451 USEA electrodes, stimulus intensities for threshold and saturation muscle forces were identified, and the 3D direction and linearity of the force recruitment curves were determined. Further, motor unit excitation independence for 198 electrode pairs was measured using the refractory technique. This study demonstrates the utility of 3D endpoint force monitoring as a simple and non-invasive metric for characterizing the muscle-activation properties of hundreds of implanted peripheral nerve electrodes, allowing for electrode and parameter selection for neuroprosthetic applications.

Characterization of a 3D optrode array for infrared neural stimulation.

This paper characterizes the Utah Slant Optrode Array (USOA) as a means to deliver infrared light deep into tissue. An undoped crystalline silicon (100) substrate was used to fabricate 10 × 10 arrays of optrodes with rows of varying lengths from 0.5 mm to 1.5 mm on a 400-µm pitch. Light delivery from optical fibers and loss mechanisms through these Si optrodes were characterized, with the primary loss mechanisms being Fresnel reflection, coupling, radiation losses from the tapered shank and total internal reflection in the tips. Transmission at the optrode tips with different optical fiber core diameters and light in-coupling interfaces was investigated. At λ = 1.55µm, the highest optrode transmittance of 34.7%, relative to the optical fiber output power, was obtained with a 50-µm multi-mode fiber butt-coupled to the optrode through an intervening medium of index n = 1.66. Maximum power is directed into the optrodes when using fibers with core diameters of 200 µm or less. In addition, the output power varied with the optrode length/taper such that longer and less tapered optrodes exhibited higher light transmission efficiency. Output beam profiles and potential impacts on physiological tests were also examined. Future work is expected to improve USOA efficiency to greater than 64%.

Coordinated, multi-joint, fatigue-resistant feline stance produced with intrafascicular hind limb nerve stimulation.

The production of graceful skeletal movements requires coordinated activation of multiple muscles that produce torques around multiple joints. The work described herein is focused on one such movement, stance, that requires coordinated activation of extensor muscles acting around the hip, knee and ankle joints. The forces evoked in these muscles by external stimulation all have a complex dependence on muscle length and shortening velocities, and some of these muscles are biarticular. In order to recreate sit-to-stand maneuvers in the anesthetized feline, we excited the hind limb musculature using intrafascicular multielectrode stimulation (IFMS) of the muscular branch of the sciatic nerve, the femoral nerve and the main branch of the sciatic nerve. Stimulation was achieved with either acutely or chronically implanted Utah Slanted Electrode Arrays (USEAs) via subsets of electrodes (1) that activated motor units in the extensor muscles of the hip, knee and ankle joints, (2) that were able to evoke large extension forces and (3) that manifested minimal coactivation of the targeted motor units. Three hind limb force-generation strategies were investigated, including sequential activation of independent motor units to increase force, and interleaved or simultaneous IFMS of three sets of six or more USEA electrodes that excited the hip, knee and ankle extensors. All force-generation strategies evoked stance, but the interleaved IFMS strategy also reduced muscle fatigue produced by repeated sit-to-stand maneuvers compared with fatigue produced by simultaneous activation of different motor neuron pools. These results demonstrate the use of interleaved IFMS as a means to recreate coordinated, fatigue-resistant multi-joint muscle forces in the unilateral hind limb. This muscle activation paradigm could provide a promising neuroprosthetic approach for the restoration of sit-to-stand transitions in individuals who are paralyzed by spinal cord injury, stroke or disease.