First-in-man study with a novel PEGylated recombinant human insulin-like growth factor-I.

OBJECTIVE: This study is a first time assessment of safety and tolerability, pharmacokinetics, and pharmacodynamics of RO5046013 in human, in comparison with unmodified rhIGF-I. DESIGN: The study was conducted as a single-center, randomized, double-blinded, placebo-controlled, single ascending dose, parallel group study in a clinical research unit in France. A total of 62 healthy volunteers participated in this clinical trial. RO5046013 was given as single subcutaneous injection, or as intravenous infusion over 48h, at ascending dose levels. The active comparator rhIGF-I was administered at 50µg/kg subcutaneously twice daily for 4days. Safety and tolerability, pharmacokinetics, and pharmacodynamics of RO5046013 were evaluated. RESULTS: PEGylation resulted in long exposure to RO5046013 with a half-life of 140-200h. Exposure to RO5046013 increased approximately dose proportionally. RO5046013 was safe and well tolerated at all doses, injection site erythema after SC administration was the most frequent observed AE. No hypoglycemia occurred. Growth hormone (GH) secretion was almost completely suppressed with rhIGF-I administration, whereas RO5046013 caused only a modest decrease in GH at the highest dose given IV. CONCLUSIONS: PEGylation of IGF-I strongly enhances half-life, reduces the negative GH feedback and hypoglycemia potential, and therefore offers a valuable alternative to rhIGF-I in treatment of relevant diseases.

Ultrasound-based nonviral gene delivery induces bone formation in vivo.

Nonviral gene delivery is a promising, safe, therapeutic tool in regenerative medicine. This study is the first to achieve nonviral, ultrasound-based, osteogenic gene delivery that leads to bone tissue formation, in vivo. We hypothesized that direct in vivo sonoporation of naked DNA encoding for the osteogenic gene, recombinant human bone morphogenetic protein-9 (rhBMP-9) would induce bone formation. A luciferase plasmid (Luc), encoding rhBMP-9 or empty pcDNA3 vector mixed with microbubbles, was injected into the thigh muscles of mice. After injection, noninvasive sonoporation was applied. Luc activity was monitored noninvasively, and quantitatively using bioluminescence imaging in vivo, and found for 14 days with a peak expression on day 7. To examine osteogenesis in vivo, rhBMP-9 plasmid was sonoporated into the thigh muscles of transgenic mice that express the Luc gene under the control of a human osteocalcin promoter. Following rhBMP-9 sonoporation, osteocalcin-dependent Luc expression lasted for 24 days and peaked on day 10. Bone tissue was formed in the site of rhBMP-9 delivery, as was shown by micro-computerized tomography and histology. The sonoporation method was also compared with previously developed electrotransfer-based gene delivery and was found significantly inferior in its efficiency of gene delivery. We conclude that ultrasound-mediated osteogenic gene delivery could serve as a therapeutic solution in conditions requiring bone tissue regeneration after further development that will increase the transfection efficiency.

Echo suppression in the human cortex is affected by the spatial and temporal proximity of the primary sound and echo.

Echo suppression in the human auditory cortex was studied with auditory middle latency evoked potentials (AMEP) using virtual reality acoustic stimuli, including distance and elevation cues, presented by earphones. The purpose of the study was to assess the effect of proximity of the source sound and echo on the degree of echo suppression. Sixteen subjects were presented with source-echo pairs in which the preceding source sound was always at the vertex, and the echo varied among ten positions on the coronal plane. Positions varied in elevation, distance and time lag between source and echo. The psychoacoustic location judgment of the fused source-echo pair was closer to the source sound (more echo suppression) the nearer the echo drew to the source in its elevation and time. The equivalent dipole magnitudes of the cortical components of AMEP were significantly reduced (more suppression) with shorter echo lags and when echo elevation was similar to that of the source sound. The distances used in this study did not significantly affect echo suppression. These results indicate that echo suppression in the auditory cortex is more pronounced the closer are the primary sound and echo in locational attributes and timing. As source sound and echo draw apart, echo suppression in the cortex decreases and the perceived localization of the fused source-echo is more biased toward the echo.

Short latency visual evoked potentials in occupational exposure to organic solvents.

OBJECTIVES: Short latency visual evoked potentials (SVEP), in response to high-intensity flashes from light emitting diodes (LED), were used to detect subclinical effects along the visual pathway in four groups of subjects with different levels of exposure to gasoline, all within legally acceptable limits. METHODS: Potentials and exposure levels were obtained from 31 subjects with different occupational exposure levels to gasoline fumes, as well as from 17 non-exposed control subjects. SVEP were recorded from four electrode sites (infra-orbital, Cz, Pz, Oz), in response to flashes presented to each eye in turn from goggle-mounted LEDs. SVEP components were defined after digital filtering, which eliminated the high-frequency oscillatory potentials and accentuated five major components: a periocular P30, attributed to the retina; a fronto-central N50, attributed to the optic nerve; centro-parietal P65 and N85, attributed to the optic tracts and radiation; and an occipital, cortical P105. RESULTS: The latencies of successive SVEP components of the exposed subjects showed a significant latency prolongation compared to controls, beginning with activity attributed to the optic nerve and increasing cumulatively with the later components. Retinal components were not affected by the exposure to organic solvents. Among the exposed groups, differences in latency prolongation corresponded to occupational exposure. CONCLUSION: The low-frequency components of SVEP were reliably measured and proved to be sensitive to subclinical effects of organic solvents on conduction along the visual pathway. These components are likely to be sensitive to other subcortical visual pathway lesions, but their clinical promise needs further verification.

Auditory brain-stem evoked potentials in patients with thyroid and parathyroid dysfunction: adaptation to chronic hormonal dysequilibrium.

Auditory Brainstem Evoked Potentials (ABEPs) and pure tone audiograms were obtained from 24 patients with parathyroid dysfunction (17 hypercalcemia and 7 hypocalcemia) and 12 patients with thyroid dysfunction (6 hyperthyroid and 6 hypothyroid) and from 10 control subjects. ABEPs were characterized by I-V interpeak latency difference at 10/sec click rate and by the effect of increasing stimulus rate to 55/sec. None of the ABEP measures were significantly affected by levels of serum calcium, thyroid hormones or their interactions. Moreover no correlation was found between biochemical and electrophysiological measures. This stability of ABEP measures contrasts with earlier reports on acute effects of calcium and thyroid hormonal levels on auditory brainstem evoked potentials. We propose that chronic calcium or thyroid hormonal homeostatic changes are associated with adaptive mechanisms resulting in normal function of the auditory brainstem.

Motor evoked potentials in the preoperative and postoperative assessment of normal pressure hydrocephalus.

Motor evoked potentials and central motor conduction time (CMCT) were examined from both upper and lower limbs in patients with normal pressure hydrocephalus to find a predictor for the success of shunting procedures. The hypotheses that walking disturbances are due to pyramidal tract compression as well as the possibility that the upper limbs are affected subclinically in these patients were also studied. The study suggests that the walking disturbances are not the result of a major pyramidal tract dysfunction but probably involve the sensorimotor integration leading to normal gait. Furthermore, CMCT measured with electromagnetic motor stimulation can help in selecting the patients that will benefit from shunting. The study does not provide electrophysiological evidence of upper limb involvement in normal pressure hydrocephalus.

Short latency visual evoked potentials to flashes from light-emitting diodes.

Short latency visual evoked potentials (SVEPs) have been described in response to high-intensity, strobe flashes. High-intensity flashes can now be generated from goggle-mounted light emitting diodes (LEDs) and the SVEPs to such flashes have been shown to be reproducible across subjects, avoiding photic spread to the examination room and acoustical artifacts from the strobe stimulator. In this study, SVEPs from multichannel records are described in terms of normative latencies and amplitudes, as well as scalp distributions, to explore their generators. Potentials were recorded from 10 young male subjects, from 16 scalp locations, in response to flashes from goggle-mounted LEDs. Flashes were presented to each eye in turn, as well as binocularly. The latencies, scalp distributions and intersubject variabilities of the LED evoked SVEPs were similar to those obtained with strobe flashes. SVEP components were divided into 3 groups, according to their latency and the electrodes at which they were recorded with the largest amplitudes: periocular (under 40 msec latency), fronto-central (40-55 msec) and parieto-occipital (55-80 msec latency). The scalp distributions observed in this study suggest subcortical generators along the visual pathway, beginning at the retina. The use of goggle-mounted LEDs should promote routine evaluation of the integrity of the visual pathway between retina and cortex using SVEPs.

Effects of myelin or cell body brainstem lesions on 3-channel Lissajous' trajectories of fast and slow components of feline auditory brainstem evoked potentials.

Auditory brainstem evoked potentials (ABEP) were recorded from 16 awake cats using three orthogonal differential electrode pairs before and during a week after inducing neuronal lesions localized to the cochlear nucleus (CN) or the superior olivary complex (SOC), or myelin lesions localized to the fibers of the trapezoid body. ABEPs were digitally filtered to include only the slow ('pedestal') component, or only the faster first to fifth components, and three-channel Lissajous' trajectories (3CLTs) of these fast and slow components of ABEP were obtained. Cell body lesions and myelin lesions induced effects on 3CLT measures of both fast and slow components of ABEP. The results suggest a primary contribution of cell body activity to the slow component, and a primary fiber tract contribution to the fast components. However, the results do not support exclusive generation of the pedestal by cell body and dendritic post-synaptic potentials and of the faster components by action potentials along fibers. The results are consistent with a set of generators for each of the slow and fast components of ABEP, consisting of both cell bodies and their output fibers, that are spatially distributed in the brainstem.

A high-intensity, goggle-mounted flash stimulator for short-latency visual evoked potentials.

The few studies that have been done on short-latency, subcortical visual evoked potentials (SVEPs) have all used stroboscopic flashes as the evoking stimulus. The dimensions of the stimulator, the acoustical artifacts and the photic spread to the examination room limited the use of SVEPs to research laboratories. With the advent of high-efficiency light-emitting diodes (LEDs), high-intensity flashes can now be generated from goggle-mounted LEDs. In this study, a goggle-mounted high-intensity stimulator was constructed and its flashes used to evoke SVEPs. The reproducibility of SVEPs across subjects and the ease of using the high-intensity LED flash stimulator make them a promising candidate for testing subcortical visual pathway function in the operating room.

The effects of digital filtering on feline auditory brain-stem evoked potentials.

The power spectrum of the feline auditory brain-stem evoked potentials (ABEPs) consists of 3 frequency bands, similar to the human wave form, but differing in range. The frequency bands in the feline spectra were separated by notches at 326 Hz and 732 Hz. Click-evoked ABEP from 15 cats were digitally filtered in 3 passbands: (1) below 326 Hz ('slow filter'), (2) between 326 and 732 Hz ('medium filter'); and (3) between 732 and 1790 Hz ('fast filter'). Filtering in each of these bands differentially affected the ABEP components. The vertex positive components are labeled by their order of appearance, i.e., 1, 2, ... 5. Peak 1 is subdivided into 2 subcomponents labeled 1a and 1b. The slow filter was associated with the loss of all components leaving a slow potential shift, i.e., the 'pedestal' peaking at the latency of peak 4. The medium filter was associated with the loss of components 1a, 1b and 2, sparing 3 and 4. The fast filter was associated with the loss of 1b and a diminution of 2. Comparing cat and human ABEP, feline components 2, 3 and 4 behaved precisely the same as the human II, III and V. In contrast to the human I, the feline first component (1a) was not detected with the medium filter. No feline component, following peak 1 in the unfiltered wave form, disappeared with the slow and medium filters, and reemerged with the fast filter (as human IV does).(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of myelin or cell body brainstem lesions on 3-channel Lissajous' trajectories of feline auditory brainstem evoked potentials.

Auditory brainstem evoked potentials (ABEP) were recorded from 16 awake cats to obtain 3-Channel Lissajous' Trajectories (3CLTs) using three orthogonal differential electrode configurations (nasion-midline nuchal ridge, left-right mastoids, vertex-midline under the mandible). Potentials, evoked by monaural 80 dBnHL (re, human threshold) clicks, were studied before, and up to 7 weeks after inducing neuronal lesions localized to the cochlear nucleus (CN) or the superior olivary complex (SOC), or myelin lesions localized to the fibers of the trapezoid body connecting these two structures. Neuronal lesions were induced by injection of kainic acid (KA), while myelin lesions were induced by injection of L-alpha-lysophosphatidylcholine (LPC). With CN neuronal lesions the major changes in 3CLT were in the time domain of 'b', 'c' and 'd' (components P2, P3 and P4 of single-channel ABEP). With SOC neuronal lesions the major changes were in 'c' and 'd' of 3CLT (P3 and P4 of ABEP). With trapezoid body lesions the major change was in 'c' (P3 of ABEP). The results are compatible with the peripheral generation of the first ABEP components (P1a and P1b). The second component (P2) is generated by ipsilateral CN neurones and their outputs. The third component (P3) is generated primarily by ipsilateral SOC neurones and their outputs, with the ipsilateral CN providing input. The The fourth component (P4) is generated bilaterally by the SOC neurones and their outputs, receiving their inputs from ipsilateral CN. The fifth ABEP component (P5) is generated by structures central to the SOCs and their immediate outputs. Neither focal neuronal nor myelin lesions were sufficient to produce obliteration of any component, consistent with a set of generators for each of the ABEP components, consisting of both cell bodies and their output fibers, that is distributed spatially in the brainstem.

Three modality evoked potentials in Charcot-Marie-Tooth disease (HMSN-1).

Sixteen patients with dominant hereditary motor-sensory neuropathy type I (HMSN I), members of 5 families, underwent trimodality evoked potential studies. All patients had clinically normal optic nerves. History of deafness was present in 3 patients and sensory-neural hearing defect was found in 5 of 7 patients in whom audiometry was obtained. In 43.7 percent of the subjects significant prolongation of P100 of the VEP was found. Prolongation of N19 of the SEP was found in all 12 subjects examined. Significant bilateral prolongation of peak I of the ABEP was found in 37.5 percent of the subjects and in 50 percent of the ears examined: these findings indicated that in addition to peripheral nerves, the myelin of the optic and cochlear nerves is also affected in HMSN type I.

Three-channel Lissajous' trajectories of auditory brainstem evoked potentials: contribution of fast and slow components to planar segment formation.

Three-Channel Lissajous' Trajectories (3CLT) of Auditory Brainstem Evoked Potentials (ABEP) to clicks were obtained after finite impulse response filtering in three frequency bands. These bands were chosen to replicate the widely used passband (100-3000 Hz) and to selectively enhance the definition of the 'pedestal' (10-240 Hz) or the first, third and fifth components (240-483 Hz). Quantitative measures of 3CLT were calculated to describe apex latencies, planar segment orientations, durations, trajectory amplitude peaks and their latencies. In addition, dipole moments at the latencies of apical points along 3-CLT were calculated. The planarity of ABEP 3-CLT segments persisted after selective enhancement of the 'pedestal' or the first, third and fifth components. These results rule out the suggestion that planarity of ABEP segments results from the interaction of the 'pedestal' with the superimposed faster components. These results demonstrate summation of 3-CLT planar segments ('a' 'c' and 'e' with the 'pedestal') to form new segments (wide-band 'a', 'c' and 'e'). With the exception of 'c', planar segments and the equivalent dipole moments associated with apexes did not change orientations across passbands. The effects of passband on the orientation of planar segment 'c' and the dipole moment of its apex are explained by its superimposition on the 'pedestal' in the wide-band records. A similar analysis of ABEP to clicks as compared to low-frequency stimuli (high-pass masked clicks) revealed no change in planarity nor in plane parameters. These results are compatible with the suggestion that the generators of the first, third and fifth ABEP components are curved fiber tracts. The planarity of the slow 'pedestal' may be due to the summation of slow synaptic potentials in auditory brainstem nuclei. These findings indicate that the generators of ABEP are composites that may be separated by selective lesion studies.

Effects of click polarity on auditory brain-stem potentials: a three-channel Lissajous' trajectory study.

Three-channel Lissajous' trajectories (3CLTs) of the auditory brain-stem evoked potentials (ABEP) were recorded from 16 adult subjects (28 ears) in response to rarefaction (R), condensation (C) and alternating polarity (A) clicks. 3CLTs were analysed and described in terms of their geometrical measures. All 3CLTs included 5 planar segments (named 'a', 'b', 'c', 'd' and 'e') whose latencies, durations, orientations, sizes and shapes were not affected by click polarity. Occasionally, planar segment 'd' was not defined, and its absence was parallelled by the absence of peak IV in the Vertex-Mastoid records. A small (0.03 microV), but significant increase was found in the trajectory amplitude of planar segment 'e' in C clicks. The effects of click polarity on 3CLT observed in this study suggest that some previously described ABEP changes with click polarity were the result of interactions between electrode positions and relative contributions of overlapping generators. The effects on the fourth and fifth ABEP components may be the results of changes in the temporal overlap of the activity of their generators.

Auditory brainstem evoked potentials peak identification by finite impulse response digital filters.

Linear phase finite impulse response (FIR) filtering can be used to differentiate auditory brainstem evoked potentials (ABEP) components. The power spectrum of ABEP at high intensities indicates that they contain 3 frequency bands that can be distinguished by applying appropriate digital filters with the following characteristics: up to 240 Hz (revealing slow components), 240-483 Hz (resulting in medium components) and above 500 Hz (leaving only fast components). The results using these filters, indicate that the medium components coincide with peaks I, III and V and that the slow filter results in a 'pedestal' whose peak coincides with peak V. These findings were used for automatic identification of ABEP peaks. A coincidence of the 'pedestal' peak with a medium component was sought and labelled peak V. The preceding medium peaks were labelled, in order of decreasing latency, III and I. Validation of this procedure was conducted on ABEP from normal subjects, using different stimulus rates and intensities, as well as from selected neurological patients with lesions affecting the brainstem. Provided the waveform included a 'pedestal', the results proved this procedure to be reliable and in very good agreement with manual identification and measurement of ABEP peaks.

Application of the three-channel lissajous trajectory of auditory brainstem-evoked potentials to the question of generators.

In studies of generators of auditory brainstem-evoked potentials (ABEP), depth recordings and clinical correlates may not reflect all the activity involved. Although the 'inverse problem' is mathematically ill-posed, a synthesis of physico-anatomical data with a three-channel Lissajous trajectory (3CLT) may indicate the actual number and identity of generators. A 3CLT of ABEP in three-dimensional voltage space, is isomorphic with the vector tip representing the momentary centrally located dipole in a uniform, spherical volume conductor. This isomorphism enables approximations of ABEP dipole equivalents to be made. 3CLT summarizes the surface activity in a single plot. It accentuates correlations within the orthogonal records, such as planarity of segments, allowing segmentation of the analysis epoch into components. These properties of 3CLT simplify the analysis of parametric effects on the surface distribution of ABEP components. The results of applying this technique indicate the experimental conditions under which the assumptions for the approximation of generators are closest to being fulfilled. Of all the points along the trajectory, apices (points with maximum absolute amplitude and curvature) have the most unique generators. The 3CLT to low-intensity and high-rate clicks, indicate more unique generators than the customarily used high-intensity slow-rate clicks, in which considerable temporal overlap of generator activity exists.

Three-channel Lissajous trajectories of auditory brainstem-evoked potentials in patients with neurological lesions affecting the brainstem: preliminary impressions.

Three-channel Lissajous trajectories (3CLT), as well as vertex-mastoid records of auditory brainstem-evoked potentials (ABEP) were obtained from 12 patients with neurological lesions localized to the auditory nerve, the medulla, the pons, the pons and midbrain and the midbrain. Patients selected had well-localized lesions at the levels of the presumed generators of ABEP (10 patients), as well as below (1 patient) and above (1 patient) those levels. The 3CLT measures used were trajectory amplitude, peak latency and amplitude, planar segment number, apex latency, duration, orientation in voltage space, as well as size and shape. Latencies and amplitudes of vertex-positive peaks I-V, as well as interpeak latency differences, were determined. Vertex-mastoid ABEP and 3CLT were compared in terms of their sensitivity to the existence of a lesion, the accuracy of lesion localization as well as their specificity to the lesion. 3CLT had an advantage in all three comparisons, with the most prominent advantage in specificity, which was over twice as high as that of single-channel ABEP. The results of this preliminary study indicate that 3CLT affords a sensitive analysis of ABEP, which is superior to single-channel analysis in sensitivity, accuracy and specificity. These results also indicate that the 'contralateral effect' of peak V latency prolongation in cerebellopontine angle tumors may be related to anatomical changes of its generators' orientation, rather than slowed conduction along the upper brainstem.

Three-channel Lissajous' trajectory of the human short latency visual evoked potentials.

Three orthogonally derived voltage-time records, when plotted simultaneously on three-dimensional voltage-voltage-voltage coordinates, produce a 3-Channel Lissajous' Trajectory (3CLT). 3CLTs of short latency visual evoked potentials (SVEP) were obtained for 8 adult humans (16 eyes) in response to monocular flash stimulation. Intersubject variability of 3CLT of SVEP was small enough to enable identification along the trajectory of comparable planar-segments of approx. 3-5 ms duration across subjects. In each planar-segment, the point at which the trajectory exhibited marked bending was noted. These points were called apices and they corresponded to maxima in the trajectory's distance from the origin (Trajectory Amplitude). Intersubject variability in apex latencies was comparable to, or smaller than, peak latency variability of single-channel records. 3CLT of SVEP consisted of 8 planar-segments in each of the latency ranges of 0-40, 40-70 and 70-100 ms. Analysis of 3CLT, combining the criteria of segment planarity, apices and trajectory amplitude peaks, enabled differentiation between components that had been considered the same in surface distribution studies. 3CLT of SVEP seemed to be influenced by the direction of propagation along the visual pathway.

Three-channel Lissajous' trajectory of human auditory brain-stem evoked potentials. III. Effects of click rate.

Three-channel Lissajous' trajectories (3-CLT) of the human auditory brain-stem evoked potentials (ABEPs) were recorded from 14 adult subjects using click rates of 10, 55 and 80/sec. The 3-CLTs were analysed and described in terms of their constituent planar segments and their trajectory amplitudes at each stimulus rate. Increasing stimulus rate resulted in an increase of planar segment duration which was more pronounced for segments 'a' and 'e', an increase in apex latency which was more pronounced the later the component and a decrease in planar segment size and peak trajectory amplitude which was more pronounced the earlier the component. These findings support the involvement of synaptic efficacy changes in the effects of stimulus rate on ABEP. The results are explained by overlapping convergence and divergence in the ascending auditory pathway. These results support the notion that the principal generator of each component is activated by the principal generator of the previous component, with some temporal overlap of their activities. Such temporal overlap may be minimized by using low intensity high rate stimuli.

Somatosensory evoked potentials to muscle percussion in humans: upper extremity.

Cortical as well as presumably subcortical potentials were recorded from 20 normal subjects in response to mechanical percussion of the thenar eminence. Nuchal-to-frontal recordings detected an initial negative component at 21 msec, followed by positivities at 29 and 36 msec. C-prime to frontal recordings produced prominent positivities at 29 and 44 msec. Recordings over the peripheral nerve did not reveal consistent potentials. Control experiments indicate that potentials are initiated by fast-adapting muscle receptors. If additional control experiments continue to support these findings, yet another constituent of electrically evoked somatosensory potentials--which may prove useful in the definition of muscle innervation disorders--will have been identified.