Effects of ELF and static magnetic fields on calcium oscillations in islets of Langerhans.

Several experimental studies have produced contradictory results on the effects of extremely low frequency (ELF) magnetic fields on cellular processes involving calcium ions. Furthermore, the few positive results have not been independently replicated. In most of these studies, isolated cells were used. Our study used mouse islets of Langerhans, in which very regular oscillations of calcium concentration can be observed at length. These oscillations are sustained by processes that imply energetic and inter-intracellular communication. Various magnetic fields were applied, either sinusoidal at different frequencies (50 Hz or multiples of the natural oscillation frequency) at 0.1 or 1 mT or static at 1 mT. Islets were also exposed to "cyclotron resonance" conditions. There was neither alteration of the fundamental oscillation frequency nor the degree of organisation under all exposure conditions. Using this sensitive model, we could not show new evidence of alterations of calcium processes under exposure to various magnetic fields.

Safety of the magnetic field generated by a neuronal magnetic stimulator: evaluation of possible mutagenic effects.

OBJECTIVE: The possible mutagenicity of a magnetic stimulus was checked using the Ames test with Salmonella typhimurium TA98 and TA100 as tester strains. METHODS: Samples of these bacteria were exposed to a pulsed magnetic field, on the order of 1 T. The magnetic pulses were generated by a neuronal magnetic stimulator with a flat coil. The magnetic stimulus was a continuous sequence of slightly damped half sinusoids at a rate of 5 pulses/s. Exposure times were 2-5 and 15 min. Exposure position was such as to maximise the magnetic field and minimise the induced electric field. Room temperature was maintained at 28.5 +/- 0.5 degrees C and the temperature was measured inside the samples. RESULTS: None of the exposure conditions showed any increase in mutation in either of the two bacterial strains. CONCLUSIONS: These results are discussed in comparison with effects found in the literature. The magnetic stimulation used under the conditions of this study does not appear to have mutagenic effects. This does not apply to cases where both strong electric and magnetic fields are present.

Changes in electrovestibular brainstem responses after aminoglycoside intoxication in guinea pigs.

OBJECTIVE: To verify the usefulness of short-latency vestibular responses evoked by a combination of round window electrical stimulation and sinusoidal rotation (electrovestibular brainstem responses; EVBRs) as a new monitoring tool of the vestibular function in animal experiments. METHODS: EVBRs were obtained before, during, and after treatment with aminoglycosides, along with compound action potential (CAP) audiograms. The changes in EVBRs were compared with morphological changes observed by scanning electron microscopy. RESULTS: EVBR amplitudes did not change in the group of guinea pigs treated with amikacin, but markedly decreased in the streptomycin and gentamicin- treated groups. CAP audiograms indicated a significant threshold elevation in the amikacin group, a moderate elevation in the gentamicin group, and no change in the streptomycin group. Under scanning electron microscopy, the loss of the sensory hair cells observed in the cristae ampullares was slight to moderate in the amikacin group, moderate to severe in the streptomycin group, and severe in the gentamicin group. CONCLUSION: EVBRs reflect overall pathological changes undergone by vestibular hair cells, and support the vestibular specificity of EVBRs.

Rate influences on tone burst summating potential amplitude in electrocochleography: clinical(a) and experimental(b) data.

Electrocochleographic recordings of action and summating potentials are widely used in the electrophysiological assessment of endolymphatic hydrops (ELH). Increased amplitudes of the summating potential (SP) in response to tone burst stimuli are indicative of positive ELH. This study reports the effect of repetition rate of tone burst stimulation on the SP amplitude. Using transtympanic electrocochleography (ECochG), the SP in response to 1 kHz tone bursts was recorded in both a Ménière and a non-Ménière population. Absolute values of the SP were systematically higher in the Ménière group. Moreover, in the Ménière and non-Ménière groups, the response amplitudes of the SP at a repetition rate of 8.4 tone bursts/s were only 66 and 32%, respectively, of the maximal response amplitude which was obtained at the rate of 37.4 tone bursts/s. Additionally, in normal guinea pigs chronically implanted with a round window electrode, the SP was recorded to 0.5-16 kHz tone burst stimulations presented at 100 dB SPL with the same different repetition rates. Similar enhancement of the SP amplitude was observed from 8.4 to 37.4 stimuli/s, whatever the frequency. This effect is interpreted as an increased asymmetry of vibration of the cochlear partition, whose mechanical operating point would not return to the normal resting position at high repetition rates, since it is permanently shifted in ELH.

Discharge rate of the auditory nerve during noise revealed by electrocochlear stimulation.

The purpose of this study was to evaluate the average discharge rate of all fibres in the whole auditory nerve (R(wn)) when a broad-band noise with steady-state effects is applied to the ear. We assessed the R(wn) parameter by detecting the state of refractoriness of the nerve during noise stimulation using an electric stimulus (ES) as a probe. The technique, applied in awake pre-implanted guinea pigs (Charlet de Sauvage et al., 1994), made it possible to obtain electro-acoustic responses (EARs), from which an estimate of the R(wn) parameter could be deduced. Negative current pulses of 100 micros duration, each followed by an identical pulse of positive polarity for charge balance, were applied between round window and indifferent vertex electrodes at intervals of 160 ms. The 120 ms wide-band noise masker started 92 ms before every other negative ES. The signal on the stimulating electrodes was averaged over a 5.12 ms window in synchrony with the negative pulse. EARs were obtained by alternately subtracting recordings during noise from those during silence. The R(wn) parameter was determined by comparing experimental and computed EAR patterns. For this purpose, a model of unit response incorporating changes in amplitude and conduction velocity during the relative refractory period was designed. The recovery function of the firing probability in response to ES was evaluated. Fibres were classified in different categories according to their background discharge rates. The probability of response of single fibres to ES in each category was calculated on the basis of their interval histograms during silence and noise. Individual spikes were combined accordingly to obtain the computed EAR waveform. R(wn) was determined by adjusting the EAR amplitude of the model in relation to that of the experimental EAR. R(wn) generally increases in a linear fashion with respect to noise intensity expressed in dB, thus following the increase in loudness perception estimated by Weber's law. At the highest noise levels, R(wn) tends to saturate. The estimated saturation rate was found to be about 380 spikes/s.

Bioelectrical cochlear noise and its contralateral suppression: relation to background activity of the eighth nerve and effects of sedation and anesthesia.

The bioelectrical activity of the cochlea, without any ipsilateral acoustic stimulation, was recorded in awake guinea pigs (GPs) between electrodes chronically implanted at the round window (RW) and the skull. Measuring its power in the band centered around 1.0 kHz (0.5-2.5 kHz) provided an indirect measure of the ensemble background (EBA) activity of the eighth nerve. Contralateral white-noise (CLWN) stimulation reduced this EBA, presumably by activation of medial olivocochlear fibers. The aim of the investigation was to validate measurements of EBA and of its contralateral suppression in order to study the medial efferent function. The first goal was to find the best conditions for recording the EBA in the absence of ipsilateral stimulation and for studying its suppression by contralateral acoustic stimulation, which implies that no noise was generated by the experimental animal. Thus recordings were compared in normal, awake GPs and in GPs under sedation with xylazine, anesthetized with a combination of xylazine and ketamine, and with and without temperature regulation. In order to monitor the effects of sedation and anesthesia, the recordings were analyzed not only in the 0.5- to 2.5-kHz frequency band but also in the other frequency bands, 5-50 Hz, 50-150 Hz, and 150-500 Hz, which presumably include general central and neuromuscular contributions. The results show that sedation with xylazine accompanied by regulation of body temperature does not affect the EBA value nor its contralateral suppression. Nevertheless, anesthesia should be avoided, even with control of body temperature. The second goal of this study was to identify the specific cochlear contribution to the raw RW signal. Thus recordings were performed in normal and deafened animals and analyzed in the frequency band 0.5-2.5 kHz and also in the other frequency bands of 5-50 Hz, 50-150 Hz, and 150-500 Hz. The results indicate that most of the cochlear activity lies in the frequency band 0.5-2.5 kHz, with also some minor contribution coming from the 150- to 500-Hz band. Analysis and comparison of power values in the different conditions indicate that specific cochlear EBA power was about 60 microV2. From a commonly accepted mean background discharge rate of 50 spikes/s (sp/s), the EBA power without CLWN should have been around 4.4 microV2 if the fibers' activity was random. This difference suggests that there is probably some degree of synchrony between individual fibers. There was a reduction of approximately 45% during CLWN stimulation. This suppression might correspond to a reduction in both discharge rate and synchrony of the fibers.

Electrical and physiological changes during short-term and chronic electrical stimulation of the normal cochlea.

In the electrical stimulation (ES) of auditory pathways, the type of stimulus and the electrode/tissue interface are critical parameters for the safety and efficacy of the protocol. In this study the influence of alternate pulses, applied between round window and vertex electrodes in chronically implanted guinea pigs, and maintained during 1 and 25 daily periods of 2 h (short-term and long-term experiments, respectively), was investigated. ES consisted of monophasic current pulses of +/- 70 microA and 300 (micro)s in duration at a rate of 167/s, with alternate polarity. Compound Action Potential (CAP) audiograms, amplitudes and latencies of click-evoked CAPs, amplitudes and latencies of electrically-evoked auditory responses (EARs), and electrode impedances, were measured periodically outside or during the ES periods. Short-term ES induced no change in CAP thresholds, amplitude and latency in response to clicks at 80 dB above normal threshold, but induced a slight latency increase and amplitude decrease of the EAR, correlated with an exponential decay of the electrode impedance. On a long-term basis, CAP audiograms and latencies did not change significantly, whereas CAP amplitudes and electrode impedances increased, in correlation with each other. In control guinea pigs receiving no ES, the same CAP amplitude and impedance changes were observed over the same long-term period. The EAR and CAP changes can be explained by a variation of the electrical impedance of the electrode/tissue interface. This is possibly due to a change in electrolytes around the electrode under the influence of the ES for the short-term variation, and to an electrode encapsulation by fibrous tissue independent of the ES for the long-term change. In itself, and under the conditions of this experiment, the ES demonstrated no adverse effects on the auditory function and can be safely used for inner-ear exploration.

Changes in CM and CAP with sedation and temperature in the guinea pig: facts and interpretation.

The influence of xylazine on the amplitude, latency and waveform of VIIIth nerve compound action potential (CAP) and cochlear microphonic (CM) in response to clicks at 95 dB SPL in normal awake preimplanted guinea pigs was investigated. The animals' temperature was monitored but no thermoregulation was exerted, except in one control experiment. Following a 0.2 ml injection of xylazine, CM showed minor variations while CAP audiograms for tone pips between 0.5 and 25 kHz remained normal. However, a progressive decrease in temperature and a strongly correlated increase in CAP amplitude and in N1 and N2 latencies were noticed. For peak N1 the changes were equivalent to linear amplitude and time expansions, and could be reproduced through CAP synthesis with convolution methods using time expanded unit response model and firing density functions. All changes were maximal after 2 h of sedation and recovered within approximately another 2 h. Whereas xylazine is known to induce hypothermia, all the changes disappeared if the animal was thermoregulated. Therefore the changes are interpreted as a result of hypothermia. The mechanism of N1 latency lengthening and increase in amplitude during hypothermia can be understood as a simultaneous increase in spike duration, hair cell/nerve synaptic delay and postsynaptic time constant. This hypothesis yielded a theoretical temperature coefficient for N1 latency (-52 microseconds/degree C) matching that measured experimentally (-55 microseconds/degree C). When compared with peak N1, peak N2 appeared relatively more expanded. Arguments about the origin of N2 are discussed.

Differential sensitivity to rotation measured on potentials evoked by electrical stimulation of the guinea-pig ear.

Responses to electrical stimulation of the ear applied between round-window and vertex electrodes were recorded in awake guinea-pigs from the same electrodes or from separate vertex/mastoid subdermal needle electrodes. They were averaged during opposite phases of sinusoidal rotation or before and after constant velocity rotation. In both cases the responses were subtracted from each other and yielded differential per- or post-rotatory "electrovestibular" responses. For comparison, responses were also recorded in the same animals and conditions of electrical stimulation during silence and during silence and during presentation of a broad-band noise. The difference yielded "electroacoustic" responses. In round-window records, electrovestibular and electroacoustic responses presented typical compound nerve action potential patterns. Electrovestibular responses could be recorded for head angular velocities as low as 3 degrees sec-1 at 0.1 Hz. Response amplitude showed a logarithmic relation to head velocity. Changes in amplitude, as a function of time after rotation, were comparable to those reported for vestibular nerve fibre responses. In vertex/mastoid records, electroacoustic responses presented a sequence of peaks similar to the click-evoked auditory brain-stem responses, and electrovestibular responses presented two peaks, presumably representing contributions of central vestibular structures. Such "electrovestibulography" permits the study of an individual ear and makes available to the investigator a large range of vestibular stimulation conditions.

[Implants for the middle ear, development of a human prototype]. / Implants d'oreille moyenne, développement d'un prototype humain.

Middle ear implants are a new type of hearing aid. Their principle is direct stimulation of the inner ear fluids, by a vibrator implanted in the middle ear. Two technologies are developed by different teams: electromagnetic vibrators, and piezoelectric vibrators. Middle ear implants should be partially implantable (P-MEI), or totally implantable (T-MEI). Authors develop, concurrently with animal experimentation an in vitro method to measure the performance date of the vibrators. The specifications of a future P-MEI have been defined, and vibrators tested with this method. First results encourage to develop this project.

[Functional study of vestibular receptors by combined electric and rotatory stimulations]. / Exploration fonctionnelle des récepteurs vestibulaires par stimulations électriques et rotatoires combinées.

After a brief review of the anatomical and functional characteristic of the inner ear, emphasizing audio-vestibular relationships, a direct method for recording peripheral vestibular potentials, the Electrovestibulogram (EGV) is described and compared with the techniques Electronystagmographic. Stimulating electrodes were implanted in the round window and vertex of a group of guinea-pigs, with recording electrodes near the internal acoustic meatus. A permanent electric stimulation was combined with an intermittent vestibular stimulation (horizontal angular acceleration) to derive afterfact-free vestibular specific responses. The characteristics of the EVG responses were analyzed and compared with the data in the literature concerning the responses of single fibres, to validate the technique. The diagnostic value of EGV is discussed.

Electrovestibulogram: first results in the guinea pig.

This paper describes a method of investigation of the peripheral vestibular system. Electrical stimulations (ES) are applied on the round window of chronically implanted guinea pigs, with and without vestibular stimulation (either per or post rotational accelerations). The whole nerve action potential recorded at the output of the internal auditory meatus, the difference between the two conditions, reveals the change in electrical excitability and thus presumably in discharge rate of vestibular fibres determined by rotational stimuli. This electrical vestibular action potential (EVAP) presents as a mainly monophasic potential with a peak latency to ES of about 0.3 ms. Right and left accelerations versus rest are shown to give responses with opposite polarity, reflecting the inhibitory and excitatory influences of these opposite accelerations, whereas the transfer function of EVAP versus acceleration amplitude appears roughly linear. These observations appear altogether coherent with published mechano-physiological data about individual discharge rates of canal fibres. Also relative amplitudes of acoustical and vestibular responses are in agreement with related number of fibres in the two systems. This EVAP could be the basis of a quantitative evaluation method for the vestibular system, the electrovestibulogram (EVG).

Clinical significance of the summating potential in Menière's disease.

Transtympanic (TT) electrocochleography (ECochG) data recorded with click stimuli and tone bursts (1, 2, 4, and 8 kHz) were evaluated in 50 patients with Meniere's disease and compared with the data from control groups of 10 sensorineural hearing impaired patients and 5 subjects with normal hearing. The mean summating potential (SP) amplitude was larger in the Meniere's disease group for 1, 2, and 8 kHz. The low frequency (1 or 2 kHz) SP decreased in 59% of the Meniere's disease patients during a glycerol dehydration test, whereas subjective hearing improved in only 29%. The use of ECochG with the monitoring of SP thus improves the sensitivity of the glycerol test for the detection of endolymphatic hydrops. There appeared no clear relation between clinical observations and the ECochG data either before or after glycerol. However, the Meniere's disease patients with a large negative SP at low frequencies also had larger action potential (AP) and often had short-term symptoms. The present report confirms the usefulness of measuring the SP in the diagnosis of Meniere's disease.

Mathematical analysis of VIIIth nerve cap with a linearly-fitted experimental unit response.

The compound action potential (CAP) recorded at the round window has for some time been investigated in order to determine the underlying temporal spike sequence: the firing probability density (FPD) function of the fibres. Charlet de Sauvage et al. [(1980) Hear. Res. 2, 343-346; (1983) J. Acoust. Soc. Am. 73, 616-627] have previously described a close estimate of the far-field unit response (UR) by presenting to the ear a transient electrical stimulation combined with an acoustical masking noise. A linear model of UR, free of noise or extrinsic potentials and based on both experimental data and on the dipole principle of Teas et al. [(1962) J. Acoust. Soc. Am. 34, 1438-1459], was proposed [(1985) Hear. Res. 18, 121-125]. Using the UR model, deconvolutions were performed on CAPs recorded in various conditions of stimulation and of cochlear pathologies in the guinea pig. The principal findings were: (1) a plateau was observed in the amplitude function around 60-70 dB above normal threshold both for click-evoked CAPs and for the corresponding FPDs; (2) FPDs resulting from the deconvolution of high-frequency derived responses showed 2 main peaks with a 0.7 ms interval followed by a second homologous pair with lower amplitude. Convolution computations confirmed the need for the second set with peaks 2 and 3 being only 0.3 ms apart. These findings are interpreted in terms of mechanical excitation pattern.

Investigation of cochlear mechanisms using combined acoustical and electrical stimulations.

Electrically-evoked VIIIth nerve CAP responses evoked by electrical stimulation and derived by masking with sounds are recorded in the guinea pig, using different masking paradigms (pure tones and high-pass filtered white noise). CAP frequency threshold curves correlate with threshold curves determined by other means, in normal as well as in pathological guinea pigs. Amplitudes of the responses as a function of pure tones frequency and intensity are also measured. Such measurements provide an estimate of the number of fibers activated by the pure tones. These responses can be further analysed using masking with high-pass filtered white noise.

Origins of eighth nerve unit response pattern in round window cap recordings.

An experimental technique based on a combined electrical and acoustical stimulation of the cochlea (R. Charlet de Sauvage et al., 1983, J. Acoust. Soc. Am. 73, 616-627) allowing to record a close estimation of single unit contribution to the VIIIth nerve CAP has been recently proposed. D.C. Teas et al.'s (1962, J. Acoust. Soc. Am. 24, 1431-1459) theory about this pattern is that, due to conduction time in the internal auditory meatus, the propagated depolarisation partly differentiates in far-field recordings, giving rise to a specifically diphasic pattern. In order to evaluate Teas's hypothesis, 83 unit waveforms collected in 10 guinea pigs are analysed. Several reproducible peaks are identified. Latency and slope measurements are performed on these peaks. This data is processed, after identifying two homologous components which could combine in accordance with Teas's theory. The schematic pattern of a primary waveform is actually determined. Its relative amplitude on the two electrodes and the delay in the meatus are inferred. Results are in good agreement with published data. This is taken as an indirect validation of Teas's hypothesis.