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The evolution of the auditory midlatency response in evaluating unconscious memory formation during general anesthesia.

The inability to objectively evaluate the amnesic status of an anesthetized patient has been a perplexing problem for the anesthesia provider. One approach thought to be effective in evaluating the amnesic status of the anesthetized patient is the auditory midlatency response (AMLR). The AMLR is an electrophysiological response that is recorded from scalp electrodes 10 to 80 ms after the auditory pathways begin to process acoustic stimuli. The response is thought to reflect the anesthetized patient's ability to consolidate an acoustic stimuli into an explicit or implicit memory. This article defines the amnesic state, describes the various components of AMLR, discusses clinical uses of the AMLR as an indicator of memory formation during the anesthetized state, and explains the clinical implications of using the AMLR in the surgical suite. Recent results have noted that the Pa waveform, the first positive deflection of the AMLR, may be the component that may serve as an intraoperative indicator of the anesthetized patient's ability to potentially consolidate an intraoperative acoustic stimuli into a memory. With the establishment of the Pa waveform of the AMLR as a reliable indicator of intraoperative memory formation, the AMLR can then be used to significantly decrease the occurrences of traumatic neurosis in the surgical patient and subsequent medicolegal consequences for the health care team. Thus, the use of the AMLR strives to promote a safer intraoperative environment for both the patient and the anesthesia provider.

Anesthesia, General↗

Leupeptin, a calpain inhibitor, protects inner ear hair cells from aminoglycoside ototoxicity.

Inner ear hair cells play a major role in the auditory pathway that converts sound stimulation into electrical signals, and then into a neural code. However this function is often lost by aminoglycoside ototoxicity. The injury of inner ear hair cells from aminoglycoside treatment is considered apoptosis, and caspase is an important participant in the apoptosis pathway in many organs. It has been reported that calpain, a calcium-dependent protease, is essential for mediation and promotion of cell death. The purpose of the present study was to investigate effects of caspase and calpain inhibitors on the inner ear hair cells after aminoglycoside treatment, and to explore the cell death pathway. Cochlea explant cultures were prepared from mice of postnatal 6 days, cultured with neomycin and/or protease inhibitors, and then stained with phalloidin-fluorescein isothiocyanate (phalloidin-FITC), which was used as a marker to identify surviving hair cells. We demonstrated that neomycin (0.1-1 mM) reduced the number of outer hair cells in a dose-dependent manner. Furthermore, we showed that leupeptin, a calpain inhibitor, significantly protects against the neomycin-induced loss of outer hair cells, whereas a caspase inhibitor was effective only against a lower concentration of neomycin (0.2 mM). Using the TdT-mediated dUTP-biotin nick and labeling method, we also found that a calpain inhibitor, but not a caspase inhibitor, prevents apoptotic DNA fragmentation after treatment with 1 mM neomycin. These results suggest that calpain, rather than caspase, may be responsible for apoptosis induced by aminoglycoside. Thus, leupeptin may prevent hearing loss from aminoglycoside ototoxity.

Animals↗

Auditory evoked potentials from auditory cortex, medial geniculate nucleus, and inferior colliculus during sleep-wake states and spike-wave discharges in the WAG/Rij rat.

OBJECTIVE: Click auditory evoked potentials (AEP) were simultaneously recorded from the auditory cortex (ACx), the medial geniculate nucleus (MGN), and the inferior colliculus (IC) in the freely moving WAG/Rij rat, to investigate state-dependent changes of the AEP in different anatomical locations along the auditory pathway. METHODS: AEPs obtained during active (AW) and passive wakefulness (PW), slow wave sleep (SWS), rapid-eye-movement sleep (REM) and generalized spike-wave discharges (SWD; a specific trait of the WAG/Rij rat, a genetic model for absence epilepsy), were compared. RESULTS: The early components in ACx, MGN and IC were stable throughout the sleep-wake cycle and SWD, apart from a slight increase in the IC during SWD. At all three locations a prominent enlargement of a later component (i.e., N32 in IC, N33 in MGN, and N44 in ACx) was found during SWS and SWD. CONCLUSIONS: The early AEP components are not modulated by the normal sleep-wake states, and are not impaired during SWD. A strong state-dependent modulation of a later AEP component occurs at all three anatomical locations investigated. This suggests that apart from the thalamic burst firing mode, additional mechanisms must exist for the enlargement of the AEP during EEG-synchronized states at the prethalamic and cortical level.

Action Potentials↗

Neuronal correlates of perceptual amplitude-modulation detection.

The goal of the present paper is to relate the coding of amplitude modulation (AM) in the auditory pathway to the behavioral detection performance. To address this issue, the detectability of AM was estimated by modelling a single neuron located in the central nucleus of the inferior colliculus (IC). The computational model is based on cochlear nucleus responses and a coincidence detection mechanism. The model replicated the main feature of the neuronal AM transfer function, namely a bandpass function. The IC-unit model was initially tuned to a 200-Hz modulation frequency. A single neurometric function for AM detection at this modulation frequency was generated using a 2-interval, 2-alternative forced-choice paradigm. On each trial of the experiments, AM was taken to be correctly detected by the model if the number of spikes in response to the modulated signal exceeded the number of spikes in an otherwise identical interval that contained an unmodulated signal. Psychometric functions for 4 human subjects were also measured under the same stimulus conditions. Comparison of the simulated neurometric and psychometric functions suggested that there was sufficient information in the rate response of an IC neuron well-tuned in the modulation-frequency domain to support behavioral detection performance.

Acoustic Stimulation↗

[Effects of lobectomy on the auditory mid-latency response in temporal lobe epileptic patients].

AIM: Auditory mid-latency response (MLR) has been studied in temporal lobe epileptic patients in order to evaluate the relationship between the different components of this evoked response and their probable generators in the most rostral part of auditory pathway. PATIENTS AND METHODS: MLR was studied in eight patients with temporal lobe epilepsy before lobectomy, and 1, 6 and 12 months after the surgical treatment, and eight healthy subjects matched for age and gender. Recording electrodes were placed in Cz, T3 and T4 according to the international 10-20 system. Comparisons were carried out in terms of peak to peak amplitude, latency and interpeak intervals of the main components (Mann-Whitney U test, Wilcoxon test). RESULTS: Epileptic patients showed statistically significant prolonged latency of all components, and increased Pa-Pb interpeak interval duration respect to control group. No significant variations were found in the amplitude of the studied components. There were no differences between left and right lobotomized patients. However, a marked tendency to the shortening of latency was observed in the left group after the surgical treatment. CONCLUSIONS: The differences between the pre-surgical records of patients and controls reflect an impaired activity in the generating areas for MLR components. There appears to be a trend to a different behaviour after right or left lobectomy, probably related to the extension of the removed tissue.

Adult↗

Acoustic feature extraction by cross-correlation in crickets?

Common concepts of acoustic feature extraction within the auditory pathway of vertebrates and insects assume temporal filters tuned to particular periodicities. Crickets respond selectively to the conspecific song pattern and reveal a bandpass characteristic, which is thought to arise from a matched filter for a restricted range of periods. Unexpectedly, females of the two sibling species Teleogryllus oceanicus and T. commodus differed in fundamental filter properties. While T. oceanicus revealed a period filter, T. commodus exhibited a pulse duration filter. This finding raises the question of how so distinct properties of homologous neuronal circuits for pattern analysis have evolved during speciation. Evidence is presented that signal analysis by cross-correlation offers a simple explanation for differences in pattern selectivity as well as for the evolutionary transition. By cross-correlation the similarity of an external pattern with an internal template is determined over a certain time window. A comparison of behavioural data and cross-correlation values suggested that both species have similar templates. However, time windows were significantly different between species (T. oceanicus: 180-400 ms, T. commodus: 90-160 ms). Consequently, solely a change in the evaluation time window is required to account for the observed differences in feature extraction that serve to maintain species isolation.

Acoustic Stimulation↗

The clinical utility of distortion-product otoacoustic emissions.

Otoacoustic emissions permit, for the first time, an unbiased means of examining the preneural elements of the peripheral auditory pathway that make the initial contribution to the perception of acoustic stimuli. Distortion-product otoacoustic emissions (DPOAEs) represent one type of evoked emission that has significant potential for becoming an important test in the audiometric evaluation of hearing capacity. In the present review, selected examples of several forms of sensorineural hearing loss demonstrate that DPOAEs have the ability to act as objective indicators of the frequency/level configuration of the conventional audiogram in cases in which hearing impairment results primarily from damage to the outer hair cells. In contrast, normal DPOAE functioning, in the presence of a significant hearing loss, indicates a locus of damage central to the region of the outer hair cells. Like the other emitted responses, DPOAEs can be measured noninvasively, are highly repeatable, under test-retest conditions, and are simple and rapid to detect using microcomputer-based instrumentation. Further, DPOAEs test both the "threshold" and suprathreshold levels of outer hair-cell activity in the form of response/growth functions, over a 30- to 40-dB stimulus range. In combination, these attributes indicate that DPOAEs can provide an objective and comprehensive assessment of the cochlear reserve of a given ear.

Audiometry, Evoked Response↗

[Early auditory evoked potentials in very small premature infants].

BACKGROUND: Brainstem auditory evoked response (BAER) is a sensitive test of the functional integrity of the auditory pathway. Latencies and amplitudes of measured waveforms reflect maturation of this brainstem pathway and may allow a prediction for neurological outcome. PATIENTS AND METHODS: BAER were measured prospectively in preterm infants with a birthweight less than 1500 g at 3 to 4 day following birth and during treatment on neonatal intensive care unit (NICU) every two weeks. Pre- and postnatal risk factors and frequency of apnea were evaluated. Outcome was scored after 1 year corrected age by neurological examination. RESULTS: There were no significant differences in latencies of waves between 5 children with severe asphyxia and/or IVH resulting in major neurological handicaps after 1 year. However, two of these children have bilateral abnormalities. The slope of decay of wave latencies decreased between 30 and 32 weeks gestational age, while the number of registered apnea increased. In follow up investigations during treatment on NICU there was a decrease in interpeak latencies wave V to wave III from 24/25 weeks gestational age to term infants and increase in amplitudes. CONCLUSION: Early performed BAER alone is not a good predictor for neurological outcome of preterm infants because of great interindividual variability of normal neonates and the anatomical site of cerebral lesions. The method contributes information on the brainstem maturation and may also reflect the different maturational stages of the respiratory system.

Asphyxia Neonatorum↗

Alteration of activator protein 1 DNA binding activity in gentamicin-induced hair cell degeneration.

Sensorineural hearing loss is often associated with damage of cochlear hair cells and/or of the neurons of the auditory pathway. This damage can result from a variety of causes, e.g. genetic disorders, aging, exposure to certain drugs such as aminoglycosides, infectious disease and intense sound overexposure. Intracellular events that mediate aspects of aminoglycoside-mediated damage to hair cells have been partially unraveled. Several independent research groups have demonstrated a crucial role of mitogen-activated protein kinase signaling in aminoglycoside-induced ototoxicity. Mitogen-activated protein kinases are important mediators of signal transduction from the cell surface to the nucleus. Jun N-terminal kinases, members of the mitogen-activated protein kinase family, are strongly activated in cell culture conditions by stress inducing stimuli, including ultraviolet light, heat shock and tumor necrosis factor; therefore they are also referred to as stress-activated protein kinases. In hair cells aminoglycoside treatment was shown to activate the Jun N-terminal kinase signaling pathway. Activation of Jun N-terminal kinase leads to phosphorylation and thereby activation of transcription factors and consequently to altered gene expression. There are many nuclear Jun N-terminal kinase substrates including c-Jun, ATF-2, and Elk-1 proteins. One of the downstream targets of Jun N-terminal kinase is the transcription factor activating protein-1. Activating protein-1 is a dimeric complex composed of members of the Fos and Jun proteins. A variety of different stimuli is known to induce activating protein-1 activity. Induction of activating protein-1 is thought to play a central role in reprogramming gene expression in response to external stimuli. In this study we have analyzed the effect of gentamicin treatment on the downstream targets of Jun N-terminal kinase. Our results demonstrate that gentamicin treatment of explants of organ of Corti results in increased activating protein-1 binding activity. The main component of these activating protein-1 complexes is the c-Fos protein. Moreover, we show that the activating protein-1 induction is transient and occurs exclusively in hair cells of rat organ of Corti explants.

Actins↗

Evidence of altered central nervous system development in infants with iron deficiency anemia at 6 mo: delayed maturation of auditory brainstem responses.

Iron deficiency anemia has long been thought to have effects on the central nervous system (CNS). Finding direct evidence of this in human infants, however, has been challenging. Auditory brainstem responses (ABRs) provide a noninvasive means of examining an aspect of the CNS that is rapidly maturing during the age period when iron deficiency is most common. ABRs represent the progressive activation of the auditory pathway from the acoustic nerve (wave I) to the lateral lemniscus (wave V). The central conduction time (CCT, or wave I-V interpeak latency) is considered an index of CNS development because myelination of nerve fibers and maturation of synaptic relays lead to an exponential reduction in the CCT from birth to 24 mo. In 55 otherwise healthy, 6-mo-old Chilean infants (29 with iron deficiency anemia and 26 nonanemic control infants), the CCT was longer in those who had been anemic at 6 mo, with differences becoming more pronounced at 12- and 18-mo follow-ups despite effective iron therapy. The pattern of results--differences in latencies but not amplitudes, more effects on the late ABR components (waves III and V), and longer CCTs (as an overall measure of nerve conduction velocity)--suggested altered myelination as a promising explanation, consistent with recent laboratory work documenting iron's essential role in myelin formation and maintenance. This study shows that iron deficiency anemia in 6-mo-old infants is associated with adverse effects on at least one aspect of CNS development and suggests the fruitfulness of studying other processes that are rapidly myelinating during the first 2 y of life.

Anemia, Iron-Deficiency↗

Towards a unifying basis of auditory thresholds: the effects of hearing loss on temporal integration reconsidered.

For signal detection and identification, the auditory system needs to integrate sound over time. It is frequently assumed that the quantity ultimately integrated is sound intensity and that the integrator is located centrally. However, we have recently shown that absolute thresholds are much better specified as the temporal integral of the pressure envelope than of intensity, and we proposed that the integrator resides in the auditory pathway's first synapse. We also suggested a physiologically plausible mechanism for its operation, which was ultimately derived from the specific rate of temporal integration, i.e., the decrease of threshold sound pressure levels with increasing duration. In listeners with sensorineural hearing losses, that rate seems reduced, but it is not fully understood why. Here we propose that in such listeners there may be an elevation in the baseline above which sound pressure is effective in driving the system, in addition to a reduction in sensitivity. We test this simple model using thresholds of cats to stimuli of differently shaped temporal envelopes and durations obtained before and after hearing loss. We show that thresholds, specified as the temporal integral of the effective pressure envelope, i.e., the envelope of the pressure exceeding the elevated baseline, behave almost exactly as the lower thresholds, specified as the temporal integral of the total pressure envelope before hearing loss. Thus, the mechanism of temporal integration is likely unchanged after hearing loss, but the effective portion of the stimulus is. Our model constitutes a successful alternative to the model currently favored to account for altered temporal integration in listeners with sensorineural hearing losses, viz., reduced peripheral compression. Our model does not seem to be at variance with physiological observations and it also qualitatively accounts for a number of phenomena observed in such listeners with suprathreshold stimuli.

Animals↗

[Circadian variation in the latency of auditory brainstem response].

The auditory brainstem response (ABR) has been found to reflect many pathological conditions within the auditory system and brainstem. And now, many neurosurgeons are using it to monitor the integrity of the auditory pathway during neurosurgical procedures. It is said that ABR shows little variation from person to person or laboratory to laboratory, nor is it easily affected by anesthesia, level of consciousness, fluctuation of blood pressure or hypoxemia. On the other hand, previous studies have shown that component waves of the ABR increase in latency and decrease in amplitude with lowered temperature. We reported here that naturally occurring circadian variations in body temperature were correlated with similar changes in the latency of the ABR. Tympanic temperature (Tty), deep forehead temperature (Thd) and ABR were recorded every 3 hours during a 24-hour period for a total of 8 recording sessions from each of 6 healthy persons (2 males and 4 females, mean age 24.3 years). The subjects were free to come and go during the day but slept overnight in the laboratory. All subjects had circadian variations in each temperature on the order of one degree. Thd had a tendency to fluctuate and its range of difference from Tty was -0.5-0.4 degree C. There was a more significant negative correlation between the latency of the ABR and Tty than that of Thd. It has become apparent that ABR latency is affected by small temperature changes such as circadian variation. The rate of a latency change in the ABR was 0.15msec per degree (C).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

[Effect of anesthesia on the intraoperative elicited stapedius reflex].

Evaluation of the intraoperative electrically evoked stapedius reflexes during cochlear implant surgery has two purposes: (1) The functioning of the device and the peripheral auditory pathways can be proven immediately and (2) The data of reflex threshold can be used for the prediction of later threshold (T)- and maximum comfortable (C) levels. This is especially useful for the later fitting of the speech processor in very young children. When trying to use the stapedius reflex data for the prediction of T- and C-levels it must be considered that during surgery the drugs used for general anaesthesia influence the values of the stapedius reflex. Depending on the drug, the stapedius reflex threshold can be increased or the reflex can even be totally blocked. This is not only due to relaxants but also to anaesthetics. To investigate this influence we first determined the acoustically evoked stapedius reflex threshold in normal hearing adults under general anaesthesia with 7 different anaesthetics without additional relaxation. It was found that especially with Dormicum and Brevimytal the stapedius threshold increased only very lightly or did not change at all. In a second investigation the electrically evoked stapedius reflex thresholds were obtained in adults who underwent a cochlear implant surgery using the same anaesthetics for anaesthesia. The intraoperatively evaluated reflex data were compared with postoperatively obtained data. It was found that both reflex thresholds were similar if evaluated in Dormicum or Brevimytal anaesthesia. This suggests that only with the use of the right anaesthetics the data of the electrically evoked stapedius reflex obtained intraoperatively can be used for predicting T- and C-levels.

Anesthesia, General↗

Temporal masking reveals properties of sound-evoked inhibition in duration-tuned neurons of the inferior colliculus.

The inferior colliculus (IC) is the first place in the central auditory pathway where duration-selective neurons are found. Previous neuropharmacological and electrophysiological studies have shown that they are created there and have led to a conceptual model in which excitatory and inhibitory inputs are offset in time so that the cell fires only when sound duration is such that onset- and offset-evoked excitation coincide; the response is suppressed by inhibition at other durations. We tested predictions from the model using paired tone stimulation and extracellular recording in the IC of the big brown bat, Eptesicus fuscus. Responses to a best duration (BD) tone were used as a probe to examine the strength and time course of inhibition activated by a nonexcitatory (NE) tone of the same frequency but differing in duration. As the relative time between the BD and NE tones was varied, the activity evoked by the BD tone was affected in ways comparable with backward, simultaneous, and forward masking. Responses to the BD tone were completely suppressed at short interstimulus intervals when the BD tone preceded the NE tone. Suppression was also seen when the stimuli temporally overlapped and summed and at intervals when the BD tone followed the NE tone. The results show that duration-selective neurons receive an onset-evoked, inhibitory input that precedes their excitatory input. The period of leading inhibition was correlated with BD and first spike latency. The results suggest how inhibition in the CNS could explain temporal masking phenomena, including backward masking.

Animals↗

[Prognostic evaluation of severe brain damage by auditory brainstem evoked potentials (BAEPs)].

Auditory brainstem evoked potentials (BAEPs) reflect activities of the brainstem auditory pathway and the influence of activities of structures surrounding the pathway. It is said that BAEPs are reliable in predicting the outcome of patients with severe brain damage. The author studied the relationship between initial BAEP findings, particularly interpeak latencies of waves I to V and waves III to V, and mortality as based on the cause of each disorder. BAEPs were sequentially recorded in 241 cases of various neurological diseases resulting in severe brain damage (GCS less than 8) during the first few days after onset. BAEPs were graded into four groups based on initial findings. Group A: all waves I to V were recorded (159 cases). Group B: waves IV & V absent (11 cases). Group C: either wave I or waves I & II only were recorded (28 cases), Group D: all waves absent. Prognosis in Group B & C was poor, only two patients surviving in a vegetative state. There were no survivors in Group D. The relationship in Group A between interpeak latencies (I-V ipl, III-V ipl) and prognosis is discussed. This group consisted of 41 cases subarachnoid hemorrhage, 29 cases hypertensive intracerebral hemorrhage, 5 cases severe cerebral infarction, 21 cases infratentorial cerebral vascular disease, 49 cases supratentorial severe head injury, and 14 cases infratentorial head injury. In deceased cases of subarachnoid hemorrhage and supratentorial head injury, there was significant prolongation of I-V ipl and III-V ipl over the mean latency of 20 normal subjects plus 2SD.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Hearing problems and hormonal disturbances in the elderly.

The most frequent hormonal diseases attracting audiological interest are hypothyroidism (myxodema) and diabetes mellitus. For many years these diseases have been considered to cause hearing disorders with the lesion located predominantly in the inner ear and central auditory pathways, resulting in sensorineural hearing loss. However, the causal relationship between hearing loss and hypothyroidism has been questioned, and thus a study was undertaken to elucidate this problem. A sample of hypothyroid patients (n = 15) with a median age of 76 years (range 61-92) underwent audiological examination before and after treatment with L-thyroxine (average 5 months; range 2-12), and re-examination (n = 13) after an observation period of average 40 months (range 32-46). No significant improvement in their hearing thresholds, speech reception thresholds or discrimination scores could be demonstrated, and the hearing ability in the hypothyroid patients did not differ significantly from that found in an age- and sex-matched population. In addition, histological investigation of the temporal bones from an 83-year-old woman with myxoedema showed no morphological changes or deposition of glycosaminoglycans. Audiological examinations in patients with insulin-dependent diabetes mellitus also show contradictory results. Therefore, the cochlear and retrocochlear hearing functions were evaluated in 20 patients with diabetic microangiopathy with a median age of 41 years (range 25-66), and in 19 patients without microangiopathy with a median age of 27 years (range 17-42). No significant differences in hearing thresholds or discrimination scores were present between the two diabetic groups, nor was any difference found between the diabetic patients and an age- and sex-matched population. In the patients with long-term insulin-dependent diabetes mellitus, brainstem audiometry revealed abnormal responses in 40%, indicating the presence of diabetic encephalopathy.

Aged↗

Auditory brainstem responses in thyroid diseases before and after therapy.

The aim of this study was to evaluate through the auditory brainstem responses (ABRs) the electrical events generated along the auditory pathway in 56 adult patients affected with hyper- and hypothyroidism. Twenty-four normal-hearing hyperthyroid patients affected with Graves' disease and 32 normal-hearing hypothyroid patients (9 with subclinical and 23 with overt hypothyroidism) were subjected to standard (clicks at 21 pps) and sensitized ABR with masking wide-band (masking noise). In addition, thyroid scintiscan and ultrasonography, free T3 and T4, total T3 and T4, TSH, antimicrosomal and antithyroglobulin antibodies, audiogram and impedance tests were performed in all the patients. This study was repeated after 6-12 months of treatment in conditions of euthyroidism. The results showed changes of ABRs both in the standard procedure as well as in the sensitized test in 6 hyperthyroid (25%) and 8 hypothyroid patients (25%). All the patients with abnormal ABRs had overt hypothyroidism (8/23; 34.7%). The ABRs became normal in 5 out of 6 Graves' patients after 6-12 months of methimazole treatment. ABRs remained abnormal in all the hypothyroid patients despite their having been on L-thyroxine treatment for 6-12 months and were euthyroid for at least 5 months before the study was repeated. These findings suggest that ABR abnormalities are indicative only of a nonspecific injury in the bulbo-ponto-mesencephalic centers. Alterations of ABRs in thyroid diseases are not specific in relation to hyper- or hypothyroidism. Lastly, there is a relationship between ABR abnormalities and the degree of hypothyroidism, even if ABR alterations are not always reversible after long-term therapy.

Adult↗

Brain stem auditory evoked responses: studies of waveform variations in 50 normal human subjects.

Brain stem auditory evoked responses (BAERs) were recorded in 50 normal adult subjects at various click rates. Attention was paid to absolute latencies, interwave latencies, interear interwave latencies, absolute amplitudes, and various amplitude ratios. The variability of waves VI and VII suggests that the clinical utility of these waves is restricted-their absence is not necessarily due to a CNS lesion. The wave IV-V complex appears with six different patterns. These variations must therefore be considered normal; none should be misconstrued as indicative of disease of the CNS. Repeated studies over a period of two to nine months showed no statistically significant changes in amplitude or latency measurements with the passage of time. Knowledge of these normal values and their variations, as a precondition for establishing criteria for abnormality, is essential to the interpretation of BAERs in clinical situations.

Acoustic Stimulation↗