Promotion of noise-induced cochlear injury by toluene and ethylbenzene in the rat.

Ethylbenzene + toluene are known individually to have ototoxic potential at high exposure levels and with prolonged exposure times generally of 4-16 weeks. Both ethylbenzene + toluene are minor constituents of JP-8 jet fuel; this fuel has recently been determined to promote susceptibility to noise-induced hearing loss. Therefore, the current study evaluates the ototoxic potential of combined exposure to ethylbenzene + toluene exposure in a ratio calculated from the average found in three laboratories. Rats received ethylbenzene + toluene by inhalation and half of them were subjected simultaneously to an octave band of noise (OBN) of 93-95 dB. Another group received only the noise exposure which was designed to produce a small, but permanent auditory impairment while an unexposed control group was also included. In two separate experiments, exposures occurred either repeatedly on 5 successive days for 1 week or for 5 days on 2 successive weeks to 4000 mg/m(3) total hydrocarbons for 6 h based upon initial pilot studies. The concentration of toluene was 400 ppm and the concentration of ethylbenzene was 660 ppm. Impairments in auditory function were assessed using distortion product otoacoustic emissions and compound action potential testing. Following completion of these tests, the organs of Corti were dissected to permit evaluation of hair cell loss. The uptake and elimination of the solvents was assessed by harvesting key organs at two time points following ethylbenzene + toluene exposure from additional rats not used for auditory testing. Similarly, glutathione (GSH) levels were measured in light of suggestions that oxidative stress might result from solvent-noise exposures. Ethylbenzene + toluene exposure by itself at 4000 mg/m(3) for 6 h did not impair cochlear function or yield a loss of hair cells. However, when combined with a 93-dB OBN exposure combined solvent + noise did yield a loss in auditory function and a clear potentiation of outer hair cell death that exceeded the loss produced by noise alone. No evidence was found for a loss in total GSH in lung, liver, or brain as a consequence of ethylbenzene + toluene exposure.

The role of deferoxamine in the prevention of gentamicin ototoxicity: a histological and audiological study in guinea pigs.

CONCLUSION: The addition of deferoxamine to gentamicin seems to confer partial functional and histological protection to the cochlea. OBJECTIVE: Aminoglycosides are known ototoxic agents. The toxicity occurs via an activation process involving the formation of an iron-gentamicin complex with free radical production. Iron chelation will supposedly limit this toxic effect. This study aimed to determine the possible cochleoprotective role of deferoxamine on the ototoxic effect of gentamicin. MATERIALS AND METHODS: Sixty healthy active guinea pigs, weighing 400-600 g, with an average age of 6 months were used. They were divided into three groups. Group 1 received intramuscular gentamicin 8 mg/kg/day, group 2 received gentamicin 8 mg/kg/day and deferoxamine 150 mg/kg twice daily for 19 days and group 3 served as a control. All animals had a baseline measurement of distortion product oto-acoustic emissions. At the end of 33 days they were submitted to another measurement and then the animals were sacrificed and their cochleas were examined histologically by light and transmission electron microscopy. RESULTS: In group 1 the mean amplitude post-injection ranged from 5.83 dB at 1001 Hz to 22.33 dB at 6348 Hz. In the deferoxamine + gentamicin group the mean amplitude post-injection ranged from 5.10 dB at 1001 Hz, to 24.45 dB at 6348 Hz. This was statistically significant. At 4004, 5042 and 6348 Hz group 2 showed less histological damage than group 1.

[Clinical and audiological findings in children with auditory neuropathy]. / Klinische und audiologische Befunde von Kindern mit auditorischer Neuropathie und ihre Versorgung mit einem Cochlea-Implantat.

BACKGROUND: Auditory neuropathy is a disorder characterised by preservation of outer hair cells function with normal otoacoustic emissions (OAEs), but with absent auditory brainstem responses (ABR). Perisynaptic synchronisation disorder is one of the possible pathogenesis underlying auditory neuropathy. In this paper we describe the clinical presentation and audiological findings in pediatric auditory neuropathy and its management. PATIENTS AND METHODS: 9 children with auditory neuropathy could be included in the study. An audiological evaluation was performed in all children including behavioural audiometry, measurement of the OAEs as well as electrocochleography (ECoG) and ABR recordings. Children who failed to get any benefit from conventional amplification received a cochlear implant. Prior to implantation the responses to electrical stimuli were examined with the promontory test and with the electrically evoked ABR. RESULTS: One child showed auditory neuropathy only on one side with normal hearing thresholds on the contralateral ear. Another child had normal hearing thresholds after the follow up period. Four children received a hearing aid. But variable hearing reactions were observed. Thus in three cases a CI is planned. In three children cochlea implantation was done. Following implantation a remarkable improvement in hearing/speech capabilities with the CI compared to conventional hearing aids were observed in all three cases. Beside, these three children developed open set speech discrimination and are using now oral language for communication. CONCLUSIONS: Auditory neuropathy is a disorder which presents with different clinical and audiological findings. Thus the management of this disorder must be an individual one. In light of our findings we support the use of cochlear implants as an option for children with auditory neuropathy in cases where conventional amplification does not work sufficiently.

Effects of intense noise exposure on the outer hair cell plasma membrane fluidity.

Outer hair cells (OHCs) play an important role in cochlear amplification via their length changes (electromotility). A noise-induced cochlear amplification loss leading to a permanent threshold shift (PTS) was observed without a significant hair cell loss in rats [Chen, G.D., Liu, Y., 2005. Mechanisms of noise-induced hearing loss potentiation by hypoxia. Hear. Res. 200, 1-9.]. Since motor proteins are inserted in the OHC lateral membrane, any change in the OHC plasma membrane may result in a loss of OHC electromotility, leading to a loss of cochlear amplification. In this study, the lateral diffusion in the OHC plasma membrane was determined in vitro in guinea pigs by fluorescent recovery after photobleaching (FRAP) after an in vivo noise exposure. The lateral diffusion in the OHC plasma membrane demonstrated a length-dependence, which increased as OHC length increased. A reduction in the lateral diffusion was observed in those OHCs with lengths of 50-70 microm after exposure to an 8-kHz octave band noise at 110 dB SPL for 3h. This membrane fluidity change was associated with the selective PTS at frequencies around 8 kHz. The reduction of the lateral diffusion in the OHC lateral wall indicated that noise could impair the micromechanics of the OHC lateral wall and might consequently impair OHC electromotility to induce threshold shift.

Influence of sympathetic fibers on noise-induced hearing loss in the chinchilla.

The influence of the sympathetic efferent fibers on cochlear susceptibility to noise-induced hearing loss is still an open question. In the current study, we explore the effects of unilateral and bilateral Superior Cervical Ganglion (SCG) ablation in the chinchilla on hearing loss from noise exposure, as measured with inferior colliculus (IC) evoked potentials, distortion product otoacoustic emissions (DPOAE), and outer hair cell (OHC) loss. The SCG was isolated at the level of the bifurcation of the carotid artery and removed unilaterally in 15 chinchillas. Another eight chinchillas underwent bilateral ablation. Twelve animals were employed as sham controls. Noise exposure was a 4kHz octave band noise for 1h at 110dB SPL. Results showed improved recovery of DPOAE amplitudes after noise exposure in ears that underwent SCGectomy, as well as lower evoked potential threshold shifts relative to sham controls. Effects of SCGectomy on OHC loss were small. Results of the study suggest that sympathetic fibers do exert some influence on susceptibility to noise, but the influence may not be a major one.

Evaluation of hearing and cochlear function by DPOAE and audiometric tests in patients with ankylosing spondilitis.

The aim of this study was to investigate cochlear functions in patients with ankylosing spondilitis (AS). Prospective, case control study. Twenty-eight AS patients (56 ears) and 25 healthy control subjects (50 ears) were included in the study. Pure-tone audiometry at 250, 500, 1,000, 2,000, 4,000, 6,000 Hz and immittance measures including tympanometry and acoustic reflex and DPOAEs (Distortion Product Otoacoustic Emission) testing were performed in the patients and controls. Pure-tone audiometry findings of the patients and controls were significantly different in all frequencies (P < 0.05). Sensorineural hearing loss was found in 10 patients (35%) that was bilateral in seven and unilateral in three patients. On DPOAE testing, there was no statistically significant difference between the levels of noise floor of the patients and controls (P > 0.05). However, the DPOAE responses of the patients and controls were significantly different in 3,000, 4,000, 5,000 and 6,000 Hz frequencies (P < 0.05). There is a damage of outer hair cells in patients with AS, and damaged outer hair cell regions mostly corresponds to the basal and mid-portions of the cochlea.

Low-dose, long-term caroverine administration attenuates impulse noise-induced hearing loss in the rat.

CONCLUSION: Physiological and morphological assessments indicated that low-dose and long-term caroverine delivery might be a new approach to protect against impulse noise-induced hearing loss. BACKGROUND: Although the exact mechanisms by which impulse noise causes hearing loss are still unclear, there is accumulating evidence that increased reactive oxygen species (ROS) production and excessive glutamate released from the inner hair cells lead to hair cell loss and consequently hearing loss. Caroverine is an antagonist of two glutamate receptors, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and N-methyl-D-aspartate (NMDA) receptors in the inner ear, as well as an antioxidant. MATERIALS AND METHODS: In this study, caroverine was delivered subcutaneously using an osmotic pump. This kind of delivery has the advantage, via continuous, long-term and low dose drug administration, of avoiding systemic side effects. RESULTS: It was shown that caroverine could significantly protect the cochlea against impulse noise trauma.

Effects of heat stress on Young's modulus of outer hair cells in mice.

Intense sound exposure causes permanent hearing loss due to hair cell and cochlear damage. Prior conditioning with sublethal stressors, such as nontraumatic sound, heat stress and restraint protects the ear from acoustic injury. However, the mechanisms underlying conditioning-related cochlear protection remain unknown. In this paper, Young's modulus and the amount of filamentous actin (F-actin) of outer hair cells (OHCs) with/without heat stress were investigated by atomic force microscopy and confocal laser scanning microscopy, respectively. Conditioning with heat stress resulted in a statistically significant increase in Young's modulus of OHCs at 3-6 h after application, and such modulus then began to decrease by 12 h and returned to pre-conditioning level at 48 h after heat stress. The amount of F-actin began to increase by 3 h after heat stress and peaked at 12 h. It then began to decrease by 24 h and returned to the pre-conditioning level by 48-96 h after heat stress. These time courses are consistent with a previous report in which heat stress was shown to suppress permanent threshold shift (PTS). In addition, distortion product otoacoustic emissions (DPOAEs) were confirmed to be enhanced by heat stress. These results suggest that conditioning with heat stress structurally modifies OHCs so that they become stiffer due to an increase in the amount of F-actin. As a consequence, OHCs possibly experience less strain when they are exposed to loud noise, resulting in protection of mammalian hearing from traumatic noise exposure.

A model of peripherally developing hearing loss and tinnitus based on the role of hypoxia and ischemia.

The incidence of sensorineural hearing loss often caused by direct damage to the cochlear hair cells is by far more frequent and more serious than disorders affecting the external ear or the middle ear. Mechanisms that are discussed to be relevant for the genesis of tinnitus and acquired hearing impairment are hair cell loss, signal transduction disturbances in the region of the outer and inner hair cells and the spiral ganglion, impairment of cochlear blood flow, mechanical disturbance, and hypoxia and ischemia. The present model surveys the possible cellular and molecular biological causes of peripherally developing hearing loss and tinnitus. In particular, the paper discusses the roles of hypoxia and ischemia in the cochlea and in the etiology of the neurosensory types of tinnitus. Peripheral origins of hearing disturbances and tinnitus may be: (a) damage to the stereocilia and the tip links, (b) dysfunction of potassium channels or (c) modification of the glutamate release. Moreover, the hypoxia inducible factor-1 may have an important role to play as a key transcription factor in the cells' adaptation to hypoxia and ischemia. An impairment of the cochlear blood flow may be induced by the expression of target genes like nitrogen monoxide synthase and endothelin-1 resulting in tinnitus. The paper discusses consequences resulting from the present model for the medical treatment of peripherally developing tinnitus and hearing loss.

Influence of leisure-time noise on outer hair cell activity in medical students.

OBJECTIVE: Noise exceeding a certain level can damage outer hair cells and thus cause hearing loss. In the past, noise-induced hearing loss was mainly caused by occupational noise. Leisure-time noise may be a promoting factor, particularly in young adults. The purpose of this study was to investigate whether transient evoked otoacoustic emissions (TEOAE) can be used to evaluate outer hair cell damage in young adults with no history of hearing complaints. The data obtained from the measurement of TEOAE were correlated with the participants' listening habits and exposure to leisure-time noise. METHODS: Eighty-eight young adults (47 women, 41 men; age 22.9+/-2.9 years) were examined. TEOAE were measured using standard ILO 88 equipment. All participants had normal hearing (hearing thresholds better than 20 dB HL; frequency range 0.125-10 kHz). None of the participants suffered from permanent tinnitus. All participants answered a questionnaire concerning their listening habits. RESULTS: On average, the participants frequented a discotheque 1.4 times a month; 25% had never visited a discotheque, 35% visited once a month and 32% twice or three times a month. Sixteen per cent reported transient tinnitus after every visit to a discotheque and 58% after nearly every visit. Eight per cent suffered from transient hearing loss after every visit to a disco and 37% after nearly every visit. Three per cent (4%) reported tinnitus (nearly) every morning after visiting a discotheque. The TEOAE level was above 6 dB in all participants [9.2+/-3.6 dB (mean +/- SD)] and reproducibility was above 60% (90+/-9%). All values matched pass criteria for normal TEOAE under clinical conditions. However, TEOAE levels and reproducibility decreased significantly with an increased number of visits to discotheques. CONCLUSION: Outer hair cell damage could be measured using TEOAE in individuals exposed to leisure-time noise, although these individuals exhibited no measurable puretone hearing loss.

A BAD link to mitochondrial cell death in the cochlea of mice with noise-induced hearing loss.

Acoustic overstimulation induces calcium overload and activation of mitochondria-mediated cell death pathways in outer hair cells (OHC) of the cochlea. However, it is not known whether these events are interrelated or independent. We have recently reported that the calcium-dependent phosphatase calcineurin is activated in OHC following noise exposure and now postulate that calcium overload triggers mitochondria-mediated death pathways through activation of Bcl-2-associated death promoter (BAD) by calcineurin. CBA/J mice were exposed to broadband noise (2-20 kHz), causing a permanent threshold shift of about 40 dB at 12 and 20 kHz, corresponding to damage in the middle and basal turns of the cochlea. Loss of OHC in the basal region was evident in surface preparations. BAD immunostaining in control animals had a cytoplasmic distribution in the cells of the organ of Corti. Five hours after acoustic overstimulation, mitochondria and BAD redistributed to the perinuclear region of OHC in the basal and middle turns but not in the apical turn. The nonapoptotic phospho-BAD (Ser 112) was up-regulated in cells undamaged by noise (supporting cells and inner hair cells) but not in OHC. These data establish a connection between calcium overload and mitochondria-mediated death pathways in OHC and also suggest a dual role for BAD. The translocation of BAD to the mitochondria in degenerating cells is indicative of the activation of its proapoptotic capacity, whereas up-regulation of phospho-BAD is consistent with a nonapoptotic role of BAD in less vulnerable cells.

Mice with altered KCNQ4 K+ channels implicate sensory outer hair cells in human progressive deafness.

KCNQ4 is an M-type K+ channel expressed in sensory hair cells of the inner ear and in the central auditory pathway. KCNQ4 mutations underlie human DFNA2 dominant progressive hearing loss. We now generated mice in which the KCNQ4 gene was disrupted or carried a dominant negative DFNA2 mutation. Although KCNQ4 is strongly expressed in vestibular hair cells, vestibular function appeared normal. Auditory function was only slightly impaired initially. It then declined over several weeks in Kcnq4-/- mice and over several months in mice carrying the dominant negative allele. This progressive hearing loss was paralleled by a selective degeneration of outer hair cells (OHCs). KCNQ4 disruption abolished the I(K,n) current of OHCs. The ensuing depolarization of OHCs impaired sound amplification. Inner hair cells and their afferent synapses remained mostly intact. These cells were only slightly depolarized and showed near-normal presynaptic function. We conclude that the hearing loss in DFNA2 is predominantly caused by a slow degeneration of OHCs resulting from chronic depolarization.

[DPOAE in tinnitus patients with cochlear hearing loss considering hyperacusis and misophonia]. / Otoemisja DPOAE u osób z szumami usznymi i niedosluchem slimakowym z uwzglednieniem nadwrazliwosci na dzwieki i misophonii.

UNLABELLED: The most probable place generating tinnitus in auditory pathway are outer hair cells (OHC) inside cochlea. To asses their activity otoacoustic emission is used. The goal of the investigation was estimation the features of otoemission DPOAE in groups with tinnitus patients with cochlear hearing loss, estimation of diagnostic value of DPOAE parameters for analysis of function of the cochlea in investigated patients emphasizing DPOAE parameters most useful in localizing tinnitus generators and estimation of hypothetic influence of hyperacusis and misophony on parameters of DPOAE in tinnitus patients with cochlear hearing loss. The material of the study were 42 tinnitus patients with cochlear hearing loss. In the control group there were 21 patients without tinnitus with the same type of hearing loss. Then tinnitus patients were divided into three subgroups--with hyperacusis, misophony and without both of them, based on audiologic findings. METHOD: after taking view on tinnitus and physical examination in all the patients pure tone and impedance audiometry, supratreshold tests, ABR and audiometric average and discomfort level were evaluated. Then otoemission DPOAE was measured in three procedures. First the amplitudes of two points per octave were assessed, in second--"fine structure" method-- 16-20 points per octave (f2/f1 = 1.2, L1 = L2 = 70 dB). Third procedure included recording of growth rate function in three series for input tones of value f2 = 2002, 4004, 6006 Hz (f2/f1= 1.22) and levels L1=L2, growing by degrees of 5dB in each series. RESULTS: DPOAE amplitudes in recording of 2 points per octave and fine structure method are very valuable parameters for estimation of cochlear function in tinnitus patients with cochlear hearing loss. Decreasing of DPOAE amplitudes in patients with cochlear hearing loss and tinnitus suggests significant role of OHC pathology, unbalanced by IHC injury in generation of tinnitus in patients with hearing loss of cochlear localization. DPOAE fine structure provides us the additional information about DPOAE amplitude recorded in two points per octave, spreading the amount of frequencies f2, where differences are noticed in comparison of two groups--tinnitus patients and control. Function growth rate cannot be the only parameter in estimation of DPOAE in tinnitus patients with cochlear hearing loss, also including subjects with hyperacusis and misophony. Hyperacusis has important influence on DPOAE amplitude, increases essentially amplitude of DPOAE in the examined group of tinnitus patients.

Age-related declines in distortion product otoacoustic emissions utilizing pure tone contralateral stimulation in CBA/CaJ mice.

One role of the medial olivocochlear (MOC) auditory efferent system is to suppress cochlear outer hair cell (OHC) responses when presented with a contralateral sound. Using distortion product otoacoustic emissions (DPOAEs), the effects of active changes in OHC responses due to the MOC as a function of age can be observed when contralateral stimulation with a pure tone is applied. Previous studies have shown that there are age-related declines of the MOC when broad band noise is presented to the contralateral ear. In this study, we measured age-related changes in CBA/CaJ mice by comparing DPOAE generation with and without a contralateral pure tone at three different frequencies (12, 22, and 37 kHz). Young (n = 16), middle (n = 10) and old-aged (n = 10) CBA mice were tested. DPOAE-grams were obtained using L1 = 65 and L2 = 50 dB SPL, F1/F2 = 1.25, using eight points per octave covering a frequency range from 5.6-44.8 kHz. The pure tone was presented contralaterally at 55 dB SPL. DPOAE-grams and ABR levels indicated age-related hearing loss in the old mice. In addition, there was an overall change in DPOAEs in the middle-aged and old groups relative to the young. Pure tone stimulation was not as effective as a suppressor compared to broadband noise. An increase in pure tone frequency from 12 to 22 kHz induced greater suppression of DPOAEs, but the 37 kHz was least effective. These results indicate that as the mouse ages, there are significant changes in the efficiency of the suppression mechanism as elicited by contralateral narrowband stimuli. These findings reinforce the idea that age-related changes in the MOC or the operating points of OHCs play a role in the progression of presbycusis - age-related hearing loss in mammals.

GATA3 haploinsufficiency causes a rapid deterioration of distortion product otoacoustic emissions (DPOAEs) in mice.

Human HDR (hypoparathyroidism, deafness and renal dysplasia)-syndrome is caused by haploinsufficiency of zinc-finger transcription factor GATA3. The hearing loss due to GATA3 haploinsufficiency has been shown to be peripheral in origin, but it is unclear to what extent potential aberrations in the outer hair cells (OHCs) contribute to this disorder. To further elucidate the pathophysiological mechanism underlying the hearing defect in HDR-syndrome, we investigated the OHCs in heterozygous Gata3-knockout mice at both the functional and morphological level. While the signal-to-noise ratios of distortion product otoacoustic emissions (DPOAE) in wild type mice did not change significantly during the first half-year of live, those in the heterozygous Gata3 mice decreased dramatically. In addition, both light microscopic and transmission electron microscopic analyses showed that the number of OHCs containing vacuoles was increased in the mutants. Together, these findings indicate that outer hair cell malfunctioning plays a major role in the hearing loss in HDR-syndrome.

Cochlear compression in listeners with moderate sensorineural hearing loss.

Psychophysical estimates of basilar membrane (BM) responses suggest that normal-hearing (NH) listeners exhibit constant compression for tones at the characteristic frequency (CF) across the CF range from 250 to 8000 Hz. The frequency region over which compression occurs is broadest for low CFs. This study investigates the extent that these results differ for three hearing-impaired (HI) listeners with sensorineural hearing loss. Temporal masking curves (TMCs) were measured over a wide range of probe (500-8000 Hz) and masker frequencies (0.5-1.2 times the probe frequency). From these, estimated BM response functions were derived and compared with corresponding functions for NH listeners. Compressive responses for tones both at and below CF occur for the three HI ears across the CF range tested. The maximum amount of compression was uncorrelated with absolute threshold. It was close to normal for two of the three HI ears, but was either slightly (at CFs < or =1000 Hz) or considerably (at CFs > or =4000 Hz) reduced for the third ear. Results are interpreted in terms of the relative damage to inner and outer hair cells affecting each of the HI ears. Alternative interpretations for the results are also discussed, some of which cast doubts on the assumptions of the TMC-based method and other behavioral methods for estimating human BM compression.

Sensitivity of distortion product otoacoustic emissions in noise-exposed chinchillas.

The present study investigates the effect of small amounts of outer hair cell (OHC) loss on distortion product otoacoustic emission (DPOAE) levels and evoked potential permanent threshold shifts (PTS) in a population of 12 noise-exposed chinchillas. The group mean DPOAE level, which decreased by up to approximately 15 dB in the presence of less than 8 dB PTS and 15% OHC loss, indicates that DPOAEs can detect an underlying cochlear pathology (i,e., OHC damage/loss) despite the presence of normal to near normal thresholds. The sensitivity of DPOAEs in detecting OHC loss makes this test measure suited for diagnosing sensorineural hearing impairment, particularly when abnormal auditory symptoms (i.e., speech discrimination problems) are associated with a normal audiogram in the clinical setting and as part of a hearing conservation program.

DPOAE-grams in patients with acute tonal tinnitus.

OBJECTIVE: To investigate cochlear outer hair cell function in patients with acute tonal tinnitus and normal or near-normal hearing threshold. STUDY DESIGN AND SETTING: Prospective controlled study in an academic tertiary health center. Distortion products of otoacoustic emissions (DPOAE)-grams of 32 ears with acute tonal tinnitus and normal hearing or minimal hearing loss were compared with those of 17 healthy nontinnitus ears. RESULTS: Tinnitus ears exhibited relatively increased amplitudes of DPOAE at high frequencies (4-6.3 kHz) when compared with the group of healthy ears and relatively decreased DPOAE amplitudes at middle frequencies (1650-2400 Hz). Statistically significant ( P < 0.01) increased mean values of DPOAE amplitudes were observed only at a frequency of f2 equal to 4.9 kHz. CONCLUSIONS AND SIGNIFICANCE: These findings suggest an altered functional state of the outer hair cells at a selected high-frequency region of the cochlea in ears with acute tonal tinnitus and normal or near-normal hearing threshold.

Acrylonitrile potentiates hearing loss and cochlear damage induced by moderate noise exposure in rats.

The diversity of chemical and drugs that can potentiate noise-induced hearing loss (NIHL) has impeded efforts to predict such interactions. We have hypothesized that chemical contaminants that disrupt intrinsic antioxidant defenses hold significant risk for potentiating NIHL. If this is true, then acrylonitrile (ACN) would be expected to potentiate NIHL. ACN, one of the 50 most commonly used chemicals in the United States, is metabolized via two pathways that are likely to disrupt intrinsic reactive oxygen species (ROS) buffering systems: (1) it conjugates glutathione, depleting this important antioxidant rapidly; (2) a second pathway involves the formation of cyanide, which can inhibit superoxide dismutase. We hypothesized that moderate noise exposure, that does not produce permanent hearing loss by itself, could initiate oxidative stress and that ACN could render the inner ear more sensitive to noise by disrupting intrinsic antioxidant defenses. Temporary and persistent effects of ACN alone (50 mg/kg, sc 5 days), noise alone (95 or 97 dB octave band noise, 4 h/day for 5 days), or ACN in combination with noise were determined using distortion product otoacoustic emissions (DPOAEs) and compound action potential (CAP) amplitudes. Histopathological damage to hair cells resulting from these treatments was also investigated using surface preparations of the organ of Corti. Individually, neither ACN nor noise exposures caused any permanent hearing or hair cell loss; only a reversible temporary threshold shift was measured in noise-exposed animals. However, when given in combination, ACN and noise induced permanent threshold shifts (13-16 dB between 7 and 40 kHz) and a decrease in DPOAE amplitudes (up to 25 dB at 19 kHz), as well as significant outer hair cell (OHC) loss (up to 20% in the first row between 13 and 47 kHz). This investigation demonstrates that ACN can potentiate NIHL at noise levels that are realistic in terms of human exposure, and that the OHCs are the main target of toxicity. While the exact mechanism is unknown, the results are consistent with the hypothesis of ROS involvement in NIHL at moderate levels.