Protective effect of lutein against acrolein-induced ototoxicity in rats.

BACKGROUND: Acrolein is a reactive aldehyde that forms during burning of wood and other fuels. It is also a product of lipid peroxidation (LPO) reactions and is present in cigarette smoke. Acrolein is known to cause oxidative stress and inflammatory nerve tissue damage. Lutein is a tetraterpenoid molecule with antioxidant and anti-inflammatory properties. There appear to be no studies on the effect of lutein on vestibulocochlear nerve damage induced by acrolein. The aim of this study was to investigate the effect of lutein on vestibulocochlear nerve damage induced by acrolein in rats using biochemical and histopathological methods. METHODS: The rats were divided into three groups (n = 6, for each group) a healthy control group (HG), an acrolein (ACR) group and a lutein and acrolein (LACR) group. In the LACR group, lutein was administered (1 mg/kg) via oral gavage. The ACR and HG groups received saline via oral gavage. Then, 1 h after the administration of lutein and saline, the LACR and ACR groups were treated with 3 mg/kg of acrolein via oral gavage. This procedure was repeated once a day for 30 days. RESULTS: The results of biochemical experiments showed that in the vestibulocochlear nerve tissues of the animals treated with acrolein, the levels of malondialdehyde, total oxidants, nuclear factor kappa b, tumor necrosis factor alpha and interleukin 1 beta significantly increased, whereas the levels of total glutathione and total antioxidants decreased as compared to those in the HG and LACR groups. In addition, severe histopathological damage was observed in vestibulocochlear nerve tissue of the acrolein group, whereas this damage was alleviated in the lutein group. CONCLUSION: Lutein protected vestibulocochlear nerve tissue from acrolein-associated oxidative and proinflammatory damage. This suggests that lutein might be useful in preventing or treating acrolein-induced ototoxicity.

The role of astrocyte mitochondria in differential regional susceptibility to environmental neurotoxicants: tools for understanding neurodegeneration.

In recent decades, there has been a significant expansion in our understanding of the role of astrocytes in neuroprotection, including spatial buffering of extracellular ions, secretion of metabolic coenzymes, and synaptic regulation. Astrocytic neuroprotective functions require energy, and therefore require a network of functional mitochondria. Disturbances to astrocytic mitochondrial homeostasis and their ability to produce ATP can negatively impact neural function. Perturbations in astrocyte mitochondrial function may accrue as the result of physiological aging processes or as a consequence of neurotoxicant exposure. Hydrophobic environmental neurotoxicants, such as 1,3-dinitrobenzene and α-chlorohydrin, cause regionally specific spongiform lesions mimicking energy deprivation syndromes. Astrocyte involvement includes mitochondrial damage that either precedes or is accompanied by neuronal damage. Similarly, environmental neurotoxicants that are implicated in the etiology of age-related neurodegenerative conditions cause regionally specific damage in the brain. Based on the regioselective nature of age-related neurodegenerative lesions, chemically induced models of regioselective lesions targeting astrocyte mitochondria can provide insight into age-related susceptibilities in astrocyte mitochondria. Most of the available research to date focuses on neuronal damage in cases of age-related neurodegeneration; however, there is a body of evidence that supports a central mechanistic role for astrocyte mitochondria in the expression of neural injury. Regional susceptibility to neuronal damage induced by aging by exposure to neurotoxicants may be a reflection of highly variable regional energy requirements. This review identifies region-specific vulnerabilities in astrocyte mitochondria in examples of exposure to neurotoxicants and in age-related neurodegeneration.

[Cochleovestibular disorders: approaches to diagnostics and treatment].

The aim of this work was to evaluate the efficacy of introduction of milgamma and milgamma compositum in the treatment of 52 patients with cochleovestibular disorders of different etiology. Thirteen patients enrolled in the study received standard therapy and 39 others were given its combination with milgamma preparations. Combined therapy with milgamma and milgamma compositum ensured faster vestibular compensation including posturographic characteristics than the standard treatment (within 3-4 weeks compared with 5 weeks in controls). The results of the study give reason to recommend milgamma and milgamma compositum as neurotropic medicines in addition to standard therapy for the management of the patients presenting with cochleovestibular disorders for the acceleration of the vestibular compensation.

The effects of sound conditioning on gentamicin-induced vestibulocochlear toxicity in gerbils.

OBJECTIVES/HYPOTHESIS: Recent studies in animal models have shown via physiologic and histologic measures that the administration of exogenous antioxidants is protective against gentamicin-induced oto-vestibulo toxicity. In addition, studies have also shown that sound conditioning increases cochlear antioxidants. The objective of this study is to determine whether sound conditioning provides protection against gentamicin in the cochlear and/or vestibular system. STUDY DESIGN: : Prospective animal study. METHODS: Three-month-old gerbils were divided into three groups (A, B, and C). The gerbils in group A were sound conditioned only (n = 2). In group B, the animals received gentamicin on the round window (n = 2). The gerbils in group C were sound conditioned first and later received gentamicin to the round window (n = 2). The animals were ultimately sacrificed and their right cochlea and posterior crista ampullaris were removed, processed, and sectioned. The specimens were analyzed for inner hair cell (IHC) and outer hair cell (OHC) loss and vestibular supporting and sensory hair cell nuclei per micrometer of vestibular epithelium. RESULTS: The sound-conditioned group (A) had no loss of cochlear hair cells. The gerbils treated with gentamicin only (B) had a 34% decrease of OHCs and 49% decrease of IHCs. The sound-conditioned plus gentamicin-treated group (C) had a 5.5% decrease in OHCs and 12% decrease in IHCs. There were no significant differences with regards to supporting cell nuclei within the posterior crista across all groups. When compared to group A, the gerbils in groups B and C did have a 23 to 42% decrease in the number of sensory cell nuclei per micrometer of vestibular epithelium. CONCLUSIONS: Sound conditioning does appear to attenuate the effects of gentamicin in the cochlea, although not significantly altering its vestibulotoxicity. An upregulation of cochlear-specific antioxidants is believed to be an important factor. As we had a small sample size, we can only note trends in the data, but future studies with more animals and measurements of antioxidant levels after sound conditioning would be useful to quantify this effect and determine if it can be exploited clinically. Laryngoscope, 2009.

Sudden bilateral sensorineural hearing loss following speedballing.

BACKGROUND: Hearing loss is an infrequently-reported consequence of recreational drug abuse. Although there are sporadic reports of hearing loss from heroin and cocaine ingested separately, there are no reports of hearing loss resulting from the combination of both drugs ingested simultaneously in the form of speedballing. PURPOSE: The purpose of this report is to document a case of bilateral sensorineural hearing loss associated with an episode of speedballing. RESEARCH DESIGN: Case Report. DATA COLLECTION AND ANALYSIS: The subject of this report was a 40-year-old man with a 20-year history of substance abuse. Data collected included a case history, pure tone audiometry, tympanometry and acoustic reflexes, and transient evoked otoacoustic emissions. RESULTS: The audiologic evaluation indicated a mild to moderate, relatively flat, bilateral sensorineural hearing loss that was worse in the right ear. CONCLUSIONS: A bilateral sensorineural hearing loss involving both cochlear and neural pathology may be a rare complication of cocaine, heroin, or the combination of the two drugs.

Episodic vestibular disruption following ablation of the inferior olive in rats: behavioral correlates.

The experiments herein investigate whether the behavioral responses to transient and episodic vestibular disruption and permanent ablation are distinct in the absence of climbing fiber input. Subjects in group 1 received an IP injection of PBS followed by an IP injection of niacinamide. Seven days later these rats received the first of 3 serial transtympanic injections of TTX on the same side with 7 days between each injection. Following each TTX injection rats displayed unilateral vestibular symptoms that persisted beyond 48h. Spontaneous barrel rolling behavior was not observed. Group 2 subjects received an IP injection of 3-acetylpyridine (3-AP)+niacinamide followed by the same TTX regimen as group 1. Following each TTX injection vestibular symptoms (severe body twisting and persistent spontaneous barrel rolling) emerged rapidly (<15min) and resolved by 72h. Group 3 subjects received an IP injection of 3-AP+niacinamide and 7 days later a single unilateral transtympanic injection of sodium arsanilate. Rats in group 3 developed vestibular symptoms similar to those observed in group 2 although there was no resolution of these symptoms. The results indicate that TTX has a rapid rate of infiltration and blockade of the VIIIth nerve that persists for >48h and then completely resolves. The contrast in vestibular symptoms between groups 1 and 2 suggest that climbing fibers are recruited soon after onset of vestibular disruption and play a role in attenuating the severity of vestibular symptoms associated with transient/episodic vestibular disruption.

A procedure to label inner ear afferent nerve endings for calcium imaging.

Characterization of synaptic transmission between the inner ear sensory cells and primary neuron dendrites has been hampered by the limited access to the postsynaptic terminals. Because direct physiological recording of postsynaptic currents are difficult to achieve, no information regarding the synaptic and dendritic events are available. This is due to the small size of the postsynaptic afferent nerve endings that do not allow a clear identification, and thus compromise direct electrophysiological recordings of the buttons. To study the physiology of afferent nerve endings, we have developed a two-photon imaging technique in cochlear and vestibular slice preparations from neonatal rats and turtles. This technique is based on a retrograde labeling of afferent nerve endings with high-affinity calcium-sensitive dyes. Dye filling was achieved by 6 h application of the dextran-amine conjugate of calcium green-1. Calcium changes were measured in afferent nerve endings in line scan and time lap mode. To address recording in a near-physiological situation, iontophoretic application of K+ was performed in the area of the stereocilia whereas glutamate was applied at the basal pole of sensory hair cells. Both types of application cause a reversible and sustained increase of Ca2+ in the button of afferent nerve fibers. Typical recordings are presented and potential interests for pharmacological studies of inner ear sensory cell synapses are discussed.

Episodic blockade of cranial nerve VIII provokes asymmetric changes in lobule X of the rat.

Although debilitating syndromes like Ménière's disease are in part characterized by recurrent or episodic vestibular disturbance the study of episodic vestibular disruption has only recently been possible with the introduction of a new model utilizing tetrodotoxin (TTX). In the present study, serial unilateral transtympanic administration of TTX produced behavioral symptoms indicative of transient vestibular disruption and novel patterns of Fos activity in the brainstem and cerebellum. Following two or three serial injections of TTX and a final survival time of 2 h, Fos immunocytochemistry revealed a distinct pattern of labeling in the brainstem that differed temporally from that observed following a single unilateral TTX injection. Specifically there was protracted expression of Fos in the beta subdivision of the inferior olive (IO) on the side ipsilateral to TTX treatment. In the cerebellum, the hallmark of episodic vestibular blockade was an asymmetric pattern of Fos labeling that involved all three layers of the cortex. In particular, there was prominent Fos labeling of Purkinje cells in the contra-TTX half of lobule X. In view of the fact that Fos labeling is not found in Purkinje cells following a single transient event or following peripheral vestibular ablation, it is suggested that Fos expression in Purkinje cells is a unique feature of episodic vestibular disruption and may represent a novel plastic response by a select population of Purkinje cells to episodic functional deafferentation.

Mild carbon monoxide exposure and auditory function in the developing rat.

We have examined the influence of chronic mild exposure to carbon monoxide (CO) on cognitive (learning) and auditory function in the developing rat. We have demonstrated that the auditory pathway is compromised at exposures less than 50 ppm, whereas learning was not influenced at 100 ppm. Artificially reared rat pups were exposed to CO during the brain growth spurt and onset of myelination. Spatial learning was assessed using the Morris Water Maze and three tests of auditory function: (1) auditory brainstem conduction times; (2) the amplitude of the eighth nerve's action potential; and (3) otoacoustic emissions carried out on rat pups (age 22- 24 days). The pups were gastrostomy-reared on a rat milk substitute and chronically exposed to CO at discrete concentrations in the range of 12-100 ppm from 6 days of age to post-weaning at 21-23 days of age. We found no difference in auditory brainstem conduction times at all CO concentrations in comparison to non-exposed controls. There was a difference in otoacoustic emissions for test and controls at CO concentrations of 50 ppm but not at lower concentrations. There was a consistent attenuation of the amplitude of the eighth nerve's action potential, even at the lowest CO exposure examined. The attenuation of the amplitude of the action potential of the eighth nerve at 50 ppm carbon monoxide exposure did not completely recover by 73 days of age. We conclude that prolonged mild exposure to carbon monoxide during development causes measurable functional changes at the level of the eighth cranial nerve.

Distribution of Fos labeling in the inferior olive following transient blockade of the VIIIth cranial nerve.

The sodium channel blocker, tetrodotoxin (TTX), is an effective tool for blockade of action potentials in neurons. Unilateral transtympanic administration of 3 mM TTX produced behavioral symptoms paralleling those previously reported following unilateral vestibular ablation. Behavioral symptoms were evident as early as 15 min post-TTX. Fos immunocytochemistry revealed an initial bilateral distribution of Fos in the inferior olive (IO) followed by an almost exclusively unilateral distribution of Fos. By 1 h, Fos was predominantly localized in subdivisions of the IO contralateral to TTX treatment. Fos labeling in the IO was most pronounced at 2- and 6-h survival times and was localized in the contralateral IOA, IOB, IOC, IOBe, and IOK subdivisions and bilaterally in the IOM and IODM. Other regions of the brainstem including the vestibular nuclei, prepositus hypoglossi, dorsal paragigantocellular reticular nucleus, nucleus of the tractus solitarius and locus coeruleus also exhibited altered patterns of Fos labeling following TTX. The finding that Fos activity in the IO is initially bilateral and then rapidly becomes unilateral has not been reported for the traditional vestibular ablation models and may be unique to the TTX model. In addition, since altered Fos activity is readily detected in the IO at time-points prior to detectable changes in Fos in the central vestibular complex it is possible that the IO is particularly sensitive to events precipitated by unilateral vestibular disturbance.

Hydrogen sulfide exposure without loss of consciousness: chronic effects in four cases.

Adverse effects of acute exposure to hydrogen sulfide (H2S) are well documented, but long-term effects of occupational exposure to low levels of the gas are not. To evaluate effects of such exposure we performed physical, neurologic, psychiatric, and chemosensory (smell and taste) examinations of four workers who were present but did not lose consciousness when the gas was accidentally released at a construction site. None of the four workers tested positive for functional problems, but all met diagnostic criteria for at least three, and up to eight, H2S-induced neuropsychiatric clinical disorders and from zero to two subclinical disorders. All four had abnormal P300 evoked responses (electrical neurophysiologic tests of brain waves). Our data indicate that exposures to even relatively low concentrations of H2S are hazardous. A rigorous epidemiologic investigation of persons who work with H2S is warranted.

Increased cell death in the developing vestibulocochlear ganglion complex of the mouse after prenatal ethanol exposure.

BACKGROUND: Previous studies have demonstrated that excessive prenatal alcohol exposure can damage the auditory and vestibular systems, in particular, cochlear hair cells. However, the direct effect of ethanol on the peripheral neurons in these pathways has not been examined. To study the effects of prenatal ethanol exposure on the developing vestibulocochlear ganglion (VCG) complex and the peripheral sensory organs, we exposed pregnant mice to ethanol and examined the levels of cell death in the inner ear. METHODS: Pregnant C57BL/6J mice were administered one of three doses of either ethanol (3.0, 4.5, and 5.5 g/kg) or isocaloric maltose/dextrin via intragastric intubation on gestational day (GD) 12.5. Embryos were dissected out of the uterus 8 hr after the intubation. Dying cells in the inner ear were stained with Nissl stain and labeled by in situ terminal dUTP nick-end labeling (TUNEL), and the percentage of dying cells was quantified. RESULTS: Ethanol exposure produced region-specific effects, with ethanol-exposed embryos exhibiting enhanced cell death only in the VCG complex, and not in the primitive saccule, cochlea, semicircular canal, or endolymphatic sac. The effects of ethanol on cell death in the VCG are dose dependent, with a significant increase in the level of cell death found only at the higher doses. CONCLUSIONS: Ethanol has a selective cytotoxic dose-dependent effect on the VCG at GD 12.5 suggesting that loss of VCG neurons may contribute to hearing and /or vestibular abnormalities in FAS children. Furthermore, the presence of TUNEL-positive cells and DNA laddering is consistent with the cells undergoing apoptotic cell death.

Multiple-site optical recording of mouse brainstem evoked by vestibulocochlear nerve stimulation.

We used optical imaging to investigate the mouse cochlear and vestibular nucleus in brainstem slices using a voltage-sensitive dye, RH 155. As a result, the spatiotemporal patterns of excitatory propagation were shown. These optical signals consisted of two components consisting of a spike-like fast signal and a long-lasting slow signal. All responses were abolished by tetrodotoxin. The slow signals were eliminated under a Ca(2+)-free solution. In addition, synaptic fatigue was also observed. The present study indicated the feasibility of optical recording for visually revealing the synaptic transmission in both the vestibular and cochlear nucleus.

Pharmacology of sensory gating in the ascending auditory system of the pigeon (Columba livia).

RATIONALE: Whether the underlying neurochemical basis of sensori(motor) gating is exclusively the result of mammalian brain evolution is not known. OBJECTIVE: The effects of ketamine (KET), benztropine (BTP), apomorphine (APO), methylphenidatehydrochloride (AMP) and haloperidol (HAL) on sensorimotor gating of the acoustic startle and gating of auditory input into the telelencephalon was assessed in a within-subject design in pigeons (Columba livia) using the prepulse inhibition (PPI) paradigm. METHODS: The startle blink reflex was recorded using EMG electrodes which were chronically implanted into the adjoining Musculus palpepralis superior et inferior, Musculus elevator palpebralis superior, and Musculus nictitantis. Thalamic gating was recorded using electrodes which were chronically implanted into the nucleus ovoidalis thalami and the neostriatum caudale (field L), respectively. RESULTS: KET, APO and AMP disrupted dose-dependently sensorimotor gating. The effect of APO and AMP was blocked by HAL. PPI disruption following BTP did not reach statistical significance. KET disrupted thalamic gating and increased prepulse-induced inhibition in field L. By contrast, AMP increased thalamic and decreased field L inhibition of field potentials when preceded by a pre-stimulus. Both effects were antagonised by HAL thus providing preliminary evidence for a D2-mediated auditory gating mechanism in the thalamus. However, while the effect of APO at the thalamic level was similar to AMP, prepulse-induced inhibition of field L activity was enhanced. This may be explained by concurrent D1-mediated telencephalic inhibition. CONCLUSION: It is concluded that thalamic gating is modulated by a dopaminergic/glutamatergic mechanism. The findings also confirm the notion of an homologous neurochemical basis of sensorimotor gating in mammals and birds.

AMPA-preferring glutamate receptors in cochlear physiology of adult guinea-pig.

1. The present study was designed to determine which glutamate (Glu) receptors are involved in excitatory neurotransmission at the first auditory synapse between the inner hair cells and the spiral ganglion neurons. 2. The Glu receptors present at the membrane level were investigated on isolated spiral ganglion neuron somata from guinea-pigs by whole-cell voltage-clamp measurements. Glu and AMPA induced a fast onset inward current that was rapidly desensitized, while kainate induced only a non-desensitizing, steady-state current. NMDA induced no detectable current. 3. To further discriminate between the AMPA and kainate receptors present, we used the receptor-specific desensitization blockers, cyclothiazide and concanavalin A. While no effect was observed with concanavalin A, cyclothiazide greatly enhanced the Glu-, AMPA- and kainate-induced steady-state currents and potentiated Glu-induced membrane depolarization. 4. To extrapolate the results obtained from the somata to the events occurring in situ at the dendrites, the effects of these drugs were evaluated in vivo. Cyclothiazide reversibly increased spontaneous activity of single auditory nerve fibres, while concanavalin A had no effect, suggesting that the functional Glu receptors on the somata may be the same as those at the dendrites. 5. The combination of a moderate-level sound together with cyclothiazide increased and subsequently abolished the spontaneous and the sound-evoked activity of the auditory nerve fibres. Histological examination revealed destruction of the dendrites, suggesting that cyclothiazide potentiates sound-induced Glu excitotoxicity via AMPA receptors. 6. Our results reveal that fast synaptic transmission in the cochlea is mainly mediated by desensitizing AMPA receptors.

Neuroaudiological effects in a case of fatal dimethylmercury poisoning.

OBJECTIVE: The audiological examination of this patient was to determine the degree and type of hearing loss that was incurred from apparent dimethylmercury poisoning. DESIGN: This was a single subject case study design. Audiologic tests were selected to help determine sensory from neural and/or central auditory system dysfunction. RESULTS: This patient demonstrated an inability to understand speech, both in formal and informal assessments, yet relatively good hearing sensitivity for pure tones bilaterally. Distortion product otoacoustic emissions showed only minimal deficits in each ear. The auditory brain stem response was abnormal bilaterally, indicating neural and/or central involvement. CONCLUSION: Dimethylmercury poisoning, in this case, resulted in compromise of the auditory neural system with little effect on the sensory (cochlea) mechanism.

Could antioxidant therapy reduce the incidence of deafness following bacterial meningitis?

Sensorineural hearing loss following acute bacterial meningitis could be caused by hydroxyl radicals generated by the inflammatory response. Obstruction of cerebrospinal fluid circulation through the tela choroidae of the choroid plexuses, with subsequent rupture of the tela choroidae, would expose the auditory nerve to selective radical damage. Acute administration of lipophilic antioxidants might provide the auditory nerve with increased protection.

Monitoring of anaesthesia in neurophysiological experiments.

Cortical activity can be substantially changed by the type of anaesthetic used, and by its dose level. For easy monitoring of depth of anaesthesia we describe the changes in electroencephalogram and electrocardiogram accompanying changes in depth of anaesthesia in the cat. Anaesthesia was induced by the volatile anaesthetic isoflurane. The high-frequency components (around 30 Hz) in the electroencephalogram disappear in deep anaesthesia. The electrocardiogram also shows substantial changes in contamination due to muscle fasciculations with anaesthesia level. Fasciculations appear as noise in the electrocardiogram. The amplitude of the electrical muscle activity contaminating the ECG can be easily used for the maintainance of a constant level of anaesthesia during a neurophysiological experiment.

Acute and chronic effects of aminoglycosides on cochlear hair cells.

The first detectable effect on the auditory system after a single high-dose injection of an aminoglycosidic antibiotic (AA) like gentamicin (GM) is the reversible blockade of medial efferent function, probably via blockade of calcium channels at the base of the outer hair cells (OHC). The kinetics of this effect are compatible with that of the molecule in perilymph. In the course of chronic treatment with lower doses, however, ototoxicity develops only after several days of treatment. Still GM can be observed inside the OHCs as soon as 24 hours after the first injection, and will be still present in some OHCs as long as 11 months after a chronic, nonototoxic 6-day treatment. In vitro, the short-term viability of isolated OHCs is not affected by exposure to AAs, but their transduction channels and their response to acetylcholine are reversibly blocked. However, developing organs of Corti in culture are highly and rapidly affected by exposure to AAs. Yet during direct intracochlear perilymphatic perfusion of GM, 2-mM solutions are not ototoxic, and with perfusion with a 20-mM solution ototoxicity develops only after several days of perfusion. From these various observations one can describe some aspects of the mechanisms of ototoxicity of AAs, from their access to perilymph and endolymph, to penetration in the hair cells, likely via endocytosis at their apical pole, and intracellular cytotoxic events.