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[Measuring beta-trace protein for detection of perilymph fistulas].

BACKGROUND: beta-trace protein is an immunological marker for cerebrospinal fluid or inner ear fluids. We present two initial cases in which a beta-trace protein nephelometric assay provided immunological proof of perilymph in samples taken from the nasopharynx. PATIENTS AND METHODS: Two cases are presented of an acute vestibulocochlear disorder following stapedotomy. Fluid samples, taken from the nasopharynx, were investigated using beta-trace protein laser nephelometry. The samples were collected using Merocel tamponades, which were left in place over night. RESULTS: In both cases, the samples were positive for beta-trace protein. Tympanoscopy was performed and the oval window was closed. After tympanoscopy a partial recovery of the hearing impairment was observed. CONCLUSION: Using beta-trace protein, it seems to be possible to diagnose a perilymphatic fistula by noninvasive means.

Adult↗

[Differential perilymph fistula diagnosis].

If sudden unilateral deafness accompanied by vertigo and spontaneous nystagmus on the afflicted side occurs during birth, a perilymph fistula with rupture of the round window membrane caused by pressure has to be considered. A 35-year-old pregnant patient was presented with the aforementioned symptoms on the day she gave birth to triplets. During examination, surditas and a paranasal hyposensitivity were found on the right side. Furthermore, a spontaneous nystagmus on the affected side occurred which was not suppressed by fixation. The nystagmus was apparent as a direction-fixed gaze nystagmus. Saccades during smooth pursuit testing as well as a lack of the auriculofacial reflex during contralateral stimulation of the deaf ear could be observed. Magnetic resonance imaging (MRI) of the skull revealed a fresh hemorrhagic cavernoma of the cerebellar pedunculi. This rare lesion caused a nerve compression of the fifth and eighth cranial nerve due to perifocal edema. Neurosurgical intervention was not necessary because of subsequent spontaneous resorption of the lesions and abatement of the deafness within a few days.

Adult↗

Total magnesium concentrations of perilymph, cerebrospinal fluid and blood in guinea pigs fed different magnesium-containing diets.

The total magnesium (Mg) concentrations of the perilymph (PL), cerebrospinal fluid (CSF), plasma and red blood cells (RBCs) of anesthetized guinea pigs separated into three groups and fed different Mg-containing diets were determined by atomic absorption spectrometry. Due consideration was given to the significant sources of error connected with the sampling procedure, particularly contamination of PL with CSF. The Mg levels of the individuals fluids differed significantly (P < 0.05/0.01) within each group. In the normal Mg group, the mean values of the PL, CSF and plasma were 0.66, 0.81 and 0.97 mmol/l, respectively, and 7.83 mmol/kg dry weight for RBCs. The analytical data were found to depend on the Mg content of the animals' diet, but to a different degree in the individual specimens (plasma > PL > CSF). A correlation was found to exist between all specimens tested (P < 0.05/0.01), except for CSF and RBCs, with the closest relation being that between plasma and PL. These findings suggest that the perilymphatic Mg equilibrates with the Mg level of plasma rather than with that of CSF. This is the first report showing Mg data of PL, CSF, plasma and RBCs obtained from the same subject, and the dependency on the Mg content of the animals' diet.

Animal Feed↗

Amino acid content of guinea pig perilymph collected under conditions of quiet or sound stimulation.

With the cochlear aqueducts blocked, guinea pig cochleas were perfused with artificial perilymph. The collected perfusates were treated with 5-(dimethylamino)-1-naphthalenesulfonyl chloride (dansyl-Cl). The derivatized constituents were analyzed by high- performance liquid chromatography using fluorometric detection. The concentration of none of the compounds measured, including the putative transmitters glutamate and aspartate, was found to increase in the perfusate in response to sound stimulation. These results do not support the suggestion that one of these compounds might be primary afferent transmitter of audition.

Acoustic Stimulation↗

Biochemical differences between perilymph, cerebrospinal fluid and blood plasma in the guinea pig.

Seven selected biochemical substances were systematically analyzed in perilymph (PL), cerebrospinal fluid (CSF) and blood plasma or serum of anesthetized guinea pigs in order to compare the composition of these fluids with regard to the origin and biochemical nature of PL. Compositional differences between the fluids were expressed by PL/CSF and PL/plasma concentration ratios of the chemical components measured. Contrary to the prevailing notion, the results suggest that the specific biochemical composition of PL corresponds neither to that of CSF nor to that of a simple ultrafiltrate of blood plasma. Furthermore, except for the total protein concentration, no significant differences were found in the composition of PL between the scala tympani and scala vestibuli.

Animals↗

Modifications of cochlear microphonic frequency responses following transient changes of hydrostatic pressure in the perilymph.

Cochlear microphonic potential was recorded with differential electrodes implanted in the various turns of the guinea-pig cochlea. Isointensity frequency responses were plotted in normal conditions and after excessive displacements of the cochlear partition. These displacements were provoked by changes of hydrostatic pressure in the perilymph of scala tympani or scala vestibuli. Typical modifications of the frequency response were observed. The most noticeable was a division in two parts of the response zone which suggested the existence of two resonance peaks. Scanning electron microscopy revealed that changes of hydrostatic pressure provoked alterations of the stereocilia in the outer rows of external hair cells, probably in relation with a decoupling of the tectorial membrane from the organ of Corti. These results are discussed in terms of possible alterations of cochlear micromechanics.

Animals↗

Uptake of horseradish peroxidase from perilymph by cochlear hair cells.

The specific mechanisms involved in the uptake of an exogenous protein, horseradish peroxidase (HRP), by cochlear hair cells were studied morphologically in light and electron microscopy. HRP was internalized by coated vesicles which formed from the plasma membrane only in the basal portion of the hair cells. Inner hair cells demonstrated relatively greater uptake than adjacent outer hair cells. The lateral plasma membrane of outer hair cells was unique in that reaction product was never bound to this portion of the membrane. Subsequent to endocytosis, HRP was transported to the Golgi and its associated system of endoplasmic reticulum and lysosomes. The exogenous protein was sequestered in a striking accumulation of secondary lysosomes and multivesicular bodies which were restricted to the infracuticular region of the hair cells and persisted for periods of at least 72 h after introduction of HRP. It is not clear whether the hair cells were slowly degrading the HRP or if the lysosomal enzymes necessary for its breakdown were lacking. The pathway demonstrated by HRP uptake and intracellular transport may represent one route by which macromolecules requisite for basic metabolic or nutritional requirements of the hair cells are supplied from the perilymph.

Animals↗

Potassium concentration in the inner sulcus is perilymph-like.

Using potassium-selective microelectrodes and an improved observation technique, it is shown that the potassium concentration in the fluid of the inner sulcus of the guinea pig is like that of perilymph. The implications for the common concepts about the role of potassium in the cochlea are discussed.

Animals↗

Intensity-dependent changes in oxygenation of cochlear perilymph during acoustic exposure.

This study examined the effects of acoustic exposure at different intensities on local oxygenation of the cochlea. The oxygen partial pressure (pO2) of perilymph in the basal scala tympani was measured polarographically in anesthetized guinea pigs exposed to either wide-band noise at 85 dB SPL or a 10 kHz pure tone at 90, 105, or 125 dB SPL for 1 h. Cochlear temperature, heart rate, arterial blood pressure and acid-base status were monitored. The cochlear microphonics (CM) and compound action potentials (CAP) were recorded before and after exposure. There were clear intensity-dependent differences in the effect of acoustic exposure on perilymphatic oxygenation. Moderate exposure intensities (85-90 dB SPL) were found to increase the pO2 by an average of about 20% of the initial level. In contrast, high intensity acoustic exposure (125 dB SPL) resulted in a mean decrease of about 20%. These changes persisted within a subsequent 30-min post-exposure period. There was no significant change in cochlear temperature and cardiorespiratory variables during and after any of the exposures as compared to the controls. CM and CAP amplitudes showed an extensive loss after acoustic overstimulation (125 dB SPL), but no permanent change with lower exposure intensities. These findings suggest that intracochlear oxygenation plays an important role in inner ear physiology during acoustic stimulation.

Acoustic Stimulation↗

The junctions of the spindle-shaped cells of the stria vascularis: a link that completes the barrier between perilymph and endolymph.

It is current opinion that the intercellular spaces of the stria vascularis represent a closed compartment isolated from the endolymph by the tight junctions of the marginal cells and from the perilymph by the junctional complexes of the basel cells. However, it has not yet been investigated whether these two barriers meet at the stria margins toward Reissner's membrane and the spiral prominence. Possible candidates for this sealing could be junctions between the spindle-shaped cells. In the present study freeze-fracture replicas of guinea pig specimens fixed in the presence of filipin were used in order to investigate the junctions of the spindle-shaped cells and to localize the cholesterol in their plasma membrane. Replicas reveal that, below the belt-like apical zonula occludens, the basolateral plasma membranes of the spindle-shaped cells adjacent to each other and to the basal cells are joined over their entire extension by a large number of junctional strands intermingled with numerous filipin-cholesterol-complexes. Gap junctions are present in the meshes formed by these junctional strands. Thus, the plasma membrane of the spindle-shaped cells shows morphological and cytochemical characteristics which indicate that they are the anatomical components completing the barrier isolating the intrastrial compartment from the surrounding fluids.

Animals↗

Adenosine 5'-triphosphate (ATP) concentrations in the endolymph and perilymph of the guinea-pig cochlea.

The concentration of adenosine 5'-triphosphate (ATP) in endolymph (EL), perilymph (PL) and cerebrospinal fluid (CSF), collected from anesthetized guinea pigs was determined using the luciferase-luciferin reaction. The cochlea was exposed by a ventrolateral approach and the bone overlying scala media of the third turn (EL) or scala vestibuli of the first turn (PL) was shaved to a thin layer and a small fenestrum made. For EL sampling, a double-barrelled pipette was inserted through the spiral ligament-stria vascularis complex. One barrel was filled with 150 mM KCl to record the endocochlear potential (EP) and upon the appearance of the positive EP, 0.12-1.22 microliter of fluid was aspirated into the other barrel by gentle negative pressure. For PL sampling, a single-barrelled pipette was advanced into scala vestibuli and 0.3-1.6 microliter of fluid was collected by capillarity. CSF (0.36-1.75 microliter) was obtained from the cisterna magna. The cochleae were removed and processed for light microscopy to determine the extent of tissue damage from the sampling procedure. ATP concentrations (mean +/- SEM, nM) for EL, PL and CSF were 12.95 +/- 2.4 (n = 10), 10.5 +/- 3.9 (n = 11) and 16.1 +/- 5.4 (n = 11) respectively. Differences in ATP concentrations among fluids were not statistically significant. To test the effect of hypoxia on ATP levels, a group of guinea pigs was subjected to a 90 s period of respiratory anoxia prior to sampling of EL, PL or CSF. ATP concentrations were 14.4 +/- 3.5 (n = 11), 20.7 +/- 4.1 (n = 10) and 13.5 +/- 4.6 (n = 4) for EL, PL and CSF, respectively; only PL ATP concentrations were statistically different (P = 0.018, Wilcoxon rank sum test) to basal conditions. This is the first study which demonstrates the presence of free ATP in cochlear fluids. The results indicate that ATP is present in cochlear fluids at concentrations close to those known to cause hair cell depolarization in vitro.

Adenosine Triphosphate↗

Separation and determination of dexamethasone sodium phosphate in cochlear perilymph fluid by liquid chromatography with ultraviolet monitoring and electrospray ionization mass spectrometry characterization.

The method for separation and determination of dexamethasone sodium phosphate (DexP) in cochlear perilymph fluid (CPF) of cavy was developed using HPLC with ultraviolet (UV) monitoring and electrospray ionization/mass spectrometry (ESI/MS) identification. The quantitative determination of DexP in CPF was achieved by HPLC with UV detection at 245 nm. The separation was carried out on a Phenomenex ODS(3) column ( 250 mm x 4.6 mm i.d., 5 microm) with the mobile phase of acetonitrile-5mmol/l ammonium acetate (23:77 (v/v)) at a flow rate of 1.0 ml/min. DexP was baseline separated from the matrices of CPF blanks within 15 min. The linearity ranged from 0.5 to 50 microg/ml. The limit of detection was 0.10 microg/ml. The recovery ranged from 98.5 to 100.8%. The relative standard deviations (R.S.D.s) of intra- and inter-day peak area were between 0.7-1.3 and 1.2-3.5%, respectively. Both full scan MS and MS2 of DexP with positive and negative polarity were obtained and elucidated. The specific ions were chosen to characterize DexP in the CPF sample. Using the proposed HPLC-UV-ESI/MS method, the concentration of DexP in CPF samples after both vein and middle ear injections were determined, and the relationships between concentration and time were obtained. This method offered reference data for clinical investigation of DexP to cure ear diseases.

Chromatography, High Pressure Liquid↗

Mass changes and dissolution of platinum during electrical stimulation in artificial perilymph solution.

The electrochemistry of platinum electrodes in artificial perilymph solution subjected to cyclic and steady-state potentials was studied by a quartz crystal electrochemical microbalance and by analysis of electrolyte for dissolved platinum. The effect of amino acid adsorption appears to be one of blocking sites for platinum oxidation and oxide reduction, a process in which the amino acid competes with chloride from phosphate-buffered saline. For amino acids such as cysteine, which are known to interact strongly with platinum, the voltammograms became nearly featureless and only a small change in mass was observed during cycling of the potential. There were no mass changes of an electrode in solution containing human serum albumen, but its presence did inhibit dissolution of platinum. The overall magnitudes of dissolved platinum found in the buffered solutions were low, remaining below 5 ppb in most cases. Dissolution was greatest in solutions containing high concentrations of cysteine. Extrapolation of the results to implanted auditory prosthesis electrodes indicated that platinum dissolution would not lead to toxic concentrations in the body.

Adsorption↗

Hyperbaric compression in the guinea pig with perilymph fistula.

OBJECTIVE: The study goal was to evaluate the effects of hyperbaric therapy on the auditory and vestibular function of guinea pigs compromised by perilymph fistula (PLF). METHODS: Twenty-four pigmented guinea pigs underwent induction of bilateral eustachian tube dysfunction before the creation of a unilateral PLF. Half of the animals were randomly assigned to receive immediate hyperbaric compression treatment of 4 "dives" each. Hearing was tested electrocochleographically, and signs of vestibulopathy were recorded before and after compression. RESULTS: After accounting for the effects of PLF, compression was not associated with significant hearing loss (P = 0.5411). Vestibulopathy was seen only in ears with PLF, and its incidence was similar for compression and noncompression groups. CONCLUSION: Hyperbaric compression does not compromise auditory or vestibular function in guinea pig ears with experimental PLF. SIGNIFICANCE: Our observations support the relative safety of recompression in patients with PLF.

Animals↗

Hydrostatic pressure measurement of endolymph and perilymph in the guinea pig cochlea.

Hydrostatic pressures of endolymph and perilymph were measured with a servo-micropipet system. The validity of the experiment was assessed by observation of endolymphatic pressure changes during and after 3 minutes of anoxia. Simultaneous recording of endocochlear DC potential confirmed the location of the electrode tip. Following verification of the efficacy of the system, measurements of endolymphatic or perilymphatic pressures were made in ten guinea pigs in which the middle ears had been filled with a saturated sodium chloride solution. Both endolymphatic and perilymphatic measurements showed gradual declines in pressure, with minimum pressure at 15 to 20 minutes and slow recovery during the next 20 minutes.

Animals↗

Perilymph fistula--45 case analysis.

PURPOSE OF THE STUDY: Though perilymph fistula (PLF) is not a rare disease, preoperative diagnosis still remains to be established. Some new diagnostic methods are challenging, but there is still no established diagnostic method except exploratory tympanotomy that verifies the occurrence of leakage. Early diagnosis of PLF is fully depending on history taking and some clinical examinations. To know the clinical features of PLF is one of the greatest helps to make both earlier and accurate diagnosis. In spite of some innovations in clinical examinations classic diagnostic procedure is thought to be still reliable. PROCEDURES: We investigated the clinical symptoms, basic tests results and therapeutic results in patients with PLF. RESULTS: From 1983 to 1998 PLF was identified in 44 patients (45 ears) with exploratory tympanotomies in our hospital. With respect to clinical history the predisposing factors such as blowing the nose, lifting heavy goods, and landing in an airplane were found in almost half of the patients, while the rest of them had no clear inducing factors. Their major symptoms included hearing loss (93%), vertigo and dizziness (91%), tinnitus (76%), and aural fullness (31%). The patients who have a clear predisposing factor tended to make diagnosis easily; on the other hand the rest of the patients who do not have clear etiology had some diagnostic difficulty. Subjective positive fistula signs were observed in 71% of patients. Vestibular symptoms improved in 80% of patients after closure of PLF. CONCLUSIONS: These results suggest that the variety of clinical manifestation make diagnosis more difficult. At the moment meticulous clinical history taking and close follow-up applying repeating fistula tests are the most important for not only earlier but also accurate diagnosis.

Adolescent↗

Estimation of perilymph concentration of agents applied to the round window membrane by microdialysis.

OBJECTIVE: The round window membrane (RWM) is known to be permeable to various biological substances. Application of biological substances to the RWM has been shown to affect inner ear fluid composition and damage hair cells, resulting in functional changes RWM instillation of gentamicin, a preferentially vestibulotoxic aminoglycoside, is used as a therapeutic treatment for patients with intractable vertigo and is gaining acceptance as a chemical vestibular ablation agent, despite considerable variations in the incidence and severity of hearing loss associated with gentamicin. Clearly, the susceptibility of vestibular and auditory hair cells to the ototoxic effects of gentamicin is not well understood. The aim of this study was to understand the kinetics of urea and methylene blue instilled into the inner ear space through the RWM and to establish a method for determining the optimal dosage for the treatment of inner ear disorders. MATERIAL AND METHODS: We used inner ear microdialysis to quantify changes in perilymph concentration of low molecular weight agents applied to the RWM in a chinchilla model. RESULTS: Preliminary results after placement of a microdialysis probe and application of a low molecular weight marker (urea) to the RWM were extrapolated from a time versus concentration plot from dialysates sampled over a 3-min interval using modifications of standard microdialysis equations for estimation of in vivo recovery. Our data suggest that inner ear microdialysis can be used to measure the pharmacokinetics of a low molecular weight agent within the perilymphatic space without the need for repeated direct sampling. CONCLUSION: Inner ear microdialysis may be a useful method for establishing a therapeutic dosage for ototoxic agents used in the treatment of inner ear disorders.

Animals↗

Hypothermia reduces glutamate efflux in perilymph following transient cochlear ischemia.

The effect of hypothermia on ischemic injury of the cochlea in gerbils was studied with particular regard to glutamate efflux in the perilymph. Under normothermic conditions interruption of the blood supply to the cochlea for 15 min caused a remarkable elevation of the compound action potential (CAP) threshold, and an increase in perilymphatic glutamate. The CAP threshold recovered to some extent with reperfusion, but not to preischemic levels. CAP thresholds, under hypothermic conditions and with reperfusion, recovered promptly to near pre-ischemic levels, while glutamate concentration did not change. These results, together with electron microscopy studies, suggest that hypothermia prevents hearing loss primarily through reduction of glutamate efflux at the synopses between inner hair cells and primary afferent auditory neurons.

Animals↗