Simulation of hearing loss can induce pitch shifts for complex tones.

Background: Most studies on pitch shift provoked by hearing loss have been conducted using pure tones. However, many sounds encountered in everyday life are harmonic complex tones. In the present study, psychoacoustic experiments using complex tones were performed on healthy participants, and the possible mechanisms that cause pitch shift due to hearing loss are discussed. Methods: Two experiments were performed in this study. In experiment 1, two tones were presented, and the participants were asked to select the tone that was higher in pitch. Partials with frequencies less than 250, 500, 750, or 1,000 Hz were eliminated from the harmonic complex tones and used as test tones to simulate low-tone hearing loss. Each tone pair was constructed such that the tone with a lower fundamental frequency (F0) was higher in terms of the frequency of the lowest partial. Furthermore, partials whose frequencies were greater than 1,300 or 1,600 Hz were also eliminated from these test tones to simulate high-tone hearing loss or modified sounds that patients may hear in everyday life. When a tone with a lower F0 was perceived as higher in pitch, it was considered a pitch shift from the expected tone. In experiment 2, tonal sequences were constructed to create a passage of the song "Lightly Row." Similar to experiment 1, partials of harmonic complex tones were eliminated from the tones. After listening to these tonal sequences, the participants were asked if the sequences sounded correct based on the melody or off-key. Results: The results showed that pitch shifts and the melody sound off-key when lower partials are eliminated from complex tones, especially when a greater number of high-frequency components are eliminated. Conclusion: Considering that these experiments were performed on healthy participants, the results suggest that the pitch shifts from the expected tone when patients with hearing loss hear certain complex tones, regardless of the underlying etiology of the hearing loss.

Modifying Deutsch's scale illusion for application in music.

Deutsch's scale illusion demonstrates that the overall pitch range is the preferred organization when in competition with both local (note-to-note) pitch proximity and laterality (differences in the input ear). Such intricate factors can make it difficult to mimic this illusion. If a note is under a condition in which grouping by the overall pitch range and the local pitch proximity do not conflict, we hypothesized that an illusion would be perceived simply as the result of the competition between pitch proximity and laterality. In this paper, we aimed to replicate such a condition by modifying Deutsch's scale illusion. Psychophysical studies were conducted with healthy subjects. In the first half of the study, the C major scale with successive tones was presented in ascending form, alternating between the right and left ears; counterpart notes were simultaneously presented to the opposite ear, and the subjects were asked to listen to these dichotic tone patterns. Several counterpart notes were applied; we found that when the sequences of counterpart notes were close in note-to-note pitch proximity and were not overlapped with the ascending scale in pitch, the subjects appeared to perceive the scale clearly. In the latter half of the study, we applied this condition in music and devised auditory illusions such that melodies of the passages of "Lightly Row," "Cherry Blossoms," and "Jingle Bells" were perceived by listening to "jagged" dichotic tone patterns. The method we described in this paper is simple, and it is possible to easily create auditory illusions in music by applying our method.

Complex tone stimulation may induce binaural diplacusis with low-tone hearing loss.

To clarify the possible mechanism causing binaural diplacusis with low-tone hearing loss, two psychoacoustic experiments were performed with 20 healthy subjects, using harmonic complex tones. In the first experiment, two tones were presented unilaterally, either from the right or left side. One of the tones presented was higher in frequency in terms of the fundamental component, but lower or equal in frequency in terms of the highest component, than the other tone. The subjects were asked which tone was higher in pitch after listening to both tones. They were also asked to compare tones in which low-tone components were eliminated. In the second experiment, the subjects heard these complex tones binaurally, with low-tone components eliminated in one ear. In the first experiment, most subjects perceived pitch direction, that is, higher or lower, in a reverse way when low-tone components were eliminated from the complex tones. In the second experiment, approximately half of all subjects heard the tones at different pitches in both ears. Under certain conditions, complex tone stimulation may induce binaural diplacusis when low-tone hearing is lost in one ear.

Development and validation of a novel tool for assessing pitch discrimination.

OBJECTIVE: To develop and validate a tool for pitch discrimination that can be easily applied in otolaryngology clinics. METHODS: Twenty healthy subjects performed three psychoacoustic tasks for assessing pitch discrimination thresholds, which were compared among the three tasks. In the pitch direction discrimination (PDirD) task, the subjects judged the pitch of two tones as higher or lower. In the pitch difference discrimination (PDiffD) task, the subjects judged the pitch of two tones as same or different. In the pitch pattern detection (PPD) task, the subjects judged the pattern of a tone complex. Using tone sets from level 0 to level 8, pitch thresholds were determined using the Parameter Estimation by Sequential Testing (PEST) staircase procedure. The tasks were performed without prior training. RESULTS: Every test was performed twice for each ear, and the better results were adopted as the threshold values. The PDirD thresholds varied considerably among the subjects and showed significantly larger deviations relative to those of the PPD thresholds. The number of below 0 thresholds, which are considered inadequate results, was significantly larger in the PDiffD task. In the PPD task, all thresholds except one were above level 6. The average total time taken to complete the PPD task was 7.7min, which was significantly lesser than that taken to complete the other two tasks. CONCLUSION: The results of our study suggest that the PPD task is a reliable and easily available tool for the assessment of pitch discrimination.

Effect of low-intensity focused ultrasound on the middle ear in a mouse model of acute otitis media.

We hypothesized that low-intensity focused ultrasound (LIFU) increases vessel permeability and antibacterial drug activity in the mouse middle ear. We determined appropriate settings by applying LIFU to mouse ears with the external auditory canal filled with normal saline and performed histologic and immunohistologic examination. Acute otitis media was induced in mice with nontypable Haemophilus influenzae, and they were given ampicillin (50, 10, or 2 mg/kg) intraperitoneally once daily for 3 days with or without LIFU (1.0 W/cm(2), 20% duty cycle, 30 s). In the LIFU(+) groups receiving the 2- and 10-mg/kg doses, viable bacteria counts, number of inflammatory cells and IL-1ß and TNF-α levels in middle ear effusion were significantly lower than in the LIFU(-) groups on the same doses. Severity of AOM also tended to be reduced more in the LIFU(+) groups than in the LIFU(-) groups. LIFU application with antibiotics may be effective for middle ear infection.

Molecular cloning of the Coch gene of guinea pig inner ear and its expression analysis in cultured fibrocytes of the spiral ligament.

CONCLUSIONS: We have cloned guinea pig Coch cDNA and the sequence information will be useful for future molecular study combined with physiological experiments. Proper Coch gene expression appears to be dependent on the unique extracellular micro-environment of the inner ear in vivo. These results provide insight into the Coch gene expression and its regulation. OBJECTIVE: To characterize the guinea pig Coch gene, we performed molecular cloning and expression analysis in the inner ear and cultured fibrocytes of the spiral ligament. METHODS: The Coch cDNA was isolated using RACE. Cochlin isofoms were studied by Western blot using three different types of mammalian inner ear. The cochlear fibrocytes were cultured and characterized by immunostaining. Coch gene expression in the fibrocytes was investigated and the influence of cytokine stimulation was evaluated. RESULTS: The full-length 1991 bp Coch cDNA that encodes a 553 amino acid protein was isolated. The sequence had significant homology with other mammals, and the sizes of the Cochlin isoforms were identical. In the cultured fibrocytes, Coch mRNA was expressed in a very small amount and the isoform production was different, compared with the results in vivo. Cytokine stimulation did not alter the level of mRNA expression or isoform formation.

Nitric oxide production from cultured spiral ligament fibrocytes: effects of corticosteroids.

CONCLUSION: This is the first report of nitric oxide (NO) production by cultured spiral ligament (SL) fibrocytes. The data suggest that dexamethasone (Dex) suppresses NO production by SL fibrocytes. Given that SL fibrocytes play a role in cochlear fluid and ion homeostasis, glucocorticoids may suppress cochlear malfunction caused by NO production in SL fibrocytes. OBJECTIVE: To investigate NO production by cultured SL fibrocytes and regulation of NO production by Dex in mouse secondary cell cultures. MATERIALS AND METHODS: Cultured SL fibrocytes were stimulated with tumor necrosis factor-alpha (TNF-alpha), and NO production was visualized with membrane-permeable 4-amino-5-methylamino-2',7'-difluorofluorescein diacetate (DAF-FM diacetate). Inducible nitric oxide synthase (NOSII) messenger RNA (mRNA) was measured by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: After TNF-alpha stimulation, the fluorescence intensity increased in a time-dependent manner. This increase in fluorescence intensity was suppressed by a nonspecific NOS inhibitor, N(G)-nitro L-arginine methyl ester (L-NAME), and Dex. After TNF-alpha stimulation, expression of NOSII mRNA was observed. Expression of NOSII mRNA was inhibited by Dex.

Effects of low-intensity focused ultrasound on the mouse submandibular gland.

Ultrasound is expected to make a considerable contribution to drug delivery systems (DDSs). We tested the hypothesis that low-intensity focused ultrasound (LIFU) increases vessel permeability in the mouse submandibular gland without causing parenchymal damage. In a preliminary study, LIFU at 3 W/cm2 with a 50% duty cycle for 2 minutes did not cause histologic damage. We therefore applied LIFU to mouse submandibular gland at these conditions before and after injecting horseradish peroxidase. Single labeling laser scanning confocal microscopy revealed positive horseradish peroxidase staining around the excretory ducts in the mucous-producing part of the gland, but absence of staining in control glands. Immunostaining for fibrinogen was positive in the same region. Fibrinogen is an intravascular protein that does not pass through intact vessels. These findings suggest that LIFU increases vessel permeability and disruption without destruction. It is anticipated that this process will be useful in establishing a DDS that uses LIFU.

Dexamethasone inhibits tumor necrosis factor-alpha-induced cytokine secretion from spiral ligament fibrocytes.

To investigate the effect of proinflammatory cytokines on spiral ligament (SL) fibrocytes and regulation of cytokines by dexamethasone (Dex), in vitro studies were performed in murine secondary cell cultures. Cultured SL fibrocytes were stimulated with tumor necrosis factor-alpha (TNF-alpha), and the secretion of various mediators was measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcribed-polymerase chain reaction (RT-PCR). After stimulation with TNF-alpha, levels of keratinocyte-derived cytokine (KC), monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-2 (MIP-2), interleukin-6 (IL-6) and soluble intercellular adhesion molecule-1 (sICAM-1) were elevated in the culture supernatant, and their corresponding messenger RNAs were detected in the cultured fibrocytes. When the cultures were incubated with both TNF-alpha and Dex, the levels of KC, MCP-1, MIP-2 and IL-6 were significantly lower than those in cultures treated with TNF-alpha alone. The data suggest that Dex suppresses the inflammatory response in SL fibrocytes. Given that SL fibrocytes play a role in cochlear fluid and ion homeostasis, glucocorticoids may suppress the cochlear malfunction caused by SL inflammation.

Cytokine expression in experimental chronic otitis media with effusion in mice.

OBJECTIVES: Although otitis media with effusion (OME) is still a common disease in children and adults, the pathogenesis is not yet fully understood. We studied the effects of intratympanic injection with endotoxin purified from nontypeable Haemophilus influenzae on the characteristics of middle ear effusion (MEE). METHODS: Murine model of OME was developed by eustachian tube (ET) blockage followed by intratympanic inoculation with endotoxin (endotoxin group) or saline (control group). The mice were decapitated and histological changes and the production of inflammatory cytokines in MEEs were examined 3 days, 2 weeks, and 2 months after injection. RESULTS: All mice showed OME until 2 months after ET blockage. Most MEEs in the control group were serous, and mucoid or pultaceous MEEs were found only in the endotoxin group. Subepithelial space of middle ear mucosa was severely thickened with the infiltration of a large number of mononuclear cells in the endotoxin group. The levels of tumor necrosis factor-alpha (TNF-alpha) in MEEs were significantly higher in the endotoxin group than in the control group at all time points. Further, in situ hybridization showed that TNF-alpha messenger RNA was expressed not only by leukocytes and macrophages in MEEs but mononuclear cells present in the subepithelial space of middle ear mucosa. CONCLUSIONS: These results indicate that ET blockage is essential for the induction of serous MEE and additional administration of endotoxin is associated with the production of mucoid MEE accompanied by histological changes with inflammatory cell infiltration and cytokine production in the tympanic cavity.

Expression of adhesion molecules by cultured spiral ligament fibrocytes stimulated with proinflammatory cytokines.

Secondary cultures from murine spiral ligament (SL) fibrocytes were stimulated with proinflammatory cytokines interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha), and expression of various adhesion molecules was investigated. Cultures without cytokine stimulation did not show positive immunostaining for vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), or mucosal addressin cell adhesion molecule-1 (MAdCAM-1). Although staining was also negative after stimulation with IL-1beta, VCAM-1 and ICAM-1 staining was observed after the cells were stimulated with TNF-alpha. Reverse transcription-polymerase chain reaction analysis showed messenger RNAs for both VCAM-1 and ICAM-1 expression to be present after fibrocytes were stimulated with TNF-alpha. These data suggest that activated fibrocytes may cause inflammatory cells to persist in the SL. Given that SL fibrocytes may play a role in homeostasis of cochlear fluid and ion concentrations, prolongation of the inflammatory response could lead to fibrocyte damage that might ultimately result in cochlear malfunction.

Localization of glucocorticoid receptors in the murine inner ear.

We performed an immunohistochemical investigation of the distribution of glucocorticoid receptors (GRs) in the murine inner ear and found that GRs were expressed extensively, but with various degrees of immunoreactivity in different regions. We observed the strongest GR expression in the type III fibrocytes of the spiral ligament. Although the immunoreactivity of the cochlear hair cells and of the vestibular sensory epithelia was weak, the neighboring cochlear supporting cells and the subepithelial regions of the vestibular sensory epithelia were immunostained. Staining for GRs was also positive in the spiral ganglia and vestibular ganglia, as well as in the endolymphatic sac. The role of GRs in the inner ear is discussed.