The multiple sclerosis susceptibility genes TAGAP and IL2RA are regulated by vitamin D in CD4+ T cells.

Multiple sclerosis (MS) is an inflammatory, demyelinating disorder of the central nervous system that develops in genetically susceptible individuals. The majority of the MS-associated gene variants are located in genetic regions with importance for T-cell differentiation. Vitamin D is a potent immunomodulator, and vitamin D deficiency has been suggested to be associated with increased MS disease susceptibility and activity. In CD4+ T cells, we have analyzed in vitro vitamin D responsiveness of genes that contain an MS-associated single-nucleotide polymorphism (SNP) and with one or more vitamin D response elements in their regulatory regions. We identify IL2RA and TAGAP as novel vitamin D target genes. The vitamin D response is observed in samples from both MS patients and controls, and is not dependent on the genotype of MS-associated SNPs in the respective genes.

Water permeability of the mammalian cochlea: functional features of an aquaporin-facilitated water shunt at the perilymph-endolymph barrier.

The cochlear duct epithelium (CDE) constitutes a tight barrier that effectively separates the inner ear fluids, endolymph and perilymph, thereby maintaining distinct ionic and osmotic gradients that are essential for auditory function. However, in vivo experiments have demonstrated that the CDE allows for rapid water exchange between fluid compartments. The molecular mechanism governing water permeation across the CDE remains elusive. We computationally determined the diffusional (PD) and osmotic (Pf) water permeability coefficients for the mammalian CDE based on in silico simulations of cochlear water dynamics integrating previously derived in vivo experimental data on fluid flow with expression sites of molecular water channels (aquaporins, AQPs). The PD of the entire CDE (PD = 8.18 × 10(-5) cm s(-1)) and its individual partitions including Reissner's membrane (PD = 12.06 × 10(-5) cm s(-1)) and the organ of Corti (PD = 10.2 × 10(-5) cm s(-1)) were similar to other epithelia with AQP-facilitated water permeation. The Pf of the CDE (Pf = 6.15 × 10(-4) cm s(-1)) was also in the range of other epithelia while an exceptionally high Pf was determined for an epithelial subdomain of outer sulcus cells in the cochlear apex co-expressing AQP4 and AQP5 (OSCs; Pf = 156.90 × 10(-3) cm s(-1)). The Pf/PD ratios of the CDE (Pf/PD = 7.52) and OSCs (Pf/PD = 242.02) indicate an aqueous pore-facilitated water exchange and reveal a high-transfer region or "water shunt" in the cochlear apex. This "water shunt" explains experimentally determined phenomena of endolymphatic longitudinal flow towards the cochlear apex. The water permeability coefficients of the CDE emphasise the physiological and pathophysiological relevance of water dynamics in the cochlea in particular for endolymphatic hydrops and Ménière's disease.

Fluctuations of MS births and UV-light exposure.

BACKGROUND: Patients with multiple sclerosis (MS) are more frequently born in spring when compared to autumn. Fluctuation of UV-light has been hypothesized to drive this phenomenon. AIM: To assess the correlation between fluctuation of sunlight and birth season in persons with MS. METHODS: For this record-linkage study, we collected from the international MSBase and the Italian MS iMed-web databases the dates of birth of 11,415 patients with MS from 36 centres from 15 countries worldwide and compared these to dates of live-births from national registries. From all participating sites, we collected data on UV-light fluctuation and assessed its correlation with seasonal fluctuation in MS births. RESULTS: Compared with the reference cohort, an increased proportion of persons with MS were born in spring and a decreased proportion in autumn (odds ratio (OR) to be born in spring versus autumn = 1.158, χ² = 36.347, P < 0.001). There was no significantly increased fluctuation of MS births with increased quartile of ambient UV-light fluctuation (Ptrend = 0.086). CONCLUSION: Seasonal fluctuation of MS births as found in this worldwide cohort of patients with MS did not correlate with variation in seasonal fluctuation of UV-light. Most likely, it results from a complex interplay between fluctuation of sunlight, behavioural factors, other environmental factors and (epi)genetic factors.

Fatigue and heat sensitivity in patients with multiple sclerosis.

OBJECTIVES: Fatigue is one of the most common and troubling symptoms of multiple sclerosis (MS), and heat is often reported as a trigger. Although it is assumed that this heat sensitivity is specific for MS, the evidence for disease specificity is limited. We studied the relationship between fatigue, heat sensitivity, and environmental temperature, and its specificity for MS. MATERIALS AND METHODS: We compared 88 MS patients with 76 patients with ulcerative colitis (UC), another chronic auto-immune disease. As most important outcome measures, heat sensitivity, physical fatigue, mental fatigue, environmental temperature, and ambient UV-light levels were determined. RESULTS: More patients with MS reported heat sensitivity for fatigue, compared to patients with UC (53.4% vs 35.5%, respectively, P = 0.016). However, heat-sensitive patients were equally fatigued as heat-insensitive patients. Climatological data, including day temperature and amount of ambient UV light, were not related to fatigue in both heat-sensitive and heat-insensitive patients with MS. CONCLUSIONS: Our findings support the assumption that heat sensitivity regarding fatigue has an MS-specific component. Although patients with MS experience a relationship between environmental temperature and fatigue, objective assessment by climatological data could not confirm this.

Vitamin D status in patients with MS is negatively correlated with depression, but not with fatigue.

BACKGROUND: Depressive symptoms and fatigue are frequent and disabling symptoms of multiple sclerosis (MS). Depression and fatigue have been associated with a poor vitamin D status, and a poor vitamin D status is often found in MS. AIM: Assess whether vitamin D status contributes to depressive symptoms and fatigue in MS. METHODS: Patients with MS that participated in previous studies in which depression and fatigue were assessed and of whom serum 25-hydroxyvitamin D (25(OH)D) levels were available within a timeframe of less than one half-life of 25(OH)D were included. Depression and fatigue were assessed with the Hospital Anxiety and Depression Scale and the Multidimensional Fatigue Inventory. RESULTS: Fifty-nine patients were included. Mean scores of fatigue and depression were 14.6 (SD 4.2) and 6.2 (SD 4.4), respectively. The mean vitamin D status was 62.3 nm (SD 27.8). Vitamin D status correlated negatively with depression (r=-0.326, P=0.006). No significant correlation was found between vitamin D status and fatigue. In a multiple regression model, vitamin D status was not a significant contributor to depression, after controlling for age Expanded Disability Status Scale score and fatigue (P=0.078). Alternatively, depression and fatigue did not contribute to vitamin D status. DISCUSSION: This study shows a negative correlation between vitamin D status and depressive symptoms in patients with MS. Although multiple confounders exist, we observed an indication that vitamin D status might contribute to the presence of depressive symptoms in MS. Therefore, further studies on vitamin D in MS should include depressive symptoms as outcome measures to confirm these findings.

Brain-derived neurotrophic factor treatment does not improve functional recovery after hair cell regeneration in the pigeon.

CONCLUSIONS: Brain-derived neurotrophic factor (BDNF) supply to the inner ear does not improve the time course or the extent of functional recovery after hair cell regeneration. Specifically it does not improve the residual threshold elevation observed after the completion of spontaneous recovery. OBJECTIVE: The avian inner ear is capable of hair cell regeneration and substantial functional recovery, but residual hearing deficits remain. We investigated whether functional recovery can be improved by intracochlear application of BDNF, which plays an important role in auditory ontogenesis and maintenance during adult life. METHODS: Hair cells in adult pigeons were destroyed by local application of gentamicin. After 3 days either BDNF or control solution was administered to the scala tympani by implanted osmotic minipumps for 8 weeks. Auditory brain stem responses (ABR) to tone pips were used to assess recovery of hearing thresholds in both groups. RESULTS: The application of gentamicin caused a frequency-dependent hearing loss that ranged from 24.8 dB SPL at low frequencies to 66.2 dB SPL at high frequencies. After day 10 substantial recovery was observed, but a significant threshold shift remained. The time course of recovery in the control and BDNF-treated groups was similar, without significant residual threshold differences in any frequency range.

Blood metal ion levels are not a useful test for adverse reactions to metal debris: A systematic review and meta-analysis.

OBJECTIVES: Alarm over the reported high failure rates for metal-on-metal (MoM) hip implants as well as their potential for locally aggressive Adverse Reactions to Metal Debris (ARMDs) has prompted government agencies, internationally, to recommend the monitoring of patients with MoM hip implants. Some have advised that a blood ion level >7 µg/L indicates potential for ARMDs. We report a systematic review and meta-analysis of the performance of metal ion testing for ARMDs. METHODS: We searched MEDLINE and EMBASE to identify articles from which it was possible to reconstruct a 2 × 2 table. Two readers independently reviewed all articles and extracted data using explicit criteria. We computed a summary receiver operating curve using a Bayesian random-effects hierarchical model. RESULTS: Our literature search returned 575 unique articles; only six met inclusion criteria defined a priori. The discriminative capacity of ion tests was homogeneous across studies but that there was substantial cut-point heterogeneity. Our best estimate of the "true" area under curve (AUC) for metal ion testing is 0.615, with a 95% credible interval of 0.480 to 0.735, thus we can state that the probability that metal ion testing is actually clinically useful with an AUC ≥ 0.75 is 1.7%. CONCLUSION: Metal ion levels are not useful as a screening test for identifying high risk patients because ion testing will either lead to a large burden of false positive patients, or otherwise marginally modify the pre-test probability. With the availability of more accurate non-invasive tests, we did not find any evidence for using blood ion levels to diagnose symptomatic patients.Cite this article: M. Pahuta, J. M. Smolders, J. L. van Susante, J. Peck, P. R. Kim, P. E. Beaule. Blood metal ion levels are not a useful test for adverse reactions to metal debris: a systematic review and meta-analysis. Bone Joint Res 2016;5:379-386. DOI: 10.1302/2046-3758.59.BJR-2016-0027.R1.

[Balance disorder and subacute hydrocephalus due to haemorrhage in a cerebral cavernous malformation]. / Evenwichtsstoornis en subacute hydrocefalus door bloeding in een cerebrale caverneuze malformatie.

A 58-year-old man presented with a balance disorder, followed by progressive memory disturbance, urinary incontinence and vomiting. MRI of the brain revealed multiple cavernous malformations, with recent haemorrhage from one of them, in the brainstem, causing a hydrocephalus. After treatment for the hydrocephalus, the symptoms resolved quickly. Cerebral cavernomas may cause epileptic seizures, haemorrhaging and progressive neurological deficits. Diagnosis is on the basis of the typical MRI pattern of a hyperintense core, surrounded by a hypointense rim of old blood. Treatment consists of surgery if the lesion is easily accessible and can be completely removed. Other options are gamma-knife surgery, stereotactic radiotherapy and conservative treatment.

Increased inflammasome related gene expression profile in PBMC may facilitate T helper 17 cell induction in multiple sclerosis.

The NLRP3 inflammasome is a macromolecular complex importantly involved in IL-1ß processing. A role for this has been described in multiple sclerosis (MS). One mechanism by which IL-1ß might be involved in MS is by inducing pathogenic Th17 cells, i.e. GM-CSF+ Th17 cells. In the present study, we show that expression of the inflammasome related genes, NLRP3, caspase-1, IL-1ß and the IL-1ß/IL-1Ra ratio, was increased in PBMC from MS patients compared to healthy controls (HC). However, in an in vitro inflammasome activity assay with PBMC, IL-1ß protein secretion and the IL-1ß/IL-1Ra protein ratio were similar in MS patients and HC. Th cells cultured in the presence of supernatant derived from LPS/ATP inflammasome activated PBMC showed increased Th17 and GM-CSF+ Th17 cell frequencies in HC and MS patients and decreased anti-inflammatory IL-10+Th cell frequency in HC compared to Th cells cultured in the presence of control supernatant. Moreover, addition of the immune modulator calcitriol to the former condition resulted in reduced frequencies of Th17 and GM-CSF+Th17 cells, and also of IL-10+ Th cells. Evidently, our data indicate that inflammasome activity can skew the Th cell population toward a more pro-inflammatory composition, an effect that might be inhibited by vitamin D, and that might be importantly involved in inflammation within the central nervous system.

GM-CSF production by CD4+ T cells in MS patients: regulation by regulatory T cells and vitamin D.

BACKGROUND/OBJECTIVE: Data from animal models of MS suggest that GM-CSF(+)CD4(+)T cells are pathogenic cells. Therefore, GM-CSF production by CD4(+)T cells of MS patients and their susceptibility to regulatory mechanisms were investigated. METHODS: Intracellular flowcytometry was performed to determine the GM-CSF(+)CD4(+)T cell fraction in PBMC and CSF of MS patients and controls. The effect of regulatory T cells (Tregs) on GM-CSF production by CD4(+)T cells was studied in MS patients using a proliferation-suppression assay. Finally, GM-CSF(+)CD4(+)T cell fraction and GM-CSF protein levels in supernatant were assessed in anti-CD3-stimulated CD4(+)T cell cultures derived from healthy controls and MS patients, in the presence or absence of the active vitamin D metabolite calcitriol. RESULTS: The GM-CSF(+)CD4(+)T cell fraction in the peripheral blood did not differ between controls and MS patients. This T cell population could also be detected in the CSF of both subjects with MS as well as subjects with another diagnosis. In the CSF, it comprised a significant fraction of the T cell population. Upon in vitro stimulation of PBMC with anti-CD3 antibody, no differences were observed in GM-CSF(+)CD4(+)T cell frequencies. GM-CSF secretion was susceptible to regulation by Treg and vitamin D. Suppression of GM-CSF secretion by vitamin D was reduced in MS patients. CONCLUSIONS: Our study showed no elevation in GM-CSF(+)CD4(+)T cell fractions in MS patients compared to controls. Furthermore, GM-CSF secretion was prone to regulation by Treg and vitamin D, the latter being less effective in MS patients.

Performance of the avian inner ear.

In spite of morphological similarities, the avian inner ear has apparently developed mechanisms of sound transduction that differ from the mammalian solution. This paper is a compilation of the present knowledge.

AMPA-type glutamate receptor subunits are expressed in the avian cochlear hair cells and ganglion cells.

The cellular localization of AMPA-type glutamate receptor subunits was examined in the pigeon inner ear using subunit specific polyclonal antibodies (GluR1-4). In the auditory ganglion cell bodies immunoreactivity for the subunits GluR2/3 and GluR4, but not for GluR1 was detected. The hair cells showed diffuse immunoreactivity for GluR4. Additionally, immunostaining for the subunits GluR2/3 and GluR4 was present below the hair cells. These results indicate that the AMPA type glutamate receptors play a role in neurotransmission at the hair cell afferent synapse in the avian auditory system.

Discharge properties of pigeon single auditory nerve fibers after recovery from severe acoustic trauma.

The time course of recovery of compound action potential (CAP) thresholds was observed in individual adult pigeons after severe acoustic trauma. Each bird had electrodes implanted on the round window of both ears. One ear was exposed to a tone of 0.7 kHz at 136-142 dB SPL for 1 hr under general anesthesia. Recovery of CAP audiograms was monitored twice a week after trauma. Single unit recordings from auditory nerve fibers were made after 3 weeks and after 4 or more months of the exposure. The CAP was abolished immediately after overstimulation in all animals. Based on the temporal patterns of functional recovery of the CAP three groups of animals were identified. The first group was characterized by fast functional recovery starting immediately after trauma followed by a return to pre-exposure values within 3 weeks. In the second group, slow functional recovery of threshold started 1-2 weeks after trauma followed by a return to pre-exposure values by 4-5 weeks. A mean residual hearing loss of 26.3 dB at 2 kHz remained. The third group consisted of animals that did not recover after trauma. Three weeks after the exposure, tuning curves of single auditory nerve fibers were very broad and sometimes irregular in shape. Their thresholds hovered around 120 dB SPL. Spontaneous firing rate and driven rate were much reduced. Four or more months after exposure, the thresholds and sharpness of tuning of many single units were almost completely recovered. Spontaneous firing rate and driven rate were comparable to those of control animals. In the slow recovery group neuronal tuning properties showed less recovery, especially at frequencies above the exposure frequency. Thresholds and sharpness of tuning were normal at frequencies below the exposure frequency, but were much poorer at frequencies above the exposure. Spontaneous firing rate was much reduced in fibers with high characteristic frequencies. In fast recovering animals, the papilla was repopulated with hair cells after 4 months. In slow recovering animals, short (abneural) hair cells were still missing over large parts of the papilla after 4 months of recovery. Residual short (abneural) hair cell loss was largest at two areas, one more basal and the other more apical to the characteristic place of the traumatizing frequency. The results show that, in adult birds, functional recovery from severe damage to both short (abneural) and tall (neural) hair cells occurs. However, the onset of recovery is delayed and the time course is slower than after destruction of short (abneural) hair cells alone. Also, recovery is incomplete, both functionally and morphologically. There is residual permanent hearing loss, and regeneration of short (abneural) hair cells is incomplete.

Hair cell regeneration after local application of gentamicin at the round window of the cochlea in the pigeon.

Hair cells in the basilar papilla of birds have the capacity to regenerate after injury. Methods commonly used to induce cochlear damage are systemic application of ototoxic substances such as aminoglycoside antibiotics or loud sound. Both methods have disadvantages. The systemic application of antibiotics results in damage restricted to the basal 50% of the papilla and has severe side effects on the kidneys. Loud sound damages only small parts of the papilla and is restricted to the short hair cells. The present study was undertaken to determine the effect of local aminoglycoside application on the physiology and morphology of the avian basilar papilla. Collagen sponges loaded with gentamicin were placed at the round window of the cochlea in adult pigeons. The time course of hearing thresholds was determined from auditory brain stem responses elicited with pure tone bursts within a frequency range of 0.35-5.565 kHz. The condition of the basilar papilla was determined from scanning electron micrographs. Five days after application of the collagen sponges loaded with gentamicin severe hearing loss, except for the lowest frequency tested, was observed. Only at the apical 20% of the basilar papilla hair cells were left intact, all other hair cells were missing or damaged. At all frequencies there was little functional recovery until day 13 after implantation. At frequencies above 1 kHz functional recovery occurred at a rate of up to 4 dB/day until day 21, beyond that day recovery continued at a rate below 1 dB/day until day 48 at the 5.6 kHz. Below 1 kHz recovery occurred up to day 22, the recovery rate was below 2 dB/day. A residual hearing loss of about 15-25 dB remained at all frequencies, except for the lowest frequency tested. At day 20 new hair cells were seen on the basilar papilla. At day 48 the hair cells appeared to have recovered fully, except for the orientation of the hair cell bundles. The advantage of the local application of the aminoglycoside drug over systemic application is that it damages almost all hair cells in the basilar papilla and it has no toxic side effects. The damage is more extensive than with systemic application.

Synchronized responses of primary auditory fibre-populations in Caiman crocodilus (L.) to single tones and clicks.

Measurements of the responses to tones and clicks were made from single primary auditory fibres of the caiman. The distribution of the amplitude and phase of the fundamental component of the response rate modulation over the best frequencies of the fibres is comparable to that reported in the cat, despite the fact that the basilar membrane in caiman is only 4.5 mm long. However, much higher intensities are needed in the caiman (75-85 dB SPL) than reported in the cat (20 dB SPL) to obtain systematic distributions of the phase of the responses, probably due to the larger scatter of the phase responses in the caiman. The slopes of the phase distributions are very similar to those in cat. Single unit phase responses as a function of stimulus frequency at 85 dB SPL can be approximated by one, or in fibres with low best frequency, two straight lines. At lower intensities the deviation of the phase-frequency responses from a straight line increases as the group delay at the best frequency becomes larger. The shortest latencies of click responses are obtained with rarefaction clicks. Group delay estimates obtained from the responses to clicks and from the straight line approximations of the phase-frequency responses are related in a way expected for linear filter systems and accurately predict the measured distributions of the phase of the responses over the neural best frequency. The obtained group delays and click latencies in the caiman are very similar to those reported by other workers in the cat, the squirrel monkey and the treefrog, despite large morphological and probably functional differences of their inner ears. The click latencies are also very similar to those in the pigeon. The results are consistent with the existence of a mechanical travelling wave reported previously on the basilar membrane of the caiman, but at the same stimulus level the phase characteristic of the present single unit responses is steeper and the wave length estimates from the neural population phase distributions are shorter than those observed directly in the motion of the basilar membrane. Since the neural responses are an indirect estimate of the basilar membrane motion it cannot be decided whether the difference between neural and mechanical data is due to deterioration of the basilar membrane responses during the direct measurements or whether the basilar membrane response is sharpened by additional tuning mechanisms.

Responses of auditory nerve fibers innervating regenerated hair cells after local application of gentamicin at the round window of the cochlea in the pigeon.

Hair cells in the basilar papilla of birds have the capacity to regenerate after injury. There is also functional recovery of hearing after regeneration of the hair cells. The present study was undertaken to determine the effect of local aminoglycoside application on the physiology of auditory nerve fibers innervating regenerated hair cells. Collagen sponges loaded with gentamicin were placed at the round window of the cochlea in adult pigeons. The local application of gentamicin-loaded collagen sponges resulted in total hair cell loss over at least the basal 62% of the basilar papilla. According to the pigeon cochlear place-frequency map (Smolders, Ding-Pfennigdorff and Klinke, Hear. Res. 92 (1995) 151-169), frequencies above 0.3 kHz are represented in this area. Physiological data on single auditory nerve fibers were obtained 14 weeks after gentamicin treatment. The response properties showed the following characteristics when compared to control data: CF thresholds (CF = characteristic frequency) were elevated in units with CF above 0.15 kHz, sharpness of tuning (Q10dB) was reduced in units with CF above 0.38 kHz, low-frequency slopes of the tuning curves were reduced in units with CF above 0.25 kHz, high frequency slopes of the tuning curves were reduced in units with CF above 0.4 kHz, spontaneous firing rate was reduced in units with CF above 0.38 kHz, dynamic range of rate-intensity functions at CF was reduced in units with CF above 0.4 kHz and the slopes of these rate-intensity functions were elevated in units with CF above 0.4 kHz. Maximum discharge rate was the only parameter that remained unchanged in regenerated ears. The results show that the response properties of auditory nerve fibers which innervate areas of the papilla that were previously devoid of hair cells are poorer than the controls, but that action potential generation in the afferent fibers is unaffected. This suggests that despite structural regeneration of the basilar papilla, functional recovery of the auditory periphery is incomplete at the level of the hair cell or the hair cell-afferent synapse.

Mechanics of the basilar membrane in Caiman crocodilus.

Vibration measurements were made at a number of positions near the proximal (basal) end of the basilar membrane, and on the columella footplate, of Caiman crocodilus using a capacitive probe. The measurements established the existence of a mechanical travelling wave in this species. They showed no significant change of mechanical tuning with temperature, and were highly significantly different from previous reports of neural temperature sensitivity (Smolders, J. and Klinke, R. (1984): J. Comp. Physiol. 155, 19-30). Thus the neural sensitivity to temperature change appears not to depend upon basilar membrane mechanics. One interpretation of this is that the basilar membrane passively precedes an active temperature-sensitive filter. It was also found that the limbus supporting the basilar membrane had a measurable, but unturned, vibration and that the effect of draining scala tympani for the measurements was to increase the basilar membrane tuning frequency by a factor of about 1.5.

Basilar membrane motion in the pigeon measured with the Mössbauer technique.

Vibration measurements were made of the basilar membrane (BM), limbi and columella footplate (CFP) of pigeon using the Mössbauer technique. Recordings were located at 0.23-1.33 mm from the basal end of the BM. The existence of a travelling wave mode, propagating from base to apex, was established for papillae in apparently good physiological condition. For these papillae the characteristic frequency (CF) of the BM isovelocity (0.08 mm X s-1) response was an exponential function of distance with a frequency mapping constant of 0.91 +/- 0.10 mm (equivalent to 0.63 +/- 0.07 mm X oct-1); BM CF at the base was 5.95 +/- 0.65 kHz. Travelling wave motion was not demonstrated for papillae in poor physiological condition; tonotopy of BM CF was still evident, although the correlation with distance was less (1.08 +/- 0.30 mm X oct-1; 4.35 +/- 0.73 kHz at the base). BM motion was linear and the isovelocity responses were less sensitive and less sharp than single unit threshold tuning curves: for papillae in good physiological condition the SPL at BM CF at 0.08 mm X s-1 was 51 +/- 6 dB SPL; Q10 dB was 1.24 +/- 0.38; high- and low-frequency slopes were 20 +/- 6 dB X oct-1 and -14 +/- 4 dB X oct-1, respectively. The response of the BM relative to the CFP for papillae in good physiological condition was reminiscent of a second order resonant system with damping constant of 0.33 +/- 0.06 and group delay at BM CF of 0.89 +/- 0.36 periods.

Mechanics of a single-ossicle ear: I. The extra-stapedius of the pigeon.

The motion of the conical peak of the tympanic membrane (TM) at the tip of the extra-stapedius (ES) and of the columella footplate (CFP) were measured in the pigeon using the Mössbauer technique. The dimensions of middle-ear structures were measured in some of the experimental animals. The averaged velocity response at the ES for frequencies of 0.25-2.378 kHz was that of a second order, mass and stiffness controlled, resonant system with resonant frequency of 1.2 kHz and Q3 dB of 1.2. The mean velocity amplitude at resonance was 3.7 mms-1 at 100 dB SPL, which is approximately equal to the theoretical value of 3.5 mms-1 required for maximum energy transfer from a uniform plane acoustic wavefront in air. For the frequency regions 0.125-0.25 kHz and 2.378-5.657 kHz, the mean amplitude slopes for the velocity at the ES were 2 dB oct-1 and -3 dB oct-1, respectively. Above 5.657 kHz there was considerable inter-animal variation in the ES velocity responses. The direction of motion at the ES was frequency dependent above 1 kHz. For frequencies up to 1 kHz the ratio of CFP to ES velocity was independent of frequency; the mechanical lever ratio was 2.7, which was attributed to the geometry of the middle ear. At these frequencies the total transformer ratio for the middle ear, expressing the ratio of fluid pressure at the CFP to sound pressure at the ES, was estimated to be 35 dB.

Mechanics of a single-ossicle ear: II. The columella footplate of the pigeon.

The motion of the columella footplate (CFP) was measured in the pigeon using the Mössbauer technique. At the upper frequency limit of the cochlea the measured CFP response exhibited anti-resonant phenomena. These high-frequency responses were dependent on the orientation of the radiation detector, in a way which could not be explained by the cosine effect. The dependence of the recorded phase response on the measurement axis implies an additional vibration mode, which was out of temporal phase and non-colinear with the presumed translational vibration mode. The anti-resonant phenomena were not observed when the cochlear labyrinth was extirpated, thus excluding an explanation in terms of extraneous vibrations in the experimental apparatus or of loading by the Mössbauer source. Intra-cochlear reflection is proposed as the origin of the interference mode.