Furin, PC1/3, and/or PC6A process rabbit, but not human, pro-lactase-phlorizin hydrolase to the 180-kDa intermediate.

Small intestinal lactase-phlorizin hydrolase (LPH) is synthesized as a large precursor (prepro-LPH) of 1926 amino acids. In the endoplasmic reticulum, prepro-LPH is split by signal protease. The resulting pro-LPH is cut to mature LPH directly (human) or via a 180-kDa intermediate (rabbit), most likely in the trans-Golgi network or in a later compartment. Antibodies directed against different regions of rabbit pro-LPH locate the cleavage site resulting in the 180-kDa intermediate between amino acid residues 79 and 286. This stretch contains the two sequences -Arg-Cys-Tyr-Arg114 approximately -Arg-Ala-Ser-Arg191 approximately, which are potential cleavage sites for subtilisin-like proprotein convertases. These sites are not conserved in human pro-LPH. By coexpression in COS 7 cells of rabbit prepro-LPH and proprotein convertases (PC 1/3, PC2, PC6A, PC6B, furin), we show that furin, PC 1/3, and PC6A generate a processing intermediate that is immunologically indistinguishable from the one observed in vivo. Furin, PC 1/3, and PC6A are all expressed in the small intestine as shown by a polymerase chain reaction-based approach and, more specifically, in enterocytes, as shown by in situ hybridization. These results suggest that furin, PC 1/3, and/or PC6A are responsible for the in vivo processing of rabbit pro-LPH to the 180-kDa intermediate.

The levels of lactase and of sucrase-isomaltase along the rabbit small intestine are regulated both at the mRNA level and post-translationally.

We determined along the small intestine of young and adult rabbits the activities of lactase (LPH) and sucrase (SI), the levels of their cognate mRNAs, and examined the in vitro biosynthesis of LPH and pro-SI. Lactase activity is low in the proximal 1/3 of the intestine, whereas the mRNA levels are high. However, the rates of biosynthesis of the LPH forms correlated well with the steady-state levels of LPH mRNA in all segments, indicating that factor(s) acting post-translationally produce a decline in brush border LPH in the proximal small intestine. These factor(s) are not involved in the processing of pro-LPH to mature LPH, since the relative amounts of the various forms of LPH are almost the same along the small intestine. Unexpectedly, we find that also for SI the ratio of activity to mRNA is low in proximal intestine. The biosynthesis of pro-SI correlates with the steady-state levels of its mRNA. Hence, the steady-state levels of LPH and SI along the small intestine are regulated both by mRNA levels and by posttranslational factor(s).

On the effect of interaural phase differences on loudness.

The loudness of four monaurally presented Gaussian shaped, 60-ms tone bursts was matched to that of four similar pulses presented binaurally. The stimuli to be matched were all presented in continuous binaural noise of three levels and, in different experiments, either the monaural or the binaural stimuli were adjusted by the observer. With 250- and 710-Hz tone bursts, there are large differences in loudness that, at low signal-to-noise ratios, depend on the interaural phase conditions in a manner consistent with the changes in masked threshold produced by the phase manipulation; there is little, if any, effect of interaural phase on loudness for 2-kHz signals. The effect of interaural phase on loudness decreases with increasing level but, at 250 Hz, remains measurable some 30-40 dB above masked threshold. The matching function for signals out-of-phase grows in proportion to the level to be matched over the entire range from masked threshold to the highest level used. In contrast, for the in-phase condition, the observers show a step at a level that depends both on frequency and on the observer from proportional growth near thresholds to parallel proportional growth some 6 to 12 dB higher.

The effect of masker spectral asymmetry on overshoot in simultaneous masking.

"Overshoot" is a simultaneous masking phenomenon: Thresholds for short high-frequency tone bursts presented shortly after the onset of a broadband masker are raised compared to thresholds in the presence of a continuous masker. Overshoot for 2-ms bursts of a 5000-Hz test tone is described for four subjects as a function of the spectral composition and level of the masker. First, it was verified that overshoot is largely independent of masker duration. Second, overshoot was determined for a variety of 10-ms masker bursts composed of differently filtered uniform masking noise with an overall level of 60 dB SPL: unfiltered, high-pass (cutoff at 3700 Hz), low-pass (cutoff at 5700 Hz), and third-octave-band-(centered at 5000 Hz) filtered uniform masking noises presented separately or combined with different bandpass maskers (5700-16000 Hz, 5700-9500 Hz, 8400-16000 Hz) were used. Third, masked thresholds were measured for maskers composed of an upper or lower octave band adjacent to the third-octave-band masker as a function of the level of the octave band. All maskers containing components above the critical band of the test tone led to overshoot; no additional overshoot was produced by masker components below it. Typical values of overshoot were on the order of 12 dB. Overshoot saturated when masker levels were above 60 dB SPL for the upper octave-band masker. The standard neurophysiological explanation of overshoot accounts only partially for these data. Details that must be accommodated by any full explanation of overshoot are discussed.

Dependence of binaural loudness summation on interaural level differences, spectral distribution, and temporal distribution.

Loudness of interaurally correlated narrow- and broadband noises was investigated using a loudness estimation paradigm (with two anchors) presented via headphones. Throughout the experiments (most performed by 12 subjects), the results from both anchors agreed very well. In the first experiment, third-octave-band noises centered around 250, 710, or 2000 Hz, as well as broadband red (-10 dB/oct), pink (-3 dB/oct), and blue (+10 dB/oct) noises, with interaural level differences of delta L = 0, 4, 10, 20, and infinity dB, were presented as test signals while the same sound presented monaurally or diotically served as anchor. The binaurally summed loudness at delta L = 0 dB was found to be larger than the loudness of a monaural signal of the same SPL by a factor of about 1.5 and decreased with increasing delta L. No dependence of this behavior on frequency, level, or spectral shape was found. In a second experiment, abutting frequency bands of varying width were alternately presented to the subject's left and right ears with the overall spectrum encompassing the whole audio range. The binaural loudness was larger for fewer but broader frequency bands. In a third experiment, uniform exciting noise was switched between the two ears at various speeds. Increasing the switching frequency gave rise to an increase in loudness of about 20%. All results are discussed from the viewpoint of the use of the standardized loudness meter. At this point, there is no evidence that any significant systematic errors due to single-channel evaluation (in contrast to the human two-channel processing) are made by measuring loudness using these meters.

On the dependence of (f2-f1) difference tones on subject and on additional masker.

For a group of eight subjects showing stronger than usual irregularities in the level dependence of the quadratic distortion product, the level and phase of the (f2-f1) difference tone were measured using the method of cancellation for three sets of primary frequencies as a function of the primary levels. An additional masker seems to "linearize" the level dependence toward regular behavior. Using data sets produced with and without the additional masker, it is possible to separate two sources of quadratic nonlinearity, one with regular behavior presumably located in the middle ear and another with irregular behavior (similar to that of cubic distortion) presumably located in the characteristics of outer hair cells. Through the "subtraction" of empirically determined patterns from idealized patterns, it is possible to approximate patterns stemming from the inner-ear source alone.

The effect of a low-frequency masker on loudness.

Intense low-frequency tones produce masking-period patterns in which the masking of a higher-frequency tone burst varies by up to 25 dB over the period of the masker. In this experiment, observers were asked to match the loudness of partially masked test-tone bursts in one ear by adjusting the level of unmasked bursts presented to the other ear. It was found that the variation in masked threshold over the period of the masker also affects loudness matches. This effect on loudness, although it decreases in size with increasing level above masked threshold, persists even 25 dB above masked threshold.

On the influence of acoustical probe impedance on evoked otoacoustic emissions.

Using two different probes representing different acoustical impedances, simultaneously evoked otoacoustic emissions, thresholds in quiet, and delayed evoked emissions were measured in the frequency region of 900 to 1100 Hz for one subject who has extremely large emissions at these frequencies. All data show that the sound pressure produced in the closed ear canal clearly depends on two contributions, that of the activity of the inner ear, and that of the acoustical impedance of the probe. These effects can be simulated in an analog model of peripheral preprocessing.

[Temporary threshold shift after nuclear magnetic resonance tomography]. / Zeitlich begrenzte Schwellenverschiebung nach Kernspin-Tomographie.

The third-octave band levels of the noises produced in a MR-imager (1.0 Tesla Magneton, Siemens) were measured. Corresponding sounds presented through earphones have been used in simulation experiments to produce in normal-hearing listeners a temporary threshold shift lasting several minutes. These effects could be verified objectively using otoacoustic emissions. The consequences especially for patients with already impaired hearing are discussed: ear protection is strongly recommended.

Psychoacoustical and ear canal cancellation of (2f1-f2)-distortion products.

Level and phase of the (2f1-f2)-difference tone were measured as a function of primary-tone level using the psychoacoustical method of cancellation and the objective method of emission cancellation for four frequency separations of f1 = 1620 Hz and f2 in four subjects. Differences between hearing- and emission-cancellation levels ranged from 60-33 dB as delta f = f2-f1 increased from 180 to 432 Hz. For smaller separations of the primaries, phase changes for emission cancellation covered a wide range and had sharp "steps," whereas for hearing cancellation, the phase varied only slightly. With wider separations of the primaries, the phase became more varied for hearing cancellation and more homogeneous for emission cancellation. Both emission- and hearing-cancellation level functions were nonmonotonic as a function of constant SL1 and varied SL2. Remarkable phase shifts always appeared near minima in level at all separations of the primaries for emission cancellation. Four sources may be contributing to the differences in results: (a) the frequency-dependent attenuation of the middle-ear transfer function, (b) the frequency-dependent mismatch of the acoustical impedances at the eardrum, (c) the frequency dependence of the microphone's sensitivity mounted within the probe, and (d) the different reaction of active nonlinear cochlear processes on the hearing- and emission-cancellation tones.

Cochlear preprocessing in analog models, in digital models and in human inner ear.

Data of spontaneous, delayed, and simultaneously evoked otoacoustic emissions produced in human subjects are compared with data produced in an analog and in a digital wave-parameter realization of our model. The high probability of the frequency distance of the emission's extreme values at 0.4 Bark, corresponding to a local distance of 0.5 mm along the basilar membrane is found also in the models. The influence of the lateral feedback coupling in the models on the frequency selectivity and on the appearance of emissions show the high quality of simulating human peripheral signal processing by the models.

Frequency selectivity and temporal resolution in patients with various inner ear disorders.

Both frequency selectivity and temporal resolution were examined in patients with various inner ear disorders. These included noise-induced hearing loss (n = 24), Menière's disease (n = 16), sudden deafness (n = 25), toxic inner ear damage (n = 14), presbyacusis (n = 38) and degenerative progressive inner ear hearing loss (n = 8). To facilitate quantitative comparison, various factors were introduced, namely, frequency resolution factor (FRF), temporal resolution factor (TRF) and a combined resolution factor (FTRF). The FRF of normal-hearing subjects in background noise conditions was found to be approximately 20% less than in comparative test conditions without noise, whereas the TRF of normal-hearing persons tested under background noise conditions showed a remarkable increase (factor 3). The frequency resolution performance and/or the temporal resolution performance were found to be impaired in all patient groups with inner ear hearing loss. This is particularly noticeable for temporal resolution in test conditions involving the addition of background noise. It can be concluded that in such cases, speech discrimination can be seriously jeopardized.

Delayed evoked otoacoustic emissions--an ideal screening test for excluding hearing impairment in infants.

A screening test is badly needed with which cochlear hearing impairment in small children and newborns can be detected. Delayed evoked otoacoustic emissions (DEOAEs) have been found in laboratory research to be a very useful tool to test the normality of cochlear preprocessing. The characteristics of a simple apparatus for clinical use to measure DEOAEs are described together with typical examples of emissions. Many case studies have already demonstrated the clinical usefulness of DEOAEs in neonates and small children. Five cases are explained in detail and the advantages of such an early screening test discussed.

Why can a decrease in dB(A) produce an increase in loudness?

Loudness measured by the method of absolute magnitude estimation is compared to loudness calculated in accordance with ISO 532 B (International Organization for Standardization, Geneva, 1966). The measured and calculated loudness functions exhibit a similar pattern of loudness growth. Both measured and calculated loudness of a complex sound composed of a 1000-Hz tone and broadband noise is a nonmonotonic function of the overall SPL of the complex. The nonmonotonic loudness-growth pattern holds over a 30-dB range from 73.5 to 103.5. To facilitate understanding of the results, a single cycle of data is analyzed in detail. The analysis shows that loudness patterns produced in the auditory system by the tone-noise complex can account for the observed effects. Moreover, they show that the A-weighting and the loudness of the complex are negatively related. This inverse relation means that the A-weighted SPL is an inappropriate and misleading indicator of the loudness of sound combinations with heterogeneous spectral envelopes. Consequently, its suitability for noise control is diminished. A loudness meter that combines the spectral shapes of different sounds to produce an overall perceived magnitude offers greater promise.

Correlation between time functions of sound pressure, masking, and OAE suppression.

Periodic sound-pressure time functions with frequency components below 50 Hz were used to measure within their period (a) the temporal course of masking, called a masking-period pattern (MPP), and (b) the temporal course of suppression of delayed evoked otoacoustic emissions, called a suppression-period pattern (SPP). Three different time functions were used: an alternating Gaussian impulse, its first integral, and its second integral. In each case, the course of the SPP is a mirror image of that of the MPP: Small masking corresponds to small suppression, while strong masking coincides with almost total suppression. Since otoacoustic emissions are assumed to have their origin in the inner ear, it can be argued that simultaneous masking, an effect including central processing, is very strongly based on peripheral processes located in the cochlea. Both MPP and SPP are closely related to the second derivative of the sound-pressure time function.

Decision rules in detection of simple and complex tones.

Detection of simple and complex tones in the presence of a 64-dB SPL uniformly masking noise was examined in two experiments. In both experiments, the signals were either pure tones (220, 1100, or 3850 Hz) or an 18-tone complex consisting of equally intense components between 110 and 7260 Hz. In experiment 1, psychometric functions were obtained for detection in a 2I, 2AFC task. Results for eight normal listeners show that the psychometric functions are parallel for simple and complex tones. As expected, the masked thresholds for the pure tones are 43-44 dB SPL independent of frequency; the masked threshold for the complex tone is about 37 dB SPL per tone. These results indicate that the simultaneous presence of signal energy in many auditory channels aids detection. In experiment 2, psychometric functions were obtained with all four signals presented in random order within a block of trials. Results for four normal listeners show that the psychometric functions are parallel to one another and to those obtained in experiment 1. The thresholds are elevated to about 46 dB for the pure tones and to 40.5 dB for the complex tone. These results are nearly, but not quite, consistent with a multiband energy-detector model using an optimum decision rule; it appears that listeners may only make an unweighted sum of decision variables across an optimum selection of channels.

A hardware cochlear nonlinear preprocessing model with active feedback.

A hardware model of the nonlinear preprocessing established in the inner ear consisting of 90 sections corresponding to a frequency range from 900 to 8000 Hz is described. The model is based on assumptions described by Zwicker [Biol. Cybern. 35, 243-250 (1979)]: The outer hair cells act as saturating nonlinear mechanical amplifiers which feed back to the vibration of the basilar membrane while only the inner hair cells transfer information towards higher centers. The model shows many effects which correlate very closely to physiological and psychoacoustical counterparts. Quantitative data on the level-dependence of frequency responses and phase responses as well as an example of suppression are outlined.

"Otoacoustic" emissions in a nonlinear cochlear hardware model with feedback.

Spontaneous, simultaneous evoked as well as delayed evoked emissions are studied in a hardware model of peripheral preprocessing with nonlinear feedback. The results suggest many very close parallels to the data found for otoacoustic emissions in man. From these parallels and the additional data measurable only in the model, it can be concluded that: the cochlea acts in a similar way as established in the model; the three kinds of emissions stem from the same source; the phase response of the cochlea's hydromechanics is responsible for the frequency distance between neighboring emissions as well as for the additional tips in suppression tuning curves; the long delay of the delayed evoked emissions is due to the many decaying contributions from the places along the basilar membrane which cancel each other in the early part but sum up to the delayed emission in the later part; and the double-peaked shape of the suppression-period patterns produced by high-level, low-frequency tones reflects the symmetrically shaped saturating nonlinearity of the feedback loops in the model which correspond to the function of the outer hair cells.

Suppression and (2f1-f2)-difference tones in a nonlinear cochlear preprocessing model with active feedback.

The two nonlinear effects of two-tone suppression and of (2f1-f2)-difference tone creation are measured in a hardware model which consists of 90 sections containing nonlinear feedback loops. The basic data are the level and phase distributions along the 90 sections produced by single tones in the linear passive system which are almost identical to those produced in the nonlinear active system at high levels. Enhancement is created at medium and low input levels resulting in more strongly peaked level-place patterns. Two-tone suppression is, therefore, described as a "de-enhancement" which is produced by the gain reduction in the saturating nonlinearity of the feedback loop in consequence of increasing input levels (that of the feedback loop in consequence of increasing input levels (that of the suppressor as well!). Characteristics of suppression are given in normalized form. The creation of (2f1-f2)-difference tones is based on the same nonlinear effects. In each section, difference-tone wavelets are created which travel--changing level and phase thereby--to their characteristic place, where they add up to a vector sum corresponding to the audible difference tone. In case of cancellation, the vector sum has to be compensated by an additional tone of the same frequency and amount but opposite phase. Based on this strategy of (2f1-f2)-difference tone development, the relevant relations are measured on the model and averaged either in normalized graphs or in equations in order to offer the possibility to simulate the hardware model on the computer. Psychoacoustically measured cancellation data are compared with data measured using the model. The two data sets agree not only in general but also in many details indicating that the model describes cochlear nonlinear preprocessing to a useful approximation.