Single-tube genotyping of MBL-2 polymorphisms using melting temperature analysis.

Recently several authors correlated MBL-2 gene polymorphisms with different pathologies and there is a growing interest for MBL-2 genotyping in a large number of individuals. We have developed a single-tube, rapid, economic, and fully automated melting temperature analysis screening method, based on ABI 7700 Sequence Detection System technology and SYBR Green I chemistry, for the detection of three polymorphisms (exon 1, codons 52, 54, 57) in the MBL-2 gene. We also developed an electronic sheet for the automatic calling of different genotypes, based on the analysis of the first derivative of ABI 7700 raw data.

Quantitative in situ detection of high-risk human papillomavirus in cytological specimens by SYBR Green I fluorescent labeling.

In this study we developed an in situ protocol for quantitative detection of high-risk human papillomavirus (HPV), based on direct in situ polymerase chain reaction (PCR) with SYBR Green I labeling and GeneAmp 5700 Sequence Detection System technology. This protocol was applied on cytological specimens of patients with cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma (SCC). We performed direct in situ quantitative PCR on cell smears, uninfected human skin fibroblasts, Hela and Caski cells. After in situ amplification, slides were counterstained with propidium iodide and analyzed under a fluorescent microscope in order to localize high-risk HPV and verify preservation of morphology. After PCR optimization, we obtained the following results. The Hela cells showed values ranging from 15 to 33 copies of high-risk HPV per cell, the Caski cell line from 220 to 300 high-risk HPV copies per cell and the cell smear (both CIN and SCC) around 20-35 copies of high-risk HPV per cell. No high-risk HPV amplification was detected in uninfected human fibroblasts, healthy controls, non-amplification control, and non-specific primer control. A positive intranuclear high-risk HPV amplification was detected in cell smears from 20 patients with CIN and 10 with SCC. In conclusion, our in situ quantitative protocol for high-risk HPV detection on cell smears combines both quantitative data and in situ localization of the target, with preservation of morphology. For this reason it could be used as a rapid screening tool when both morphological and quantitative results are requested on the same slide.

A rapid and quantitative mass spectrometry method for determining the concentration of acylcarnitines and aminoacids in amniotic fluid.

We describe a quantitative, rapid, sensitive and reproducible tandem mass spectrometry (MSMS) method for the one-step detection of aminoacid (AAs) and acylcarnitine (ACs) concentrations in amniotic fluid. This technology is quicker and more sensitive than other methods used to date since it is possible to determine very low AA and AC concentrations in samples simultaneously in a single run. The high degree of automation allows a large number of pregnancies to be screened for metabolic defects in a very short time.

Neurobiological foundations for the theory of harmony in western tonal music.

Basic principles of the theory of harmony reflect physiological and anatomical properties of the auditory nervous system and related cognitive systems. This hypothesis is motivated by observations from several different disciplines, including ethnomusicology, developmental psychology, and animal behavior. Over the past several years, we and our colleagues have been investigating the vertical dimension of harmony from the perspective of neurobiology using physiological, psychoacoustic, and neurological methods. Properties of the auditory system that govern harmony perception include (1) the capacity of peripheral auditory neurons to encode temporal regularities in acoustic fine structure and (2) the differential tuning of many neurons throughout the auditory system to a narrow range of frequencies in the audible spectrum. Biologically determined limits on these properties constrain the range of notes used in music throughout the world and the way notes are combined to form intervals and chords in popular Western music. When a harmonic interval is played, neurons throughout the auditory system that are sensitive to one or more frequencies (partials) contained in the interval respond by firing action potentials. For consonant intervals, the fine timing of auditory nerve fiber responses contains strong representations of harmonically related pitches implied by the interval (e.g., Rameau's fundamental bass) in addition to the pitches of notes actually present in the interval. Moreover, all or most of the partials can be resolved by finely tuned neurons throughout the auditory system. By contrast, dissonant intervals evoke auditory nerve fiber activity that does not contain strong representations of constituent notes or related bass notes. Furthermore, many partials are too close together to be resolved. Consequently, they interfere with one another, cause coarse fluctuations in the firing of peripheral and central auditory neurons, and give rise to perception of roughness and dissonance. The effects of auditory cortex lesions on the perception of consonance, pitch, and roughness, combined with a critical reappraisal of published psychoacoustic data on the relationship between consonance and roughness, lead us to conclude that consonance is first and foremost a function of the pitch relationships among notes. Harmony in the vertical dimension is a positive phenomenon, not just a negative phenomenon that depends on the absence of roughness--a view currently held by many psychologists, musicologists, and physiologists.

Development of speechreading supplements based on automatic speech recognition.

In manual-cued speech (MCS) a speaker produces hand gestures to resolve ambiguities among speech elements that are often confused by speechreaders. The shape of the hand distinguishes among consonants; the position of the hand relative to the face distinguishes among vowels. Experienced receivers of MCS achieve nearly perfect reception of everyday connected speech. MCS has been taught to very young deaf children and greatly facilitates language learning, communication, and general education. This manuscript describes a system that can produce a form of cued speech automatically in real time and reports on its evaluation by trained receivers of MCS. Cues are derived by a hidden markov models (HMM)-based speaker-dependent phonetic speech recognizer that uses context-dependent phone models and are presented visually by superimposing animated handshapes on the face of the talker. The benefit provided by these cues strongly depends on articulation of hand movements and on precise synchronization of the actions of the hands and the face. Using the system reported here, experienced cue receivers can recognize roughly two-thirds of the keywords in cued low-context sentences correctly, compared to roughly one-third by speechreading alone (SA). The practical significance of these improvements is to support fairly normal rates of reception of conversational speech, a task that is often difficult via SA.

A method to determine the speech transmission index from speech waveforms.

A method for computing the speech transmission index (STI) using real speech stimuli is presented and evaluated. The method reduces the effects of some of the artifacts that can be encountered when speech waveforms are used as probe stimuli. Speech-based STIs are computed for conversational and clearly articulated speech in several noisy, reverberant, and noisy-reverberant environments and compared with speech intelligibility scores. The results indicate that, for each speaking style, the speech-based STI values are monotonically related to intelligibility scores for the degraded speech conditions tested. Therefore, the STI can be computed using speech probe waveforms and the values of the resulting indices are as good predictors of intelligibility scores as those derived from MTFs by theoretical methods.

Effects of token variability on our ability to distinguish between vowels.

Even when the speaker, context, and speaking style are held fixed, the physical properties of naturally spoken utterances of the same speech sound vary considerably. This variability imposes limits on our ability to distinguish between different speech sounds. We present a conceptual framework for relating the ability to distinguish between speech sounds in single-token experiments (in which each speech sound is represented by a single wave form) to resolution in multiple-token experiments. Experimental results indicate that this ability is substantially reduced by an increase in the number of tokens from 1 to 4, but that there is little further reduction when the number of tokens increases to 16. Furthermore, although there is little relation between the ability to distinguish between a given pair of tokens in the multiple- and the 1-token experiments, there is a modest correlation between the ability to distinguish specific vowel tokens in the 4- and 16-token experiments. These results suggest that while listeners use a multiplicity of cues to distinguish between single tokens of a pair of vowel sounds, so that performance is highly variable both across tokens and listeners, they use a smaller set when distinguishing between populations of naturally produced vowel tokens, so that variability is reduced. The effectiveness of the cues used in the latter case is limited more by internal noise than by the variability of the cues themselves.

Consistency among speech parameter vectors: application to predicting speech intelligibility.

Previous researchers interested in physical assessment of speech intelligibility have largely based their predictions on preservation of spectral shape. A new approach is presented in which intelligibility is predicted to be preserved only if a transformation modifies relevant speech parameters in a consistent manner. In particular, the parameters from each short-time interval are described by one of a finite number of symbols formed by quantizing the output of an auditory model, and preservation of intelligibility is modeled as the extent to which a one-to-one correspondence exists between the symbols of the input to the transformation, and those of the output. In this paper, a consistency-measurement system is designed and applied to prediction of intelligibility of linearly filtered speech and speech degraded by additive noise. Results were obtained for two parameter sets: one consisting of band-energy values, and the other based on the ensemble interval histogram (EIH) model. Predictions within a class of transformation varied monotonically with the amount of degradation. Across classes of transformation, the predicted effect of additive-noise transformations was more severe than typical perceptual effects. With respect to the goal of achieving predictions that varied monotonically with human speech-perception scores, performance was slightly better with the EIH parameter set.

Speaking clearly for the hard of hearing IV: Further studies of the role of speaking rate.

The contribution of reduced speaking rate to the intelligibility of "clear" speech (Picheny, Durlach, & Braida, 1985) was evaluated by adjusting the durations of speech segments (a) via nonuniform signal time-scaling, (b) by deleting and inserting pauses, and (c) by eliciting materials from a professional speaker at a wide range of speaking rates. Key words in clearly spoken nonsense sentences were substantially more intelligible than those spoken conversationally (15 points) when presented in quiet for listeners with sensorineural impairments and when presented in a noise background to listeners with normal hearing. Repeated presentation of conversational materials also improved scores (6 points). However, degradations introduced by segment-by-segment time-scaling rendered this time-scaling technique problematic as a means of converting speaking styles. Scores for key words excised from these materials and presented in isolation generally exhibited the same trends as in sentence contexts. Manipulation of pause structure reduced scores both when additional pauses were introduced into conversational sentences and when pauses were deleted from clear sentences. Key-word scores for materials produced by a professional talker were inversely correlated with speaking rate, but conversational rate scores did not approach those of clear speech for other talkers. In all experiments, listeners with normal hearing exposed to flat-spectrum background noise performed similarly to listeners with hearing loss.

Recognition of amplitude-modulation patterns in the presence of a distractor. I. Effects of correlation and frequency relation.

Listeners' ability to compare the amplitude modulation pattern of 200- and 500-Hz targets when distractors that were also amplitude modulated were presented simultaneously was evaluated. The amplitude modulations of the distractors were either uncorrelated, partially correlated, or fully correlated with the amplitude modulations of the comparisons. Relative to the case of no distractor, performance tended to decrease when a distractor was present, and the degree of interference increased as the modulation correlation between the comparisons and distractors decreased. Although the interference was greater when the comparisons and distractors were separated by 50 Hz, there was also significant interference when the separation was 300 Hz. Whether the comparison was higher or lower in frequency than the distractor had no overall effect. However, the effect of modulation correlation was greater when comparisons were higher than distractors rather than lower. Patterns of interference are compared to those found in studies of modulation detection and discrimination interference, and implications for the use of multiple-band signals that carry the amplitude envelopes from different spectral regions of a speech signal to convey speech are discussed.

Recognition of amplitude-modulation patterns in the presence of a distractor. II. Effects of dichotic presentation and unmodulated distractors.

Listeners indicated which of two comparisons had the same pattern of amplitude modulation as a target signal when distractors were presented simultaneously with the comparisons. The frequency separation between distractors and comparisons was either narrow or wide. Distractors were either modulated independently of the comparisons, comodulated with the comparisons, or unmodulated. At wide frequency separations, none of the distractors interfered significantly with modulation comparison, relative to performance with no distractors. At narrow frequency separations, comodulated distractors produced less interference than did independently modulated distractors. Unmodulated distractors also produced some interference. There was no difference between diotic presentation and dichotic presentation, in which distractors were presented to the opposite ear from targets and comparisons. Implications for the presentation of multiple envelope signals derived from different spectral regions of a speech signal to convey speech to hearing-impaired listeners are discussed.

Effect of frequency transposition on the discrimination of amplitude envelope patterns.

The effects of systematic training on listeners' ability to compare the amplitude envelopes of signals differing in frequency was tested. Listeners indicated which of two comparison signals had the same amplitude envelope as the target signal. During training, center frequencies of comparison signals were gradually increased or decreased relative to target signal center frequencies of 500, 1600, and 3160 Hz. After training, performance was still worse when target and comparison signals were at different frequencies rather than at the same frequency, except possibly when comparison signals were higher in frequency than the 1600-Hz target. Thus the amplitude envelopes of signals do not appear to be perceived independently of the signal itself. Listeners who received no training performed similarly to the trained listeners, except that their performances declined when comparison signals were higher in frequency than the 1600-Hz target. Training did not reduce interlistener differences in overall performance or in the extent of the decline in performance when frequency differences between the target and comparison signals were introduced. The effects of frequency lowering on amplitude envelope discrimination do not appear to be related to the reduced efficacy of frequency-lowered speech-derived amplitude envelopes in supplementing speechreading.

Intelligibility of conversational and clear speech in noise and reverberation for listeners with normal and impaired hearing.

The effect of articulating clearly on speech intelligibility is analyzed for ten normal-hearing and two hearing-impaired listeners in noisy, reverberant, and combined environments. Clear speech is more intelligible than conversational speech for each listener in every environment. The difference in intelligibility due to speaking style increases as noise and/or reverberation increase. The average difference in intelligibility is 20 percentage points for the normal-hearing listeners and 26 percentage points for the hearing-impaired listeners. Two predictors of intelligibility are used to quantify the environmental degradations: The articulation index (AI) and the speech transmission index (STI). Both are shown to predict, reliably, performance levels within a speaking style for normal-hearing listeners. The AI is unable to represent the reduction in intelligibility scores due to reverberation for the hearing-impaired listeners. Neither predictor can account for the difference in intelligibility due to speaking style.

Auditory supplements to speechreading: combining amplitude envelope cues from different spectral regions of speech.

Many listeners with severe-to-profound hearing losses perceive only a narrow range of low-frequency sounds and must rely on speechreading to supplement the impoverished auditory signal in speech recognition. Previous research with normal-hearing subjects [Grant et al., J. Exp. Psychol. 43A, 621-645 (1991)] demonstrated that speechreading was significantly improved when supplemented by amplitude-envelope cues that were extracted from various spectral regions of speech and presented as amplitude modulations of carriers with frequencies at or below the speech band from which the envelope was derived. This experiment assessed the benefit to speechreading provided by pairs of such envelope cues presented simultaneously. In general, greater improvements in speechreading scores were observed for pairs than for single envelopes when the carrier signals were chosen appropriately. However, when pairs of envelope signals were transposed to low frequencies, the benefit to speechreading was no better than the most effective single-band envelope signal tested, or for a low-pass-filtered speech signal with the same overall bandwidth. Suggestions for improving the efficacy of frequency-lowered envelope cues for hearing-impaired listeners are discussed.

Automatic speech recognition to aid the hearing impaired: prospects for the automatic generation of cued speech.

Although great strides have been made in the development of automatic speech recognition (ASR) systems, the communication performance achievable with the output of current real-time speech recognition systems would be extremely poor relative to normal speech reception. An alternate application of ASR technology to aid the hearing impaired would derive cues from the acoustical speech signal that could be used to supplement speechreading. We report a study of highly trained receivers of Manual Cued Speech that indicates that nearly perfect reception of everyday connected speech materials can be achieved at near normal speaking rates. To understand the accuracy that might be achieved with automatically generated cues, we measured how well trained spectrogram readers and an automatic speech recognizer could assign cues for various cue systems. We then applied a recently developed model of audiovisual integration to these recognizer measurements and data on human recognition of consonant and vowel segments via speechreading to evaluate the benefit to speechreading provided by such cues. Our analysis suggests that with cues derived from current recognizers, consonant and vowel segments can be received with accuracies in excess of 80%. This level of performance is roughly equivalent to the segment reception accuracy required to account for observed levels of Manual Cued Speech reception. Current recognizers provide maximal benefit by generating only a relatively small number (three to five) of cue groups, and may not provide substantially greater aid to speechreading than simpler aids that do not incorporate discrete phonetic recognition. To provide guidance for the development of improved automatic cueing systems, we describe techniques for determining optimum cue groups for a given recognizer and speechreader, and estimate the cueing performance that might be achieved if the performance of current recognizers were improved.

Intensity perception. XIV. Intensity discrimination in listeners with sensorineural hearing loss.

Intensity discrimination of pulsed tones (also called level discrimination) was measured as a function of level in 13 listeners with sensorineural hearing impairment of primarily cochlear origin, one listener with a vestibular schwannoma, and six listeners with normal hearing. Measurements were also made in normal ears presented with masking noise spectrally shaped to produce audiograms similar to those of the cochlearly impaired listeners. For unilateral impairments, tests were made at the same frequency in the normal and impaired ears. For bilateral-sloping impairments, tests were made at different frequencies in the same ear. The normal listeners showed results similar to other data in the literature. The listener with a vestibular schwannoma showed greatly reduced intensity resolution, except at a few levels. For listeners with recruiting sensorineural impairments, the results are discussed according to the configuration of the impairment and are compared across configurations at equal SPL, equal SL, and equal loudness level. Listeners with increasing hearing losses at frequencies above the test frequency generally showed impaired resolution, especially at high levels, and less deviation from Weber's law than normal listeners. Listeners with decreasing hearing loss at frequencies above the test frequency showed nearly normal intensity-resolution functions. Whereas these trends are generally present, there are also large differences among individuals. Results obtained from normal listeners who were tested in the presence of masking noise indicate that elevated thresholds and reduced dynamic range account for some, but not all, of the effects of recruiting sensorineural impairment on intensity resolution.

Intelligibility of frequency-lowered speech produced by a channel vocoder.

Frequency lowering is a form of signal processing designed to match speech to the residual auditory capacity of a listener with a high frequency hearing loss. A vocoder-based frequency-lowering system similar to one studied by Lippmann was evaluated in the present study. In this system, speech levels in high frequency bands modulated one-third-octave bands of noise at low frequencies, which were then added to unprocessed speech. Results obtained with this system indicated, in agreement with Lippmann, that processing improved the recognition of stop, fricative, and affricate consonants when the listening bandwidth was restricted to 800 Hz. However, results also showed that processing degraded the perception of nasals and semivowels, consonants not included in Lippmann's study. Based on these results, the frequency-lowering system was modified so as to suppress the processing whenever low frequency components dominated the input signal. High and low frequency energies of an input signal were measured continuously in the modified system, and the decision to process or to leave the signal unaltered was based on their relative levels. Results indicated that the modified system maintained the processing advantage for stops, fricatives, and affricates without degrading the perception of nasals and semi-vowels. The results of the present study also indicated that training is an important consideration when evaluating frequency-lowering systems.

Analytic study of the Tadoma method: improving performance through the use of supplementary tactual displays.

Although results obtained with the Tadoma method of speechreading have set a new standard for tactual speech communication, they are nevertheless inferior to those obtained in the normal auditory domain. Speech reception through Tadoma is comparable to that of normal-hearing subjects listening to speech under adverse conditions corresponding to a speech-to-noise ratio of roughly 0 dB. The goal of the current study was to demonstrate improvements to speech reception through Tadoma through the use of supplementary tactual information, thus leading to a new standard of performance in the tactual domain. Three supplementary tactual displays were investigated: (a) an articulatory-based display of tongue contact with the hard palate; (b) a multichannel display of the short-term speech spectrum; and (c) tactual reception of Cued Speech. The ability of laboratory-trained subjects to discriminate pairs of speech segments that are highly confused through Tadoma was studied for each of these augmental displays. Generally, discrimination tests were conducted for Tadoma alone, the supplementary display alone, and Tadoma combined with the supplementary tactual display. The results indicated that the tongue-palate contact display was an effective supplement to Tadoma for improving discrimination of consonants, but that neither the tongue-palate contact display nor the short-term spectral display was highly effective in improving vowel discriminability. For both vowel and consonant stimulus pairs, discriminability was nearly perfect for the tactual reception of the manual cues associated with Cued Speech. Further experiments on the identification of speech segments were conducted for Tadoma combined with Cued Speech. The observed data for both discrimination and identification experiments are compared with the predictions of models of integration of information from separate sources.