Nonlinear viscoelastic characterization of human vocal fold tissues under large-amplitude oscillatory shear (LAOS).

Viscoelastic shear properties of human vocal fold tissues were previously quantified by the shear moduli (G' and G″). Yet these small-strain linear measures were unable to describe any nonlinear tissue behavior. This study attempted to characterize the nonlinear viscoelastic response of the vocal fold lamina propria under large-amplitude oscillatory shear (LAOS) with a stress decomposition approach. Human vocal fold cover and vocal ligament specimens from eight subjects were subjected to LAOS rheometric testing with a simple-shear rheometer. The empirical total stress response was decomposed into elastic and viscous stress components, based on odd-integer harmonic decomposition approach with Fourier transform. Nonlinear viscoelastic measures derived from the decomposition were plotted in Pipkin space and as rheological fingerprints to observe the onset of nonlinearity and the type of nonlinear behavior. Results showed that both the vocal fold cover and the vocal ligament experienced intercycle strain softening, intracycle strain stiffening, as well as shear thinning both intercycle and intracycle. The vocal ligament appeared to demonstrate an earlier onset of nonlinearity at phonatory frequencies, and higher sensitivity to changes in frequency and strain. In summary, the stress decomposition approach provided much better insights into the nonlinear viscoelastic behavior of the vocal fold lamina propria than the traditional linear measures.

Vocal Range Variations Elicited With a Standard Mandarin Reading Passage in Individuals With Voice Disorders.

PURPOSE: This study examined whether the "Three Bears Passage" (TB), a standard Mandarin reading passage, could elicit significant vocal range variations in individuals with voice disorders. Relative sensitivity of TB versus another existing standard reading passage, "Passage in Mandarin" (PM), for differentiating between individuals with and without voice disorders was also evaluated. METHOD: Forty-two individuals with normal voice and 30 individuals with voice disorders participated in the study. Maximum fundamental frequency (f0), minimum f0, mean f0, f0 range, maximum vocal intensity, minimum intensity, mean intensity, and intensity range of all participants reading aloud the two passages were measured with Praat to construct speech range profiles (SRPs). RESULTS: Significantly larger vocal range was found for TB than for PM in individuals with voice disorders, including significantly higher maximum f0, mean f0, maximum intensity, mean intensity, and significantly larger f0 range and intensity range. Significantly more limited vocal range was observed in individuals with voice disorders than those without, with more obviously restricted SRPs while reading aloud TB compared to PM. Receiver operating characteristic analysis suggested that TB was more sensitive than PM in distinguishing between individuals with and without voice disorders. CONCLUSIONS: Our findings supported the potential of TB as a standard clinical assessment tool for evaluating pathological changes in vocal range. Future studies should explore if therapeutic approaches based on the passage or variations of it could be developed for overcoming functional limitations and restrictions in vocal range for specific voice disorders.

The anisotropic hyperelastic biomechanical response of the vocal ligament and implications for frequency regulation: a case study.

One of the primary mechanisms to vary one's vocal frequency is through vocal fold length changes. As stress and deformation are linked to each other, it is hypothesized that the anisotropy in the biomechanical properties of the vocal fold tissue would affect the phonation characteristics. A biomechanical model of vibrational frequency rise during vocal fold elongation is developed which combines an advanced biomechanical characterization protocol of the vocal fold tissue with continuum beam models. Biomechanical response of the tissue is related to a microstructurally informed, anisotropic, nonlinear hyperelastic constitutive model. A microstructural characteristic (the dispersion of collagen) was represented through a statistical orientation function acquired from a second harmonic generation image of the vocal ligament. Continuum models of vibration were constructed based upon Euler-Bernoulli and Timoshenko beam theories, and applied to the study of the vibration of a vocal ligament specimen. From the natural frequency predictions in dependence of elongation, two competing processes in frequency control emerged, i.e., the applied tension raises the frequency while simultaneously shear deformation lowers the frequency. Shear becomes much more substantial at higher modes of vibration and for highly anisotropic tissues. The analysis was developed as a case study based on a human vocal ligament specimen.

A Vocal Hygiene Program for Mitigating the Effects of Occupational Vocal Demand in Primary School Teachers.

PURPOSE: Teachers are occupational voice users with significant vocal demand. This study examined if a vocal hygiene program could mitigate the effects of occupational vocal demand in primary school teachers across 1 month. METHOD: Sixty female teachers participated, with 30 in an experimental group receiving vocal hygiene education plus daily home practice for 1 month and 30 in a control group with no intervention. Their vocal changes across the month were quantified with (a) acoustic measures on fundamental frequency (fo), vocal intensity, jitter and shimmer, harmonics-to-noise ratio, and smoothed cepstral peak prominence and (b) Voice Handicap Index (VHI-10) and Vocal Fatigue Index (VFI) scores. RESULTS: Analysis of covariance showed significantly larger changes (significant decreases) in conversational fo and in jitter for the experimental group relative to the control group. Post hoc pairwise comparisons following repeated-measures analysis of variance showed significant decreases in conversational fo and in jitter across the month for the experimental group. No significant differences in VHI-10 and VFI scores were found between the groups. CONCLUSIONS: Vocal demand-related changes in acoustic measures could be partially mitigated with the vocal hygiene program. Future studies with a more refined intervention program and more long-term follow-up are recommended to better understand the long-term benefits of vocal hygiene programs on primary school teachers.

Mandarin Chinese version of the Aging Voice Index.

Objectives: This study developed the Mandarin Chinese version of the Aging Voice Index (AVI), with preliminary validation of the scale for potential clinical applications. Study Design: Scale development. Methods: The experimental procedure involved: (1) cross-cultural adaptation of the original AVI into the Mandarin Chinese version (CAVI); (2) evaluation by expert panel; (3) back translation; (4) pilot testing; (5) development of the final CAVI; (6) scale validation with 68 older adults of 60-89 years old (29 females and 39 males), 34 with voice disorders and 34 age-matched with normal voice. Internal consistency reliability, test-retest reliability, content validity, criterion-related validity, and discriminatory ability (diagnostic accuracy) of the CAVI were evaluated. Results: There were high internal consistency (Cronbach's alpha = 0.9733), high test-retest reliability (intraclass correlation coefficient = 0.9578, p < 0.01), high content validity (content validity index = 0.9710), high criterion-related validity (Pearson's r = 0.9439, p < 0.01 between CAVI and Voice Handicap Index-10; r = 0.8070, p < 0.01 between CAVI and voice-related quality of life [V-RQOL]), and significant difference in CAVI scores between the two groups with huge effect size (t(34.69) = -11.59, Cohen's d = 2.81, p < 0.001). Receiver operating characteristic analysis revealed a high diagnostic accuracy of the CAVI, with an area under the curve of 0.9974 (p < 0.001) and a cut-off score of 12.0 with 100% sensitivity and 97.1% specificity. Conclusion: Our findings suggested that the CAVI could be a reliable and valid standardized self-assessment questionnaire tool for clinical evaluation of the impact of voice problems specifically for Mandarin-speaking older adults. Further studies should explore a full-scale validation of the CAVI for being a standard clinical tool, including for older adults in Mainland China. Level of evidence: 3b (case-control study).

A 4-Week Straw Phonation in Water Exercise Program for Aging-Related Vocal Fold Atrophy.

PURPOSE: This study evaluated the efficacy of a 4-week straw phonation in water (SPW) exercise program on aging-related vocal fold atrophy (VFA), with a secondary objective to examine the immediate effects of SPW exercises. METHOD: Thirty-eight older adults aged 60 years and above formally diagnosed with aging-related VFA were randomly assigned into an experimental group undergoing SPW exercises with an 8-cm depth of straw submersion into water for 4 weeks plus vocal hygiene practice (n = 20), and a control group with only vocal hygiene practice (n = 18). Outcome measures included laryngeal endoscopic measures of glottal gap, auditory-perceptual ratings of voice quality, acoustic measures, aerodynamic measures, and standardized self-assessment questionnaire scores. An additional round of acoustic and aerodynamic assessment following 20 min of SPW exercises was conducted to examine the immediate effects. RESULTS: Significant improvements in normalized glottal gap area, perceptual rating of breathiness, smoothed cepstral peak prominence, harmonics-to-noise ratio (HNR), mean oral airflow, subglottal pressure and laryngeal airway resistance at comfortable loudness, Voice-related Quality of Life scores, and Chinese Vocal Fatigue Index Factor 3 scores were observed in the experimental group relative to the control group. There were also significant immediate effects for HNR, mean oral airflow, subglottal pressure, and laryngeal airway resistance. CONCLUSIONS: These findings suggested significant immediate improvements in vocal function following SPW exercises, with additional significant improvements in vocal function as well as significant improvements in quality of life following the 4-week SPW exercise program. Further studies with more long-term follow-up are recommended to better understand the efficacy of SPW exercises with deep levels of straw submersion into water as an effective clinical option for the management of hypofunctional dysphonia associated with aging-related VFA.

Validation of a High-Sensitivity Assay for Detection of Chimeric Antigen Receptor T-Cell Vectors Using Low-Partition Digital PCR Technology.

Although in vivo engraftment, expansion, and persistence of chimeric antigen receptor (CAR) T cells are pivotal components of treatment efficacy, quantitative monitoring has not been implemented in routine clinical practice. We describe the development and analytical validation of a digital PCR assay for ultrasensitive detection of CAR constructs after treatment, circumventing known technical limitations of low-partitioning platforms. Primers and probes, designed for detection of axicabtagene, brexucabtagene, and Memorial Sloan Kettering CAR constructs, were employed to validate testing on the Bio-Rad digital PCR low-partitioning platform; results were compared with Raindrop, a high-partitioning system, as reference method. Bio-Rad protocols were modified to enable testing of DNA inputs as high as 500 ng. Using dual-input reactions (20 and 500 ng) and a combined analysis approach, the assay demonstrated consistent target detection around 1 × 10-5 (0.001%) with excellent specificity and reproducibility and 100% accuracy compared with the reference method. Dedicated analysis of 53 clinical samples received during validation/implementation phases showed the assay effectively enabled monitoring across multiple time points of early expansion (day 6 to 28) and long-term persistence (up to 479 days). CAR vectors were detected at levels ranging from 0.005% to 74% (vector versus reference gene copies). The highest levels observed in our cohort correlated strongly with the temporal diagnosis of grade 2 and 3 cytokine release syndrome diagnosis (P < 0.005). Only three patients with undetectable constructs had disease progression at the time of sampling.

Acoustic Analysis of Taiwanese Tones in Esophageal Speech and Pneumatic Artificial Laryngeal Speech.

PURPOSE: This study examined the acoustic properties of Taiwanese (Southern Min) lexical tones produced in esophageal speech (ES) and pneumatic artificial laryngeal speech (PAL), including onset fundamental frequency (F0), slope of F0 contour, duration, and amplitude (intensity) of the vowel portion of syllables carrying seven Taiwanese tones. METHOD: Seven ES speakers, seven PAL speakers, and 18 normal laryngeal (NL) speakers (all male) read aloud target syllables carrying the tones in monosyllables, disyllables, and sentences. Acoustic features were measured with Praat. RESULTS: Acoustic analysis revealed (a) no significant effects of linguistic level on acoustic parameters except for duration, which generally significantly decreased from monosyllables to disyllables to sentences for Tones 1 and 5; (b) significant differences across the speaker groups in onset F0 (PAL > ES > NL), duration (PAL > NL and PAL > ES for all tones), and mean amplitude (PAL > NL > ES); and (c) no significant difference in mean slope of F0 contour across the speaker groups. CONCLUSIONS: These preliminary data on acoustic properties of Taiwanese tones produced by ES and PAL speakers could facilitate the design of therapeutic strategies for improving the intelligibility of alaryngeal lexical tones. Further studies are recommended to examine the relative perceptual importance of the acoustic parameters.

The Reliability and Validity of the Mandarin Chinese Version of the Vocal Fatigue Index: Preliminary Validation.

PURPOSE: This study attempted to develop and to preliminarily validate the Mandarin Chinese version of the Vocal Fatigue Index (VFI) as a standardized self-assessment questionnaire tool for potential clinical applications. METHOD: The experimental procedure involved (a) cross-cultural adaptation of the VFI into the Mandarin Chinese version (CVFI), (b) evaluation by an expert panel, (c) back translation, (d) pilot testing, and (e) validation of the questionnaire by three participant groups: 50 with voice disorders, 50 occupational voice users (at-risk group), and 50 with normal voice (control group). Internal consistency, test-retest reliability, content validity, and convergent validity of the CVFI were examined, and discriminatory ability (diagnostic accuracy) for distinguishing between the groups was evaluated. RESULTS: Results showed high internal consistency (Cronbach's alpha ≥ .8817 for the total CVFI scores for all groups), high test-retest reliability (intraclass correlation coefficients ≥ .9072, p < .001 for the total CVFI scores for all groups), high content validity (total content validity index = 0.9368), and high convergent validity (Pearson r ≥ .8155, p < .001 between the total CVFI scores and Factors 1 and 2 scores). Significant differences between the three groups were found in all scores. Receiver operating characteristic analysis revealed a high diagnostic accuracy for distinguishing between the disorders group and the normal group (area under the curve ≥ 0.927, p < .001 for the total CVFI scores and Factors 1 and 2 scores), with cutoff scores of ≥ 36 (total CVFI score), ≥ 23.5 (Factor 1 score), ≥ 7.5 (Factor 2 score), and ≤ 6.5 (Factor 3 score). CONCLUSIONS: These findings suggested that the CVFI could be a reliable and valid self-assessment tool for the clinical evaluation of vocal fatigue in Mandarin Chinese-speaking populations. A full-scale validation study of the CVFI is recommended to verify these results.

Clinical Utility and Performance of an Ultrarapid Multiplex RNA-Based Assay for Detection of ALK, ROS1, RET, and NTRK1/2/3 Rearrangements and MET Exon 14 Skipping Alterations.

Several kinase fusions are established targetable drivers in lung cancers. However, rapid and comprehensive detection remains challenging because of diverse partner genes and breakpoints. We assess the clinical utility and performance of a rapid microfluidic multiplex real-time PCR-based assay for simultaneous query of fusions involving ALK, ROS1, RET, and NTRK1/2/3, as well as MET exon 14 skipping, using a 3-hour automated process. Dual analytic strategies were utilized: fusion-specific amplification and 3' to 5' expression imbalance. One-hundred and forty-three independent, formalin-fixed, paraffin-embedded tumor samples (112 surgical specimens, 31 cytologic cell blocks) were analyzed: 133 with known kinase gene alterations and 10 negative samples based on clinically validated next-generation sequencing. Testing was successful in 142 (99%) cases. The assay demonstrated a sensitivity of 97% (28/29), 100% (31/31), 92% (22/24), 81% (22/27), and 100% (20/20) for ALK, RET, ROS1, and NTRK1/2/3 rearrangements and MET exon 14 skipping alterations, respectively, with 100% specificity for all. Concordant results were achieved in specimens aged up to 5 years, with >10% tumor, and inputs of at least 9 mm2 (surgical specimens) and 9000 cells (cytologic cell blocks). The assay enables rapid screening for clinically actionable kinase alterations with quicker turnaround and lower tissue requirements compared with immunohistochemistry and molecular methods, while also circumventing the infrastructure dependencies associated with next-generation sequencing and fluorescence in situ hybridization.

Effect of Training With Anchors on Auditory-Perceptual Evaluation of Dysphonia in Speech-Language Pathology Students.

Purpose Auditory-perceptual evaluation is essential for the clinical assessment of voice disorders. Unstable perceptual voice evaluation has been shown for inexperienced listeners as compared to expert listeners. We examined the effects of perceptual training with external auditory anchors with and without immediate feedback on the evaluation of roughness and breathiness of natural, nonsynthesized speech stimuli (reading of a standard passage) in speech-language pathology students. Method Perceptual voice evaluation and training with anchors using a visual analog scale was implemented with a computer software. Forty-eight speech-language pathology students were randomly assigned into three groups, feedback group (Group F), no feedback group (Group NF), and control group (Group C), attending one training session and four assessment sessions (before training, immediately after training, and 1 and 7 weeks after training). Group F received training with anchors with immediate feedback, Group NF received training without immediate feedback, and Group C received sham training (exposure session). Results Training with anchors significantly increased the rating accuracy (agreement with expert ratings) on both roughness and breathiness for Group F, with the effects lasting for 7 weeks. No significant changes in rating accuracy with training were observed for Group NF and Group C. No improvements in intra- and interrater reliability as well as intrarater agreement were observed in all three groups, whereas interrater agreement on breathiness (but not roughness) significantly increased for all groups, with the effect lasting for 7 weeks only for Group F. Conclusions These findings suggested that perceptual training with external auditory anchors and the use of immediate feedback could be effective for facilitating the development of perceptual voice evaluation skills in speech-language pathology students. Further studies involving more extensive training with stimuli covering a full range of dysphonia severity categories and improvements in design of the training protocol are recommended to verify these results.

A Mandarin Chinese Reading Passage for Eliciting Significant Vocal Range Variations.

Purpose There is a lack of standardized Mandarin reading text material that could potentially elicit significant variations in fundamental frequency (F0) and in vocal intensity for clinical voice evaluation. In this study, a phonetically balanced "Three Bears Passage" was developed based on the classical "Goldilocks" story for this purpose. The hypothesis was that the vocal range (F0 range and Intensity range) elicited while reading aloud the passage could be similar to that of the voice range profile (VRP) obtained with sustained vowel production and significantly different from that while reading an existing standard Mandarin passage. Method One hundred twenty young adults (60 men and 60 women, aged 25.22 ± 3.74 years) without vocal pathologies and vocal training participated in the study. Vocal range data, including F0-related measures and intensity-related measures, were obtained with Praat in order to construct VRPs for the sustained /a/ and to construct speech range profiles for passage reading. Results F0 range and intensity range for the new passage were significantly larger than those for the existing Mandarin passage for both male and female participants. In particular, the F0 range for the passage was not significantly different from that of the VRP for male participants. Conclusions These findings suggested that the new passage could allow individuals without vocal training to demonstrate a large vocal range in both F0 and vocal intensity, in a passage reading setting in a relatively short time. Further studies should be pursued with the passage to investigate the vocal range and vocal potential of individuals with voice disorders.

Phonation Threshold Pressure/Flow for Reflecting Glottal Closure in Unilateral Vocal Fold Paralysis.

OBJECTIVE: Previous theoretical analysis predicted that phonation threshold flow (PTF) could be a more sensitive aerodynamic measure than phonation threshold pressure (PTP) for reflecting glottal incompetence. This study investigated the feasibility of whether PTP and PTF may differentiate subjects with unilateral adductor vocal fold paralysis and paresis (UAVFP) from those without, and whether PTP and PTF could reflect the extent of incomplete glottal closure associated with UAVFP. METHODS: PTP and PTF were quantified for 13 subjects with UAVFP and 21 control subjects with normal voice, and the normalized glottal gap area (NGGA) based on videostroboscopic image analysis was quantified for subjects with UAVFP. RESULTS: Significant differences in both PTP and PTF were found between subjects with UAVFP and control subjects. Receiver operating characteristic analysis indicated a higher discriminatory ability of PTP for differentiating subjects with UAVFP from those without (area under the curve of 0.905 for PTP, 0.678 for PTF), yet a significant positive correlation was found between PTF and NGGA (Spearman's ρ = 0.571) but not between PTP and NGGA (ρ = -0.364). CONCLUSION: Results supported the feasibility of using PTP and PTF as potential diagnostic indicators for reflecting glottal closure in UAVFP, with PTP potentially more sensitive for differentiating subjects with and without incomplete glottal closure. These preliminary findings were limited by the small sample size, with further studies needed to verify whether PTF could be more sensitive for reflecting the extent of incomplete glottal closure, as predicted theoretically. LEVEL OF EVIDENCE: 3 Laryngoscope, 131:E1598-E1604, 2021.

Arytenoid adduction for correcting vocal fold asymmetry: high-speed imaging.

OBJECTIVES: We hypothesized that high-speed digital imaging provides a quantitative method to evaluate the effect of arytenoid adduction for the correction of asymmetric and irregular vocal fold vibration in unilateral vocal fold paralysis. METHODS: Six subjects with unilateral vocal fold paralysis participated in the study (4 male, 2 female; mean [-SD] age, 52.5 +/- 21.3 years). Videokymographic and laryngotopographic methods for image analysis were performed for high-speed recordings of vocal fold vibration for visualizing the glottal vibratory patterns, and for quantifying the frequency of vibration of each vocal fold, respectively. Comparisons of the paralyzed and the normal vocal folds were made before and after arytenoid adduction. RESULTS: Analysis of the laryngotopographs revealed 2 distinct frequencies of vibration for the paralyzed and the contralateral vocal folds for all subjects before surgery. After arytenoid adduction, the vibration frequencies became identical or nearly identical in all subjects. CONCLUSIONS: Asymmetric vibration in vocal fold paralysis was exemplified by differences in vibration frequency between the vocal folds. The present data showed that after arytenoid adduction the vibration frequencies and the vibratory patterns of the contralateral vocal folds approached symmetry. This surgical procedure could improve the functional symmetry of the larynx for phonation.

Collagen injection for correcting vocal fold asymmetry: high-speed imaging.

OBJECTIVES: We hypothesized that high-speed digital imaging with videokymographic and laryngotopographic analysis would provide a quantitative method to evaluate the effect of collagen injection for the correction of asymmetric and irregular vocal fold vibration in unilateral vocal fold paralysis. METHODS: Videokymographic and laryngotopographic analysis was performed for high-speed digital recordings of vocal fold vibration for visualizing the glottal vibratory patterns, and for quantifying the frequency of vibration of each vocal fold, respectively, including comparisons between the paralyzed and normal vocal folds before and after surgery. This included prospective observations of 11 subjects with unilateral vocal fold paralysis (4 male, 7 female; mean +/- SD age, 67.1 +/- 12.0 years) using high-speed digital image analysis before and after collagen injection. RESULTS: Analysis of the laryngotopographs revealed 2 distinct frequencies of vibration for the paralyzed and contralateral vocal folds for 8 of the 11 subjects before surgery. After collagen injection, the vibration frequencies became identical, despite asymmetric vibration amplitudes. Asymmetric vibration amplitudes were also observed in the other 3 subjects before surgery, but the amplitudes became symmetric after collagen injection, despite a persistent phase shift. CONCLUSIONS: Asymmetric vibration in vocal fold paralysis was exemplified by differences in vibration frequency and amplitude between the vocal folds. The present study showed that after collagen injection, these aspects of vibratory patterns improved toward symmetry. This surgical procedure could improve the functional symmetry of the larynx for phonation.

Effects of a 6-Week Straw Phonation in Water Exercise Program on the Aging Voice.

Purpose Semi-occluded vocal tract (SOVT) exercises with tubes or straws have been widely used for a variety of voice disorders. Yet, the effects of longer periods of SOVT exercises (lasting for weeks) on the aging voice are not well understood. This study investigated the effects of a 6-week straw phonation in water (SPW) exercise program. Method Thirty-seven elderly subjects with self-perceived voice problems were assigned into two groups: (a) SPW exercises with six weekly sessions and home practice (experimental group) and (b) vocal hygiene education (control group). Before and after intervention (2 weeks after the completion of the exercise program), acoustic analysis, auditory-perceptual evaluation, and self-assessment of vocal impairment were conducted. Results Analysis of covariance revealed significant differences between the two groups in smoothed cepstral peak prominence measures, harmonics-to-noise ratio, the auditory-perceptual parameter of breathiness, and Voice Handicap Index-10 scores postintervention. No significant differences between the two groups were found for other measures. Conclusions Our results supported the positive effects of SOVT exercises for the aging voice, with a 6-week SPW exercise program being a clinical option. Future studies should involve long-term follow-up and additional outcome measures to better understand the efficacy of SOVT exercises, particularly SPW exercises, for the aging voice.

Ultrarapid EGFR Mutation Screening Followed by Comprehensive Next-Generation Sequencing: A Feasible, Informative Approach for Lung Carcinoma Cytology Specimens With a High Success Rate.

INTRODUCTION: For patients with advanced NSCLC, cytologic samples may be the only diagnostic specimen available for molecular profiling. Although both rapid and comprehensive assessment are essential in this setting, an integrated multitest approach remains an important strategy in many laboratories, despite the risks and challenges when working with scant samples. In this study, we describe our experience and high success rate in using a multitest approach, focusing on the clinical validation and incorporation of ultrarapid EGFR testing using the Idylla system followed by comprehensive next-generation sequencing (NGS). METHODS: Cytology samples received for routine molecular testing were included in this study. The performance characteristics of the EGFR Idylla assay were assessed; tissue suitability parameters and interpretation criteria to supplement automated mutation calling were established. The assay performance was monitored for 1 year, comparing the results with those of concurrent NGS testing by MSK-IMPACT (primarily) or MSK-AmpliSeq and MSK-Fusion solid panel in a subset of cases. RESULTS: Overall, 301 samples were studied; 83 samples were included in validation (60.2% [50 of 83] were positive for EGFR mutations). Concordance with the reference method was 96.4% (80 of 83) of the samples with excellent reproducibility. The limit of detection was variable depending on the total tissue input and the specific mutation tested. Unextracted tissue inputs that maintained total EGFR cycle of quantification at less than 23 allowed all mutations to be detected if present at greater than 5% variant allele frequency. Mutations could be detected at 1% variant allele frequency with total EGFR cycle of quantification of 18. During the clinical implementation phase, 218 NSCLC samples were tested by Idylla (24.3% [53 of 218] were EGFR mutation positive). Concurrent NGS testing was requested on 165 samples and successfully performed on 96.4% (159 of 165) of the samples. The Idylla automated results were concordant with those obtained by NGS in 96.2% (153 of 159) of cases and improved to 98.7% (157 of 159) after incorporation of manual review criteria to supplement automated calling, resulting in a diagnostic sensitivity of 95.6% (95% confidence interval: 84.9%-99.5%). In general, 9% (14 of 159) of the cases tested by NGS had EGFR mutations not covered by the Idylla assay, primarily insertions in exon 19 and 20 and minor mutations cooccurring with canonical sensitizing mutations. CONCLUSIONS: Comprehensive molecular testing is feasible and has a high success rate in NSCLC cytology samples when using a multitest approach. Testing with the Idylla system enables rapid and accurate determination of the EGFR status without compromising subsequent NGS testing.

Ovine Vocal Fold Tissue Fatigue Response to Accumulated, Large-Amplitude Vibration Exposure at Phonatory Frequencies.

Purpose The contribution of tissue mechanical response to vocal fatigue is poorly understood. This study investigated the fatigue response of vocal fold tissues to large-amplitude vibration exposure at phonatory frequencies, using an ex vivo ovine model. Method Twelve sheep vocal fold mucosal specimens were subjected to sinusoidal, simple-shear deformation for prolonged cycles, under a large but physiological shear strain (46%) in a frequency range of 100-230 Hz. The duration of shear varied from a critical vibration exposure limit of 1,040 s to 4 times the limit (4,160 s). Tissue viscoelastic response was quantified by the elastic shear modulus (G'), viscous shear modulus (G″), and damping ratio (G″/G'). Results Distinct response patterns were observed at different frequencies. G' and G″ generally decreased with vibration exposure at 100 Hz, whereas they generally increased with vibration exposure at 200 and 230 Hz. Statistically significant differences were found for G″ increasing with vibration exposure at 200 Hz and damping ratio decreasing with vibration exposure at 200 Hz. Significant increases with frequency were also found for all viscoelastic functions. Results suggested that the contribution of tissue viscoelastic response to vocal fatigue could be highly frequency dependent. In particular, increases in G″ with vibration exposure could lead to high phonation threshold pressures and difficulty sustaining phonation at higher frequencies following prolonged vocalization. Conclusion These preliminary findings may help us better understand vocal fatigue and recovery and should be corroborated by studies with human vocal fold tissues.

Collagen injection as a supplement to arytenoid adduction for vocal fold paralysis.

OBJECTIVES: Dysphonia associated with vocal fold paralysis can persist even after successful medialization procedures, including arytenoid adduction. It is hypothesized that laryngeal collagen injection could improve phonation following arytenoid adduction in selected patients. Our objective was to evaluate how collagen injection could result in measurable improvements in vocal function and voice quality. METHODS: Forty patients with unilateral vocal fold paralysis who had undergone arytenoid adduction underwent transoral injection of non-cross-linked bovine dermal collagen by means of indirect laryngoscopy and a curved injection device. A control group of 40 patients underwent arytenoid adduction but not collagen injection. The patients' voice quality was assessed perceptually with the GRBAS scale, and vocal function was assessed by acoustic and aerodynamic measures (maximum phonation time and transglottal DC flow). The relative glottal area was also assessed by videostroboscopy. RESULTS: Significant improvements in vocal function and voice quality were observed with collagen injection for those patients who did not achieve satisfactory glottal competence with arytenoid adduction alone. Glottal area measurements revealed that glottic insufficiency was significantly reduced after arytenoid adduction as well as after collagen injection. CONCLUSIONS: The findings suggest that collagen injection could be an effective supplementary treatment for improving voice following arytenoid adduction. It has the advantage of being a minimally invasive outpatient office procedure. The long-term efficacy of the procedure should be explored.

A simple-shear rheometer for linear viscoelastic characterization of vocal fold tissues at phonatory frequencies.

Previous studies reporting the linear viscoelastic shear properties of the human vocal fold cover or mucosa have been based on torsional rheometry, with measurements limited to low audio frequencies, up to around 80 Hz. This paper describes the design and validation of a custom-built, controlled-strain, linear, simple-shear rheometer system capable of direct empirical measurements of viscoelastic shear properties at phonatory frequencies. A tissue specimen was subjected to simple shear between two parallel, rigid acrylic plates, with a linear motor creating a translational sinusoidal displacement of the specimen via the upper plate, and the lower plate transmitting the harmonic shear force resulting from the viscoelastic response of the specimen. The displacement of the specimen was measured by a linear variable differential transformer whereas the shear force was detected by a piezoelectric transducer. The frequency response characteristics of these system components were assessed by vibration experiments with accelerometers. Measurements of the viscoelastic shear moduli (G' and G") of a standard ANSI S2.21 polyurethane material and those of human vocal fold cover specimens were made, along with estimation of the system signal and noise levels. Preliminary results showed that the rheometer can provide valid and reliable rheometric data of vocal fold lamina propria specimens at frequencies of up to around 250 Hz, well into the phonatory range.