Expression, purification, and spectral tuning of RhoGC, a retinylidene/guanylyl cyclase fusion protein and optogenetics tool from the aquatic fungus Blastocladiella emersonii.

RhoGC is a rhodopsin (Rho)-guanylyl cyclase (GC) gene fusion molecule that is central to zoospore phototaxis in the aquatic fungus Blastocladiella emersonii It has generated considerable excitement because of its demonstrated potential as a tool for optogenetic manipulation of cell-signaling pathways involving cyclic nucleotides. However, a reliable method for expressing and purifying RhoGC is currently lacking. We present here an expression and purification system for isolation of the full-length RhoGC protein expressed in HEK293 cells in detergent solution. The protein exhibits robust light-dependent guanylyl cyclase activity, whereas a truncated form lacking the 17- to 20-kDa N-terminal domain is completely inactive under identical conditions. Moreover, we designed several RhoGC mutants to increase the utility of the protein for optogenetic studies. The first class we generated has altered absorption spectra designed for selective activation by different wavelengths of light. Two mutants were created with blue-shifted (E254D, λmax = 390 nm; D380N, λmax = 506 nm) and one with red-shifted (D380E, λmax = 533 nm) absorption maxima relative to the wild-type protein (λmax = 527 nm). We also engineered a double mutant, E497K/C566D, that changes the enzyme to a specific, light-stimulated adenylyl cyclase that catalyzes the formation of cAMP from ATP. We anticipate that this expression/purification system and these RhoGC mutants will facilitate mechanistic and structural exploration of this important enzyme.

Purification and Characterization of RhoPDE, a Retinylidene/Phosphodiesterase Fusion Protein and Potential Optogenetic Tool from the Choanoflagellate Salpingoeca rosetta.

RhoPDE is a type I rhodopsin/phosphodiesterase gene fusion product from the choanoflagellate Salpingoeca rosetta. The gene was discovered around the time that a similar type I rhodopsin/guanylyl cyclase fusion protein, RhoGC, was shown to control phototaxis of an aquatic fungus through a cGMP signaling pathway. RhoPDE has potential as an optogenetic tool catalyzing the hydrolysis of cyclic nucleotides. Here we provide an expression and purification system for RhoPDE, as well as a crystal structure of the C-terminal phosphodiesterase catalytic domain. We show that RhoPDE contains an even number of transmembrane segments, with N- and C-termini both located on the cytoplasmic surface of the cell membrane. The purified protein exhibits an absorption maximum at 490 nm in the dark state, which shifts to 380 nm upon exposure to light. The protein acts as a cGMP-selective phosphodiesterase. However, the activity does not appear to be modulated by light. The protein is also active with cAMP as a substrate, but with a roughly 5-7-fold lower kcat. A truncation consisting solely of the phosphodiesterase domain is also active with a kcat for cGMP roughly 6-9-fold lower than that of the full-length protein. The isolated PDE domain was crystallized, and the X-ray structure showed the protein to be a dimer similar to human PDE9. We anticipate that the purification system introduced here will enable further structural and biochemical experiments to improve our understanding of the function and mechanism of this unique fusion protein.

Relocating the Active-Site Lysine in Rhodopsin: 2. Evolutionary Intermediates.

The visual pigment rhodopsin is a G protein-coupled receptor that covalently binds its retinal chromophore via a Schiff base linkage to an active-site Lys residue in the seventh transmembrane helix. Although this residue is strictly conserved among all type II retinylidene proteins, we found previously that the active-site Lys in bovine rhodopsin (Lys296) can be moved to three other locations (G90K, T94K, S186K) while retaining the ability to form a pigment with retinal and to activate transducin in a light-dependent manner [ Devine et al. ( 2013 ) Proc. Natl. Acad. Sci. USA 110 , 13351 - 13355 ]. Because the active-site Lys is not functionally constrained to be in helix seven, it is possible that it could relocate within the protein, most likely via an evolutionary intermediate with two active-site Lys. Therefore, in this study we characterized potential evolutionary intermediates with two Lys in the active site. Four mutant rhodopsins were prepared in which the original Lys296 was left untouched and a second Lys residue was substituted for G90K, T94K, S186K, or F293K. All four constructs covalently bind 11-cis-retinal, form a pigment, and activate transducin in a light-dependent manner. These results demonstrate that rhodopsin can tolerate a second Lys in the retinal binding pocket and suggest that an evolutionary intermediate with two Lys could allow migration of the Schiff base Lys to a position other than the observed, highly conserved location in the seventh TM helix. From sequence-based searches, we identified two groups of natural opsins, insect UV cones and neuropsins, that contain Lys residues at two positions in their active sites and also have intriguing spectral similarities to the mutant rhodopsins studied here.

Relocating the active-site lysine in rhodopsin and implications for evolution of retinylidene proteins.

Type I and type II rhodopsins share several structural features including a G protein-coupled receptor fold and a highly conserved active-site Lys residue in the seventh transmembrane segment of the protein. However, the two families lack significant sequence similarity that would indicate common ancestry. Consequently, the rhodopsin fold and conserved Lys are widely thought to have arisen from functional constraints during convergent evolution. To test for the existence of such a constraint, we asked whether it were possible to relocate the highly conserved Lys296 in the visual pigment bovine rhodopsin. We show here that the Lys can be moved to three other locations in the protein while maintaining the ability to form a pigment with 11-cis-retinal and activate the G protein transducin in a light-dependent manner. These results contradict the convergent hypothesis and support the homology of type I and type II rhodopsins by divergent evolution from a common ancestral protein.

Combined type IIIB with bilateral type I thyroplasty for pitch lowering with maintenance of vocal fold tension.

To evaluate type IIIB thyroplasty using the excised larynx bench apparatus and determine how altering vocal fold contour by performing bilateral medialization of the inferior vocal fold affects phonation. This procedure could be performed in patients for whom pitch lowering is desirable, such as female-to-male transsexuals or male patients with mutational falsetto in whom intensive voice therapy was insufficient. Aerodynamic, acoustic, and high-speed videokymographic data were collected for nine larynges at three subglottal pressure inputs for each of three conditions: normal; type IIIB thyroplasty; and combined type IIIB with modified bilateral type I thyroplasty intended to create a more rectangular glottal configuration. Each larynx served as its own control. Phonation threshold flow (p = 0.005), phonation threshold power (p = 0.031), and airflow varied across conditions with highest values for type IIIB thyroplasty and lowest for the combined procedure. Fundamental frequency was significantly different (p < 0.001), decreasing by approximately 100 Hz from control to type IIIB trials, and then by approximately 15 Hz from IIIB to combined procedure trials. Vibratory amplitudes and intrafold phase difference were highest for type IIIB trials. Addition of bilateral inferior medialization to type IIIB thyroplasty provided some further decrease in frequency, but mostly served to increase tension, reduce airflow, and produce a vibratory pattern which more closely mirrored control trials. Exploration of this combined procedure in patients may be warranted if not completely satisfied with the results from type IIIB thyroplasty alone.

A clinical study of serum lipid disturbance in Chinese patients with sudden deafness.

BACKGROUND: Pathological levels of blood lipids could be one of the causes of sudden sensorineural hearing loss (SSNHL). The objective of this manuscript is therefore to evaluate the relationship between blood lipid content and sudden sensorineural hearing loss (SSNHL). METHODS: The correlation between serum lipid parameters, including total cholesterol (TC), triglycerides, high-density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), apolipoprotein AI (apo AI), apolipoprotein B (apo B), and lipoprotein A (Lp(a)), and the onset of SSNHL was analyzed from a data set of 250 patients and an age, gender and weight matched control group of 250 subjects. RESULTS: The result of single factor logistic regression shows that TC (p<0.01), LDL-C (p<0.01), and apo B (p=0.03) of SSNHL group were significantly higher than those of the control group. The odds ratio of TC, LDL, and apo B are higher than 1, while the confidence intervals of the odds rations do not include 1. No significant difference was found with the prevalence of hypertension (P=0.818), diabetes (P=0.869) and smoking habits (P=0.653) between SSNHL group and control group. CONCLUSION: Total cholesterol, low density lipoprotein cholesterol and apolipoprotein B concentrations may be important factors in the pathogenesis of sudden sensorineural hearing loss, and should be assessed during the investigation of patients with this condition.

Cost-Effectiveness of Depression Screening for Otolaryngology-Head and Neck Surgery Residents.

OBJECTIVES: This study aims to determine the cost-effectiveness of screening and treating otolaryngology-head and neck surgery residents for depression. METHODS: A Markov model was built using TreeAgePro, version 2019 (TreeAge Software Inc.; Williamstown, MA) to assess the cost-effectiveness of five potential treatment algorithms: 1) treat all residents with psychotherapy, 2) screen and treat depressed residents with psychotherapy, 3) screen and treat depressed residents with pharmacotherapy, 4) screen and treat depressed residents with combination psychotherapy/pharmacotherapy, and 5) no intervention. A Monte Carlo probabilistic sensitivity analysis (PSA), consisting of 1 thousand simulations over a cumulative 5-year period, was performed to evaluate both base case values and a range of values for model variables. RESULTS: Screening residents for depression and treating with combination psychotherapy/pharmacotherapy was cost-effective and the optimal strategy at a willingness-to-pay threshold of $50 thousand per quality-adjusted life year (QALY). This option demonstrated an incremental cost-effectiveness ratio of $27,578 per QALY for base case values. PSA confirmed these results and demonstrated that screening residents for depression and treating with either combination pharmacotherapy/psychotherapy, pharmacotherapy alone, or psychotherapy alone were cost-effective options in 94.9% of simulations. CONCLUSION: Depression and burnout remain crucial issues among resident physicians. This study demonstrates that actively screening residents for depression is cost-effective. Based on these results, residency programs may consider trialing standardized depression screening protocols. LEVEL OF EVIDENCE: I and II. Laryngoscope, 131:502-508, 2021.

Quantitative second harmonic generation imaging of leporine, canine, and porcine vocal fold collagen.

OBJECTIVES/HYPOTHESIS: Vocal fold collagen composition is an important determinant of material properties and mucosal wave propagation. Collagen alignment and straightness are quantitatively characterized by second harmonic generation (SHG) imaging. We examined leporine, canined and porcine vocal folds showing collagen composition variation that is species, location, and strain specific. STUDY DESIGN: Animal model. METHODS: Leporine (n = 5), canine (n = 5), and porcine (n = 5) larynges were harvested and fixed in situ. Samples were transversely sectioned, and SHG images were collected for two inferior-superior sections along five anterior-posterior locations. Additional porcine samples were fixed and imaged under tensile strain (0%, 5%, 10%, 15%, 20%, n = 5 per group). Two-way repeated measures (RM) analysis of variance (ANOVA) tested for section and location differences in each species. Multiway RM-ANOVA tested for section, location, and strain differences in porcine samples. RESULTS: Alignment and straightness were higher inferiorly in the porcine (P = .0047, P = .002) and canine (P = .0011, P < .001) vocal folds, but not in leporine samples (P = .67652, P = .4831). There were significant interactions between elongation and superior-inferior section for both alignment (P = .0047) and straightness (P = .0371). CONCLUSIONS: Our results correspond well to findings in the literature that the inferior vocal fold lip is stiffer in porcine and canine larynges. The absence of a collagen gradient in the leporine vocal fold is notable because rabbits are less vocal animals, indicating the collagen gradient may be a result of voice use and an important consideration in model selection when extracellular matrix is of interest. Strain results were also consistent with the role of collagen in strain stiffening behavior of vocal fold tissue. LEVEL OF EVIDENCE: NA Laryngoscope, 129:2549-2556, 2019.

Oropharyngeal squamous cell carcinoma incidence and mortality trends in the United States, 1973-2013.

OBJECTIVE: To analyze oropharyngeal squamous cell carcinoma incidence and mortality trends in the United States for the years 1973 through 2013. STUDY DESIGN: Cross-sectional study using a large population-based cancer database. METHODS: Data on incidence and mortality rates were extracted from the Surveillance, Epidemiology, and End Results (SEER) 9 Database. Annual percentage change in rates was calculated using Joinpoint regression analysis (National Cancer Institute, Bethesda, MD). RESULTS: Incidence rates increased (annual percent change [APC]; 1.52, 95% confidence interval [CI] 0.17 to 2.88) from 1973 to 1983, remained stable (APC -0.52, 95% CI -1.30 to 0.26) from 1983 to 1997, and increased (APC 1.32, 95% CI 0.83 to 1.81) from 1997 to 2013. Overall, incidence rates increased for males (APC 0.73, 95% CI 0.22 to 1.25) but not females (APC -0.77, 95% CI -0.68 to 0.82). Incidence rates increased in the white population (APC 0.79, 95% CI 0.33 to 1.25) but decreased in the black population (APC -0.72, 95% CI -1.41 to -0.02). The incidence rates increased for tongue-base tumors (APC 1.17, 95% CI 0.42 to 1.92) and tonsil tumors (APC 0.47, 95% CI 1.10 to 4.96) but decreased for other sites. Incidence-based mortality decreased (APC -0.78, 95% CI -1.13 to -0.42) from 1993 to 2013. CONCLUSION: Oropharyngeal squamous cell carcinoma incidence rates increased in a nonlinear fashion from 1973 to 2013, whereas mortality rates declined. This, along with variation in trends by demographic and tumor factors, suggest that human papilloma virus is the main driver of the recent rise in incidence. LEVEL OF EVIDENCE: 2b. Laryngoscope, 128:1582-1588, 2018.

Excised larynx evaluation of subthyroid cartilage approach to medialization thyroplasty.

OBJECTIVES/HYPOTHESIS: To describe an alternative approach to medialization thyroplasty involving dissection underneath the thyroid cartilage with placement of a Gore-Tex implant, and to evaluate its effect on a range of phonatory measures using an excised canine larynx model. STUDY DESIGN: Animal model. METHODS: On each of eight excised canine larynges, the conditions of normal, paralysis, medialization thyroplasty by standard transthyroid cartilage approach, and medialization thyroplasty by experimental subthyroid cartilage approach were performed. Aerodynamic, acoustic, and mucosal wave parameters were measured for each condition. RESULTS: Compared to the vocal fold paralysis state, both the transthyroid and subthyroid approaches for Gore-Tex insertion resulted in significant decreases in phonation threshold pressure and phonation threshold flow. Both approaches also significantly decreased percent jitter, decreased percent shimmer, and improved signal-to-noise ratio. The mucosal wave was preserved after insertion of the Gore-Tex implant for both approaches. For all the phonatory measures except phonation threshold flow, there were no significant differences between the transthyroid and subthyroid approaches. CONCLUSIONS: Gore-Tex implantation via a subthyroid approach in an excised canine larynx model can produce effective medialization, preserve the mucosal wave, and significantly improve aerodynamic and acoustic parameters without meaningful difference compared to a traditional transthyroid approach. The subthyroid approach does not require creation of a thyroid cartilage window and could be a potentially valuable alternative method of performing medialization thyroplasty. LEVEL OF EVIDENCE: NA. Laryngoscope, 128:675-681, 2018.

Photodynamic therapy induces antifibrotic alterations in primary human vocal fold fibroblasts.

OBJECTIVES/HYPOTHESIS: Photodynamic therapy (PDT) is a promising treatment modality for laryngeal dysplasia, early-stage carcinoma, and papilloma, and was reported to have the ability to preserve laryngeal function and voice quality without clinical fibrotic response. We aimed to investigate the mechanism behind the antifibrotic effects of PDT on primary human vocal fold fibroblasts (VFFs) in vitro. STUDY DESIGN: In vitro analysis from one human donor. METHODS: Cell viability of VFFs in response to varying doses of PDT was investigated by the Cell Counting Kit-8 method. Sublethal-dose PDT (SL-PDT) was used for the following experiments. Expression of genes related to vocal fold extracellular matrix formation was analyzed by real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blotting. Effects of PDT on cell migration, collagen contraction, and transforming growth factor ß-1 (TGF-ß1)-induced myofibroblast differentiation were also analyzed. RESULTS: PDT affects the viability of VFFs in a dose-dependent manner. SL-PDT significantly changed the expression profile of VFFs with antifibrotic effects. It also inhibited cell migration, reduced collagen contraction, and reversed the fibroblast-myofibroblast differentiation induced by TGF-ß1. CONCLUSIONS: SL-PDT induces antifibrotic alterations in VFFs. This could explain the low incidence of vocal fold scar associated with PDT. Moreover, PDT may be useful in treating existing vocal fold scars. Further studies should focus on the in vivo effect of PDT on vocal fold wound healing and scar remodeling. LEVEL OF EVIDENCE: NA Laryngoscope, 128:E323-E331, 2018.

Over-vibration induced blood perfusion and vascular permeability changes may lead to vocal edema.

OBJECTIVES/HYPOTHESIS: To observe blood perfusion and vascular permeability changes under varying vibration frequency exposures. STUDY DESIGN: Animal model. METHODS: Blood perfusion was measured using laser Doppler flowmetry in eight rabbit auricular vessels (four rabbits) under nonvibration, and 62.5-Hz/1-mm, 125-Hz/1-mm, and 250-Hz/0.5-mm vibration frequency/amplitude exposures. Another 12 rabbits were randomly divided into vibration only and vibration with histamine groups. After 3 hours of continuous 125-Hz, 1-mm amplitude vibration of the auricle, vascular permeability was analyzed by absorbance of Evans blue-albumin complex. RESULTS: Significantly lower blood perfusion was observed in the vibration group, compared with no vibration exposure controls. Blood perfusion decreased 29 ± 16% as the vibration frequency was increased from 62.5 Hz to 125 Hz with the vibration amplitude constant at 1 mm. When the frequency was increased from 125 Hz to 250 Hz, while the amplitude was decreased from 1 mm to 0.5 mm, blood flow perfusion further decreased 29 ± 29%, and the decline tendency in blood perfusion showed no significant difference (P = .992). Meanwhile, in the vibration with histamine group, vascular permeability of the vibrated ears increased significantly compared to the nonvibrated ears (P = .005). CONCLUSIONS: Overvibration of the vocal folds due to voice overuse or abuse may significantly reduce blood perfusion, and increase vascular permeability in the vocal fold in inflammatory situations, which may lead to the formation of vocal edema. LEVEL OF EVIDENCE: NA Laryngoscope, 127:148-152, 2017.

Parameters From the Complete Phonatory Range of an Excised Rabbit Larynx.

OBJECTIVE: This study aims to collect data throughout the complete phonatory range using rabbit larynges. STUDY DESIGN: This is a methodological excised rabbit larynx study. METHODS: Seven rabbit larynges were dissected and mounted on a modified excised laryngeal apparatus. Phonation was initiated at phonation threshold pressure (PTP) and airflow was increased by consistent increments until phonation instability pressure (PIP) was reached. At each airflow level, aerodynamic measurements, acoustic recordings, and high-speed videos were recorded. This procedure was repeated at multiple elongation conditions to further explore the parameters. Data were then compared across subjects and elongation conditions. RESULTS: At PTP, subglottal pressure, fundamental frequency, and sound pressure level were found to increase significantly as elongation was increased. As elongation was increased at PIP, airflow was found to significantly decrease, whereas fundamental frequency was found to significantly increase. Vibratory amplitude decreased at both PTP and PIP as elongation increased. Also, as elongation increased, the range of all parameters was found to decrease significantly. CONCLUSIONS: The results obtained, combined with the similarities of the histologic structure of the vocal fold lamina propria between rabbits and humans, validate the rabbit larynx as an effective and reliable model for tissue inflammation studies.

An Excised Canine Model of Anterior Glottic Web and Its Acoustic, Aerodynamic, and High-speed Measurements.

OBJECTIVES: This study aims to build an excised anterior glottic web (AGW) model and study the basic voice-related mechanisms of the AGW through investigating the acoustic, aerodynamic, and vibratory properties. STUDY DESIGN AND METHODS: Overall, four conditions were tested for each of the eight canine larynges used. At baseline, 10%, 20%, and 33% occlusion (as determined by the placement of the suture), acoustic, aerodynamic, and high-speed video data were collected while each larynx was phonated in a soundproof booth. RESULTS: The phonation threshold pressure (PTP) and the phonation threshold flow significantly increased as percent occlusion increased (P < 0.001). There were significant increases in jitter % and shimmer % from baseline group to AGW model groups at PTP, 1.25 PTP, and 1.5 PTP (P = 0.039, P < 0.001, P < 0.001, P < 0.001, P < 0.001, and P = 0.001, respectively). The fundamental frequency significantly increased as percent occlusion increased at all given pressures (P < 0.001). Correlation dimension (D2) was significantly higher in the AGW model groups than in the baseline group at PTP, 1.25 PTP, and 1.5 PTP (P = 0.002, P < 0.001, P = 0.01, respectively). High-speed videos revealed that, the left phase shift in the AGW model groups compared with the baseline at 1.25 PTP was significant (P = 0.027) and right phase shift at 1.5 PTP (P < 0.001). CONCLUSIONS: We presented an anatomically similar model of a type 1 AGW and confirmed its validity through aerodynamic, acoustic, and high-speed video analysis in our study. We observed and investigated the glottic web movement, which may be a new explanation for the pathologic voice-related mechanism of AGW.

Evaluation of type II thyroplasty on phonatory physiology in an excised canine larynx model.

OBJECTIVES/HYPOTHESIS: Type II thyroplasty is an alternative treatment for spasmodic dysphonia, addressing hyperadduction by incising and lateralizing the thyroid cartilage. We quantified the effect of lateralization width on phonatory physiology using excised canine larynges. METHODS: Normal closure, hyperadduction, and type II thyroplasty (lateralized up to 5 mm at 1-mm increments with hyperadducted arytenoids) were simulated in excised larynges (N = 7). Aerodynamic, acoustic, and videokymographic data were recorded at three subglottal pressures relative to phonation threshold pressure (PTP). One-way repeated measures analysis of variance assessed effect of condition on aerodynamic parameters. Random intercepts linear mixed effects models assessed effects of condition and subglottal pressure on acoustic and videokymographic parameters. RESULTS: PTP differed across conditions (P < .001). Condition affected percent shimmer (P < .005) but not percent jitter. Both pressure (P < .03) and condition (P < .001) affected fundamental frequency. Pressure affected vibratory amplitude (P < .05) and intrafold phase difference (P < .05). Condition affected phase difference between the vocal folds (P < .001). CONCLUSIONS: Hyperadduction increased PTP and worsened perturbation compared to normal, with near normal physiology restored with 1-mm lateralization. Further lateralization deteriorated voice quality and increased PTP. Acoustic and videokymographic results indicate that normal physiologic relationships between subglottal pressure and vibration are preserved at optimal lateralization width, but then degrade with further lateralization. The 1-mm optimal width observed here is due to the small canine larynx size. Future human trials would likely demonstrate a greater optimal width, with patient-specific value potentially determined based on larynx size and symptom severity. LEVEL OF EVIDENCE: NA Laryngoscope, 2016 127:396-404, 2017.

A Preliminary Case Report of a High-Quality Cost-effective Rigid Laryngoscopy Setup.

OBJECTIVE: To evaluate a cost-effective modified rigid laryngoscopy setup with a portable light source and high-resolution commercially available digital camera for use in smaller otolaryngology and family practice clinics. METHODS: The modified setup was used to obtain images of the larynx using both a traditional light source and a portable light source. Varying shutter speeds and ISOs were evaluated, and the optimal settings were determined for the modified setup. RESULTS: Picture quality was adequate, and the portable light source was bright enough. ISO from 640 to 1600 with shutter speeds ranging from 1/60 to 1/160 are ideal under the normal light source, while it is better to set the ISO between 4000 and 10 000 with shutter speeds from 1/60 to 1/100 under the portable light source. Picture quality was adequate with a resolution of 2768 pixels × 1848 pixels with 350 dpi × 350 dpi. CONCLUSIONS: Results show that the modified setup obtains images of adequate quality for use in the clinic. Additionally, since the larynx requires the most illumination for endoscopic imaging, a similar setup would work for imaging the ear and nose. This setup may make laryngoscopic exams more accessible to patients at smaller laryngoscopy clinics or family practice providers.

The Potential Role of Subglottal Convergence Angle and Measurement.

OBJECTIVE: This study aimed to explore subglottal convergence angle measurement. We hypothesize that the angle will change with glottis closure condition. Changes to the angle may alter vocal fold stress distribution, which could result in vocal fold pathologies. METHODS: Three vocal fold positions were evaluated by the degree of glottal closure and divided into three groups. Neck computed tomographic images of the subjects were gathered, and three-dimensional (3D) models of the vocal folds and the airway were reconstructed using Mimics software, through which the subglottal convergence angle was measured. Using one-way factor analysis of variance, we compared the angle among three groups. We explored the effect of the angle on vibration based on the material oblique section stress analysis. RESULTS: The subglottal convergence angle was effectively measured from 3D models. The angle changed with degree of glottal closure, with statistically significant differences among the three groups (P < 0.01). Changes to the angle alter subglottal stress distribution in the subglottal shear or normal direction by the material oblique section stress analysis theory. Larger subglottal convergence angle might reduce subglottal pressure in the shear direction below the vocal folds, which would make vibration and vocal fold mucosal wave generation more difficult. CONCLUSION: 3D model generation from computed tomographic data is an effective method of measuring the subglottal convergence angle, which changes with the degree of glottis closure, and may affect subglottal pressure distribution in the subglottal shear or normal direction and influence vocal fold vibration.

Extension and Application of High-Speed Digital Imaging Analysis Via Spatiotemporal Correlation and Eigenmode Analysis of Vocal Fold Vibration Before and After Polyp Excision.

OBJECTIVES/HYPOTHESIS: To evaluate the spatiotemporal correlation of vocal fold vibration using eigenmode analysis before and after polyp removal and explore the potential clinical relevance of spatiotemporal analysis of correlation length and entropy as quantitative voice parameters. We hypothesized that increased order in the vibrating signal after surgical intervention would decrease the eigenmode-based entropy and increase correlation length. STUDY DESIGN: Prospective case series. METHODS: Forty subjects (23 males, 17 females) with unilateral (n = 24) or bilateral (n = 16) polyps underwent polyp removal. High-speed videoendoscopy was performed preoperatively and 2 weeks postoperatively. Spatiotemporal analysis was performed to determine entropy, quantification of signal disorder, correlation length, size, and spatially ordered structure of vocal fold vibration in comparison to full spatial consistency. The signal analyzed consists of the vibratory pattern in space and time derived from the high-speed video glottal area contour. RESULTS: Entropy decreased (Z = -3.871, P < .001) and correlation length increased (t = -8.913, P < .001) following polyp excision. The intraclass correlation coefficients (ICC) for correlation length and entropy were 0.84 and 0.93. CONCLUSION: Correlation length and entropy are sensitive to mass lesions. These parameters could potentially be used to augment subjective visualization after polyp excision when evaluating procedural efficacy.