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Objective: Laryngomalacia is the most common pediatric laryngeal anomaly. The pathophysiology of laryngomalacia is not well defined; the leading hypothesis suggests weak laryngeal tone and neuromuscular discoordination. Only a few studies explored the histopathology of the laryngeal submucosal nerves, with reported nerve hypertrophy. Our study aims to describe the histopathology of submucosal nerves in specimens obtained from children with severe laryngomalacia compared to pediatric cadaveric controls., Study Design: Prospective study., Settings: Tertiary care children's hospital., Methods: Histologic and immunohistochemical sections of supraglottic tissue from 26 children with severe laryngomalacia and six pediatric autopsies were digitally scanned and assessed with image analysis software (QuPath), resulting in the identification and measurement of 4561 peripheral nerves and over 100,000 foci of neurofilaments., Results: Chronic inflammation was noted in all patients. Eosinophils were rare. The mean nerve area and perimeter were significantly smaller for patients with laryngomalacia compared to the control group (1594.0 ± 593.2 μm^2 vs. 2612.1 ± 2824.0 μm^2, p < 0.0001, and 158.8 ± 30.3 μm vs. 217.6 ± 165.0 μm, p < 0.0001). Nerve-per-area unit was significantly greater for patients with laryngomalacia compared to controls (1.39E-05 vs. 6.19 E-06, p = 0.009). The mean area and the number of neurofilaments per total nerve area were similar. Immunohistochemistry for calretinin, a marker for intestinal ganglion cells in Hirschsprung disease, was absent from all specimens., Conclusions: This series includes a comparison of all identifiable nerve fibers obtained from children with severe laryngomalacia and shows that the mucosal nerves are smaller on average than controls. These findings fail to provide support for significant morphologic peripheral nerve pathology in laryngomalacia., Competing Interests: Declaration of competing interest none., (Copyright © 2024. Published by Elsevier B.V.)

Objective: This study aimed to evaluate the role of pepsin inhibitors in the inflammatory response and their effects on laryngeal mucosal integrity during gastroesophageal reflux (GERD) under in vivo conditions., Methods: A surgical model of GERD was used, in which mice were treated with pepstatin (0.3 mg/kg) or darunavir (8.6 mg/kg) for 3 days. On the third day after the experimental protocol, the laryngeal samples were collected to assess the severity of inflammation (wet weight and myeloperoxidase activity) and mucosal integrity (transepithelial electrical resistance and paracellular epithelial permeability to fluorescein)., Results: The surgical GERD model was reproduced. It showed features of inflammation and loss of barrier function in the laryngeal mucosa. Pepstatin and darunavir administration suppressed laryngeal inflammation and preserved laryngeal mucosal integrity., Conclusion: Pepsin inhibition by the administration of pepstatin and darunavir improved inflammation and protected the laryngeal mucosa in a mouse experimental model of GERD., Level of Evidence: NA Laryngoscope, 134:3080-3085, 2024., (© 2024 The American Laryngological, Rhinological and Otological Society, Inc.)

An additional microscopic diagnostic sign has been identified for verification of asphyxial type of drowning. In white non-linear male rats (age 2 months) subjected to free drowning, significant hyperplasia of argyrophilic and morphofunctional activity of serotonin-containing APUD-cells of the laryngeal mucosa were revealed under conditions of acute anoxia in comparison with the intact control. These changes promote the development of laryngospasm, which prevents water penetration into the airways and lungs in asphyxial type of drowning. The presence of statistically significant hyperplasia of argyrophilic and morphofunctional activity of serotonin-containing APUD-cells of the laryngeal mucosa under conditions of acute anoxia can be used as an additional diagnostic criterion for asphyxial type of drowning in experimental studies., (© 2024. Springer Science+Business Media, LLC, part of Springer Nature.)

Objective: The laryngeal tissue carries most of the heat during inhalation injury. This study aims to explore the heat transfer process and the severity of injury inside laryngeal tissue by horizontally studying the temperature rise process at various anatomical layers of the larynx and observing the thermal damage in various parts of the upper respiratory tract., Methods: The 12 healthy adult beagles were randomly divided into four groups, and inhaled room temperature air (control group), dry hot air of 80 °C (group I), 160 °C (group II), and 320 °C (group III) for 20 min, respectively. The temperature changes of the glottic mucosal surface, the inner surface of the thyroid cartilage, the external surface of the thyroid cartilage, and subcutaneous tissue were measured every minute. All animals were immediately sacrificed after injury, and pathological changes in various parts of laryngeal tissue were observed and evaluated under a microscope., Results: After inhaling hot air of 80 °C, 160 °C and 320 °C, the increase of laryngeal temperature in each group was ΔT = 3.57 ± 0.25 °C, 7.83 ± 0.15 °C, 11.93 ± 0.21 °C. The tissue temperature was approximately uniformly distributed, and the difference was not statistically significant. The average laryngeal temperature-time curve showed that the laryngeal tissue temperature in group I and group II showed a trend of "first decrease and then increase", except that the temperature of group III directly increased with time. The prominent pathological changes after thermal burns mainly concluded necrosis of epithelial cells, loss of the mucosal layer, atrophy of submucosal glands, vasodilatation, erythrocytes exudation, and degeneration of chondrocytes. Mild degeneration of cartilage and muscle layers was also observed in mild thermal injury. Pathological scores indicated that the pathological severity of laryngeal burns increased significantly with the increase of temperature, and all layers of laryngeal tissue were seriously damaged by 320 °C hot air., Conclusions: The high efficiency of tissue heat conduction enabled the larynx to quickly transfer heat to the laryngeal periphery, and the heat-bearing capacity of perilaryngeal tissue has a certain degree of protective effect on laryngeal mucosa and function in mild to moderate inhalation injury. The laryngeal temperature distribution was in accordance with the pathological severity, and the pathological changes of laryngeal burns provided a theoretical basis for the early clinical manifestations and treatment of inhalation injury., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Purpose: To explore the role played by Glut-1 and H + /K + -ATPase in pepsin-induced, mouse laryngeal epithelial proliferation, growth, and development., Methods: We established a mouse model of laryngopharyngeal reflux and measured Glut-1 and H + /K + -ATPase expression levels in mouse laryngeal epithelium treated with artificial gastric juice containing pepsin., Results: Artificial pepsin-containing gastric juice induced significant hyperplastic changes in mouse laryngeal epithelium compared to control mice at 15, 30, and 45 days. Inhibition of Glut-1 expression by 2-DG significantly suppressed such hyperplasia compared to mice exposed to artificial gastric juice containing pepsin at 15, 30, and 45 days. After treatment with pepsin-containing artificial gastric juice, RT-PCR and Western blotting showed that the levels of Glut-1 and H + /K + -ATPase α, β increased significantly., Conclusions: Pepsin-containing artificial gastric juice promoted mouse laryngeal epithelial hyperplasia associated with abnormal expression of Glut-1 and H + /K + -ATPase α, β., (© 2021. The Author(s).)

Stevens-Johnson syndrome (SJS) is a rare but severe mucocutaneous epidermolysis commonly triggered by medications. SJS is characterized by mucocutaneous lesions of the trunk, face, and limbs, as well as the oral cavity, gastrointestinal tract, and respiratory tract. Although uncommon, laryngeal involvement in SJS can lead to severe respiratory, phonatory and deglutitive complications. Providers caring for patients with SJS should maintain a high index of suspicion for laryngeal involvement and low threshold to solicit Otolaryngology consultation. Laryngeal complications can be more expediently managed when anticipated early in the course of disease. Laryngoscope, 131:2519-2522, 2021., (© 2021 The American Laryngological, Rhinological and Otological Society, Inc.)

Objectives/hypothesis: Cigarette smoke (CS) is a primary risk factor for the development of numerous benign and malignant laryngeal diseases. The epithelium and mucus lining the vocal folds (VF) are the first barriers against CS. The primary objective of this study was to investigate epithelial and mucus barrier changes in the mouse laryngeal mucosa upon exposure to subacute CS. The secondary objective was to compare mucus barrier changes in mice and human smokers and nonsmokers. Study Design Animal model., Methods: Mice were exposed to CS for 4 weeks for 4 hours (N = 12, high dose [HD]) or 1 hour (N = 12, low dose [LD]) per day. Air-exposed mice were used as a control group (N = 10). Larynges were harvested and VF epithelial barrier integrity was evaluated including cellular proliferation and expression of cell junctions. We also investigated mucus production by examining mucus cell area and mucin expression in mice and human smokers and nonsmokers., Results: HD CS increased VF epithelial cellular proliferation but did not alter the expression of cell junctions. HD CS also induced hypertrophy of the mucus-producing submucosal glands. However, only LD CS increased MUC5AC gene expression. MUC5AC staining appeared elevated in laryngeal specimens from smokers, but this was not significant as compared to nonsmokers., Conclusions: These findings help us identify potential adaptive mechanisms to CS exposure as well as set the foundation for further study of key aspects of epithelial and mucus barrier integrity that may be implicated in laryngeal disease development following prolonged smoking., Level of Evidence: NA Laryngoscope, 131:2530-2539, 2021., (© 2021 The American Laryngological, Rhinological and Otological Society, Inc.)

Objectives: Idiopathic subglottic stenosis (iSGS) is an inflammatory process leading to fibrosis and narrowing of the laryngotracheal airway. There is variability in patient response to surgical intervention, but the mechanisms underlying this variability are unknown. In this pilot study, we measure expression of candidate targets at the mucosal surface of the subglottis in iSGS patients. We aim to identify putative biomarkers for iSGS that provide insights into the molecular basis of disease progression, yield a gene signature for the disease, and/or predict a response to therapy., Study Design: In vitro comparative study of human cells., Methods: Levels of candidate transcripts and proteins were measured in healthy and stenotic laryngotracheal tissue specimens taken from the mucosal surface in 16 iSGS patients undergoing endoscopic balloon dilation. Pre- and post-operative pulmonary function test and patient reported voice and breathing outcomes were also assessed. Unsupervised clustering was used to define patient subgroups based on expression profile., Results: Pulmonary function and voice and breathing outcome metrics demonstrated significant post-operative improvement. Transcript levels of αSMA, CCL2, COL1A1, COL3A1, FN1, IFNG, and TGFB1 and protein levels of CCL2, IFNG, and IL-6 were significantly upregulated in stenotic as compared to healthy tissues. Marked heterogeneity was observed in the patterns of expression of candidate markers across individuals and tissue types. Patient subgroups defined by expression profile did not show a statistically significant difference in dilation interval., Conclusion: Pro-inflammatory and pro-fibrotic pathways are significantly upregulated along the mucosal surface of stenotic laryngotracheal tissues, and CCL2 and IFNG merit further investigation as potential iSGS biomarkers., Level of Evidence: 4 Laryngoscope, 131:342-349, 2021., (© 2020 The American Laryngological, Rhinological and Otological Society, Inc.)

Objectives/hypothesis: The objectives of this study were to evaluate laryngeal inflammation and mucosal integrity in a murine model of reflux disease and to assess the protective effects of topical agents including alginate, hyaluronic acid, and cashew gum., Study Design: Animal study., Methods: A surgical murine model of reflux disease was evaluated at 3 or 7 days postsurgery, and laryngeal samples were collected to measure inflammation (wet weight and myeloperoxidase [MPO]) and mucosal integrity (transepithelial resistance [TER] and mucosal permeability to fluorescein). Additional groups of animals were administered one of several topical agents (alginate, hyaluronic acid, or cashew gum) daily, and laryngeal inflammation and mucosal integrity were evaluated at 3 days postsurgery., Results: At 3 days, and not 7 days postsurgery, we observed increased laryngeal wet weight and MPO, decreased laryngeal TER, and increased laryngeal mucosa permeability. Alginate partially decreased laryngeal inflammation (wet weight and not MPO) and dramatically improved laryngeal mucosal integrity. Conversely, hyaluronic acid eliminated the inflammation; however, it had no effect on laryngeal mucosal integrity impairment. Cashew gum eliminated laryngeal inflammation as well as the impairment in laryngeal mucosal integrity., Conclusions: This study shows that a surgical model of reflux disease induced laryngeal inflammation and impairment in laryngeal barrier function. These observed alterations were partially attenuated by alginate and hyaluronic acid and completely reversed by cashew gum., Level of Evidence: NA Laryngoscope, 2020., (© 2020 The American Laryngological, Rhinological and Otological Society, Inc.)

Objectives/hypothesis: Voice outcomes of cordectomy for early glottic cancer are often poor due to vocal fold scarring and tissue defects. Improvements in this aspect could make cordectomy a more acceptable treatment option than radiotherapy. We hypothesized that a polyglycolic acid (PGA) sheet could be used to cover vocal fold defects. The present study aimed to prevent vocal fold scarring after cordectomy using the PGA sheet., Study Design: Animal experiment., Methods: Nine male beagles were divided into three groups including a control group (n = 3). Following cordectomy, the vocal fold defect was covered with the PGA sheet plus fibrin glue (PGA group; n = 3) or with the PGA sheet plus fibrin glue containing basic fibroblast growth factor (bFGF; the PGA-bFGF group, n = 3). Vocal folds were chronologically observed, and larynges were removed 6 months after surgery. Mucosal amplitude was measured using a high-speed camera, and histological analysis was performed., Results: The re-epithelialization process was delayed in the PGA and PGA-bFGF groups compared with the control group. The mucosal amplitude was significantly more normalized and the thickness ratio significantly higher in the PGA and PGA-bFGF groups compared with the control group. The PGA-bFGF group had the highest elastic fiber density, followed by the PGA group and then the control group, with a significant difference between the PGA-bFGF and control groups., Conclusions: The PGA sheet plus fibrin glue could serve as an effective regenerative scaffold for reconstructing vocal fold morphology and function after cordectomy, with the potential benefit of establishing an endoscopic sealing method for vocal fold defects., Level of Evidence: NA Laryngoscope, 130:E436-E443, 2020., (© 2019 The American Laryngological, Rhinological and Otological Society, Inc.)

Objectives: The clinical significance of the interarytenoid mucosal height (IAMH) in pediatric dysphagia, ranging from normal anatomy to a laryngeal cleft, is unknown. This study seeks to evaluate a cohort of patients who underwent evaluation of their IAMH during microdirect laryngoscopy (MDL) for associations between IAMH and dysphagia as diagnosed on preoperative videofluoroscopic swallow study (VFSS)., Methods: A retrospective case series of 1,351 patients who underwent MDL between 2011 and 2016 were reviewed for intraoperative evaluation of IAMH using our interarytenoid assessment protocol. After exclusions, 182 patients were divided into three groups: 1) thickened diet: VFSS with recommendation for thickened liquids (n = 82 of 182; 45.1%), 2) normal diet: VFSS with allowance of thin liquids (n = 19 of 182; 10.4%), and 3) control: no VFSS performed (n = 81 of 182; 44.5%)., Results: There was no difference in IAMH between groups (P = 0.35). Power analysis was able to achieve > 80% power to detect an effect size of ≥ 0.5 (1-5 mucosal height scale). The majority of patients in each group had an IAMH above the false vocal folds (thickened diet: 57.3%, normal diet: 57.9%, control: 64.2%). There were similar percentages of patients in each group with an IAMH at or below the true vocal folds (thickened diet: 4.9%, normal diet: 5.3%, control: 6.1%)., Conclusion: There was no significant association between IAMH and preoperative thickened liquid recommendation in this cohort. This data fails to support the hypothesis that the IAMH is an independent etiological factor for pediatric pharyngeal dysphagia. Further studies comparing IAMH with outcomes after feeding therapy and surgery may better clarify this relationship between anatomy and physiology., Level of Evidence: 4. Laryngoscope, 129:2588-2593, 2019., (© 2019 The American Laryngological, Rhinological and Otological Society, Inc.)

Objectives/hypothesis: We analyzed the results of 10 patients treated by unilateral autologous temporal fascia graft, comparing the pre and post surgery results, as well as the appearance of the wave and the intervals until the re-appearance of it., Study Design: Retrospective, clinical case series., Methods: Subjective and objective evaluation was preoperatively and postoperatively performed, using laryngovideostroboscopy, the Voice Handicap Index-10, GRBAS (grade, roughness, breathiness, asthenia, and strain), harmonics to noise ratio, Jitter, Shimmer, phonatory range, maximum phonation time, and S/Z ratio in all patients. Postoperative visits were scheduled after 1 week, 1 month, at 3 months, and at 6 months after the surgery., Results: After surgery, we found significant differences in the maximum phonation time and the S/Z ratio (P <0.05). Most parameters of GRBAS also improved significantly, both in general and in the subscales, except for Breathiness and Asthenia (P <0.05). There was an average decrease in Voice Handicap Index-10 of 11.2 (P <0.05). At laryngovideostroboscopy, the mucous wave reappeared in all patients treated 1 month after surgery, and increased in amplitude until 6 months postoperative., (Copyright © 2018 The Voice Foundation. Published by Elsevier Inc. All rights reserved.)

Background: Removal of Reinke's edema may result in moderate to large-sized mucosal defect on the vocal fold, which heals by secondary intention. Microsuturing this defect may lead to primary wound healing with fastened recovery and less scar, but costs extra time and effort. Exploring methods that can shorten microsuture time is helpful for the wider application of this technology., Study Design: Retrospective., Methods: 57 patients with Reinke's edema, who were admitted from November 2010 to June 2018, were enrolled as research subjects for the retrospective analysis. 27 patients were the knot pusher group (from November 2010 to March 2015), and 30 patients were the two-handed tying group (from April 2015 to June 2018). Evaluation indicators include the number of knots, the average time for suturing and tying the knot for each patient, and the occurrence of complications, subjective and objective voice assessments., Results: All patients underwent successful operation. The average time for making knots in the knot pusher group and two-handed tying group was 668.40 ± 173.73 s and 328.73 ± 121.0 s, respectively, and there was a statistically significant difference between the two groups (p < 0.001). No significant difference was noted in the mucosal avulsion, overall incidence of complications between the groups, and no significant difference was found between the two groups in terms of the preoperative and 3-month postoperative subjective and objective indicators., Conclusion: Microsuturing of Reinke's edema microflaps using the two-handed tying technique can achieve the similar effect with the knot pusher method, and save operation time while the surgeon is well trained., Level of Evidence: 4.

Objectives: The aim of this study was to assess the usefulness of 24-h pharyngeal pH monitoring in the diagnosis of laryngopharyngeal reflux (LPR) in children with voice disorders., Methods: The study was conducted on 68 children aged 3-18 years old. In all cases, pH monitoring was successful; no child removed the probe from the nasal cavity or reported discomfort or other complaints. The following definitions were adopted: LPR in the upright position if Ryan Score upright >9.41; LPR in the supine position if there was at least one supine episode below pH 5.0 (equal to a Ryan Score supine >2.91)., Results: In 43 children (63%), vocal fold edema was strongly related to recorded reflux episodes, especially in the upright position. In 18 children (26%), vocal nodules were observed, but their occurrence did not significantly affect the Ryan Score, either upright or supine. The selected potential LPR markers were significantly correlated to all the pH monitoring variables and individual elements on the Reflux Symptoms Index and the Reflux Finding Score questionnaire., Conclusion: Our findings indicate that, together with vocal fold edema, laryngeal edema and posterior commissure mucosal hypertrophy are important determinants of paediatric LPR. In fact, if LPR is suspected in a child, 24-h pharyngeal pH monitoring appears to be a valuable and welltolerated diagnostic tool. Vocal fold edema observed in laryngeal endoscopy can be considered a probable sign of LPR. The Reflux Finding Score appears to be helpful in diagnosing LPR in children, especially if a cut-off value of 4/5 is adopted., (Copyright © 2019 The Author(s). Published by Elsevier B.V. All rights reserved.)

Objectives: The aim of this paper is to investigate electronic cigarettes (e-cigarettes) from the otorhinolaryngologic point of view., Methods: The authors searched Central Database of Kirikkale University Library, Google, PubMed, and Proquest and Google Scholar., Results: An electronic cigarette or e-cigarette is a battery-powered device that vaporizes a liquid, generally including nicotine. Nowadays, e-cigarettes are used for smoking cessation or to reduce the consumption of conventional tobacco cigarettes. First generation e-cigarette devices were similar to conventional tobacco cigarettes in terms of shape and size and expressed as "cigalikes." Differently from traditional cigarettes in which tobacco is burned to generate smoke, electronic cigarettes contain a tank filled with liquid. It was found that e-cigarette liquids contained different types of chemical compounds which were either previously known carcinogens or probably prove to be carcinogenic to humans in the near future. It seems that the use of electronic cigarette does not harm the oral cells. However, the use of e-cigarette for 4 weeks led to metaplasia and hyperplasia of the laryngeal mucosa in rats. Furthermore, e-ciagarettes produce some adverse effects on the nasal mucosa, supressing the immune system., Conclusion: It should not be considered that e-cigarettes are safer unless their effects on the mucosa of the ear, nose, and throat are more precisely clarified.

Objective: This study aims to investigate the expression level of long non-coding RNA (lncRNA) SNHG20 in laryngeal squamous cell carcinoma (LSCC), and to explore further whether it can promote the development of LSCC by regulating microRNA-140 (miR-140)., Patients and Methods: Expression levels of SNHG20 in 56 pairs of LSCC tissues and adjacent normal tissues were measured by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The relationship between SNHG20 expression with pathological parameters and the prognosis of LSCC was analyzed. Besides, the SNHG20 expression in LSCC cells was also analyzed by qRT-PCR. The SNHG20 knockdown and overexpression model were constructed by lentivirus transfection in AMC-HN-8 and Hep-2 cells. Cell counting kit-8 (CCK-8) and 5-Ethynyl-2'-deoxyuridine (EdU) assay were used to analyze the effect of SNHG20 on the biological function of LSCC cells. Finally, the dual-luciferase reporter gene assay was performed to explore the potentials of SNHG20 and miR-140 in LSCC., Results: The SNHG20 expression in LSCC tissues or cells remarkably increased than controls, and the difference was statistically significant. The LSCC patients with the high expression level of SNHG20 were more likely to develop advanced tumor compared with patients with low expression of SNHG20. Moreover, the LSCC patients with the high expression level of SNHG20 had a shorter overall survival than those with low level. The cell proliferation ability significantly decreased in the SNHG20 knockdown group, while notably increased in SNHG20 overexpression group. MiR-140 was negatively correlated with SNHG20 in LSCC tissues and cells. Dual-luciferase reporter gene assay showed that SNHG20 could be targeted by miR-140 through a certain binding site. The cell rescue experiment also indicated that there was a mutual regulation between SNHG20 and miR-140, which could together affect the malignant progression of LSCC., Conclusions: We showed that the expression levels of SNHG20 in LSCC tissues or cell lines significantly increased and was associated with advanced tumor staging and undesirable prognosis of LSCC. In addition, SNHG20 could promote the malignant progression of LSCC.

Objectives/hypothesis: Probe-based confocal laser endomicroscopy (pCLE) is a novel technique allowing real-time evaluation of the histological features of tissues in vivo at the cellular level. This study aimed to evaluate the feasibility of using pCLE in the diagnosis of nasopharyngeal carcinoma (NPC)., Study Design: Feasibility study., Methods: In this study, the pCLE images of the lesions, as well as the surrounding or contralateral normal mucosa of the lesions, were acquired in vivo from each patient after intravenous injecting of 2.5 mL fluorescein. Biopsy specimens were collected at the imaged sites followed by a histopathological diagnosis by the pathologists, which was used as the gold standard. The pCLE images were compared to histopathological diagnosis of visualized sites by using sensitivity, specificity, negative predictive value (NPV), and positive predictive value (PPV)., Results: Diagnoses based on pCLE images correlated well with the gold standard diagnoses based on tissue histology. The overall sensitivity, specificity, PPV, and NPV for diagnosis of carcinoma versus nondysplasia were 93.8% (67.7%-99.7%), 90.5% (68.2%-98.3%), 88.2% (62.3%-97.9%), and 95.0% (73.1%-99.7%), respectively, and the four indices for pCLE diagnosis of dysplasia versus nondysplasia were 60.0% (17.0%-92.7%), 80.9% (57.4%-93.7%), 42.9% (11.8%-79.8%), and 89.5% (65.5%-98.2%), respectively. The overall sensitivity for diagnosis of carcinoma versus dysplasia was 93.8% (67.7%-99.7%), specificity was 40% (7.3%-83.0%), PPV was 83.3% (57.7%-95.6%), and the NPV was 66.7% (12.5%-98.2%)., Conclusions: CLE is a suitable and valid method for otolaryngologists to diagnose of NPC in vivo., Level of Evidence: NA Laryngoscope, 129:897-902, 2019., (© 2018 The American Laryngological, Rhinological and Otological Society, Inc.)

Objectives:: The aim of this study is to describe a clinical entity the authors term "Shar Pei larynx," characterized by redundant supraglottic and postcricoid mucosa that the authors hypothesize coexists in patients with obstructive sleep apnea, laryngopharyngeal reflux, and obesity. By exploring this hypothesis, the authors hope to set the foundation for future research with the goal of identifying whether Shar Pei larynx is a marker for untreated sleep apnea or other diseases., Study Design:: Retrospective chart review., Setting:: Two tertiary care academic institutions., Methods:: Data were collected from a 5-year period by querying for patients described to have "Shar Pei larynx" or "posterior supraglottic and/or postcricoid mucosal redundancy" on laryngoscopic findings. Relevant demographic and clinical characteristics were analyzed, with a focus on associations with obesity, sleep apnea, and laryngopharyngeal reflux., Results:: Thirty-two patients were identified with physical findings consistent with Shar Pei larynx. Twenty-six patients (81.3%) were obese; 16 (50%) were morbidly obese. Twenty-two patients (68.8%) either had an existing diagnosis of obstructive sleep apnea or were diagnosed on polysomnography performed after initial evaluation. Sixteen patients (50%) had type 2 diabetes mellitus, and 87.5% of these patients were obese. Twenty-eight patients (87.5%) noted histories of reflux, with a median reflux symptom index of 27 of 45. Five patients underwent procedures to reduce mucosal redundancy related to Shar Pei larynx., Conclusions:: This pilot study confirms that the majority of patients diagnosed with Shar Pei larynx also had diagnoses of obesity, obstructive sleep apnea, and reflux disease. The demonstrated association is strong enough to warrant further study.

Current knowledge of laryngeal neuroendocrine cells in man and other vertebrates is reviewed. Particular attention is paid to differences in the distribution of neuroendocrine cells between squamous and respiratory laryngeal mucosa, foetal versus post-natal spatial arrangements, relation to the laryngeal cavity and nerve fibres, and immunoreactivities of these cells. Methodological deficiencies and gaps in knowledge are outlined. Comparisons with neuroendocrine cells in lung and gut are drawn, caution with regard to existing histogenetic models of laryngeal neuroendocrine neoplasia is advised and lines of future research are suggested., (Crown Copyright © 2018. Published by Elsevier GmbH. All rights reserved.)

Objective: To study the cluster of differentiation 8 population in the laryngeal mucosa of patients with laryngeal carcinoma. To our knowledge this is the first paper to address this issue., Methods: The study group included 40 patients with known laryngeal cancer who were scheduled for laryngectomy. The control groups included 10 smokers and 10 non-smokers who were scheduled for microlaryngeal surgery. Specimens from the three groups were processed for histopathological and histochemical evaluation., Results: In patients without cancer of the larynx, the number of cluster of differentiation 8 lymphocytes was greater in smokers than non-smokers. The number of cluster of differentiation 8 lymphocytes was greatest in smokers with laryngeal cancer, and the difference between this group and the two control groups was statistically significant., Conclusion: The study showed that smoking increased the number of cluster of differentiation 8 T-lymphocytes in the laryngeal mucosa. The increase was greatest in patients who had developed laryngeal cancer.

Objective: To investigate the morphological and pathological changes of the larynx after severe laryngeal burn in dogs and their relationship with laryngostenosis. Methods: Eighteen healthy, male beagle dogs were assigned into control group, immediately after injury group, and 2, 4, 6, and 8 weeks after injury groups according to the random number table, with 3 dogs in each group. Dogs of injury group inhaled saturated steam through mouth for 5 seconds to reproduce severe laryngeal burn. Tracheotomy and intubation were performed immediately after injury, and 400 000 U/d penicillin was intravenously infused for 1 week. The feeding, activity, and vocalization of dogs in each group after injury were observed until they were sacrificed. Immediately after injury and 2, 4, 6, and 8 weeks after injury, the laryngeal morphology of the dogs in corresponding time point groups were observed by endoscope. After the observation, the dogs in each injury group were sacrificed, and the laryngeal tissue was taken. The epiglottis, glottis, and cricoid cartilage were collected to make full-thickness tissue slice, respectively, and their pathological changes were observed with hematoxylin and eosin staining. The dogs of control group were not specially treated, and their life activities, laryngeal morphological and pathological changes were observed. Results: (1) The dogs of control group had normal feeding, activities, and vocalization. All the dogs in injury group survived until they were sacrificed, and their feeding, activities, and vocalization were obviously reduced after injury compared with those of control group. The dogs of 2, 4, 6 and 8 weeks after injury groups ate and moved normally 2 weeks after injury but vocalized abnormally in frequency and volume compared with those of control group, which lasted until they were sacrificed. (2) The dog's laryngeal mucosa in control group was complete and pink, without obvious exudation. The laryngeal mucosa of the dog in immediately after injury group was pale and edematous, with obvious exudation, local ulceration, necrosis, and exfoliation, and dilated microvessels on the surface. The laryngeal mucosa of the dogs in 2 weeks after injury group was pale, edematous, and oozed less than that of immediately after injury group, and the glottis was blocked by an obviously extruding mass. The paleness and edema of laryngeal mucosa were significantly reduced in the dogs of 4 weeks after injury group compared with those of 2 weeks after injury group, without dilated microvessel, and the glottic extruding mass was obviously smaller than that of 2 weeks after injury group. The sizes of glottic mass were similar between the dogs of 6 and 8 weeks after injury groups, which were obviously smaller than that in 4 weeks after injury group. (3) In the dogs of control group, the epithelial cells of epiglottis, glottis, and cricoid cartilage were normal in morphology, the proper glands were visible in the intrinsic layer, and the muscle fibers and the chondrocytes were normal in morphology. In the dogs of immediately after injury group, large sheets of epiglottis epidermis exfoliated, the epithelial cells were swollen and necrotic, the intrinsic glands were atrophic and necrotic, and the chondrocytes were degenerated and necrotic. The epidermis of the glottis partially exfoliated, the epithelial cells were swollen and necrotic, the intrinsic glands were atrophic and necrotic, the muscle fibers were partially atrophic and fractured, and the vacuolar chondrocytes were visible. The cricoid cartilage epidermis was ablated, the epithelial cells were swollen, the intrinsic layer and submucosal layer were slightly edematous, and the morphological structure of glands, chondrocytes, and muscle fibers were normal. In the dogs of 2 weeks after injury group, the epiglottis epidermis was completely restored, a small amount of glands in the intrinsic layer were repaired, and obsolete necrotic chondrocytes and new chondrocytes could be seen. A large number of fibroblasts, new capillaries, and inflammatory cells infiltration were observed in the epidermis of glottis, and intrinsic layer glands were repaired. The cricoid cartilage epidermis was repaired intactly, and there was no edema in the intrinsic layer. In the dogs of 4 weeks after injury group, the epiglottis intrinsic layer glands were further repaired compared with those of 2 weeks after injury group, and new chondrocytes were seen in the submucosa of the glottis. The condition of cricoid cartilage was consistent with that of control group. The dog's epiglottis, glottis, and cricoid cartilage were similar between the 6 and 8 weeks after injury groups, and no significant change was observed compared with those of 4 weeks after injury group. Conclusions: The morphological changes of larynx after severe laryngeal burn in dogs include mucosa detachment and necrosis, and mass blocking glottis. Pathological changes include epidermis shedding and necrosis, gland atrophy and necrosis, vascular congestion and embolism, chondrocytes degeneration, necrosis and proliferation, even local granulation tissue formation and cartilaginous metaplasia. These results may be the cause of laryngostenosis after laryngeal burn.

Objective: The aim of this study was to evaluate the early histopathological changes of gastroesophageal reflux and irradiation on laryngeal mucosa in rats., Study Design: Animal study., Setting: Experimental animal laboratory, tertiary referral center., Subject and Method: Twenty-four adult female Wistar Albino rats were grouped as: control (n = 6), reflux and irradiation (n = 10), and irradiation (n = 8). Rats were operated to create a reflux model 30 days before irradiation. Ionizing radiation was administered in a single fraction of a 20 Gy to the larynx. Laryngeal tissue samples were taken at the 4th day of irradiation and all specimens underwent histopathological examination., Results: Edema and vascular dilation in lamina propria were higher in the reflux and irradiation, and irradiation groups than control group. Inflammation was higher in the reflux and irradiation group than the control group. Inflammation in squamous epithelium was higher in the reflux and irradiation and irradiation groups compared to the control group. Inflammation in the squamous epithelium of the irradiation group was higher than the reflux and irradiation group. In the respiratory tract epithelium, inflammation was higher in the reflux and irradiation group; additionally, a significant loss of cilia was present in the reflux and irradiation and irradiation groups while pseudostratification was higher in the reflux and irradiation group., Conclusion: Ionizing radiation-induced inflammation may increase on previously inflammated area due to gastroesophageal reflux. Therefore, it may be helpful to investigate and treat the reflux in laryngeal cancer patients that will receive ionizing radiation.

Videolaryngostroboscopy is a useful investigation required for a correct diagnosis of laryngeal diseases and voice disorders. We present a form for the collection of basic laryngostroboscopic findings, which provides for the evaluation of the classical six parameters codified by Hirano (symmetry and periodicity of glottic vibration, glottic closure, profile of vocal fold edge, amplitude of vocal fold vibration, mucosal wave) and six other parameters which we have included in the form for an essential and complete laryngostroboscopic evaluation (supraglottic framework behaviour, seat of phonatory vibration, vocal fold morphology and motility, level of the vocal fold, stops of vocal fold mucosa vibration). This form was created in 2002 during the elaboration of the protocol for the assessment of dysphonia of the Italian Society of Phoniatrics and Logopedics, which follows the guidelines of the European Laryngological Society published in 2001. We used this form for 15 years in our daily laryngological practice with great satisfaction. We propose a more detailed version of this form, which provides for drawings which show the various videolaryngostroboscopic findings, helping the laryngologist in the collection of videolaryngostroboscopic examination basic findings.

Purpose: Aberrant fibroblast growth factor receptor (FGFR) expression is thought to contribute to the development of many types of cancer. As yet, however, their impact on the course and prognosis of head and neck cancer remains to be determined. Here, we aimed to investigate the effects of expression of the FGFR family members FGFR1 and FGFR3, as well as their downstream PI3K/AKT signal-regulated kinases, on the aggressiveness and prognosis of laryngeal cancer., Methods: In total 137 surgically removed squamous cell laryngeal cancer (SCLC) and 100 matched non-cancerous laryngeal mucosa (NCLM) samples were assessed for mRNA expression using quantitative real-time PCR. The corresponding proteins were analyzed by Western blotting. SLUG expression was assessed by immunohistochemistry. The expression data were subsequently related to tumor front grading (TFG), local/nodal recurrences, prognosis and overall survival., Results: The FGFR1, FGFR3 and PI3K/AKT kinase mRNA and protein levels were found to be significantly higher in the SCLC than the NCLM samples (p < 0.05). A high FGFR1 mRNA/protein expression level was found to be associated with an increased invasion rate, according to TFG scale and SLUG level, a high local/nodal recurrence rate and a poor prognosis (p < 0.05). Similarly, we found that a high FGFR3 mRNA/protein expression level was associated with a shorter survival time (p < 0.05). In addition, we found that high PI3K/AKT kinase mRNA/protein levels were associated with a high TFG (p < 0.05). We also found that FGFR1/3 mRNA and FGFR1 protein levels were inversely associated with overall survival (log-rank test: FGFR1 mRNA p = 0.03, FGFR3 mRNA p = 0.04, FGFR1 protein p = 0.03). Subsequent multivariate analyses revealed that high FGFR3 mRNA expression may serve as an independent poor prognostic factor (HR 2.32, 95% CI 1.03-6.59; p = 0.04). We also found that the p-PI3K regulatory kinase protein level was significantly associated with survival in the cohort studied (HR 1.78, 95% CI 0.64-8.53; p = 0.03)., Conclusions: From our data we conclude that FGFR1 and FGFR3, as well as its downstream regulatory PI3K/AKT kinases, may serve as potential biomarkers for the invasiveness and prognosis of laryngeal cancer. The expression of FGFR1/3-PI3K/AKT regulatory pathway members may be instrumental for the identification of patients at risk for an unfavorable clinical outcome.

Objective: Subglottic stenosis remains a clinical challenge. The aim of this study was to evaluate the effect of human adipose tissue-derived mesenchymal stem cells (hAMSCs) in rat model of subglottic stenosis., Subjects and Methods: Ninety-six 13-week-old male rats were enrolled in this study. They were divided into 3 groups as normal control (NC) group, a subglottic injury and media injection (SM) group, and a subglottic injury and media-stem cell injection (SMSC) group. The hAMSCs were immediately injected into subglottis after injury. Histologic characteristics of subglottis; the mRNA expressions of interleukin-1β, cyclooxygenase-2, tumor growth factor-β and basic fibroblast growth factor; and hAMSCs' survival were evaluated., Results: The hAMSCs survived in the subglottis of the rat until 10 days after implantation. The NC and SMSC groups had a significantly wider subglottic lumen and thinner lamina propria than the SM group at 56 days after injury. Collagen intensity of subglottis was significantly higher in the SM group than in the NC and SMSC groups at 28 days after injury. Gene expression didn't show significant difference between the SM group and the SMSC group., Conclusions: The hAMSCs injection was found to be helpful for preventing subglottic stenosis in a rat model.

Objectives: To evaluate CD163+ tumor-associated macrophages (TAMs), Ki-67, and cyclin D1 to differentiate laryngeal dysplasia in the 2017 World Health Organization classification., Methods: Immunohistochemistry for CD163, Ki-67, and cyclin D1 was performed using paraffin-embedded specimens. CD163+ TAMs infiltrating the epithelium were estimated. Ki-67 and cyclin D1 were evaluated in four parts of the epithelium-basal, parabasal, middle third, and upper third layers., Results: In total, 133 specimens were analyzed, including low-grade dysplasia (n = 31), high-grade dysplasia (n = 49), carcinoma in situ (n = 23), and normal mucosa (n = 30). CD163+ TAMs infiltrating the epithelium were significantly higher in high-grade dysplasia than in low-grade dysplasia. In the basal layer, Ki-67+ and cyclin D1+ cells were overexpressed in high-grade dysplasia (P < .0001). The area under the curve was 0.958 for Ki-67 and 0.909 for CD163+ TAMs (P < .0001)., Conclusions: CD163+ TAMs infiltrating the epithelium and Ki-67 overexpression in the basal layer may serve as biomarkers to differentiate low-grade dysplasia from high-grade dysplasia of the larynx. A symmetric proliferative pattern was observed during laryngeal carcinogenesis following Ki-67 overexpression., (© American Society for Clinical Pathology, 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com)

Background: Lymphatic metastasis contributes to the poor prognosis of laryngeal squamous cell carcinoma (LSCC). This study aimed to investigate the roles of two metastasis suppressor genes, KAI1 and nm23, in lymphangiogenesis and lymph metastasis of LSCC., Methods: A total of 45 LSCC patients were enrolled in this study. The positive expression rates of KAI1 and nm23 protein were detected via immunohistochemistry in 45 LSCC and 22 normal laryngeal mucosa adjacent to LSCC. Micro-lymphatic vessel density (MLVD) was detected via immunohistochemistry with the specific antibody D2-40. Associations between KAI1 and nm23 expression and clinical characteristics of LSCC were then evaluated., Results: The positive expression rates of KAI1 and nm23 were significantly lower in LSCC than normal laryngeal mucosa (P < 0.05). Significantly lower positive rates of KAI1 and nm23 were found in LSCC with lymphatic metastasis than those without lymphatic metastasis (P < 0.05), whereas MLVD was negatively correlated with the expression of KAI1 and nm23 (P < 0.05). However, no significant associations were found between KAI1 and nm23 expression and clinical characteristics of LSCC (sex, age, disease position, differentiation, and T-stage)., Conclusions: Both KAI1 and nm23 can inhibit lymphangiogenesis and lymphatic metastasis in LSCC.

Background and Aim: Autoimmune bullous disease (ABD) is induced by autoantibodies against cell adhesion molecules, and blistering may occur on the mucous membranes of the eyes, nose, mouth, oral cavity, laryngopharynx, and esophagus. Endoscopic prevalence and features of ABD-associated esophageal lesions are not well known. We conducted the present study to assess the endoscopic prevalence of ABD-associated mucosal lesions., Methods: Endoscopic prevalence of mucosal lesions, particularly laryngopharyngeal and esophageal lesions, was used as the primary endpoint to assess the significance of upper gastrointestinal endoscopy, and clinical and endoscopic features were secondary endpoints., Results: Of 123 ABD patients, 50.4% had apparent oral or laryngopharyngeal lesions and 30.8% had laryngopharyngeal lesions. Esophageal lesions were detected through normal observation in 16.8% of affected patients, whereas 40.6% exhibited epidermolysis or blood blisters by mechanical inducement, regardless of esophageal mucosal lesion detection by normal observation. Additionally, 56.0% exhibited the Nikolsky sign with mechanical inducement. Of the 123 patients, 29.2% did not have exposed skin lesions. Of these patients, 77.7% had oral cavity or laryngopharyngeal lesions, 36.1% had esophageal lesions, and 58.3% exhibited the Nikolsky sign on esophageal mucosa., Conclusion: It is important to determine the endoscopic characteristics and findings of ABD. ABD can be suspected from endoscopic findings., (© 2017 Japan Gastroenterological Endoscopy Society.)

Competing Interests: Interessenkonflikt: Die Autoren geben an, dass kein Interessenkonflikt besteht.

Objectives/hypothesis: A custom-designed probe was developed to measure vocal fold surface resistance in vivo. The purpose of this study was to demonstrate proof of concept of using vocal fold surface resistance as a proxy of functional tissue integrity after acute phonotrauma using an animal model., Study Design: Prospective animal study., Methods: New Zealand White breeder rabbits received 120 minutes of airflow without vocal fold approximation (control) or 120 minutes of raised intensity phonation (experimental). The probe was inserted via laryngoscope and placed on the left vocal fold under endoscopic visualization. Vocal fold surface resistance of the middle one-third of the vocal fold was measured after 0 (baseline), 60, and 120 minutes of phonation. After the phonation procedure, the larynx was harvested and prepared for transmission electron microscopy., Results: In the control group, vocal fold surface resistance values remained stable across time points. In the experimental group, surface resistance (X% ± Y% relative to baseline) was significantly decreased after 120 minutes of raised intensity phonation. This was associated with structural changes using transmission electron microscopy, which revealed damage to the vocal fold epithelium after phonotrauma, including disruption of the epithelium and basement membrane, dilated paracellular spaces, and alterations to epithelial microprojections. In contrast, control vocal fold specimens showed well-preserved stratified squamous epithelia., Conclusions: These data demonstrate the feasibility of measuring vocal fold surface resistance in vivo as a means of evaluating functional vocal fold epithelial barrier integrity. Device prototypes are in development for additional testing, validation, and for clinical applications in laryngology., Level of Evidence: NA Laryngoscope, 127:E364-E370, 2017., (© 2017 The American Laryngological, Rhinological and Otological Society, Inc.)

The use of narrow band imaging (NBI) and further technological achievements concerning the resolution and magnification of endoscopic images have revolutionized laryngology in the past 10 years. The diagnosis and therapy of dysplasia and early laryngeal carcinoma have become significantly easier. There are also clear benefits for benign laryngeal lesions. Central to these techniques is the assessment of epithelial, connective tissue and vascular changes caused by diverse diseases.

Objective: The aim of this study was to investigate effects of smoke produced by electrocautery on the laryngeal mucosa., Materials and Methods: We used 16 healthy, adult female Wistar albino rats. We divided the rats into two groups. Eight rats were exposed to smoke for 60 min/d for 4 weeks, and eight rats were not exposed to smoke and served as controls. The experimental group was maintained in a plexiglass cabin during exposure to smoke. At the end of 4 weeks, rats were sacrificed under high-dose ketamine anesthesia. Each vocal fold was removed. An expert pathologist blinded to the experimental group evaluated the tissues for the following: epithelial distribution, inflammation, hyperplasia, and metaplasia. Mucosal cellular activities were assessed by immunohistochemical staining for Ki67. Results taken before and after effect were compared statistically., Results: There was a significant difference in the extent of inflammation between the experimental group and the control group. Squamous metaplasia was detected in each group, but the difference was not significant. None of the larynges in either group developed hyperplasia., Conclusions: We showed increased tissue inflammation due to irritation by the smoke., (Copyright © 2017 The Voice Foundation. Published by Elsevier Inc. All rights reserved.)

Background: Mucosal melanoma of the upper aerodigestive tract (MM-UADT) occurs in a complex anatomic region. It represents a small number of tumors of the head and neck and a small number of melanoma cases., Methods: Search strategies initially identified 600, 11 of which were included in this study., Results: All studies involved surgery and radiotherapy. None were randomized, and all were assessed as having a high risk of selection and performance bias. No studies reported quality of life, treatment-related mortality, or morbidity. The results indicate that the addition of radiotherapy to surgery reduces the rate of locoregional recurrence (hazard ratio [HR], 0.60; 95% confidence interval [CI], 0.42-0.87). There was no statistically significant difference in overall survival (HR, 1.16; 95% CI, 0.98-1.37)., Conclusion: Surgical resection with postoperative radiotherapy remains the optimal treatment strategy for locoregional control. More robust studies and the use of molecular targeted therapies need to be undertaken to improve overall survival. © 2016 Wiley Periodicals, Inc. Head Neck 39: 819-825, 2017., (© 2016 Wiley Periodicals, Inc.)

Cigarettes contain toxic and carcinogenic substances. In this context, cigarette smoking, and similar activities, are associated with numerous pathologies, being considered a risk factor in up to 10% of the total number of deaths in adults. Recent evidence suggests that the exposure of children to smoking in the early days of their development causes many diseases. Using light microscopy, this study aims to analyze the possible histopathological effects of an experimental model of chronic inhalation of cigarette smoke (passive smoking) on the laryngeal and tracheal mucosa of young Wistar rats. A total of 24 young Wistar rats were studied for a period of 120 days. The animals were divided into two groups: passive smoking (n = 16) and control (n = 8). The level of exposure to cigarette smoke was evaluated from the urinary cotinine level. Although no cancerous lesions were identified, histopathological analysis in the laryngeal and tracheal mucosa of all the animals in the experimental group showed that the proportion of moderate and focal inflammation was higher in animals exposed to chronic inhalation of cigarette smoke (P = 0.041). Histopathologic analysis revealed moderate and focal inflammatory lesions in the region of the infraglottic mucosa in exposed animals, although without dysplastic or neoplastic lesions in the laryngeal and tracheal mucosa., (Copyright © 2017 The Voice Foundation. Published by Elsevier Inc. All rights reserved.)

Objective: Diagnosis of laryngopharyngeal reflux (LPR) in clinical practice is generally made subjectively based on history, symptoms, and endoscopic assessment. This study presents the diagnostic role of the digital photographic assessment of the laryngopharyngeal region for LPR., Methods: Seventy-two patients with LPR symptoms and 35 healthy volunteers were evaluated using the Reflux Finding Scores (RFS) and the Reflux Symptom Index (RSI). The results of these scales were recorded in 72 patients representing group 1 with LPR diagnosis; they returned after 1 and 2 months of treatment as groups 2 and 3, respectively. Thirty-five volunteers represented group 4. Laryngopharyngeal regions of all subjects were examined and photographed endoscopically. Red, green, and blue (RGB) values of particular oropharyngeal and laryngeal points were measured., Results: RSI and RFS values of group 1 were significantly different from the other three groups, as were the scores of group 2; however, the comparison of the RSI and RFS values of groups 3 and 4 did not reveal a statistically significant difference. Laryngopharyngeal RGB values also duplicated statistical significance as above., Conclusion: Measurement of RGB values can be a cheap and easy-to-use method to quickly provide objective and corroborative information to help in the diagnosis of LPR in conjunction with subjective methods., (Copyright © 2017 The Voice Foundation. Published by Elsevier Inc. All rights reserved.)

Few cases of primary mucosal melanoma of the larynx have been documented in the literature, so only a limited amount of data exists regarding its diagnosis and treatment. The prognosis is poor, as patients often present at a late stage with regional or distant metastases. We describe the case of a 66-year-old man who presented with hoarseness and dysphagia. Laryngoscopy identified a dark discoloration of the supraglottic larynx and incomplete mobility of the right vocal fold; an excisional biopsy confirmed the diagnosis. We discuss the epidemiology, clinical features, diagnosis, interpretation of imaging findings, and management of this rare malignant melanoma.

Recent evidence indicates the involvement of calpains (CAPNs), a family of cysteine proteases, in cancer development and progression, as well as the insufficient response to cancer therapies. The contribution of CAPNs and regulatory calpastatin (CAST) and ERK1/2 kinases to aggressiveness, disease course, and outcome in laryngeal cancer remains elusive. This study was aimed to evaluate the CAPN1/2-CAST-ERK1/2 enzyme system mRNA/protein level and to investigate whether they can promote the dynamic of tumor growth and prognosis. The mRNA expression of marker genes was determined in 106 laryngeal cancer (SCLC) cases and 73 non-cancerous adjacent mucosa (NCLM) controls using quantitative real-time PCR. The level of corresponding proteins was analyzed by Western Blot. SLUG expression, as indicator of pathological advancement was determined using IHC staining. Significant increases of CAPN1/2-CAST-ERK1/2 levels of mRNA/protein were noted in SCLC compared to NCLM (p < 0.05). As a result, a higher level of CAPN1 and ERK1 genes was related to larger tumor size, more aggressive and deeper growth according to TFG scale and SLUG level (p < 0.05). There were also relationships of CAPN1/2 and ERK1 with incidences of local/nodal recurrences (p < 0.05). An inverse association for CAPN1/2, CAST, and ERK1/2 transcripts was determined with regard to overall survival (p < 0.05). In addition, a higher CAPN1 and phospho-ERK1 protein level was related to higher grade and stage (p < 0.05) and was found to promote worse prognosis. This is the first study to show that activity of CAPN1/2- CAST-ERK1/2 axis may be an indicator of tumor phenotype and unfavorable outcome in SCLC.

Bullous pemphigoid is an autoimmune bullous disease characterized by skin lesions, with or without oral lesions. The occurrence of pharyngolaryngeal lesions is very rare in affected patients. We conducted a study to investigate the characteristics of oral and pharyngolaryngeal lesions in bullous pemphigoid. Our study population was made up of 6 consecutively presenting outpatients-2 men and 4 women, aged 40 to 83 years (mean: 68.2)-who had been referred to our department over an 11-year period. Presenting symptoms included sore throat in all 6 patients and oral pain in 3. The sites of mucosal lesions included the soft palate, epiglottis, gingiva, hypopharynx, tongue, nasal cavity, and buccal mucosa. These lesions appeared as erosions, erosions with white coating, erythematous patches, and/or blisters. Mucosal lesions preceded skin lesions in 2 patients, appeared after skin lesions in 1 patient, and appeared simultaneously with skin lesions in 3 patients. We conclude that bullous pemphigoid sometimes involves the mucosa, such as that of the laryngopharynx and the oral cavity, and it can manifest as skin lesions. In the differential diagnosis of refractory pharyngolaryngeal lesions, bullous pemphigoid should be considered.

Objectives: To evaluate the degree of agreement between the clinical and the pathological diagnosis in patients undergoing laryngeal microsurgery due to epithelial or edematous lesions of the vocal folds., Study Design: This is a retrospective study., Methods: The study was developed in the ear, nose, and throat clinic of a tertiary hospital, through chart review. We included all patients who underwent microsurgery of the larynx, whose videolaryngoscopic tests showed vocal fold lesions, from January 2003 to August 2014. During the study period, we identified 48 patients with epithelial and edematous lesions. The patients were divided into two groups. In group A, patients with edematous lesions with clinical diagnosis of polyps and Reinke edema were included. Group B comprised patients with epithelial lesions, as leukoplakic injuries. A correlation between histopathological findings and clinical hypothesis between these two groups of lesions was performed., Results: In group B, there was agreement in 88.9% of cases between the clinical and pathological diagnosis. In group A, compatibility occurred in only 46.4% of cases. We observed a statistically significant difference between the compatibility of the clinical and pathological diagnosis just in edematous lesions (P = 0.029)., Conclusions: The study showed the limitation of the pathological examination on edematous laryngeal lesions. On the other hand, on the epithelial lesions, there was more agreement between those diagnosis., (Copyright © 2016 The Voice Foundation. Published by Elsevier Inc. All rights reserved.)

Objectives/hypothesis: Sulcus vocalis is an epithelial invagination adherent to deep tissues of the vocal fold. Traditionally, dysphonia is believed to result from attenuation or absence of lamina propria and consequent alteration of mucosal dynamics. This conception does not account for several clinical features of the lesion, most notably inflammation. The goal of this study is to elucidate the clinical nature of sulcus by re-examination of its histopathology., Study Design: Retrospective review., Methods: Clinical features, including stroboscopic examination, and hematoxylin and eosin sections of 19 lesions in 15 patients who underwent surgery were reviewed., Results: Epithelial change was found uniformly in all specimens, consisting of parakeratosis (78.9%), epithelial thickening (77.8%), dyskeratosis (63.2%), basement membrane thickening (47%), epithelial inflammation (52.6%), and retention of keratin debris (36.8%). In contrast, submucosal findings were less prevalent. Seven patients (36.8%) had essentially normal subepithelial tissues. Submucosal inflammation was present in seven (36.8%), and other submucosal changes in four of 17 (23.5%). Clinical characteristics demonstrated general improvement after surgical intervention in most cases., Conclusions: Sulcus vocalis appears to have an important component of epithelial pathology, with especially high prevalence of parakeratosis, dyskeratosis, and epithelial thickening. Clinical changes may result from prominent perilesional inflammation related to epithelial changes instead of or in addition to any alteration of the lamina propria. Surgical treatment, when necessary, should refocus on removal of pathologic epithelium as a source of inflammation independent of reconstructive considerations., Level of Evidence: 4 Laryngoscope, 126:1397-1403, 2016., (© 2015 The American Laryngological, Rhinological and Otological Society, Inc.)

Unlabelled: Gastroduodenal refluxate found in the upper aerodigestive tract is not clinically uncommon. We recently demonstrated the neoplastic potential of gastroduodenal fluids (GDF) on hypopharyngeal mucosa, via NF-κB, using in vitro and in vivo models. Here we will explore the in vivo effect of GDF on laryngeal mucosa (LM) to induce early preneoplastic lesions related to NF-κB activation, along with deregulation of specific microRNA (miRNA) markers previously linked to laryngeal cancer. We used histological, immunohistochemical, automated quantitative analysis and quantitative polymerase chain reaction to examine LM from 35 C57Bl/6J mice previously treated with topical GDF against corresponding controls (4 experimental and 3 control groups; 5 mice/group). Our analysis showed that GDF produced early preneoplastic lesions in treated LM related to NF-κB activation. LM treated by acid and bile combination demonstrated significantly higher expression of the analyzed cell proliferation markers (Ki67, CK14, ∆Np63), oncogenic p-STAT3, and changes of cell adhesion molecules (E-cadherin, ϐ-catenin) versus untreated LM or LM exposed to acid alone (P < .0005). Furthermore, acidic bile but not neutral bile appeared to accelerate the expression of "oncomirs" miR-21, miR-155, and miR-192 (acidic bile versus neutral bile, P < .0001), while reducing tumor suppressor miR-375 (acidic bile versus neutral bile, P = .0137), previously linked to NF-κB and laryngeal cancer. Finally, acidic bile induced reduction of miR-34a, miR-375, and miR-451a, exhibiting an inverse correlation with NF-κB activation., Significance: Bile in combination with acid has a selective tumorigenic effect on LM, inducing deregulation of "oncomirs" and tumor suppressor miRNAs, produced by NF-κB activation with molecular and early histopathological alterations linked to neoplastic transformation. Systematic acid suppression may in part convey a protective role., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)

Aim: To determine the methylation pattern of the promoter region of the O6-methylguanine-DNA methyltransferase (MGMT) gene in laryngeal cancer and normal laryngeal mucosa samples using pyrosequencing, and to determine the relationship between the methylation pattern of MGMT, and tumor stage, survival, recurrence, and chemosensitivity in patients with laryngeal cancer., Materials and Methods: Laryngeal cancer and normal laryngeal mucosa specimens were obtained from our paraffin block archives, and then subjected to pyrosequencing. Different cut-off values were used to detect methylation. Clinicopathological data for the patients that provided specimens were obtained from archive records., Results: When 5% was used as the cut-off value, 78% of the laryngeal cancer specimens (64 of 82), and 27.3% of normal laryngeal mucosa specimens (3 of 11) were considered methylated. When 10% was used as the cut-off value, 47% of the laryngeal cancer specimens (39 of 82), and none of the normal laryngeal mucosa specimens were considered methylated. There was not a significant relationship between the methylation status of MGMT, and clinicopathological parameters, including age, tumor stage, histopathological differentiation, chemoradiotherapy protocol used, recurrence, or disease-free survival., Conclusion: Pyrosequencing is a reliable semiquantitative technique that can be used to detect the methylation pattern. Methylation was common in the laryngeal cancer specimens, but there was not a significant relationship between the methylation status of MGMT and clinicopathological parameters., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

Objective: The aim of this study was to measure the effects of endotracheal intubation on innate immune response within the pig laryngeal mucosa., Study Design: Prospective controlled basic science study., Setting: The animal experiments and analyses were conducted at the University of Bristol., Samples and Methods: Eighteen pigs, matched at the major histocompatibility complex (MHC), were used in the study. The pigs were divided into 9 pairs. One of each pair (9 pigs in total) was intubated with an endotracheal tube under general anesthesia for 90 minutes. Two days later, pinch biopsies were taken from the supraglottis (specifically the false cords) and subglottis of both pigs. The experiment was repeated 8 more times. Based on quantitative immunohistochemistry, percentage areas of positive staining for CD172a, CD163, MHC class II, CD14, and CD16 were calculated separately for the epithelium and lamina propria of each biopsy., Results: Total areas of laryngeal mucosa (epithelium and lamina propria) expressing CD172a and coexpressing CD163 and CD172a were significantly reduced at 2 days following endotracheal intubation (P = .039 and P = .037, respectively). MHC class II expression and MHC class II coexpression with CD172a were similarly reduced following intubation (P = .003 and P = .005, respectively). In the supraglottis, MHC class II coexpression with CD16 and CD14 was also reduced following endotracheal intubation (P = .037)., Conclusions: Our results indicate that endotracheal intubation reduces the number of innate immune cells within the upper airway mucosa. This may be an important first step in a cascade leading to chronic wound and scar formation causing airway stenosis., (© American Academy of Otolaryngology-Head and Neck Surgery Foundation 2015.)

Objective: We aimed to assess the effects of electronic nicotine delivery system (ENDS) or also termed electronic cigarette vapor on the laryngeal mucosa of rats., Materials and Methods: Sixteen female Wistar albino rats were divided into two groups. The study group was exposed to ENDS vapor for 1 hour/day for 4 weeks. The control group was not subjected to any chemical or physical stimulus. The vocal folds of the study and control group rats were evaluated histopathologically by hematoxylin and eosin staining and immunohistochemically by Ki67 staining. Epithelial distribution, inflammation, hyperplasia, and metaplasia were evaluated., Results: Epithelial distribution and inflammation did not differ between the two groups. Two cases of hyperplasia were detected in the study group but there was no hyperplasia in the control group. Four cases of metaplasia were detected in the study group and one case in the control group. Statistical analysis revealed no significant difference between the study and control groups (P = 0.131 and 0.106, respectively)., Conclusions: Exposure to ENDS for 4 weeks caused hyperplasia and metaplasia of the laryngeal mucosa of rats but this was not significant statistically. These results implemented that further studies with larger cohort and longer duration are required to evaluate long-term effects., (Copyright © 2015 The Voice Foundation. Published by Elsevier Inc. All rights reserved.)

Objectives: The larynx is susceptible to irradiation, which causes significant vocal fold (VF) edema and dehydration shortly after radiotherapy for head and neck cancers. However little is known about radiation-induced damage to VF liquid homeostasis. To evaluate the effects of irradiation on VF hydration and lubrication, we investigated changes in water transporters (aquaporins [AQPs]) and mucin production in vivo and ex vivo, as well as morphometric changes in the laryngeal mucosa and glands of irradiated rat larynges., Study Design: Animal study., Materials and Methods: Local irradiation at 18 Gy was delivered to rat larynges. Histologic changes in laryngeal mucosa and glands were observed by light microscopy, and the distributions of AQPs and mucin were investigated by immunofluorescence staining 3 months after irradiation. Early effects on gene regulation of AQPs and mucin were evaluated by quantitative real-time polymerase chain reaction of the extirpated VFs and subglottic laryngeal mucosa at 12, 24, and 72 hours after irradiation., Results: Laryngeal glands exhibited severe atrophic changes and showed decreased density throughout the irradiated larynx. The expression of AQP1, 4, 5, and mucin in VFs, as well as AQP5 and mucin in submucosal laryngeal glands, decreased significantly 3 months after irradiation. An ex vivo study revealed that the gene expression of AQP5 in VF tissues was significantly downregulated at 12 hours postirradiation., Conclusion: Laryngeal irradiation induces damage in laryngeal mucosal barriers and alters laryngeal liquid homeostasis, which may be one reason for vocal dysfunction following irradiation., Level of Evidence: N/A., (© 2015 The American Laryngological, Rhinological and Otological Society, Inc.)

Unlabelled: The aim of this prospective study was to look at the laryngeal findings and acoustic changes in light cigar smokers in comparison to nonsmokers, in the setting of a voice clinic. A total of 22 cigar smokers and 19 nonsmokers used as controls were enrolled in the study. Demographic data included age, number of years smoking, number of cigars per week, history of allergy, and history of reflux. The confounding effects of allergy and reflux were accounted for in the control group. Subjects underwent laryngeal endoscopy and acoustic analysis. On laryngeal endoscopy, the most common laryngeal finding was thick mucus. There was no significant difference in the prevalence of any of the laryngeal findings in cigar smokers vs., Controls: In comparison with the control group, both the fundamental frequency and habitual pitch were significantly lower in cigar smokers (p value = 0.034 and 0.004, respectively). We conclude that cigar smokers have lower fundamental frequency and habitual pitch compared to nonsmokers.