Predictive significance of computed tomography in bilateral sinonasal respiratory epithelial adenomatoid hamartoma.

KEY POINTS: Respiratory epithelial adenomatoid hamartoma (REAH) is easily confused with nasal polyps (NP). The typical manifestation of REAH on CT is the enlargement of bilateral olfactory clefts (OCs). The widening of the OCs in the CT scan is a biomarker for diagnosing REAH associated with NP.

Visualizing immune responses of the airway mucosa.

The airway mucosa is the primary tissue site exposed to inhaled particulate matter, which includes pathogens and allergens. While most inhaled particles are eliminated from the airways via mucociliary clearance, some pathogens may penetrate the mucosal epithelial barrier and an effective activation of the mucosal immune system is required to prevent further pathogen spread. Similarly, inhaled environmental allergens may induce an aberrant activation of immune cells in the airway mucosa, causing allergic airway disease. During the last years, several investigators employed advanced microscopic imaging on both intravital and tissue explant preparations to observe the dynamic behavior of various immune cells within their complex tissue environment. In the respiratory tract, most imaging studies focused on immune responses of the alveolar compartment in the lung periphery. However, equally important immunological events occur more proximally in the mucosa of the conducting airways, both during infection and allergic responses, calling for a more detailed imaging analysis also at this site. In this review, I will outline the technical challenges of designing microscopic imaging experiments in the conducting airways and summarize our recent efforts in understanding airway mucosal immune cell dynamics in steady-state conditions, during infection and allergy.

Immunological Features of Paranasal Sinus Mucosa in Patients with Graves' Orbitopathy.

Background: Previous studies showed that patients with Graves' orbitopathy (GO) had concomitant mucosal abnormality within the paranasal sinuses. It remains unknown whether the immunological reactions in sinus mucosa affect the orbit inflammation in GO. Methods: Patients with GO underwent sinus computed tomography (CT) scans for sinus mucosal disease by two independent reviewers using the Lund-MacKay systems. Ethmoid mucosal samples were collected during orbital decompression surgeries for patients with GO and correction surgeries for patients with old orbital fractures as controls. Histological analysis and immunofluorescence were performed in all sinus mucosa tissues. Flow cytometry analysis was used to examine the immunological features of sinus mucosa in both GO and control groups. Results: Immunohistochemistry showed that the paranasal sinus mucosa of patients with GO grew swelling, with goblet cell and small vessel proliferation, endothelial cell swelling, and inflammatory cell infiltration. The number of T helper (Th)1, Th17, and gamma-delta T cells in nasal sinus mucosa of patients with GO increased significantly compared with those from controls. Further, the proportion of Th1 cells was significantly correlated with clinical activity score. In addition, there was a decreased number of regulatory T cells in patients with GO. The number of Th2 cells showed no significant difference between the two groups. Finally, the proportion of interleukin-22-producing cell subsets in gamma-delta T cells of patients with GO was significantly increased compared with those from controls. Conclusions: Our observations illustrated a potential pathogenic role of mucosal-infiltrating T cells, which may have the possibility to aggravate inflammatory responses in GO.

Novel automated vessel pattern characterization of larynx contact endoscopic video images.

PURPOSE: Contact endoscopy (CE) is a minimally invasive procedure providing real-time information about the cellular and vascular structure of the superficial layer of laryngeal mucosa. This method can be combined with optical enhancement methods such as narrow band imaging (NBI). However, these techniques have some problems like subjective interpretation of vascular patterns and difficulty in differentiation between benign and malignant lesions. We propose a novel automated approach for vessel pattern characterization of larynx CE + NBI images in order to solve these problems. METHODS: In this approach, five indicators were computed to characterize the level of vessel's disorder based on evaluation of consistency of gradient and two-dimensional curvature analysis and then 24 features were extracted from these indicators. The method evaluated the ability of the extracted features to classify CE + NBI images based on the vascular pattern and based on the laryngeal lesions. Four datasets were generated from 32 patients involving 1485 images. The classification scenarios were implemented using four supervised classifiers. RESULTS: For classification of CE + NBI images based on the vascular pattern, polykernel support vector machine (SVM), SVM with radial basis function (RBF), k-nearest neighbor (kNN), and random forest (RF) show an accuracy of 97%, 96%, 96%, and 96%, respectively. For the classification based on the histopathology, Polykernel SVM showed an accuracy of 84%, 86% and 84%, RBF SVM showed an accuracy of 81%, 87% and 83%, kNN showed an accuracy of 89%, 87%, 91%, RF showed an accuracy of 90%, 88% and 91% for classification between benign histopathologies, between malignant histopathologies and between benign and malignant lesions, respectively. CONCLUSION: These promising results show that the proposed method could solve the problem of subjectivity in interpretation of vascular patterns and also support the clinicians in the early detection of benign, pre-malignant and malignant lesions.

In vitro model for predicting acute inhalation toxicity by using a Calu-3 epithelium cytotoxicity assay.

INTRODUCTION: As the current methods to predict the inhalation toxicity of chemicals using animal models are limited, alternative methods are required. We present a new in vitro prediction method for acute inhalation toxicity using the Calu-3 epithelial cytotoxicity assay applicable for water-soluble inhalable chemicals. METHOD: To confirm the characteristics of the optimal Calu-3 epithelium, tight-junction formation, morphology, and mucus secretion were verified using scanning electron microscopy, transepithelial electrical resistance analysis, and immunofluorescence after growth in an air-liquid interface (ALI). Sixty chemicals, including 38 positive and 22 negative for acute inhalation toxicity, were selected from the European Chemical Agency chemical database. The cell viability of the exposed cells was assessed using an MTT assay to predict the acute inhalation toxicity by calculating the area under the receiver operating characteristic (ROC) curve and accuracy. RESULTS: When cultivated in an ALI, the epithelium was thicker and secreted more mucin than that under submerged cultivation, characteristic of the in vivo respiratory epithelium. The areas under the ROC curve were 0.75 and 0.78 when exposed to chemicals at concentrations of 2.5 and 5%, respectively. The highest accuracy of the methods was 68 and 78% at cut-off values of 85 and 40% cell viability, respectively. DISCUSSION: The in vitro model was moderately accurate with good prediction. It is replicable because of its advantages, i.e., the use of cultured cells and the simplicity of the method. Overall, the Calu-3 epithelial cytotoxicity assay may be a useful and simple approach to identify substances that cause acute inhalation toxicity.

Molecular subtyping reveals immune alterations associated with progression of bronchial premalignant lesions.

Bronchial premalignant lesions (PMLs) are precursors of lung squamous cell carcinoma, but have variable outcome, and we lack tools to identify and treat PMLs at risk for progression to cancer. Here we report the identification of four molecular subtypes of PMLs with distinct differences in epithelial and immune processes based on RNA-Seq profiling of endobronchial biopsies from high-risk smokers. The Proliferative subtype is enriched with bronchial dysplasia and exhibits up-regulation of metabolic and cell cycle pathways. A Proliferative subtype-associated gene signature identifies subjects with Proliferative PMLs from normal-appearing uninvolved large airway brushings with high specificity. In progressive/persistent Proliferative lesions expression of interferon signaling and antigen processing/presentation pathways decrease and immunofluorescence indicates a depletion of innate and adaptive immune cells compared with regressive lesions. Molecular biomarkers measured in PMLs or the uninvolved airway can enhance histopathological grading and suggest immunoprevention strategies for intercepting the progression of PMLs to lung cancer.

Redox imaging and optical coherence tomography of the respiratory ciliated epithelium.

Optical coherence tomography (OCT) is an emerging technology for in vivo airway and lung imaging. However, OCT lacks sensitivity to the metabolic changes caused by inflammation, which drives chronic respiratory diseases such as asthma and chronic obstructive pulmonary disorder. Redox imaging (RI) is a label-free technique that uses the autofluorescence of the metabolic coenzymes NAD(P)H and flavin adenine dinucleotide (FAD) to probe cellular metabolism and could provide complimentary information to OCT for airway and lung imaging. We demonstrate OCT and RI of respiratory ciliated epithelial function in ex vivo mouse tracheae. We applied RI to measure cellular metabolism via the redox ratio [intensity of NAD(P)H divided by FAD] and particle tracking velocimetry OCT to quantify cilia-driven fluid flow. To model mitochondrial dysfunction, a key aspect of the inflammatory process, cyanide was used to inhibit oxidative metabolism and reduce ciliary motility. Cyanide exposure over 20 min significantly increased the redox ratio and reversed cilia-driven fluid flow. We propose that RI provides complementary information to OCT to assess inflammation in the airway and lungs.

Point-of-care endoscopic optical coherence tomography detects changes in mucosal thickness in ARDS due to smoke inhalation and burns.

BACKGROUND: The prevalence of acute respiratory distress syndrome (ARDS) in mechanically ventilated burn patients is 33%, with mortality varying from 11-46% depending on ARDS severity. Despite the new Berlin definition for ARDS, prompt bedside diagnosis is lacking. We developed and tested a bedside technique of fiberoptic-bronchoscopy-based optical coherence tomography (OCT) measurement of airway mucosal thickness (MT) for diagnosis of ARDS following smoke inhalation injury (SII) and burns. METHODS: 16 female Yorkshire pigs received SII and 40% thermal burns. OCT MT and PaO2-to-FiO2 ratio (PFR) measurements were taken at baseline, after injury, and at 24, 48, and 72h after injury. RESULTS: Injury led to thickening of MT which was sustained in animals that developed ARDS. Significant correlations were found between MT, PFR, peak inspiratory pressure (PIP), and total infused fluid volume. CONCLUSIONS: OCT is a useful tool to quantify MT changes in the airway following SII and burns. OCT may be effective as a diagnostic tool in the early stages of SII-induced ARDS and should be tested in humans.

Optical coherence tomography for observation of the olfactory epithelium in mice.

OBJECTIVE: Optical coherence tomography (OCT) is an imaging tool that exploits the coherence of infrared light and is clinically utilized in the field of ophthalmology and dermatology. This study aimed to examine the feasibility of using OCT for diagnosing degeneration and regeneration of the olfactory epithelium in mice. METHODS: The olfactory and respiratory epithelia in excised nasal septa of adult mice were observed using OCT. Subsequently, histological assessments were performed with hematoxylin and eosin (H-E) staining. The thicknesses of the olfactory or respiratory epithelia were measured in both OCT images and H-E-stained paraffin sections. The ability of OCT to distinguish olfactory epithelia from respiratory epithelia in normal mice was compared with that of H-E staining. The feasibility of using OCT assessments for detecting changes in the thickness of olfactory epithelia was tested in a mouse model of the degeneration and regeneration of olfactory epithelia. RESULTS: OCT allowed visualization of the gross morphology of the olfactory and respiratory epithelium in normal mice, although it was limited in terms of visualizing cellular components. OCT-based measurements of epithelial thickness helped to distinguish olfactory epithelia from respiratory epithelia. Similar to H-E staining, OCT also clarified changes in the olfactory epithelium thickness after methimazole application. CONCLUSIONS: These findings indicate the utility of OCT for assessment of olfactory epithelial thickness and its potential for clinical evaluation of human olfactory epithelia.

Functional Anatomic Imaging of the Airway Surface.

The airway surface functional microanatomy, including the ciliated airway epithelium and overlying mucus layer, is a critical component of the mucociliary escalator apparatus, an innate immune defense that helps to maintain a clean environment in the respiratory tract. Many genetic and acquired respiratory diseases have underlying pathophysiological mechanisms in which constituents of the airway surface functional microanatomy are defective. For example, in cystic fibrosis, mutations in the cystic fibrosis transmembrane conductance regulator gene, which normally produces a secretory anion channel protein, result in defective anion secretion and consequent dehydrated and acidic mucosal layer overlying the airway epithelium. This thick, viscous mucus results in depressed ciliary beating and delayed mucociliary transport, trapping bacteria and other pathogens, compromising host defenses and ultimately propagating disease progression. Thus, developing tools capable of studying the airway surface microanatomy has been critical to better understanding key pathophysiological mechanisms, and may become useful tools to monitor treatment outcomes. Here, we discuss functional imaging tools to study the airway surface functional microanatomy, and how their application has contributed to an improved understanding of airway disease pathophysiology.

Uncovering the regional localization of inhaled salmeterol retention in the lung.

Treatment of respiratory disease with a drug delivered via inhalation is generally held as being beneficial as it provides direct access to the lung target site with a minimum systemic exposure. There is however only limited information of the regional localization of drug retention following inhalation. The aim of this study was to investigate the regional and histological localization of salmeterol retention in the lungs after inhalation and to compare it to systemic administration. Lung distribution of salmeterol delivered to rats via nebulization or intravenous (IV) injection was analyzed with high-resolution mass spectrometry imaging (MSI). Salmeterol was widely distributed in the entire section at 5 min after inhalation, by 15 min it was preferentially retained in bronchial tissue. Via a novel dual-isotope study, where salmeterol was delivered via inhalation and d3-salmeterol via IV to the same rat, could the effective gain in drug concentration associated with inhaled delivery relative to IV, expressed as a site-specific lung targeting factor, was 5-, 31-, and 45-fold for the alveolar region, bronchial sub-epithelium and epithelium, respectively. We anticipate that this MSI-based framework for quantifying regional and histological lung targeting by inhalation will accelerate discovery and development of local and more precise treatments of respiratory disease.

Intubation-free in vivo imaging of the tracheal mucosa using two-photon microscopy.

The mucosal layer of conducting airways is the primary tissue exposed to inhaled microorganisms, allergens and pollutants. We developed an in vivo two-photon microscopic approach that allows performing dynamic imaging studies in the mouse trachea, which is a commonly used in vivo model of human small-diameter bronchi. By providing stabilized access to the tracheal mucosa without intubation, our setup uniquely allows dynamic in vivo imaging of mucociliary clearance and steady-state immune cell behavior within the complex airway mucosal tissue.

Biomechanical Flow Amplification Arising From the Variable Deformation of the Subglottic Mucosa.

OBJECTIVE: This study mapped the variation in tissue elasticity of the subglottic mucosa, applied these data to provide initial models of the likely deformation of the mucosa during the myoelastic cycle, and hypothesized as to the impact on the process of phonation. STUDY DESIGN: Six donor human larynges were dissected along the sagittal plane to expose the vocal folds and subglottic mucosa. A linear skin rheometer was used to apply a controlled shear force, and the resultant displacement was measured. These data provided a measure of the stress/strain characteristics of the tissue at each anatomic point. A series of measurements were taken at 2-mm interval inferior of the vocal folds, and the change in elasticity was determined. RESULTS: It was found that the elasticity of the mucosa in the subglottic region increased linearly with distance from the vocal folds in all 12 samples. A simple deformation model indicated that under low pressure conditions the subglottic mucosa will deform to form a cone, which could result in a higher velocity, thus amplifying the low pressure effect resulting from the Venturi principle, and could assist in maintaining laminar flow. CONCLUSIONS: This study indicated that the deformation of the subglottic mucosa could play a significant role in the delivery of a low pressure airflow over the vocal folds. A large scale study will now be undertaken to secure more data to evaluate this hypothesis, and using computational fluid dynamics based on actual three-dimensional structure obtained from computed tomography scans the aerodynamics of this region will be investigated.

[Nasopharyngeal cyst of the respiratory epithelium in a 9-year-old Yorkshire terrier]. / Nasopharyngeale Zyste des respiratorischen Epithels bei einem 9 Jahre alten Yorkshire Terrier.

In a 9-year-old Yorkshire terrier a cyst of the respiratory epithelium of the nasopharynx was diagnosed. A complete obstruction of the nasopharynx leading to dyspnea was detected by computed tomography and endoscopy. A minimally invasive ablation of the cystic wall was performed under endoscopic guidance, followed by a pathohistological examination. Immediately after resection of the cyst, the clinical symptoms resolved. The follow-up endoscopical examination 3 months postoperatively was unremarkable. In the presented case the minimally invasive endoscopic ablation of the cystic wall was a successful treatment method.

Radioiodine sinus uptake related to mucosal thickening or aspergilloma: a case series of an unrecognized event well evidenced by SPECT/CT.

BACKGROUND: False-positive radioiodine (RAI) uptake related to chronic sinusitis and mucocele has only rarely been reported in patients with differentiated thyroid cancer (DTC) even with the recent use of single photon emission tomography with computed tomography (SPECT/CT) acquisition. No other etiology of sinus RAI uptake has been mentioned to date. OBJECTIVES: We report five cases of DTC patients with sinus RAI uptake on post-RAI scintigraphy. SPECT/CT clearly localized RAI uptake either in the sphenoid, the maxillary or the frontal sinus and highly suspected mucosal thickening in four patients and sinus aspergilloma in one patient. CONCLUSION: These data confirm the possibility of false-positive sinus RAI uptake, provide a new cause of such benign uptake, i.e. sinus aspergilloma, and demonstrate the clinical relevance of head and neck SPECT/CT acquisition in the diagnosis of such uptake. Nuclear medicine physicians should be aware of this pitfall when interpreting post-RAI scintigraphy.

Amplitude of Glottal Mucosal Wave After Vocal Fold Microflap With or Without Fibrin Glue.

INTRODUCTION: The vocal fold microflap technique is the ideal to remove benign vocal fold pathology. Our objective is to compare the amplitudes of the mucosal wave before and after the closure of microflap defect with fibrin glue, and when microflap is left to heal by secondary intention. MATERIALS AND METHODS: The present study is a retrospective series, including 32 patients treated by intracordal phonosurgery, with closure of the microflap either with fibrin glue or by healing by secondary intention. They all had both preoperative and 6-month postoperative track records to allow voice analysis, a subjective Voice Handicap Index 10 (VHI-10), and a good image quality strobe. RESULTS: After selecting the patients was found that the mean overall preoperative VHI-10 was 26.6, and improved up to 10.5 after surgery, a statistical differences (P = 0.03). When comparing both groups, with or without fibrin glue, fibrin glue did not improved results in VHI-10. On the contrary, there was a significant difference in the improvement of the open glottal phase after surgery (P = 0.03), showing a much higher improvement when fibrin glue was used. CONCLUSIONS: The use of fibrin glue after a vocal fold microflap for advanced pathology, such as sulcus vocalis in pocket, vergeture, or vocal fold scar, increases the amplitude of the mucosal wave of the vocal folds, but does not improve the VHI-10 results in our cohort of female patients. So far, patient-reported outcome shows that healing by secondary intention continues to provide excellent voice results.