Modeling gap junction beta 2 gene-related deafness with human iPSC.

There are >120 forms of non-syndromic deafness associated with identified genetic loci. In particular, mutation of the gap junction beta 2 gene (GJB2), which encodes connexin (CX)26 protein, is the most frequent cause of hereditary deafness worldwide. We previously described an induction method to develop functional CX26 gap junction-forming cells from mouse-induced pluripotent stem cells (iPSCs) and generated in vitro models for GJB2-related deafness. However, functional CX26 gap junction-forming cells derived from human iPSCs or embryonic stem cells (ESCs) have not yet been reported. In this study, we generated human iPSC-derived functional CX26 gap junction-forming cells (iCX26GJCs), which have the characteristics of cochlear supporting cells. These iCX26GJCs had gap junction plaque-like formations at cell-cell borders and co-expressed several markers that are expressed in cochlear supporting cells. Furthermore, we generated iCX26GJCs derived from iPSCs from two patients with the most common GJB2 mutation in Asia, and these cells reproduced the pathology of GJB2-related deafness. These in vitro models may be useful for establishing optimal therapies and drug screening for various mutations in GJB2-related deafness.

Presence of anaerobic bacteria and symptoms supports diagnosis of odontogenic sinusitis.

PURPOSE: The present study aimed to investigate the diagnostic indicators of odontogenic sinusitis other than computed tomography (CT) findings and the history of dental treatment such as detected bacteria and symptoms. MATERIALS AND METHODS: We performed a retrospective analysis of 87 patients who underwent surgery for unilateral sinusitis between 2016 and 2020 (n = 87). Patients with cysts and fungal sinusitis were excluded from the study. We analyzed the relationship between the presence/absence of CT findings such as periapical lesions and oroantral fistulas; anaerobic bacteria; and symptoms in patients with unilateral sinusitis. RESULTS: There was a significant correlation between the detection of anaerobes and CT findings. Peptostreptococcus sp., Prevotella sp., Streptococcus anginosus group, and Fusobacterium sp. were the most commonly isolated species. Detection of these anaerobic bacteria supports the diagnosis of odontogenic sinusitis. Moreover, ODS often present with symptoms, such as a foul smell and facial pain. CONCLUSIONS: Our results suggest that not only the presence of CT findings such as PAL and OAF, but the detection of anaerobic bacteria, and the presence of any symptoms may aid in the diagnosis of ODS.

Clinical, polysomnographic, and cephalometric features of obstructive sleep apnea with AHI over 100.

PURPOSE: Obstructive sleep apnea (OSA) is induced by a sleep-related collapse of the upper airway in association with multiple factors. The severity of OSA is determined by the apnea-hypopnea index (AHI). Although obesity and sex differences are common factors in OSA, the level of the AHI varies to the same degree according to the age and sex and degree of obesity. However, only a few studies have evaluated AHI over 100/h, those reports did not describe why they set the AHI cutoff at 100/h. The purpose of this study was to elucidate the pathogenesis of "very" severe OSA, defined as having an AHI > 100/h. METHODS: AHI > 100/h was set as very severe OSA (VS-OSA) in this study. As controls, moderate to severe OSA patients, matched with VS-OSA for age, sex, and body mass index (BMI), were enrolled. The findings of polysomnography and cephalography between VS-OSA and controls were compared. RESULTS: Eleven patients in the VS-OSA group (mean AHI 110.2/h) and 22 patients in the control group (mean AHI 41.6/h) were compared (mean age 50.2 vs 50.6, male:female 5:6 vs 10:12, mean BMI 35.4 kg/m2 vs 34.5 kg/m2). There were no significant differences in the clinical characteristics. In the polysomnographic parameters, the VS-OSA group showed apnea predominance, the mean percutaneous oxygen saturation (SO2) was significantly lower in all sleep stages, and the minimum SO2 was significantly lower (49.0% vs 77.5%, p = 0.002). A similar apnea duration and rather shorter hypopnea duration were shown. The time of apnea-to-arousal was significantly earlier (- 0.1 s vs 0.9 s, p = 0.003). Lung-to-finger circulation time showed no differences. The cephalometric findings showed no significant differences. CONCLUSIONS: VS-OSA patients were more likely to have apnea predominance, desaturation when sleeping despite a similar apnea duration, and rather shorter hypopnea duration, and arousals were evoked significantly earlier.

Effects of endoscopic sinus surgery on nasal spray deposition using dye-based methods for humans and a human silicone sinonasal cavity model.

OBJECTIVE: We have evaluated that the deposition patterns of corticosteroid nasal spray in the sinonasal cavity of both post-operated human cases, which were further compared with a computed tomography-based sinonasal airway model. METHODS: Fifty-one patients with chronic rhinosinusitis following an endoscopic sinus surgery were enrolled in this study. Nasal spray mometasone furoate hydrate (Nasonex®) containing 0.1% indigocarmine was applied to the patients' nasal cavities and the sinonasal cavity was observed by endoscopy and video documentation. A single plaster sinonasal model was used to quantify the sinonasal deposition of nasal sprays containing 10% red ink solution using 12 round paper strips. RESULTS: The predominant areas of the spray deposition of the operated sinonasal cavities were recognized in the ethmoid sinus and the olfactory cleft in the human study. The droplets were mainly deposited in the inferior turbinate followed by the posterior part of the ethmoid sinus, the olfactory cleft, and anterior part of the ethmoid sinus in a sinonasal model. CONCLUSION: The corticosteroid nasal spray efficiently reached the olfactory cleft and the ethmoid sinus in post-operative conditions, which was demonstrated by post-operated human cases and a computed tomography-based sinonasal airway model.

A case of perilymphatic fistula caused by surgical drilling with preserved normal hearing.

We introduce our horrible experience of lateral semicircular canal exposure due to unintended drilling during left facial nerve decompression. Nearly half of the canal was drilled-out, however, the membranous labyrinth was preserved and the defect was covered with temporal fascia. Immediately after surgery, the patient complained of vertigo with right beating nystagmus. However, the patient could hear an audible tuning fork sound and the Weber-test showed left-sided deviation. The vertigo gradually subsided and the facial palsy was completely recovered 3 months after the surgery. One and half years later, the patient spent a normal life with normal hearing nevertheless after this terrifying episode.

Possible reversibility between epithelioid and sarcomatoid types of mesothelioma is independent of ERC/mesothelin expression.

BACKGROUND: Mesothelioma is histologically divided into three subgroups: epithelioid, sarcomatoid, and biphasic types. The epithelioid or sarcomatoid type is morphologically defined by polygonal or spindle-like forms of cells, respectively. The biphasic type consists of both components. It is not yet understood how histological differentiation of mesothelioma is regulated. ERC/mesothelin is expressed in most cases of the epithelioid type, but not in the sarcomatoid type of mesothelioma. Consequently, its expression is well correlated to the histological subtype. We hypothesized that ERC/mesothelin expression influences the histological differentiation of mesothelioma, and tested this hypothesis. METHODS: We performed studies using the overexpression or knockdown of ERC/mesothelin in mesothelioma cells to examine its effect on cellular morphology, growth kinetics, or migration/invasion activity, in vitro. We then transplanted ERC/mesothelin-overexpressing and control cells into the intraperitoneal space of mice. We examined the effect of ERC/mesothelin overexpression on mouse survival and tumor phenotype. RESULTS: In vitro cell culture manipulations of ERC/mesothelin expression did not affect cellular morphology or proliferation, although its overexpression enhanced cellular adhesion and the migration/invasion activity of mesothelioma cells. The survival rate of mice following intraperitoneal transplantation of ERC/mesothelin-overexpressing mesothelioma cells was significantly lower than that of mice with control cells. The histological evaluation of the tumors, however, did not show any morphological difference between two groups, and our hypothesis was not validated. Unexpectedly, both groups (ERC/mesothelin-overexpressing and control) of mesothelioma cells that were morphologically monophasic and spindle-like in vitro differentiated into a biphasic type consisting of polygonal and spindle-like components in the transplanted tumor, irrespective of ERC/mesothelin expression. CONCLUSIONS: These results suggested that the histological transition of mesothelioma between epithelioid and sarcomatoid types may be reversible and regulated not by ERC/mesothelin, but by other unknown mechanisms.

The seventh nationwide surveillance of six otorhinolaryngological infectious diseases and the antimicrobial susceptibility patterns of the isolated pathogens in Japan.

The Japanese Three Academic Societies Joint Antimicrobial Susceptibility Surveillance Committee conducted a nationwide surveillance on six otorhinolaryngological diseases and investigated the antimicrobial susceptibility patterns and isolation rates of the causative pathogens. The surveillance program was conducted in the otorhinolaryngological departments of 12 universities, and 36 affiliated hospitals and clinics. Patients with acute otitis media (children under six years old), chronic otitis media, acute nasal sinusitis, chronic nasal sinusitis, acute tonsillitis, and peritonsillar abscess (over 20 years old) between December 2015 and June 2017 were investigated. The collected swab or incision samples were cultivated for microbial identification, and the antimicrobial susceptibility of the detected bacteria was measured at the Kitasato University Research Center for Infections and Antimicrobials. The surveillance focused on three gram-positive bacteria (Streptococcus pneumoniae, Streptococcus pyogenes, and Staphylococcus aureus), three gram-negative bacteria (Haemophilus influenzae, Moraxella catarrhalis, and Pseudomonas aeruginosa), and three anaerobic bacteria (anaerobic gram-positive cocci, Prevotella spp., Porphyromonas spp., and Fusobacterium spp.). Bacterial susceptibility to 40 antimicrobial agents was investigated. We were unable to completely mitigate the rise in the occurrence of resistant bacteria, such as methicillin-resistant S. aureus, penicillin-resistant S. pneumoniae, penicillin-intermediate resistant S. pneumoniae, beta-lactamase non-producing ampicillin-resistant H. influenzae, and beta-lactamase producing ampicillin-resistant H. influenzae. We suggest promoting the proper usage of antimicrobial agents to prevent the spread of these bacteria. We also suggested that immunization with pneumococcal vaccines is useful for decreasing the occurrence of otorhinolaryngological infectious diseases caused by pneumococci.

Cryosurgical ablation for treatment of common warts on the nasal vestibule.

BACKGROUND: Common warts rarely occur on the nasal vestibule. It should be treated more carefully than other skin lesions because scar contracture results in the narrowing of the external naris, causing nasal obstruction and reducing the patient's quality of life. In this report, we introduce our method for common warts on the nasal vestibule using handheld cryoablation devise. PATIENTS AND OPERATIVE METHODS: We performed a series of four cryosurgical ablation procedures to treat common warts on the nasal vestibule. All the procedures employed a reusable handheld cryoablation device (CRY-AC®, Brymill Cryogenic Systems, Ellington, CT). RESULT: All patients were successfully treated for four to six sessions without any adverse event. CONCLUSION: Cryotherapy for common warts on nasal vestibular is easily performed in an office setting under no anesthesia without bleeding. We believe this easy and safe procedure is suitable as the first line therapy for common wart on nasal vestibular.

Efficacy of the additional effect of hyperbaric oxygen therapy in combination of systemic steroid and prostaglandin E1 for idiopathic sudden sensorineural hearing loss.

PURPOSE: The efficacies of hyperbaric oxygen therapy (HBO), systemic steroid, prostaglandin E1, or the combination of any two modalities have been reported in patients with idiopathic sudden sensorineural hearing loss (ISSNHL). However, little is known about the combined efficacy of HBO, systemic steroid, and prostaglandin E1 for this disorder. We aimed to investigate the efficacy of HBO combined with systemic steroids and prostaglandin E1 as triple therapy in patients with ISSNHL. MATERIALS AND METHODS: We retrospectively evaluated the records of 67 patients with ISSNHL who were treated with systemic steroid and prostaglandin E1, with (n = 38) or without (n = 29) HBO. The inclusion criteria included a diagnosis of ISSNHL within 14 days of symptom onset, age ≥15 years, treatment according to the protocol, and clinical follow-up of at least 1 month. The patients' hearing levels were evaluated 1 month after hearing loss onset. The primary outcome was hearing improvement on pure tone audiometry. We also evaluated the demographic profiles of patients. RESULTS: Patients treated with triple therapy showed significantly greater hearing improvement (p < 0.01) than those treated without HBO, despite some differences between the two treatment groups. Multivariate logistic regression analysis revealed a significant positive correlation between pure tone audiometry improvement and hyperbaric oxygen therapy, after adjustment for confounding factors (odds ratio = 7.42; 95% and confidence interval = 2.37-23.3; p = 0.001). CONCLUSION: HBO with systemic steroid and prostaglandin E1 administration conferred significant therapeutic benefits for ISSNHL. Therefore, routine use of triple therapy is recommended for patients with ISSNHL.

Perinatal Gjb2 gene transfer rescues hearing in a mouse model of hereditary deafness.

Hearing loss is the most widespread sensory disorder, with an incidence of congenital genetic deafness of 1 in 1600 children. For many ethnic populations, the most prevalent form of genetic deafness is caused by recessive mutations in the gene gap junction protein, beta 2, 26 kDa (GJB2), which is also known as connexin 26 (Cx26). Despite this knowledge, existing treatment strategies do not completely recover speech perception. Here we used a gene delivery system to rescue hearing in a mouse model of Gjb2 deletion. Mice lacking Cx26 are characterized by profound deafness from birth and improper development of cochlear cells. Cochlear delivery of Gjb2 using an adeno-associated virus significantly improved the auditory responses and development of the cochlear structure. Using gene replacement to restore hearing in a new mouse model of Gjb2-related deafness may lead to the development of therapies for human hereditary deafness.

Nationwide surveillance of 6 otorhinolaryngological infectious diseases and antimicrobial susceptibility pattern in the isolated pathogens in Japan.

The Japanese Three Academic Societies Joint Antimicrobial Susceptibility Surveillance Committee has conducted a nationwide surveillance on antimicrobial susceptibility patterns and rates of isolation in 6 otolaryngological diseases. The surveillance program was conducted in the otorhinolaryngological departments of 29 universities, and their 26 affiliated hospitals. Patients suffering from acute otitis media, chronic otitis media, acute nasal sinusitis, chronic nasal sinusitis, acute tonsillitis, and peritonsillar abscess between January 2011 and June 2012 were investigated. The collected swab or incision samples were cultivated for microbial identification, and the drug susceptibility of detected bacteria was measured at the Kitasato University Research Center for Infections and Antimicrobials. The surveillance focused on three gram-positive bacteria (Streptococcus pneumoniae, Streptococcus pyogenes, and Staphylococcus aureus), three gram-negative bacteria (Haemophilus influenzae, Moraxella Catarrhalis, and Pseudomonas aeruginosa), and three anaerobic bacteria (Peptostreptococcus spp., Prevotella spp., and Fusobacterium spp.). Bacterial susceptibility to 39 antimicrobial drugs was investigated. We compared bacterial isolation ratio of each disease in this surveillance from those of past 4 times surveillance which we performed formerly, and we also compared percentage of main drug resistant strains from those of past 4 times surveillance. The age composition between this time and former surveillances was not statistically significant by student-t test. We were unable to completely resolve the rise in resistant bacteria, such as methicillin-resistant S. aureus, penicillin-resistant S. pneumoniae, penicillin-intermediate resistant S. pneumoniae, beta-lactamase non-producing ampicillin-resistant H. influenzae, beta-lactamase producing ampicillin-resistant H. influenzae, and beta-lactamase producing amoxicillin clavulanic acid-resistant H. influenzae. We suggest promoting the proper usage of antimicrobial drugs in order to avoid the spread of these bacteria.

Comparative bactericidal activity of four fluoroquinolones against Pseudomonas aeruginosa isolated from chronic suppurative otitis media.

BACKGROUND: The aim of the present study was to evaluate the bactericidal activity of four new fluoroquinolones against current isolates of Pseudomonas aeruginosa from the patients with chronic suppurative otitis media (CSOM). METHODS: We examined bactericidal activity of four types of fluoroquinolones, garenoxacin (GRNX), levofloxacin (LVFX), ciprofloxacin (CPFX) and sitafloxacin (STFX) against current isolates of P. aeruginosa (50 strains). RESULTS: STFX exhibited the most potent activity of both MIC50 and MIC90, followed by CPFX, LVFX, and GRNX. The number of GRNX-resistant strains was significantly greater than those of LVFX, CPFX, and STFX (P < 0.05). CONCLUSION: STFX showed the most potent activity against P. aeruginosa for recent pathogens recovered from CSOM as compared with the others, suggesting that the clinical application of topical STFX would be useful to prevent the emergence of resistant mutants of P. aeruginosa.

Comparative analysis of cytokine release from epithelial cell cultures of the upper airway.

INTRODUCTION: Upper airway epithelial cells show a multi-potential ability to produce a variety of cytokines/chemokines in the steady-state and under external stimuli. OBJECTIVE: To compare various cytokines/chemokines released from primary cultures of human nasal epithelial cells (HNECs) derived from healthy controls and subjects with allergic rhinitis (AR), chronic rhinosinusitis with nasal polyps (CRSwNPs) in non- stimulated and IL-17A-stimulated conditions. METHODS: The supernatants derived from HNECs of healthy control, AR, CRSwNPs were used to measure 20 of cytokines/chemo- kines in the non-stimulated and IL-17A-stimulated conditions. RESULTS: AR and CRSwNPs showed significant up-regulation in the release of IL-6, IL-33, and thymic stromal lymphopoietin (TSLP), and the release of IL-6, TSLP, granulocyte macrophage colony-stimulating factor (GM-CSF), and tumor necrosis factor α (TNFα) in comparison with normal controls, respectively. Secretion of GM-CSF and TNFα were enhanced in patients with nasal polyps as compared with AR. Stimulation with IL-17A enhanced the secretion of IL-8 and granulocyte-colony stimulating factor (G-CSF) in the normal control, IL-6 and IL-8 in AR, and IL-6, TSLP, G-CSF, GM-CSF and TNFα in nasal polyps. CONCLUSION: Epithelial cells derived from AR and CRSwNPs showed up-regulation of secretion of several cytokines/chemokines both in the steady state and after IL-17A stimulation, which may contribute to the inflammatory responses of AR and CRSwNPs.

Spatiotemporal expression of TRPM4 in the mouse cochlea.

The present study was conducted to elucidate the presence of the transient receptor potential cation channel subfamily M member 4, TRPM4, in the mouse inner ear. TRPM4 immunoreactivity (IR) was found in the cell body of inner hair cells (IHCs) in the organ of Corti in the apical side of marginal cells of the stria vascularis, in the apical portion of the dark cells of the vestibule, and in a subset of the type II neurons in the spiral ganglion. Subsequently, changes in the distribution and expression of TRPM4 in the inner ear during embryonic and postnatal developments were also evaluated. Immunohistochemical localization demonstrated that the emergence of the TRPM4-IR in IHCs occurs shortly before the onset of hearing, whereas that in the marginal cells happens earlier, at the time of birth, coinciding with the onset of endolymph formation. Furthermore, semiquantitative real-time PCR assay showed that expressions of TRPM4 in the organ of Corti and in the stria vascularis increased dramatically at the onset of hearing. Because TRPM4 is a Ca(2+) -activated monovalent-selective cation channel, these findings imply that TRPM4 contributes to potassium ion transport, essential for the signal transduction in IHCs and the formation of endolymph by marginal cells.

Dominant negative connexin26 mutation R75W causing severe hearing loss influences normal programmed cell death in postnatal organ of Corti.

BACKGROUND: The greater epithelial ridge (GER) is a developmental structure in the maturation of the organ of Corti. Situated near the inner hair cells of neonatal mice, the GER undergoes a wave of apoptosis after postnatal day 8 (P8). We evaluated the GER from P8 to P12 in transgenic mice that carry the R75W + mutation, a dominant-negative mutation of human gap junction protein, beta 2, 26 kDa (GJB2) (also known as connexin 26 or CX26). Cx26 facilitate intercellular communication within the mammalian auditory organ. RESULTS: In both non-transgenic (non-Tg) and R75W + mice, some GER cells exhibited apoptotic characteristics at P8. In the GER of non-Tg mice, both the total number of cells and the number of apoptotic cells decreased from P8 to P12. In contrast, apoptotic cells were still clearly evident in the GER of R75W + mice at P12. In R75W + mice, therefore, apoptosis in the GER persisted until a later stage of cochlear development. In addition, the GER of R75W + mice exhibited morphological signs of retention, which may have resulted from diminished levels of apoptosis and/or promotion of cell proliferation during embryogenesis and early postnatal stages of development. CONCLUSIONS: Here we demonstrate that Cx26 dysfunction is associated with delayed apoptosis of GER cells and GER retention. This is the first demonstration that Cx26 may regulate cell proliferation and apoptosis during development of the cochlea.

Craniofacial anatomical risk factors in men with obstructive sleep apnea and heart failure: a pilot study.

PURPOSE: Obstructive sleep apnea (OSA) is complicated with heart failure (HF); however, the reason for this is not well understood. Craniofacial anatomic risk factors may contribute to OSA pathogenesis in HF patients. However, there are no data about cephalometric findings among OSA patients with HF. METHODS: Consecutive patients with HF and OSA (defined as total apnea-hypopnea index (AHI) ≥15/h) were enrolled. As controls, OSA patients without HF but matching the test group in age, BMI, and obstructive AHI were also enrolled. RESULTS: Overall, 17 OSA patients with HF and 34 OSA patients without HF were compared. There are no significant differences in the characteristics or polysomnographic parameters between 2 groups. In the cephalometric findings, compared with patients without HF, patients with HF showed a significantly greater angle between the line SN to point "A" (SNA) and a longer inferior airway space and greater airway area. However, the tongue area of patients with HF was more than those without HF. CONCLUSIONS: The craniofacial structures of OSA patients with HF were different from those without HF. OSA patients with HF had an upper airway anatomy that is more likely to collapse when sleeping while recumbent, despite having a larger airway space.