Innate immune responses and neuroepithelial degeneration and regeneration in the mouse olfactory mucosa induced by intranasal administration of Poly(I:C).

The pathogenesis of postviral olfactory disorder (PVOD) has not been fully elucidated. We investigated morphological changes and innate immune responses in the mouse olfactory mucosa induced by intranasal administration of polyinosinic-polycytidylic acid [Poly(I:C)], a synthetic analog of viral double-stranded RNA. Mice received three administrations of saline with or without Poly(I:C), once every 24 h. The olfactory mucosa was harvested at various intervals after the first administration (8 h, 3, 9 and 24 days). In the Poly(I:C) group, the number of apoptotic cells in the olfactory neuroepithelium had increased at 8 h. At 9 days, the olfactory neuroepithelium had severely degenerated and behavioral tests demonstrated that the mice showed signs of olfactory deterioration. At 24 days, the structure of the neuroepithelium had regenerated almost completely. Regarding the innate immune responses, many neutrophils had infiltrated the olfactory neuroepithelium at 8 h and had exuded into the nasal cavity by 3 days. Macrophages had also infiltrated the olfactory neuroepithelium at 8 h although to a lesser extent, but they still remained in the neuroepithelium at 24 days. Poly(I:C)-induced neuroepithelial damage was significantly inhibited by a neutrophil elastase inhibitor and was suppressed in neutropenic model mice. These findings suggest that the secondary damage caused by the neutrophil-mediated innate immune response plays an important role in the pathogenesis of PVOD.

Non-invasive objective evaluation of radiotherapy-induced dry mouth.

BACKGROUND: Dry mouth is a common complaint in patients undergoing radiotherapy. Here, we employed the oral moisture meter Mucus III to evaluate dry mouth in head and neck tumor patients before and after they underwent radiotherapy. METHODS: We recruited 17 newly diagnosed patients with pharyngeal squamous cell carcinoma or unknown primary squamous cell carcinoma, who received head and neck radiation therapy at Tokyo University Hospital in 2008-2010. The primary sites were the epipharynx (n = 1), oropharynx (n = 6), or hypopharynx (n = 5); it was unknown in five cases. Salivary function was assessed by a dry mouth questionnaire, resting saliva test, chewing gum test, and Mucus III, before (n = 17), immediately after radiotherapy (n = 10), and at 3 (n = 9) and 12 months after radiotherapy (n = 11). RESULTS: The questionnaire, resting saliva test, and chewing gum test at 3 and 12 months after radiotherapy indicated a significantly decreased resting and stimulated whole saliva flow rate than prior radiotherapy (P < 0.05 and P < 0.001). In contrast, Mucus III results showed significant worsening of xerostomia at 12 months after radiotherapy (P < 0.05). CONCLUSION: Mucus III has been proven to be an objective diagnostic tool for patients with serious dry mouth, such as in patients with Sjogren's syndrome. However, we did not find a perfect correlation between Mucus III and other objective (resting saliva and chewing gum) and subjective (questionnaire) measures of dry mouth. To precisely diagnose radiotherapy-induced dry mouth, further improvement to the method is needed.

[Blood flow measurement and clinical usefulness of the temporal fascial scar tissue flap and the periosteal scar tissue flap in posterior canal wall reconstructed tympanoplasty for the mastoid cavity problem in the postoperative ear].

In the postoperative ear, following reconstructive tympanoplasty for a mastoid cavity problem, a very important key is to maintain a stable reconstructed posterior canal wall with the bone plate and cartilage in the posterior canal wall. The authors manage reconstruction of the posterior canal wall with the temporal fascial scar tissue flap (TFSF) and the temporal periosteal scar tissue flap (TPSF) to ensure obtaining a stable posterior canal wall and a tympanic membrane graft. Well-vascularized TFSF and TPSF enable us to acquire a solid reconstructed posterior canal wall because of the secure blood supplies to the flaps. In order to investigate the blood supplies of TFSF and TPSF, we employed laser Doppler blood flowmeters and measured blood flow in the flaps in 20 cases of posyoperative ears treated for a mastoid cavity problem. The blood supplies to both flaps were good, with the blood supply to the TFSF being statistically better than in the case of the TPSF. These findings suggested that the TFSF and TPSF were a reliable source of local well-vascularized tissue which were pliable and could facilitate the creation of a stable posterior canal wall. Furthermore it seems the good blood supply was linked to the prompt postoperative healing, the avoidance of postoperative infection, and good hearing improvement postoperatively.

Nanogel-based PspA intranasal vaccine prevents invasive disease and nasal colonization by Streptococcus pneumoniae.

To establish a safer and more effective vaccine against pneumococcal respiratory infections, current knowledge regarding the antigens common among pneumococcal strains and improvements to the system for delivering these antigens across the mucosal barrier must be integrated. We developed a pneumococcal vaccine that combines the advantages of pneumococcal surface protein A (PspA) with a nontoxic intranasal vaccine delivery system based on a nanometer-sized hydrogel (nanogel) consisting of a cationic cholesteryl group-bearing pullulan (cCHP). The efficacy of the nanogel-based PspA nasal vaccine (cCHP-PspA) was tested in murine pneumococcal airway infection models. Intranasal vaccination with cCHP-PspA provided protective immunity against lethal challenge with Streptococcus pneumoniae Xen10, reduced colonization and invasion by bacteria in the upper and lower respiratory tracts, and induced systemic and nasal mucosal Th17 responses, high levels of PspA-specific serum immunoglobulin G (IgG), and nasal and bronchial IgA antibody responses. Moreover, there was no sign of PspA delivery by nanogel to either the olfactory bulbs or the central nervous system after intranasal administration. These results demonstrate the effectiveness and safety of the nanogel-based PspA nasal vaccine system as a universal mucosal vaccine against pneumococcal respiratory infection.

Trial of Chinese medicine Wu-Ling-San for acute low-tone hearing loss.

PURPOSE: We used new criteria to elucidate the demographics of acute low-tone sensorineural hearing loss (ALHL) and tested the Chinese medicine Wu-Ling-San as a treatment for ALHL. PROCEDURES: We reviewed the medical records of patients with ALHL seen at the outpatient clinic of the Social Insurance Central General Hospital in Tokyo from April 2006 through August 2011. Patients were treated with an oral steroid, a diuretic, or Wu-Ling-San; alone or in combination. RESULTS: We identified 130 definite and 48 probable ALHL cases. The mean age and male-to-female ratio in probable cases were significantly higher than those in definite cases (p < 0.05). The steroid-Wu-Ling-San combination was significantly more effective (100% recovery) than the diuretic alone (59%), Wu-Ling-San alone (62%), or the steroid-diuretic combination (60%, p < 0.05). CONCLUSIONS: ALHL can develop in older patients more frequently than we expected. The steroid-Wu-Ling-San combination is a possible new treatment for ALHL.

Recurrent hoarseness due to inflammatory vocal fold lesions in a patient with Crohn's disease.

Crohn's disease is a chronic inflammatory bowel disease characterized by discontinuous chronic inflammation that may affect virtually all organs, including the head and neck. Laryngeal involvement in Crohn's disease is very rare, and only 9 cases have been reported. All 9 patients complained of difficulty in breathing due to edema and ulceration from the larynx to the hypopharynx. The present patient was a 31-year-old woman who had experienced the intestinal symptoms of Crohn's disease starting 20 months earlier and complained of hoarseness, sore throat, and odynophagia. The hoarseness worsened gradually because of limited ulceration of the vocal fold without edema. We describe the first case in which limited ulceration occurred on the vocal fold without airway involvement.

Objective and non-invasive evaluation of dry mouth.

OBJECTIVE: This study was designed to evaluate the moisture checker (MucusIII), a new device for measuring moisture of the oral submucosa. METHODS: Defective salivary secretion was induced by sialoadenectomy (n=8), while the remaining five guinea pigs underwent sham surgery (control group). We measured the unstimulated salivary flow rate, wetness of the oral mucosa using the oral tester (L-SALIVO) and moisture of the submucosa of the tongue using MucusIII. All tests were performed before (baseline) and 1 month after surgery. RESULTS: Sialoadenectomy significantly reduced both the salivary flow rate and the tester reading at 10 and at 30s (p<0.01). The vale due to the MucusIII as significantly reduced in animals with sialoadenectomy (p<0.01). CONCLUSION: The MucusIII is a device for objective evaluation of the moisture of the oral cavity.

Identification and Analysis of Natural Killer Cells in Murine Nasal Passages.

BACKGROUND: Natural killer (NK) cells in the upper respiratory airways are not well characterized. In the current study, we sought to characterize and functionally assess murine nasal NK cells. METHODS: Using immunohistochemistry and flow cytometry, we compared the nasal NK cells of Ncr1GFP/+ knock-in mice, whose NK cells produced green fluorescent protein, with their splenic and pulmonary counterparts. In addition, we functionally analyzed the nasal NK cells of these mice in vitro. To assess the in vivo functions of nasal NK cells, C57BL/6 mice depleted of NK cells after treatment with PK136 antibody were nasally infected with influenza virus PR8. RESULTS: Immunohistochemical analysis confirmed the presence of NK cells in the lamina propria of nasal mucosa, and flow cytometry showed that these cells were of NK cell lineage. The expression patterns of Ly49 receptor, CD11b/CD27, CD62L and CD69 revealed that nasal NK cells had an immature and activated phenotype compared with that of their splenic and pulmonary counterparts. Effector functions including degranulation and IFN(interferon)-γ production after in vitro stimulation with phorbol 12-myristate-13-acetate plus ionomycin or IL(interleukin)-12 plus IL-18 were dampened in nasal NK cells, and the depletion of NK cells led to an increased influenza virus titer in nasal passages. CONCLUSIONS: The NK cells of the murine nasal passage belong to the conventional NK cell linage and characteristically demonstrate an immature and activated phenotype. Despite their hyporesponsiveness in vitro, nasal NK cells play important roles in the host defense against nasal influenza virus infection.

Nasal Administration of Cholera Toxin as a Mucosal Adjuvant Damages the Olfactory System in Mice.

Cholera toxin (CT) induces severe diarrhea in humans but acts as an adjuvant to enhance immune responses to vaccines when administered orally. Nasally administered CT also acts as an adjuvant, but CT and CT derivatives, including the B subunit of CT (CTB), are taken up from the olfactory epithelium and transported to the olfactory bulbs and therefore may be toxic to the central nervous system. To assess the toxicity, we investigated whether nasally administered CT or CT derivatives impair the olfactory system. In mice, nasal administration of CT, but not CTB or a non-toxic CT derivative, reduced the expression of olfactory marker protein (OMP) in the olfactory epithelium and olfactory bulbs and impaired odor responses, as determined with behavioral tests and optical imaging. Thus, nasally administered CT, like orally administered CT, is toxic and damages the olfactory system in mice. However, CTB and a non-toxic CT derivative, do not damage the olfactory system. The optical imaging we used here will be useful for assessing the safety of nasal vaccines and adjuvants during their development for human use and CT can be used as a positive control in this test.

Effect of edaravone on acute brainstem-cerebellar infarction with vertigo and sudden hearing loss.

We report 2 cases with acute brainstem and brainstem-cerebellar infarction showed improvement of their signs and symptoms after administration of edaravone. Case 1, a 74-year-old woman who experienced sudden vertigo, also had dysarthria and left hemiplegia. Magnetic resonance imaging (MRI) showed an abnormal region in the right ventrolateral medulla oblongata. The patient's vertigo and hemiplegia improved completely after treatment. Case 2, a 50-year-old man who experienced sudden vertigo and sensorineural hearing loss (SNHL), developed dysarthria after admission. MRI revealed acute infarction in the right cerebellar hemisphere. Magnetic resonance angiography revealed dissection of the basilar artery and occlusion of the right anterior inferior cerebellar artery. The patient's vertigo and hearing remarkably improved. We have described 2 patients whose early symptoms were vertigo and sudden SNHL, but who were later shown to have ischemic lesions of the central nervous system. Edaravone is neuroprotective drug with free radical-scavenging actions. Free radicals in the ear are responsible for ischemic damage. Edaravone, a free radical scavenger, may be useful in the treatment of vertigo and SNHL.

Craniofacial mucosal immune system: importance of its unique organogenesis and function in the development of a mucosal vaccine.

Mucosa-associated lymphoid tissues (MALT) play a critical role as inductive sites for the initiation of antigen-specific protective immunity against pathogens penetrating the mucus membranes. Nasopharynx-associated lymphoid tissue (NALT), situated at the bottom of the rodent nasal cavity, is thought to be an important site for the induction of antigen-specific immune response to inhaled antigens. In addition, we have recently shown that tear duct-associated lymphoid tissue (TALT), present in the murine tear duct bridging the ocular and nasal cavities, is involved in the induction and regulation of both nasal and ocular immunity. Interestingly, cellular requirements for the organogenesis of NALT and TALT are quite different from those of other MALT (e.g. Peyer's patches; PPs) and peripheral lymphoid tissues. Moreover, mucosal imprinting molecules of NALT and TALT inducer cells are totally independent of currently known chemokines and adhesion molecules in PPs and lymph nodes, such as the CXCR5-CXCL13, α4ß1 integrin-vascular cell adhesion molecule-1 (VCAM1), and CCR9-CCL25 axes. NALT and TALT lymphocytes are also independent of these tissue-specific migration molecules. Together with already-characterized conjunctiva-associated lymphoid tissue (CALT ), which has been demonstrated to play a critical role in ocular defense, the MALT associated with the head region seems to be coordinately organizing the unique craniofacial mucosal immune system of the ocular, nasal, oral-pharynx mucus membranes. Clarification of the immunological network of this unique craniofacial immune system will facilitate the development of a safe and effective mucosal vaccine against respiratory and ocular infections.

Benign mass lesions deep inside the temporal bone: imaging diagnosis for proper management.

CONCLUSION: Among mass lesions inside the temporal bone, benign tumors and cholesteatomas can be differentiated by contrast enhancement in T1-weighted images (T1WI) and by diffusion-weighted images (DWI). Moreover, DWI will also facilitate discrimination between cholesteatomas accompanied by granulation and other non-neoplastic lesions such as mucoceles and cholesterol granulomas. OBJECTIVES: To review the imaging characteristics of mass lesions inside the temporal bone and to investigate pertinent imaging modalities for differential diagnosis, which is crucial for appropriate treatment planning. PATIENTS AND METHODS: This was a retrospective case series study of six patients seen between 2002 and 2005 with mass lesions deep inside the temporal bone. RESULTS: One patient had facial schwannoma, two had glomus jugulare tumor, and three had cholesteatoma. Plain high resolution CT gave few clues to the nature of the mass lesions. MRI study provided us with better clues: contrast enhancement on T1WI was observed only in benign tumors and only cholesteatomas showed high intensity on DWI. With the assistance of neurosurgeons, surgery was successfully performed in all cases.