Oral Microbial Profile Analysis in Patients with Oral and Pharyngeal Cancer Reveals That Tumoral Fusobacterium nucleatum Promotes Oral Cancer Progression by Activating YAP.

The incidence of oral cancer has recently been increasing worldwide, particularly among young individuals and women. The primary risk factors for head and neck cancers, including oral and pharyngeal cancers, are smoking, alcohol consumption, poor oral hygiene, and repeated exposure to mechanical stimuli. However, approximately one-third of the patients with oral and pharyngeal cancers are neither smokers nor drinkers, which points to the existence of other mechanisms. Recently, human microbes have been linked to various diseases, including cancer. Oral pathogens, especially periodontal pathobionts, are reported to play a role in the development of colon and other types of cancer. In this study, we employed a series of bioinformatics analyses to pinpoint Fusobacterium nucleatum as the predominant oral bacterial species in oral and pharyngeal cancer tissue samples. We successfully isolated Fn. polymorphum from the saliva of patients with oral cancer and demonstrated that Fn. polymorphum indeed promoted oral squamous cell carcinoma development by activating YAP in a mouse tongue cancer model. Our research offers scientific evidence for the role of the oral microbiome in oral cancer progression and provides insights that would help in devising preventative strategies against oral cancer, potentially by altering oral bacterial profiles.

A Case of a Congenital Cholesteatoma Without Growth for a Long Term.

Congenital cholesteatoma is typically an expanding cystic mass of keratinizing squamous epithelium located medial to the intact tympanic membrane in patients with no prior history of perforation, otorrhea and ear. It is generally thought to be a progressive disease and is usually surgically removed upon detection as the first-choice treatment. As such, it is rare to be observed for a long term without progression. Here we report a rare case of congenital cholesteatoma that remained in an undetectable size and did not deteriorate mild hearing loss for 12 years. A seven years old boy was referred to us with right hearing impairment. Pure-tone audiometry found conductive hearing loss with an air-bone gap of 25 dB and a high-resolution computed tomography (CT) scan found the eroded long process of incus but did not detect any soft tissue density indicating congenital cholesteatoma. He initially did not wish to undergo surgery. His hearing level and image finding remained virtually unchanged during the next 12 years of the follow-up period. Twelve years later, endoscopic ear surgery was performed, which revealed a very small cholesteatoma mass, an eroded long process of the incus and ossicular chain discontinuities. We suspect that the cholesteatoma was originally larger, partially eroded the incus, then regressed to a very small size, and remained small for at least 12 years under our observation.

Risk factors for adverse events associated with endoscopic submucosal dissection for superficial pharyngeal cancer.

BACKGROUND: Superficial pharyngeal cancer can be treated with curative intent while preserving function using minimally invasive peroral endoscopic resection techniques such as endoscopic submucosal dissection (ESD). However, severe adverse events occasionally occur, such as laryngeal edema requiring temporary tracheotomy and fistula formation. Therefore, we investigated the risk factors for adverse events associated with ESD for superficial pharyngeal cancer. METHODS: This retrospective observational study was conducted at a single institution, and 63 patients who underwent ESD were enrolled. The primary outcome was the risk factors for adverse events associated with ESD. The secondary outcomes were adverse events associated with ESD and their frequency. RESULTS: The overall adverse event rate was 15.9% (10/63). The incidence of laryngeal edema requiring prophylactic temporary tracheotomy was 11.1%, while laryngeal edema requiring emergency temporary tracheotomy, postoperative bleeding, aspiration pneumonia, fistula, abscess, and stricture formation occurred in 1.6% of patients, respectively. Logistic regression analyses showed that a history of radiotherapy for head and neck cancer was a risk factor for adverse events (odds ratio [OR], 16.67; 95% confidence interval [CI], 3.04-91.34; p = 0.001). After adjusting the model for differences in the baseline risk factors using the inverse probability of treatment weighting method, the adverse events were found to increase in association with a history of radiotherapy for head and neck cancer (OR, 39.66; 95% CI,5.85-268.72; p < 0.001). CONCLUSION: History of radiotherapy for head and neck cancer is an independent risk factor for adverse events associated with ESD for superficial pharyngeal cancer. Among adverse events, laryngeal edema requiring prophylactic temporary tracheotomy was particularly high.

Sequestration of RBM10 in Nuclear Bodies: Targeting Sequences and Biological Significance.

RBM10 is an RNA-binding protein that regulates alternative splicing (AS). It localizes to the extra-nucleolar nucleoplasm and S1-1 nuclear bodies (NBs) in the nucleus. We investigated the biological significance of this localization in relation to its molecular function. Our analyses, employing deletion mutants, revealed that RBM10 possesses two S1-1 NB-targeting sequences (NBTSs), one in the KEKE motif region and another in the C2H2 Zn finger (ZnF). These NBTSs act synergistically to localize RBM10 to S1-1 NBs. The C2H2 ZnF not only acts as an NBTS, but is also essential for AS regulation by RBM10. Moreover, RBM10 does not participate in S1-1 NB formation, and without alterations of RBM10 protein levels, its NB-localization changes, increasing as cellular transcriptional activity declines, and vice versa. These results indicate that RBM10 is a transient component of S1-1 NBs and is sequestered in NBs via its NBTSs when cellular transcription decreases. We propose that the C2H2 ZnF exerts its NB-targeting activity when RBM10 is unbound by pre-mRNAs, and that NB-localization of RBM10 is a mechanism to control its AS activity in the nucleus.

Stress and Nasal Allergy: Corticotropin-Releasing Hormone Stimulates Mast Cell Degranulation and Proliferation in Human Nasal Mucosa.

Psychological stress exacerbates mast cell (MC)-dependent inflammation, including nasal allergy, but the underlying mechanisms are not thoroughly understood. Because the key stress-mediating neurohormone, corticotropin-releasing hormone (CRH), induces human skin MC degranulation, we hypothesized that CRH may be a key player in stress-aggravated nasal allergy. In the current study, we probed this hypothesis in human nasal mucosa MCs (hM-MCs) in situ using nasal polyp organ culture and tested whether CRH is required for murine M-MC activation by perceived stress in vivo. CRH stimulation significantly increased the number of hM-MCs, stimulated both their degranulation and proliferation ex vivo, and increased stem cell factor (SCF) expression in human nasal mucosa epithelium. CRH also sensitized hM-MCs to further CRH stimulation and promoted a pro-inflammatory hM-MC phenotype. The CRH-induced increase in hM-MCs was mitigated by co-administration of CRH receptor type 1 (CRH-R1)-specific antagonist antalarmin, CRH-R1 small interfering RNA (siRNA), or SCF-neutralizing antibody. In vivo, restraint stress significantly increased the number and degranulation of murine M-MCs compared with sham-stressed mice. This effect was mitigated by intranasal antalarmin. Our data suggest that CRH is a major activator of hM-MC in nasal mucosa, in part via promoting SCF production, and that CRH-R1 antagonists such as antalarmin are promising candidate therapeutics for nasal mucosa neuroinflammation induced by perceived stress.

Th2 cells and macrophages cooperatively induce allergic inflammation through histamine signaling.

Histamine, which is mainly produced by mast cells and basophils, participates in various allergic symptoms, and some studies have reported that macrophages also produce histamine. Moreover, recent studies have revealed that macrophages, especially alternatively activated macrophages (M2) induced by T helper 2 (Th2) cytokines, such as interleukin (IL)-4 and IL-13, participate in the pathogenesis of allergic diseases. The major source of Th2 cytokines is antigen-specific Th2 cells. To elucidate the relationship between histamine, macrophages, and Th2 cells in allergic inflammation, we established a macrophage-Th2 cell co-culture model in vitro and an antigen-specific Th2 cell transfer mouse model of rhinitis. In vitro analyses indicated that macrophages produce histamine by interacting with antigen-specific Th2 cells through the antigen. Furthermore, Th2 cells and macrophages cooperatively elicited rhinitis in the mouse model. We determined that histamine induces Th2- and macrophage-elicited sneezing responses through H1 receptor signaling, whereas it induces nasal eosinophil infiltrations through H4 receptor signaling. Collectively, these results indicate a novel histamine production mechanism by macrophages, in which Th2 cells and macrophages cooperatively induce nasal allergic inflammation through histamine signaling.

Decrease in number of mast cells in resected nasal polyps as an indicator for postoperative recurrence of chronic rhinosinusitis.

BACKGROUND: In the clinical setting, chronic rhinosinusitis with nasal polyps (CRSwNP) is usually divided into eosinophilic-CRS (ECRS) and non-ECRS (NECRS) in Japan. Patients with the former are believed to be at risk for postoperative recurrence of CRS. However, some patients have been missed according to these phenotypic classifications due to the low number of infiltrating eosinophils in polyp tissues. OBJECTIVE: In the present study, we attempted to identify cellular or molecular candidate markers to predict nasal polyp recurrence. METHODS: Nasal polyps were collected from 32 patients with CRSwNP who had undergone an endoscopic sinus surgery. These patients were divided into ECRS and NECRS groups in accordance with the Japanese Epidemiological Survey of Refractory Eosinophilic Chronic Rhinosinusitis (JESREC) scoring system and the number of eosinophils in polyp tissues. Unclassifiable patients were referred to as the unknown group. RESULTS: Eosinophil infiltration in resected nasal polyps was most evident in the ECRS group. However, the number of mast cells and tryptase-positive cells in nasal polyps were significantly lower in ECRS and unknown groups compared with the NECRS group. A significant positive correlation was detected between the JESREC score and number of eosinophils. The numbers of mast cells and tryptase-positive cells were negatively correlated with the JESREC score in all included samples. Significant positive correlations were detected between the number of transforming growth factor ß1-positive cells and the number of mast cells, tryptase-positive cells, and chymase-positive cells mast cells. CONCLUSIONS AND CLINICAL RELEVANCE: These findings indicated that the enumeration of mast cells in resected polyps may be another approach to predict postoperative polyp recurrence in CRSwNP patients.

Assessment of Meniere's disease from a radiological aspect - saccular otoconia as a cause of Meniere's disease?

CONCLUSION: Significant reduced visualization of the reuniting duct (ductus reuniens; RD), saccular duct (SD) and endolymphatic sinus (ES) in Meniere's disease (MD) compared with normal control ears on three-dimensional (3D) CT imaging suggests the blockage of endolymphatic flow there with radiodense substances, which may be explained by dislodged otoconia from the saccule. These structures could be involved in the pathogenesis of MD. OBJECTIVE: This study was designed to visualize and assess the RD, SD and ES in patients with MD using 3D CT. METHODS: Sixty-two patients with a definite diagnose of unilateral MD, based on criteria proposed by the Committee on Hearing and Equilibrium of the American Academy of Otolaryngology-Head and Neck Surgery (AAO-HNS), were compared with contralateral ears and normal controls (26 ears) using 3D CT. The RD, SD and ES were scrutinized for patency on 3D CT images. RESULTS: MD ears showed loss of continuity of the RD, SD and ES based on evaluation of 3D CT images, and differed significantly from normal healthy control ears (p < 0.01).

Loss of mammal-specific tectorial membrane component carcinoembryonic antigen cell adhesion molecule 16 (CEACAM16) leads to hearing impairment at low and high frequencies.

The vertebrate-restricted carcinoembryonic antigen gene family evolves extremely rapidly. Among their widely expressed members, the mammal-specific, secreted CEACAM16 is exceptionally well conserved and specifically expressed in the inner ear. To elucidate a potential auditory function, we inactivated murine Ceacam16 by homologous recombination. In young Ceacam16(-/-) mice the hearing threshold for frequencies below 10 kHz and above 22 kHz was raised. This hearing impairment progressed with age. A similar phenotype is observed in hearing-impaired members of Family 1070 with non-syndromic autosomal dominant hearing loss (DFNA4) who carry a missense mutation in CEACAM16. CEACAM16 was found in interdental and Deiters cells and was deposited in the tectorial membrane of the cochlea between postnatal days 12 and 15, when hearing starts in mice. In cochlear sections of Ceacam16(-/-) mice tectorial membranes were significantly more often stretched out as compared with wild-type mice where they were mostly contracted and detached from the outer hair cells. Homotypic cell sorting observed after ectopic cell surface expression of the carboxyl-terminal immunoglobulin variable-like N2 domain of CEACAM16 indicated that CEACAM16 can interact in trans. Furthermore, Western blot analyses of CEACAM16 under reducing and non-reducing conditions demonstrated oligomerization via unpaired cysteines. Taken together, CEACAM16 can probably form higher order structures with other tectorial membrane proteins such as α-tectorin and ß-tectorin and influences the physical properties of the tectorial membrane. Evolution of CEACAM16 might have been an important step for the specialization of the mammalian cochlea, allowing hearing over an extended frequency range.

Blockage of reuniting duct in Meniere's disease.

CONCLUSION: The bony saccular orifice to the bony groove of the ductus reuniens (reuniting duct) could not be visualized in the Meniere's ear with significantly greater frequency compared with normal subjects, which suggests that the reuniting duct is affected by radio-opaque substances in CT findings. OBJECTIVE: This study investigated a more specific, objective, and simpler strategy to diagnose Meniere's disease by assessing the reuniting duct. SUBJECTS AND METHODS: We examined the ears of 12 patients with definitely diagnosed unilateral Meniere's disease in stage 3 based on Meniere's disease criteria proposed by the Committee on Hearing and Equilibrium of the American Academy of Otolaryngology-Head and Neck Surgery (AAO-HNS), and the ears of 12 normal control subjects using three-dimensional (3D) cone beam CT. RESULTS: The bony groove of the reuniting duct between the saccule and cochlea was visualized in all control subjects. However, the bony saccular orifice to the bony groove could not be visualized in the lesional ear of Meniere's patients with significantly greater frequency compared with those of the contralateral non-lesional ears and control ears (p < 0.01). This orifice was not patent in 66.7% (8 of 12 ears) on the lesional side but all contralateral non-lesional ears of the patients and normal control ears were patent.

[A case of Churg-Strauss syndrome following corticosteroid withdrawal].

A 56-year-old man with bilateral hearing impairment who had taken betamethasone combined with dexchlorpheniramine maleate for 7 years to treat chronic sinusitis developed a dry cough after discontinuing this medication and was diagnosed with asthma, and after which he sensed impaired bilateral hearing. Based on the presence of numerous eosinophilic leukocytes in otorrehea, we made a diagnosis of eosinophilic otitis media, and he was prescribed predonisolone to control the asthma, but discontinued it on his own. He then developed fever, maniphalanx stiffness, testicular pain, and facial hyperesthesia, eruptions, and the lower-limb numbness. The detection of a positive serum reaction for MPO-ANCA and evaluated of eosinophilic leukocyte levels yielded a definitive diagnosis of CSS, for which the man was treated with predonisolone and cyclophosphamide. His symptoms were relieved, even though the onset of neutropenia, necessitated the discontinuation of cyclophosphamade administration.

Cartilage ossiculoplasty by lever method.

CONCLUSION: We invented a new ossicular chain reconstruction by lever methods (OCRLM) in tympanoplasty type III with the canal wall down technique and investigated the postoperative hearing results of the 24 patients who had undergone this surgery. Postoperative air-bone gaps (ABGs) followed up for 2.1-3.2 years gave satisfactory improvement audiometrically compared with former reports. OBJECTIVE: To present newly devised methods applied from the viewpoint of mechanical dynamics for ossicular chain reconstruction in tympanoplasty. MATERIALS AND METHODS: A total of 24 cholesteatoma patients who underwent tympanoplasty with mastoidectomy using the canal wall down technique and ossicular chain reconstruction with OCRLM between May 2003 and April 2004 were investigated. The postoperative ABG of a four-frequency (0.5, 1, 2, and 4 kHz) average of OCRLM followed up for 2 years or more was assessed. RESULTS: Patients who underwent OCRLM showed good postoperative hearing results. Average postoperative ABGs of less than 10, 15, 20, 25, and 30 dB were 4.2, 37.5, 70.8, 91.6, and 100%, respectively. A gap of >30 dB was absent and the mean postoperative ABG was 17.2dB. The OCRLM method was satisfactory to improve hearing and effectively reduce the ABG postoperatively over the period of observation.

Disregard of cholesteatoma in congenital aural stenosis.

Congenital aural stenosis (CAS) is one type of congenital aural atresia (CAA) and is assumed to be a relatively mild type. Although CAS may be associated with cholesteatoma in the external ear canal, little attention has been paid to this association. We present two cases of CAS with middle ear infection due to destructive cholesteatoma. Both had been followed for microtia by a plastic surgeon. These patients had already exhibited cholesteatoma on CT examination, although it had been missed before the emergence of ear symptoms, otorrhea, and otalgia. We present our cases with successive changes in CT findings over time and discuss the problems of CAS with cholesteatoma.

Effects of cigarettes and alcohol consumption in benign paroxysmal positioning vertigo.

CONCLUSION: Smoking was associated with a lower incidence of benign paroxysmal positioning vertigo (BPPV). A larger study is required to establish the role of smoking in BPPV. OBJECTIVE: To evaluate the effect of cigarette and alcohol consumption on BPPV. PATIENTS AND METHODS: One hundred and fifty-six patients with BPPV and 155 age- and sex-matched normal subjects were compared according to their cigarette and alcohol consumption. Patients with BPPV who had had a recurrence of the disease and those who had not were also compared as to their cigarette and alcohol consumption. The question of whether the length of time until recovery was influenced by cigarette or alcohol consumption was also investigated. RESULTS: Control subjects smoked significantly more often than BPPV patients, and patients without recurrence more frequently than patients with recurrence. Alcohol consumption was also more common in control subjects than in BPPV patients, but there was no difference between patients with recurrence and without recurrence. There was a tendency for smoking patients to recover sooner than non-smoking patients. Alcohol consumption did not affect the length of time until recovery.

Fibrous dysplasia of the temporal bone with cholesteatoma.

Fibrous dysplasia is an uncommon benign bone disorder of unknown origin. It rarely affects the temporal bone. We report a case of fibrous dysplasia of the temporal bone with cholesteatoma in the external auditory canal, the first complaint of which was left otorrhea. Fibrous dysplasia invaded the external ear canal, stenosis of which causes a cholesteatoma. Mastoidectomy, tympanoplasty, and meatoplasty were performed. We discuss the characteristics of this condition, its differential diagnosis and its treatment.

Concurrent chemoradiotherapy with pirarubicin and 5-fluorouracil for resectable oral and maxillary carcinoma.

OBJECTIVE: We present the response rate and adverse effects of our regimen of concurrent chemoradiotherapy with pirarubicin (THP) and 5-fluorouracil (5-FU) for oral and maxillary carcinoma. PATIENTS AND METHODS: Fifteen patients with oral (10 cases) or maxillary (5 cases) squamous cell carcinoma who underwent our concurrent chemoradiotherapy with the combination of intraarterial pirarubicin, intravenous continuous 5-fluorouracil, and radiation between March 2001 and February 2003 in our department were entered in this study. THP (5 mg/day) was infused into the lingual or maxillary artery one hour before radiation on days 1-5 and 8-12, while intravenous 5-FU (150 mg/m2/day) was instilled continuously on days 1-5, 8-12, 15-19, and 22-26 in accordance with the radiation schedule (2 Gy/day). Consequently, total doses of THP, 5-FU, and radiation were 50 mg, 3000 mg/m2 and 40 Gy, respectively. After the treatment series, response rate and adverse effects were evaluated. RESULTS: Response rate achieved 100% (12 cases exhibited a complete response and the remaining 3 a partial response). Notably, all 10 patients with oral carcinoma exhibited complete response. The main adverse effects were leucopenia (6/15) and mucositis (6/15), both of which were acceptable. CONCLUSIONS: This concurrent chemoradiotherapy is very useful for oral and maxillary carcinoma as a preoperative modality with remarkably high response rate and acceptable adverse events.

A possible case of Werner syndrome presenting with multiple cancers.

The treatment of a man with six metachronous primary cancers is described. The primary lesions were in the soft palate, both edges of the tongue, the hard palate, the esophagus, and the right ureter. Pathologically, all of the first five tumors in the head and neck and esophagus were proven to be squamous cell carcinoma with various grades of differentiation, and the last one was transitional cell carcinoma. The cancers were found in the early clinical stage, and were completely controlled one by one except for the ureteral tumor under treatment. His characteristic medical history and physical findings, i.e. bilateral cataracts, short stature, baldness, diabetes mellitus, high-pitched voice, and multiple malignancies, met the clinical criteria for possible Werner syndrome, a genetic premature aging disorder, though the possibility of phenocopy of this syndrome has not been ruled out. We have followed him carefully because he might be vulnerable to malignant tumor formation.

A case of fatal respiratory failure after surgery for advanced supraglottic laryngeal carcinoma.

Pulmonary lymphangitic carcinomatosis (PLC) is an uncommon type of pulmonary metastasis from laryngeal carcinoma. We encountered a case of fatal respiratory failure after surgery for supraglottic laryngeal carcinoma with multiple cervical lymph node metastases. On day 25 postoperatively, the patient suddenly developed fever and noted slowly progressive difficulty in breathing. Laboratory data, radiological findings, and the clinical course of this patient were strongly suggestive of PLC. The patient refused fiberoptic bronchoscopy and glucocorticoid therapy, leading to rapid deterioration due to increasing respiratory distress, and died of respiratory failure on postoperative day 32. PLC should be considered a possible presentation of pulmonary metastasis associated with head and neck carcinoma. Chest CT may be needed to detect subclinical PLC, especially in cases of advanced head and neck carcinomas.

[Metastatic carcinoma of the neck from unknown primary sites].

Encountering a metastatic carcinoma of the neck from an unknown primary site is not unusual, despite intensive examinations of the entire body. In previous reports, the pathological diagnosis of these carcinomas was usually squamous cell carcinomas and rarely adenocarcinoma. We treated eight patients with metastatic carcinoma of the neck from unknown primary sites, including 4 cases of squamous cell carcinoma, 2 cases of adenocarcinoma, one case of small cell carcinoma, and one case of clear cell carcinoma, during a 10-year period from January 1992 to December 2001. We clinically examined these eight cases, and focusing on the two cases of metastatic cervical adenocarcinoma from unknown primary sites. The 8 cases consisted of 5 cases of N2 and 3 cases of N3 disease. Three of the 5 N2 patients underwent a neck dissection, but all three of the N3 cases were judged to be inoperable. Disease-free survival was achieved in all 3 patients who underwent surgery. In this paper, we review 36 reports on metastatic carcinomas of the neck from unknown primary sites and statistically analyze 1454 cases. Pathologically, the majority of them (81.1%) were squamous cell carcinoma; adenocarcinomas accounted for only 7.6% of the cases. Notably, 65.0% of the patients with cervical metastatic adenocarcinomas were confirmed to have primary lesions outside the head and neck region. The prognosis of primary unknown metastatic cervical adenocarcinoma is reportedly poor, and the optimal treatment is still unclear, although surgery is recommended for primary unknown metastatic cervical squamous cell carcinoma. However, we suggest that intensive treatment, including surgery, radiotherapy, and chemotherapy, of metastatic lesions of the neck may play a key role in improving patient prognosis.