Yuichi Nakano

Immunohistochemical analysis of the olfactory mucosa by use of antibodies to brain proteins and cytokeratin.

The present study deals with the immunohistochemical detection of four brain-derived proteins and cytokeratin in the normal olfactory mucosa of humans and guinea pigs. Neurofilament protein immunoreactivity was found in the olfactory vesicles, dendrites, and perikaryon of receptor cells, and in thick nerve bundles located deep in the lamina propria. The antiserum to neuron-specific enolase (NSE) selectively stained olfactory receptor cells throughout the length of the bundles. The NSE immunoreactivity also was recognized in nerve bundles of various sizes throughout the lamina propria. Glia-specific S-100 protein immunoreactivity was present in Bowmans glands as well as in all nerve bundles in the lamina propria, but not in any cellular elements constituting the olfactory epithelium. Immunoreactivity for spot-35 protein, which was considered to be specific for cerebellar Purkinje cells, was found in flasklike cells (microvillar cells) occurring near the free surface of the epithelium. The basal cells in the olfactory and respiratory epithelium were stained positively with a cytokeratin antiserum.

Neuron-specific enolase, neurofilament protein and S-100 protein in the olfactory mucosa of human fetuses

SummaryImmunohistochemical examination for neuronspecific enolase (NSE), neurofilament protein (NFP), and S-100 protein was performed in the olfactory mucosa of human fetuses. NSE and NFP immunoreactivities were found in the olfactory receptor cells, while no S-100 immunoreactive cells were recognized within the olfactory epithelium. The anti-NSE serum stained various types of nerve bundles in the lamina propria mucosae; a population of the NSE-positive nerve bundles was also immunoreactive for NFP. The anti-S-100 serum clearly demonstrated Schwann cells associated with the nerve fibers in the lamina propria mucosae. These findings 1) suggest a possibility of NSE and NFP as new marker substances for olfactory cells and 2) indicate that immunohistochemistry is a useful tool to analyse the cellular components of the olfactory organs in normal and pathological conditions.

The effect of chronic middle ear inflammation on the pneumatization of the tympanic bulla in pigs.

The effect of chronic middle ear inflammation on the pneumatization of the tympanic bulla was investigated in piglets. The pig tympanic bulla has an air cell system which is divided by trabeculae and closely resembles the human mastoid air cell system. The tympanic bulla and its air cell system in normal ears were well developed because of the bone formation and the bone resorption inside the cortex, whereas the tympanic bulla affected by chronic otitis media in the early stages of life exhibited retardation of pneumatization arising from the disturbed bone resorption by inflammatory stimulus. It was concluded that affliction with chronic middle ear inflammation in the early stages of life causes inhibition of pneumatization by hindering the development of the air cell system.

Distribution of calcium binding proteins in sensory organs of the ear, nose and throat

Distributions of spot-35 protein (S-35), calbindin (CaB), and parvalbumin (PaV), three types of calcium-binding protein, were examined immunohistochemically in sensory organs of the ear, nose and throat in guinea pigs and rats. Immunoreactivity of S-35 and CaB was found in the outer hair cells and in vestibular sensory cells situated at the top of the ampulla, and in some cells in the macula. Microvillar cells in the olfactory epithelium, periglomerular cells, and small numbers of cells in the mitral cell layer in the olfactory bulb reacted to anti-S-35 and anti-CaB antisera. In taste buds, most gustatory receptor cells reacted to anti-CaB, although a few reacted to anti-S-35 antiserum. Neuron-like cells in the upper respiratory tract reacted similarly to these antisera. No PaV-immunoreactivity was found in any region. These results indicate that S-35 and CaB play important roles in the special kinds of mechanoreceptor and chemoreceptor cells found in the otolaryngeal area.

Pneumatization of the Tympanic Bulla after Blockage of the Ventilation Route through the Eustachian Tube in the Pig

To examine whether gas exchange occurs in the middle ear air cell system independent of the eustachian tube (ET), we occluded the middle ear clefts of piglets, whose tympanic bullae closely resemble the human mastoid air cell system, and investigated subsequent changes in the air cell system. We anticipated that pneumatization and development of the air cell system would continue if gas were exchanged through the mucosa lining the air cell system. If, on the other hand, mastoid air cells depend on the ET, mastoid development would be impaired or arrested. In noninflamed ears, pneumatization was maintained, and development of the air cell system continued after the middle ear cleft had been occluded with an acrylate adhesive and the bullar air cell system was thus excluded from any communication with the ET. These results indicate that the normal middle ear air cell system has the capacity to perform gas exchange independently of the ET, with gas exchange appearing to occur through the submucosal capillary network.

Course of IL-1β, IL-6, IL-8, and TNF-α in the Middle Ear Fluid of the Guinea Pig Otitis Media Model Induced by Nonviable Haemophilus Influenzae

To characterize the local response in acute otitis media, courses of interleukin (IL)–1β, IL-6, IL-8, and tumor necrosis factor (TNF) α in middle ear fluid (MEF) of the guinea pig otitis media model induced by nonviable Haemophilus influenzae were investigated with enzyme-linked immunosorbent assay (ELISA) kits. The IL-1β concentration in H influenzae–inoculated ears peaked 24 hours after inoculation. The IL-8 concentration was significantly higher in H influenzae–inoculated ears than in controls 48 and 96 hours after inoculation. The TNF-α concentration in H influenzae–inoculated ears had an initial peak 6 hours after inoculation and had significant late increases 48 and 96 hours after inoculation. The results suggest that IL-1β and TNF-α were produced by middle ear mucosa in the early stage of the experiment by stimulation of bacterial inoculation, which caused subsequent inflammatory cell accumulation, and that IL-8 and TNF-α were produced in the late stage by accumulating inflammatory cells.

Prognosis of Otitis Media with Effusion in Children, and Size of the Mastoid Air Cell System

We studied 334 ears in 176 patients who were chosen from children diagnosed as having otitis media with effusion (OME) at our clinic. These 176 patients were re-examined about 5 years and 10 months after the first visit and categorized into three groups according to otological findings at the time of review: group A (normal), group B (sequelae), group C (OME). For this study, 168 ears of 86 persons were examined by roentgenography both at the time of the first visit and at review, and the size of mastoid pneumatization and differences in size between the three groups were compared. The results of the investigation can be summarized as follows. The size of mastoid pneumatization in group A was large at the time of the first visit and the increase in size was also greater than in the other two groups. These results led us to conclude that the degree of mastoid pneumatization is an important prognostic sign of the clinical course of OME and provides important information about treatment.

Radiological findings of adenolymphoma (Warthin's tumor)

The radiological findings of adenolymphomas (Warthins tumor) treated in six hospitals between 1985 and 1994 were compared with the operative and histological findings and the usefulness of the radiological examinations was evaluated. The total number of patients was 72. The mean age was 61.8 years; 61 were males and 11 were females. All tumors developed in the parotid gland. Tc-99m-pertechnetate salivary gland scanning was performed in 13 patients and an increased uptake of the isotope was observed in only six patients. Even if Tc-99m-pertechnetate salivary gland scanning does not reveal intense accumulation, this tumor should not be ruled out. By computed tomography (CT), ultrasonography (US), and magnetic resonance imaging (MRI), the margin of all tumors was evident; however, the contents of the tumor varied. The contents and multiplicity of the tumors were well demonstrated by MRI, which was found to be the most accurate imaging modality.

Immunohistochemical Study of the Fourth Cell Type in the Olfactory Epithelium in Guinea Pigs and in a Patient

The mammalian olfactory epithelium consists of supporting cells, olfactory receptor cells, basal cells and a fourth cell type, which has recently been discovered. In this study we examine this fourth cell type using immunohistochemical techniques. Anti-Purkinje-specific spot-35 protein (S-35), anti-S-100 protein (S-100), anti-neuron-specific enolase (NSE), antichromogranin A and antisynaptophysin antisera were used for the immunostaining. The fourth cell type immunoreacted only to anti-S-35 antiserum and did not react to other antisera in guinea pigs. On the other hand, in the human S-100 immunoreactivity was seen in the fourth cell type as well as weak reactivity to S-35. NSE immunoreactivity was found only in the olfactory receptor cells of guinea pigs and the human. From these results it is assumed that this fourth cell type is a second chemoreceptor different from the olfactory receptor cells because S-35 and S-100 are Ca(++)-binding proteins.

Suppressed Mastoid Pneumatization in Cholesteatoma

Mastoid pneumatization size was studied by X-ray in 289 patients with otitis media (134 with cholesteatoma and 155 with chronic suppurative otitis media (COM) and 73 patients with traumatic tympanic membrane perforation (controls). The results demonstrated that mastoid pneumatization in the diseased ear of cholesteatoma patients was greatly suppressed. In these patients, the contralateral ear also showed significant suppression compared with controls; mastoid pneumatization size in the healthy ear contralateral to cholesteatoma was similar to that in patients cured of otitis media with effusion (OME). Hence, we conclude that cholesteatoma is a sequela of OME in childhood.