Tatsuya Fujiyoshi

Antibodies against a 30 kilodalton cochlear protein and type II and IX collagens in the serum of patients with inner ear diseases.

Collagen molecules are major extracellular matrix proteins involved in the development and support of delicate auditory sensory organs. Type II collagen is widely distributed within inner ear tissues, while type IX is found only within the labyrinthine membrane and dense fibers of the tectorial membrane. Antibody specific for type II collagen has been shown to be elevated in some patients with hearing loss due to several presumably autoimmune illnesses (including Menieres disease, otosclerosis, chronic progressive sensorineural hearing loss, and relapsing polychondritis). Purified human type II and LX collagens and an extract of human cochlear tissue were subjected to isolation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to nitrocellulose. The sera of 21 patients with inner ear disease were examined for the presence of anticollagen and anticochlear antibodies; the sera were used to probe Western blots of purified human collagens U, LX, and XI, and cochlear protein extract with peroxidase-conjugated goat anti human polyvalent immunoglobulin as the second antibody. Anti-type II collagen antibodies were seen in 12 of 21 (57%) patients, while 13 of 21 (62%) had anti-type IX antibodies detectable by Western blot. A previously unreported 30 kd (probably noncollagen) protein was found by SDS-PAGE of human cochlear tissue extracts, with 3 patients, all with Menieres disease, having antibody activity to this protein detected by Western blot. Anti-type II and anti-type LX antibodies were found in a high percentage of patients with Menieres disease, otosclerosis, and strial atrophy. Six patients (29%), and all control patients, had no detectable antibodies to these proteins by our assay.

Functional architecture of the nasopharyngeal tonsil

The exact architecture of the normal nasopharyngeal tonsil remains obscure because most histopathologic investigations have been based on surgically removed adenoids. We compared enlarged adenoids and normal nasopharyngeal tonsils under both light and electron microscopes. The marked features of clinically enlarged adenoids were a large extension of the reticular epithelium and increased germinal centers. A tendency toward increased stratified squamous epithelium and decreased ciliated epithelium was apparent in enlarged adenoids, possibly due to inflammatory conditions. One type of nonciliated cell seemed to transport foreign material into underlying lymphocytes, as do the M cells of gut-associated lymphoid tissue. This type of nonciliated cell was rarely found in the extended reticular epithelium of enlarged adenoids. These findings suggest a disturbance of the antigen-trapping system and surface protections in adenoidal enlargement.

Distribution of immunocompetent cells in normal nasal mucosa : comparisons among germ-free, specific pathogen-free, and conventional mice

To better understand the role of immunocompetent cells in the defense mechanism of the upper respiratory tract against microbial invasions, the distribution patterns of those cells were investigated in nasal mucosa of mice maintained in three different conditions: Germ-free (GF), specific pathogen-free (SPF), and conventional (CV) conditions. Immunostaining by the indirect peroxidase method and toluidine blue staining were employed for the detection of immunocompetent cells and mast cells. For immunostaining, anti-IgG, -IgA, and -IgM polyclonal antibodies and anti-Lyt-1, -Lyt-2, and -Mac-1 monoclonal antibodies were used as primary antibodies. In nasal mucosa of CV mice, Mac-1+ cells, mast cells, and all cell types of lymphocyte subsets were present. In nasal mucosa of SPF mice, all cell types were also positive, but fewer in number than those of CV mice. In nasal mucosa of GF mice, IgG+, IgA+, and Lyt-2+ cells were rare, although IgM+ and Lyt-1+ cells were present in small numbers. An electron microscopic study revealed that follicle-like lymphocyte aggregates with high endothelial venules were present in nasal mucosa close to the mucosal epithelia. These findings suggest that lymphocytes are mobilized to nasal mucosa, responding to continuous antigenic stimuli, and play an important role in the local defense mechanism of the upper respiratory tract.

Restoration of brain stem auditory-evoked potentials by gene transfer in shiverer mice.

We studied the shiverer mouse as a model for correcting hearing disorders resulting from genetic abnormalities of the central nervous system (CNS). Shiverer mice are homozygous for an autosomal recessive mutation (deletion) in the gene for myelin basic protein (MBP), a major protein component of the myelin sheath in the CNS. Under electron microscopic observation of the cochlear nerve, the CNS portion in shiverer mice showed hypomyelination, but the peripheral portion, including spiral ganglion cells, was normal. We produced MBP-transgenic mice by microinjection of an MBP cosmid clone into the pronucleus of fertilized eggs from shiverer mice. The transgenic mice were found to recover MBP levels up to 25% of normal. A greater number of axons in the transgenic mice were myelinated than in the shiverer mice, but the myelin sheath was not as thick as in normal controls. Every interpeak latency of brain stem auditory-evoked potentials was prolonged in the shiverer mice and improved in the transgenic mice. This study provides an example of gene therapy for hearing disorders caused by a CNS abnormality. We discuss some strategies for researching genetic hearing impairment or deafness in both animals and humans.

Secretory IGA and Bacterial Adherence to Nasal Mucosal Cells

The ability of Streptococcus pyogenes to adhere to nasal mucosal cells was investigated by an in vitro assay system in order to clarify the actual role of secretory IgA in mucosal immunity in the nose. The number of bacteria adhering to isolated mucosal cells was significantly larger in patients with chronic sinusitis than in normal control subjects. The ability of bacteria to adhere, however, was significantly lower in the group of subjects having specific secretory IgA antibody activity to the M protein than in those having no secretory IgA activity. Findings demonstrated that mucosal immunity preventing or blocking the adherence of bacteria is impaired in patients with chronic sinusitis.

Molecular Basis of Type II Collagen Autoimmune Disease: Observations of Arthritis, Auricular Chondritis and Tympanitis in Mice

A type II collagen autoimmune response results in arthritis, auricular chondritis and tympanosclerosis in humans and animals. The purpose of this study is to further define the molecular and pathogenic events involved in these lesions in rodents. Type II collagen fragment CB11-specific monoclonal antibodies induced lesions in joints, ear lobes and tympanic membranes. In immunized mice, the thickness of tympanic membranes increased to two- to fourfold normal size. Electron micrography showed that the arrangement of collagen fibers is irregular in both radial and auricular layers, containing fibroblasts, a homogeneous material resembling low-density cholesterol crystals and cell infiltration. The mice with auricular chondritis had lymphocytes expressing V beta-8 T cell receptor (TCR) in arthritic joints and lymphocytes expressing V beta-6 TCR in ear lobe lesions. A monoclonal antibody specific to the TCR V beta-8 subfamily suppressed the onset of arthritis. Sequence analyses of the V beta structure of TCR involved in the lesions confirm the immunohistologic study.

Latissimus dorsi myocutaneous-iliac bone flap for reconstruction of massive defects of mandible and oral basis†

Recent advances in reconstructive surgery for head and neck cancer have improved the cure rate of advanced carcinoma, and the function of the organ. However, it still remains difficult to repair the mandible and oral floor. We devised a combined flap of myocutaneous latissimus dorsi and iliac bone, and applied it to two patients with advanced carcinoma of the oral cavity that invaded to the mandible (T4N3M0). Each patient received preoperative irradiation, totalling 30 Gy and 40 Gy. Two weeks before the extensive resection, a sufficient bony mass for the presumed mandibular defect was taken from the iliac crest and transplanted beneath the latissimus dorsi muscle. Defects of the mandible and oral floor were reconstructed using this combined flap immediately after the resection. The patients began to eat 2 weeks after surgery.

MULTIPLE FOREIGN BODIES (FISH BONES) IN THE ESOPHAGUS AND RECTUM

Multiple foreign bodies (fish bones) in the esophagus and rectum, seen in a 64-year-old man, are reported. This patient suffered from severe pain on swallowing after he had eaten fried fish. Five days later, he was referred to us, and underwent removal of a piece of fish bone under direct metal esophagoscopy. After surgery, fever and leukocytosis still continued, even though pain on swallowing disappeared. Periesophageal abscess was suspected; but no leakage or abnormal shadow was found by esophagography. Since the patient complained of severe anal pain five days after the endoscopic procedure, a romanoscopy was performed: a fish bone in the rectum and periproctal abscess were detected. Removal of the foreign body and incision of the periproctal abscess caused fever and leukocytosis to subside. A review of the literature revealed only a very few cases of multiple foreign bodies in the esophagus and lower digestive tract.

Reconstructive Surgery of Head and Neck Cancer Using Various Pedicle Flaps

Various pedicle flaps were used for the reconstruction of large defects following extirpation of head and neck malignant tumors of 55 patients. Although there are several complications, myocutaneous (MC) island flap surpassed the deltopectoral (DP) flap in the reconstruction of the pharyngo-esophagus, tongue, oral cavity, mandible, and of a massive defect. However, DP flap is still valuable material because of its steadiness in pharyngo-esophageal reconstruction in patients who have background disorders, such as serious heart disease and diabetes. Forehead or DP flap is valuable in the reconstructive surgery of full-thickness defect of the cheek. Reconstruction of extremely large defect of the head and neck could be accomplished by use of latissimus dorsi flap and/or combination with DP flap. Selection of the flap should be made in accordance with the region of the reconstruction, size of region, sex, and individual considerations.

Influence of Middle Ear Effusion on Perilymph

To investigate the influence of middle ear effusion (MEE) on perilymph (PL), an experimental otitis media with effusion (OME) was provoked in chinchillas by injecting the tympanic cavity with immune complexes. MEE remained for up to 9 days after the injection of immune complexes. PL was aspirated on the 4th, 10th, and 21st day after the inoculation. The mean concentrations of albumin, IgG, and prostaglandin E2 (PGE2) were significantly greater in PL from OME induced ears than in that form normal control ears. 3H-PGE2 placed on the round window membrane of pathologically affected ears passed into PL in significantly greater amounts than in normal controls. The findings of the present study indicate that MEE affects the biochemical environment of PL. However, this influence was transient and reversible.