Laryngeal type I allergy has rarely been reported, although it is well established that type I allergy can be induced by various antigens in the nasal cavity and bronchus.
This study was designed to determine whether type I allergic reaction occurs in the laryngeal mucosa.
We determined the following in a sensitized guinea pig model developed by Sohma et al :
1) the degree of eosinophilic infiltration in sections stained with hematoxylin-eosin (HE),
2) the number of mast cells in sections stained with toluidine blue (TB),
3) the concentration of histamine in laryngeal mucosal secretions.
Sections obtained from the larynx in sensitized guinea pigs were stained with HE or TB.
Examination of the sections stained with HE revealed a high degree of infiltration by eosinophils and edematous change in the laryngeal mucosa.
Tissue sections stained with TB were examined light microscopically in high-power fields. The number of mast cells in the subglottic epithelial layer of sensitized guinea pigs was higher than that of nonsensitized guinea pigs.
Fourteen sensitized guinea pigs underwent tracheostomy, and the larynx was washed with saline. Histamine concentration was measured by radioimmunoassay in lavage fluid collected before and after topical ovalbumin challenge. Topical challenge immediately induced a significant increase in histamine concentration.
The findings noted above indicated that antigen challenge induced a type I allergic reaction in the larynx of sensitized guinea pigs.

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The midbrain periaqueductal gray (PAG) region where electrical stimulation induces vocalization receives both excitatory inputs from the motor cortex and the limbic structures and inhibitory in-puts which contain GABA as a transmitter substance. To know whether the substantia nigra pars reticulata (SNr) is one of origin of the GABA inhibitory system, effects of GABA antagonist (bicuculline methiodide (BIC)), agonist (muscimol) or local anesthetics (lidocaine) into the SNr on the PAG threshold for vocalization were investigated in ketamine anesthetized cats.
After injection of BIC into the SNr, the PAG threshold for vocalization increased by about 20 to 100% for 60 to 90 min. With the higher stimulus intensity, vocalization was induced with similar motor pattern to that was observed before the injection, although the spontaneous respiratory cycle tended to be longer after BIC injection. This suggests that the BIC injection altered the vocalization threshold not at medulla but at midbrain.
While, injection of GABA A agonist, muscimol, or local anesthetics decreased the PAG threshold for vocalization.
These results indicate that the SNr is a plausible origin of the GABA A inhibitory system which regulates the motor outflow for vocalization, which comes from the limbic structures to the lower brain structures via the PAG area.

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The laryngeal and lingual taste buds (TBs) with their neural components in human were investigated by immunohistochemical technique and compared with each other. The antisera used in this study were against PGP 9.5, NSE, S-100 protein, calbindin-D and cytokeratin intermediate filaments.
Laryngeal TBs were observed on the laryngeal surface of the epiglottis and extended caudally, reaching peak density at the laryngeal side of the arytenoid tubercle. Their size is a little smaller than that of the tongue.
The PGP 9.5 and NSE immunoreactivities was recognized in some taste bud cells as well as nerve fibers both within the TBs and in the subepithelial connective tissue. S-100 protein immunoreactivities were found in none of cells in the TBs, but exclusively in the subepithelial neural elements. The Calbindin-D immunoreactivity was confined to a part of neural elements which was very thin. These immunohistochemical findings are essentially identical with lingual TBs.
Immunoreactivities of cytokeratin filaments with low molecular weight were typically recognized in a part of or whole TB cells.
With these results, one of the physiologic functions of the laryngeal TBs was considered to be that of a receptor of chemical stimulation like lingual TBs. Their cellular components were thought to be derived from a kind of epithelial cells.

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To investigate whether GABAergic inhibition plays a major role in controlling vocalization at the periaqueductal gray (PAG), we investigated changes of the stimulus thresholds of electrically induced vocalization following injections of a GABA antagonist, bicuculline methiodide (BIC) and GABA-a agonist, muscimol into the caudal portion of the ventrolateral PAG. BIC injections into the PAG decreased the stimulus thresholds of the pontine call site (PCS) and the injection sites for about 1 hr with a similar time course, while muscimol injections increased the stimulus thresholds. BIC or muscimol injection into the areas between the PAG and the PCS, where electrical stimulation could induce vocalization, did not affect the stimulus threshold of both the PAG and the PCS. These results suggested that GABAergic inhibition modulates the mechanism of producing vocalization in the ventrolateral PAG and that the descending fibers for vocalization from the PAG passed through the PCS.

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Silicone has been used for soft-tissue augmentation in Japan, however injectable collagen is taking over silicone because of latest reports on the carcinogenesis of silicone. This study examined histological changes over time (up to 180 days) of three types of injectable collagen (3% non crosslinked, 6.5% non cross-linked and 6.5% cross-linked), which were injected into hip muscles of mice. Each type of collagen was not absorbed and remained in a mass among muscle tissue at any time of the study. 3.5% non cross-linked type showed mild degeneration and foreign body reaction. 6.5% non cross-linked type showed severe degeneration with carcification and many foreign body giant cells in the implant. This might suggest possibility of further granuloma formation. 6.5% crosslinked type showed little degeneration and foreign body reaction and it seemed that wound healing process finished earlier than that for the other two types. Cross-linked collagen was shown in this experiment to persist as a stable bioimplant.

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Using a silver colloid staining technique, the argyrophilic nucleolar organizer region (AgNOR) was examined in tissue specimens taken from 79 laryngeal carcinomas and 53 hypopharyngeal carcinomas. The average number of AgNOR per nucleus (AgNOR number) was 4.30 in laryngeal carcinoma and 3.80 in hypopharyngeal carcinoma. In laryngeal carcinomas, the AgNOR number of advanced T groups (T3, T4) or N positive groups (N1, N2, N3) was statistically higher than T1, T2 or NO tumors. There was also a significant difference of the AgNOR number between early stages (I, II) and advanced stages (III, IV). Histologically, statistical difference of AgNOR number was present between well and moderately or between moderately and poorly differentiated squamous carcinoma. In contrast, there was no significant difference of the AgNOR number between each T, N, stage and histological differentiation in hypopharyngeal carcinomas.
As a statistically significant difference of survival curve was noted between each group with low, intermediate and high AgNOR number, AgNOR seems to be associated with better prognosis of laryngeal cancer patients.

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During the past 17 years we have been performing the tracheoesophageal (TE) fistulization for voice reconstruction following total laryngectomy. The principle of this technique is to divert the exhaled air through the TE fistula into the esophagus and vibrate hypopharyngeal mucosa. Radiographical examination and manometry during TE phonation reveal that the thyropharyngeal muscle forms the retropharyngeal prominence on which the neoglottis is located. It has been alreadly reported that the glottis is open for voiceless sounds with laryngeal articulatory adjustment in laryngeal speech. Although TE speakers have lost the larynx, they produce voiceless and voiced sounds with articulatory adjustment. This strongly suggests that neoglottic adjustment exists in TE speech. This study was designed to clarify its mechanism.
Ten speakers were subjected to this study consisting of the fiberoptic examination of the neoglottis, and aerodynamic and electromyographic (EMG) studies with the test words of voiceless or voiced plosives.
Fiberoptic examination revealed a transient neoglottal opening for voiceless sound production. The EMG activity of the thyropharyngeal muscle decreased for voiceless sound production. The EMG activity reduction started prior to the increase of the supraneoglottal pressure. These findings indicate that the neoglottic adjustment plays an important role to open the neoglottis in TE speech.

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Twenty eight laryngectomy specimens from patients with glottic carcinoma were histopathologically investigated by means of whole-organ serial sectioning, with paying special attention to microvascular invasion in the immediate vicinity of primary lesions.
1. Microvascular invasion was found in 18 out of 28 patients (64.3%) and was found in 5 out of 6 patients with clinically positive cervical nodes (83.3%) and found in 8 out of 10 patients with histologically proved cervical node metastasis (80.0%).
2. Microvascular invasions were not found in 2 cases, treated with preoperative irradiation and chemotherapy, followed by cervical node metastasis.
3. Although statistically significant correlation was found between microvascular invasion at the primary site and lymph node metastasis in glottic carcinoma, the presence of microvascular invasion may cause an increased incidence of cervical lymph node metastasis in glottic carcinoma.

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Laryngeal Leukoplakia lesions often have complicated morphological characteristics. The degree of DNA-instability as revealed by an immunohistochemical staining with anti-single-stranded DNA antibody after acid hydrolysis (DNA-instability test) was used as a marker of malignancy (Fukuda et al, 1986, 1993). The cases to which this was applied are as follows : 8 laryngeal leukoplakia, 14 oral leukoplakia, 19 squamous cell carcinoma, and 12 normal squamous epithelium. Ordinary paraffin sections taken from each of the cases were hydrolyzed for the denaturation of unstable DNA, and successively stained with anti-single-stranded DNA antiserum immunohistochemistry. The level of proliferative activity and the polarity of the proliferative cell distribution were evaluated by PCNA-immunohistochemistry. Quantitative analyses of the number, mean and largest size, and maximum shape-irregularity evaluated by AgNORs staining in the nucleus were performed. Eight laryngeal leukoplakia and 12 oral leukoplakia cases with dysplasia (90%) were positively and diffusely stained by the DNA-instability test indicating their malignancy. Reflecting their malignant character, these cases showed a remarkable increase in PCNA-index with the loss of polarity of PCNA-positive cell distribution, and also increased maximum shape-irregularity of AgNORs. In conclusion, the majority of leukoplakia occurrences should be regarded the same as in-situ cancer.

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A clinical and histological study was conducted from 1971 to 1992 on 51 cases (44 males and 7 females) who, on their first visit to the clinics of Kitasato University Hospital or Kitasato Institute Medical Center Hospital, were diagnosed as having epithelial hyperplasia of the larynx. The patients were classified into three groups based on subsequent histological findings : 1. Hyperplasia with atypia (7 cases), 2. Hyperplasia without atypia (39 cases), and 3. Carcinoma (5 cases). As for incidence by age groupings, epithelial hyperplasia was seen most frequently in subjects in their fifties, followed by those in their forties and sixties thereafter. The chief complaints were hoarseness (in 46 out of 51), difficulty in expectrating sputum, throat discomfort and coughing. The lesion was most commonly observed in the anterior 1/3 of the vocal cord, although diffuse involvement of the vocal cord was not seldomly seen. There was no positive relationship between the frequency of recurrence after surgical removal of the lesion and the degree of atypism. Subepitherial inflammation was often observed similarly in the two groups of epitherial hyperplasia. In those cases who were diagnosed as having carcinoma, subepitherial inflammatory reaction was relatively severe.

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Using 38 guinea pigs with unpaired arytenoid muscle (AR), the innervation pattern of the AR was studied viewed from reinnervation process following unilateral (left) recurrent laryngeal nerve (RLN) transection. Light and electron microscopic observation, glycogen depletion experiment, and histochemical study using the adenosine triphosphatase (ATPase) stain were carried out.
Seven days after transection, both myelinated and unmyelinated fibers were noted in the intramuscular nerve funiculus. Thus, the anastomosis of the arytenoid branches between both RLNs was found in the belly of AR. Three types of the neuromuscular junctions (NMJs) were observed; normal, degenerated, and regenerated NMJs. Five weeks after transection, however, neither degeneration nor regeneration of the NMJ were seen.
Until three weeks after transection, half of the AR fibers were stained with PAS staining following electrical stimulation of the contra lateral (right) RLN. Five weeks after transection, all the AR fibers were unstained. Therefore, all the AR fibers were considered to have been reinnervated by the contra lateral RLN.
ATP stain of the AR showed that type 2 fibers were predominant regardless of the period following unilateral denervation.
The present study indicates that transection of the unilateral RLN facilitates collateral sprouting from the contra lateral RLN, and that these collateral sprouting reinnervated the denervated muscle fibers before degeneration occurs. Therefore the AR, as a whole, receives a specific motor innervation from the bilateral RLNs, although each muscle fiber is innervated by the unilateral RLN.

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The hypopharyngeal constrictor muscle consists of the thyropharyngeal (TP) and cricopharyngeal (CP) muscles. These two muscles cooperatively and, in a sense, competitively work at the pharyngeal swallowing stage. Contraction of the TP muscle drives bolus into the esophagus, while the CP muscle ralaxes and allows the bolus to pass through the upper esophageal sphincter which is made by a contraction of the CP muscle.
We investigated histochemical properties of these two muscles. Acetylcholine esterase stain showed that the neuromuscular junctions of the CP muscle were scattered in the horizontal direction compaired to those of the TP muscle. Glycogen depletion test showed that the glycogen-negative muscle fibers were distributed uniformly througout the TP muscle, while those of the CP muscle decreased in number from the stimulated side toward the unstimulated side. These results suggested that the CP muscle fibers terminate in the belly of the muscle, and that length of the CP muscle fiber is not uniform. Actomyosin ATPase stain revealed that type 2B fiber was predominant in the TP muscle, while type 1 fiber was predominant in the CP muscle. The mean diameter of the TP muscle fibers was larger than that of the CP muscle.
In conclusion, the histochemical properties of the TP and CP muscle showed a great contrast to each other. But these different properties gradually change around the border of the two muscles.

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Recently, therapeutic outcome of laryngeal cancer has been improved, but some cases develop local recurrences. Therefore, we investigated the causes of recurrences. We previously reported that one of the causes was multicentric cancerization in the larynx, which was frequent in heavy smokers.
In this study, we investigated laryngeal double cancer, especially that coexisting hypopharyngeal cancer, as a cause of local recurrence. The following results were obtained :
1) In one hundred thirty previously untreated laryngeal cancer patients and thirty-six previously untreated hypopharyngeal cancer patients who was referred to the ENT Department of Nara Medical University during the period from 1987 to 1993, there were three double cancer cases in which laryngeal and hypopharyngeal cancer developed simultaneously.
2) All were advanced cancers, and one case demonstrated triple cancer in the larynx, hypopharynx and esophagus.
3) Comparing double cancer patients with _??_?single_??_? laryngeal cancer patients, we found that the Sake index was significantly higher (p<0.05 unpaired Wilcoxon's test) in the double cancer patients.
4) Comparing double cancer patients with _??_?single_??_? hypopharyngeal cancer patients, we also found that the Brinkman index was slightly higher in the former group.
5) From the data in this study, we propose treating heavy alcohol drinkers with laryngeal cancer as high risk patients for double cancer involving the hypopharynx.

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A 57-year-old housewife complaining of choking was admitted to our hospital. Fiberscopic examination revealed a white tumor with a smooth surface and a slight hypervascularity below the vocal cords. A cervical X-ray examination taken from the lateral view revealed that the tumor was located between the area just beneath the vocal cords and the fourth ring of the trachea. Emergent tracheostomy and biopsy of the tumor were performed. Histological diagnosis indicated that the tumor was a small, lymphocytic type B-cell malignant lymphoma (diffuse). No other lesions were detected in the systemic evaluation. After three courses of combined chemotherapy (CHOP), the tumor disappeared. The clinical and pathological characteristics of this case were similar to those of malignant lymphoma of mucosa-associated lymphoid tissue (MALT) which arises in the stomach, salivary glands. lungs. and thyroid.

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A 26-year old female suffered from dyspnea immediately after inhalation of a strong acid by mistake during a chemical experiment, and was admitted to a life-saving center. The acid was trifluoromethane sulfonic acid. Intratracheal intubation was performed promptly to prevent chemical bronchitis. When the tube was removed 3 days later, she didn't complain of dyspnea any more, although sight cough remained. She was discharged 12 days after the admission. After one and a half month after the discharge, dispnea took place again. She was referred to our institute because a large mass was found in her larynx by a doctor. The tumor originated from the right vocal fold was resected microsurgically. The histopathological examination revealed that the tumor was a nonspecific granuloma. No recurrence has been observed after the surgery for 4 months. The inhalation of acid, the intratracheal intubation, and sustaining cough were considered to be the cause of the laryngeal granuloma of this patient.

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