Doxorubicin ototoxicity is induced in mice by combination treatment with cyclosporin A.

Although doxorubicin [adriamycin (ADM)] ototoxicity has not been detected to date, it has been reported that neurotoxicity in the central nervous system was induced by chemotherapy with ADM in patients receiving chronic cyclosporin A (CsA) treatment. ADM ototoxicity may be induced by combination therapy with CsA because extrusion of ADM from the inner ear by p-glycoprotein (p-gp), which acts as an extrusion pump and is expressed on the surface of endothelial cells of capillary blood vessels, might be inhibited by CsA. resulting in significant accumulation of ADM in the inner ear. ADM (10 mg/kg) was administered to FVB mice either with or without CsA (200 mg/kg). Auditory brainstem responses (ABRs) were recorded before and after treatment. ABR changes were not observed in mice treated with either ADM or CsA alone. Threshold elevation, elongation of wave I-V latencies and interpeak latencies of waves I-II, I-III, I-IV and I-V were detected in mice treated with ADM in combination with CsA. These changes reached their peak values 3 weeks after treatment, and then recovered to pre-treatment levels. In normal mice, ADM is extruded by p-gp from the inner ear and auditory pathway, thus preventing hearing disorder. However, ADM ototoxicity was induced by combination therapy with CsA, indicating that CsA has an inhibitory action on p-gp function in the auditory pathway, including the inner ear. After organ transplantation, therefore, clinical administration of ADM in combination with CsA should be performed with caution.

Intraoperative identification of regenerated chorda tympani nerve and its relationship to recovered taste function.

This study demonstrated a simple method of repairing the severed chorda tympani nerve and a method of intraoperative identification of regenerated nerves, and evaluated taste function of regenerated nerves. Seven patients who underwent staged tympanoplasty and whose chorda tympani nerve was severed during primary surgery were evaluated. When the chorda tympani nerve was severed during primary surgery, proximal and distal stumps were anastomosed or approximated almost in the original position and fixed with fibrin glue on the temporal muscle fascia used to reconstruct the eardrum by the underlay method. During primary surgery, end-to-end anastomosis was possible in 3 patients but nerve gap defects remained in the other 4 patients. In all 7 patients, regenerated nerves were identified during secondary surgery not in the tympanic cavity but in the submucosal layer of the previously reconstructed eardrum. In all patients, complete or incomplete recovery of taste perception was observed by both the filter paper disk method and electrogustometry, suggesting that the regenerated nerves had actual taste function. From these results, it was concluded that the severed chorda tympani nerve could regenerate in the reconstructed eardrum even if nerve gap defects remained between the proximal and distal cut ends, when repair or approximation of the nerve was properly completed.

Homozygous disruption of the mdrla P-glycoprotein gene affects blood-nerve barrier function in mice administered with neurotoxic drugs.

We investigated the expression and function of mdr1a p-glycoprotein in peripheral nerves, including the VIIth and VIIIth nerves, using mdr1a p-glycoprotein gene knockout mice [mdr1a(-/-) mice] and wild-type mdr1a(+/+) mice. P-glycoprotein expression in capillary endothelial cells of the peripheral nerve tissues was detected by immunohistochemical and RT-PCR analyses in mdr1a(+/+) mice but not in mdr1a(-/-) mice. Pharmacokinetic analyses indicated that, compared to mdr1a(+/+) mice, mdr1a(-/-) mice showed a significantly higher accumulation of p-glycoprotein substrate drugs such as vinblastine and doxorubicin, which are neurotoxic. Tissue concentrations of vinblastine and doxorubicin were lower in the order of the brain, peripheral nerves and most other organs. However, increased accumulation was not detected after administering another neurotoxic drug, cisplatin, indicating that p-glycoprotein is selective at extruding drugs. These data indicate that mdr1a p-glycoprotein, which acts as an efflux pump, might play an important role in the blood-nerve barrier to prevent side effects induced by neurotoxic p-glycoprotein substrate drugs. The participation of p-glycoprotein in the blood-nerve barrier is considered to represent a new functional mechanism of this barrier.

Cyclosporin A inhibits the extrusion pump function of p-glycoprotein in the inner ear of mice treated with vinblastine and doxorubicin.

Cyclosporin A (CsA) inhibits the membrane transport protein p-glycoprotein (p-gp) and can enhance the accumulation of vinblastine (VBL) and doxorubicin (Dx) in the inner ear of mice. In mice pretreated with 200 mg/kg of CsA, there were significantly increased VBL and Dx concentrations detected in the inner ear tissue and other organs, with a small but significant increase in the brain. Furthermore, hearing thresholds measured by auditory brainstem responses were significantly elevated 3 weeks after VBL or Dx treatment in combination with CsA. However, the altered thresholds recovered to pretreatment levels 8 weeks after treatment. Pharmacokinetic and functional alterations observed in this study suggest caution in applying these combinations in clinical practice.

Expression of p-glycoprotein is associated with that of multidrug resistance protein 1 (MRP1) in the vestibular labyrinth and endolymphatic sac of the guinea pig.

Expression of p-glycoprotein (p-gp) and multidrug resistance protein 1 (MRP1) was detected in the vestibular labyrinth and endolymphatic sac (ES) of the guinea pig by immunohistochemical staining using anti-p-gp monoclonal antibody (mAb) C219 and anti-MRP mAb MRPr1. P-gp was detected in capillary endothelial cells of the crista ampullaris, utricle, saccule and ES. MRP1 was detected in the epithelial lining of the crista ampullaris, utricle, saccule, and epithelial cells of the ES. Since p-gp and MRP1 act as extrusion pumps, they may coordinate with each other in vestibular organs and ES and play an important role in the blood-labyrinth barrier.

Recent outcome of tympanoplasty in the elderly.

OBJECTIVE: To investigate the outcome of tympanoplasty in the elderly (patients older than 60 years) compared with younger patients. PATIENTS AND STUDY DESIGN: Retrospective review of 87 (28.3%) older patients among a total of 307 patients with chronic otitis media with or without cholesteatoma who were surgically treated at a university hospital by the senior author. Follow-up was systematically provided at the same institution. INTERVENTIONS: Surgery included tympanoplasty with mastoidectomy performed as the primary procedure in 358 ears. Tympanoplasty was performed with canal-wall-up or canal-wall-down with canal wall reconstruction, ossiculoplasty with autologous or homologous ossicle interposition or columella. Mean follow-up was 30 months (range, 12-70 months). MAIN OUTCOME MEASURES: Pre- and postoperative air- and bone-conduction thresholds were calculated as an average of three speech frequencies (0.5, 1, and 2 kHz). Analysis was subsequently carried out on the postoperative air-bone gap, hearing gain, and postoperative problems such as elevation of the bone-conduction threshold, delayed epithelialization, and reperforation of the eardrum. Statistical analysis was performed by chi-square or Student's t-test. A p value less than 0.05 was considered significant. RESULTS: Compared with results from younger patients, there was no particular disadvantage in postoperative hearing results and complications in the elderly, although preoperative bone-conduction thresholds were gradually worsened with age. CONCLUSIONS: There is no contraindication for tympanoplasty in older patients if their physical status is the same or better than what is normal for their chronological age.

Expression of multidrug resistance protein 1 (MRP1) in the rat cochlea with special reference to the blood-inner ear barrier.

Expression of multidrug resistance protein 1 (MRP1) was detected in the rat cochlea by RT-PCR and immunohistochemistry using anti-MRP monoclonal antibody MRPr1. Use of primers specific for rat mrp1 gene resulted in the amplification of an expected 394 bp fragment prepared from brain and cochlear tissues. Immunohistochemically, MRP was found in the choroid plexus, stria vascularis, spiral ligament, spiral prominence and cochlear nerve in the modiolus. From these results, it was suggested that MRP in the rat cochlea might function as an extrusion pump and play an important role in the blood-inner ear barrier.

Long-term follow-up results of electrogustometry and subjective taste disorder after middle ear surgery.

OBJECTIVES: The present study compares the long-term follow-up results of electrogustometry with patient reports of taste dysfunction after middle ear surgery. STUDY DESIGN: Retrospective review of 371 patients who underwent middle ear surgery. METHODS: Patients were divided into the following groups depending on the degree of manipulation or surgical damage to the chorda tympani nerves: the no-touch group (group 1 [n = 109]); the touch group (group 2 [n = 149]); and the severed nerve group (group 3 [n = 113]). Electrogustometry was periodically performed over the course of several years. RESULTS: The incidences of postoperative subjective taste disorder in groups 1, 2, and 3 were 2.8%, 25.5%, and 38.9%, respectively. Although the subjective taste disorder usually recovered within 1 to 2 years after surgery in all groups, it persisted for more than 2 years in 2.7% of the touch group and 5.3% of the severed nerve group. Concerning postoperative electrogustometric results, in the no-touch group, 8.3% of patients showed threshold elevation on electrogustometry, but the elevated thresholds completely recovered in all cases. In the touch group, 45% of patients exhibited elevated electrogustometric thresholds on their first postoperative test, including 32.9% who subsequently had complete electrogustometric recovery, 10.1% who subsequently had incomplete recovery, and 2% who failed to recover during the follow-up period. In the severed nerve group, none of the patients was responsive to the electrical stimulus on the first postoperative test, including 8.8% of patients who subsequently exhibited complete electrogustometric recovery, 32.7% who later had incomplete electrogustometric recovery, and 58.4% who never recovered any electrogustometric responsiveness. Nerve repair in the severed nerve group produced better recovery, as measured electrically. CONCLUSIONS: The incidence of postoperative subjective taste disorder was low, although inconsistent with the high incidence of threshold elevation on electrogustometry, especially in the severed nerve group. Preservation or repair of the chorda tympani nerve is recommended in order to maintain or recover gustatory function.

Morphological and functional study of regenerated chorda tympani nerves in humans.

It is still unclear whether the chorda tympani nerves in humans regenerate after being severed during middle ear surgery, although functional studies have demonstrated recovery of taste 1 to 2 years after surgery. To date, 12 cases of regenerated chorda tympani nerves have been found in our series of patients during secondary surgery. The regenerated nerves of 3 cases of the 12 were removed as samples during secondary surgery to detect regenerated myelinated axons. All regenerated nerves were in the submucosal connective tissue layer of the reconstructed eardrum. In the regenerated nerves, myelinated nerve fibers existed in a small fascicle or in connective tissue, but the number of myelinated axons was low compared with that in normal subjects ( 1.752 +/- 78; n = 3), and the distribution was sparse. The total number of regenerated myelinated axons varied from 141 (8.3%) to 979 (55.9%). From a functional study using electrogustometry, incomplete recovery of electrogustation was observed in all 3 cases before secondary surgery, suggesting that chorda tympani nerves actually regenerate in the middle ear and do function.

[Efficacy of middle ear surgery for patients with hearing aids and middle ear disease].

Of 356 cases that underwent middle ear surgery for hearing improvement, 30 (8.4%) with air conduction hearing aids and middle ear disease were evaluated pre- and postoperatively. All surgeries were performed by the same surgeon. Diagnoses included 22 chronic otitis media, 5 chronic otitis media with cholesteatoma, 1 otosclerosis and 2 ossicular anomaly. Chief complaints at the first visit to Fukui Medical University Hospital were otorrhea (17 cases), hard of hearing (28 cases), dizziness (2 cases) and tinnitus (2 cases). Nineteen patients underwent surgery on both ears and eleven on one ear including five ears that showed better hearing preoperatively. Surgical procedures were tympanoplasty type I (15 cases), modified type III (8 cases), modified type IV (3 cases), stapedotomy (2 cases) and implantable hearing device (2 cases). After surgery, 16 patients (group 1) did not need hearing aids, while 14 (group 2) still needed hearing aids. Preoperative hearing was 64.1dB (n = 30) on average and average hearing one year after surgery in group 1 (35.4 +/- 14.1dB) was significantly better than in group 2 (58.1 +/- 18.4dB). After surgery, otorrhea stopped in all cases (100%), subjective hearing loss improved in 82% of the patients, vertigo improved in 100% and tinnitus improved in 50%. These results emphasize the benefits of surgical therapy, and the reasons why it should be recommended to patients with hearing aids and middle ear disease, such as to improve hearing disorders, to stop otorrhea and to prevent progressive sensorineural hearing loss.

P-glycoprotein expression in capillary endothelial cells of the 7th and 8th nerves of guinea pig in relation to blood-nerve barrier sites.

Expression of P-glycoprotein (P-gp) was detected by immunohistochemical staining and Western blot analysis in the peripheral nerves (7th and 8th nerves) of the guinea pig using anti-P-gp monoclonal antibody C219. P-gp was detected in the capillary endothelial cells of these nerves. Immunoreactivity in these nerves was similar to that in the brain. Besides these nerves, positive staining was observed in the sciatic nerve, although immunoreactivity was somewhat lower than that of the 7th and 8th nerves. The present investigation suggested that P-gp in the endothelial cells of the peripheral nerves might play a very important role as a part of the blood-nerve barrier function, since P-ag acts as an extension pump in these cells.

Similar pharmacokinetics and differential ototoxicity after administration with cisplatin and transplatin in guinea pigs.

Transplatin is a transisomer of cisplatin. Although cisplatin exhibits strong ototoxicity, there is no report concerning the ototoxicity of transplatin. To evaluate differences in pharmacokinetics and ototoxicity, cisplatin (7.5 mg/kg) and transplatin (30 mg/kg) were administered to guinea pigs twice at an interval of 5 days. The N1 threshold of the compound action potential was significantly elevated after administration of cisplatin. Cochleogram of the cisplatin-treated group showed severe losses of outer hair cells essentially at the basal and second turns. Transplatin, however, did not induce any detectable functional or morphological changes. Furthermore, blood urea nitrogen and creatinine levels in serum were not elevated after administration of transplatin, whereas cisplatin showed strong nephrotoxicity. The serum and perilymphatic concentrations of platinum up to 24 h after administering an equimolar dose of cis- or transplatin (7.5 mg/kg) were almost similar. As has been reported by many investigators, transplatin has no anti-tumor activity because stereochemical limitations preclude transplatin from forming intra- and interstrand closs-links with nuclear or mitochondrial DNA. From these results, it was concluded that the stereochemical structure of the platinum compound and steric interaction with target molecules such as mitochondrial DNA in the cochlear outer hair cells might be important to the ototoxic mechanism of platinum compounds.

Prevention of persistent ear drum perforation after long-term ventilation tube treatment for otitis media with effusion in children.

The effect of tape patch technique using Steri-Strip tape in combination with freshening of the perforation edge after removal of long-term ventilation tubes for preventing permanent ear drum perforation was evaluated. The longer the tubes remained in place, the higher the incidence of persistent perforation after tube removal. The perforation rate after Goode T-tube treatment was 4.0% in the spontaneous extrusion group and 14.3% in the intentional removal group. In the ears treated by tape patch application, none of the perforations persisted after removal of the Goode T-tube. After removal or extrusion of Paparella Type II tube, perforations did not close in 13.2% of the group without tape patch application. When a tape patch was applied, only one perforation (3.3%) did not close. From these results, tape patch technique in combinations with freshening of the perforation edge at the time of tube removal was useful to promote healing and prevent persistent ear drum perforation.

The trajectory of the sympathetic nerve fibers to the rat cochlea as revealed by anterograde and retrograde WGA-HRP tracing.

Wheat germ agglutinin-horseradish peroxidase conjugate was injected in the unilateral superior cervical ganglion (SCG), and the projection pathways of postganglionic sympathetic nerve fibers innervating the cochlea were traced in the rat. The labeled axons advanced along the internal carotid artery (ICA), and a few advanced caudally in the major petrosal nerve (MPN) and entered the facial nerve, while the majority ran rostral to the pterygopalatine ganglion at the point where they crossed the MPN in the carotid canal. The rest of the labeled fibers remained on the surface of the ICA and advanced to the cranial cavity. Most of the labeled fibers along the facial nerve joined the cochlear nerve and finally reached the osseous spiral lamina through the spiral ganglion. Some of the labeled fibers ran along the anterior inferior cerebellar artery from the basilar artery which was previously thought to have been the only pathway. We could not find any labeled fiber on the modiolar artery from anterior inferior cerebellar artery in the cochlea. These observations are consistent with our hypothesis that the sympathetic fibers innervating the neural tissues or related structures follow nerve fibers and meninges as matrices of projection pathways rather than arteries.