Hearing and tinnitus in head and neck cancer patients after chemoradiotherapy.

Head and neck cancer patients treated with high-dose cisplatin and radiotherapy will suffer from hearing deficits. The current low-dose regimen seldom causes hearing threshold decrease. Tinnitus in this patient population has not been investigated earlier. We aimed to evaluate the possible ototoxicity of low-dose (40 mg/m(2)) weekly administered cisplatin with concomitant radiotherapy. Twenty-two patients with locally advanced head and neck cancer were prospectively recruited to participate the study after treatment recommendation for chemoradiotherapy with low-dose cisplatin and intensity-modulated radiotherapy. They filled in a Tinnitus Handicap Inventory and undertook audiologic evaluations before and after treatment. Ototoxicity was determined by >10 dB threshold shift at frequencies 4 and 8 kHz or in pure tone average. A historical cohort of nine patients treated with high-dose (100 mg/m(2)) cisplatin and radiotherapy was used for comparison. After treatment, study patients demonstrated no significant changes in their hearing over frequencies 0.5-4 kHz, and the threshold shifts were minor at 4 and 8 kHz. More than 50 % of patients reported no tinnitus after treatment and the remainder only had slight to moderate tinnitus causing no interference with their daily activities. In contrast, five of the nine patients having received high-dose cisplatin reported disturbing tinnitus. Further, changes in pure tone averages were exhibited in three of these patients and six had significant threshold shifts at 4 and 8 kHz. Head and neck cancer patients treated with concomitant intensity-modulated radiotherapy and low-dose cisplatin seem to experience only minor audiological sequelae and therefore, these patients appear to require no routine audiological monitoring. Such evaluation could be performed only when needed.

Hyperbaric oxygen therapy seems to enhance recovery from acute acoustic trauma.

CONCLUSION: The average recovery of hearing and cessation of tinnitus was significantly better after hyperbaric oxygen therapy (HBOT) than after normobaric oxygen therapy (NBOT). HBOT can be valuable adjuvant therapy for patients with acute acoustic trauma (AAT). OBJECTIVES: AAT was one of the early indications for the use of HBOT. The rationale of administering oxygen to patients with AAT is based on experimental studies showing that noise exposure results in cochlear hypoxia, which could be compensated by HBOT. The aim of this study was to investigate the efficacy of HBOT in patients with AAT. PATIENTS AND METHODS: We compared the recovery from hearing impairment and tinnitus in 60 ears treated with HBOT with 60 ears treated with NBOT. The HBOT was given daily for 1-8 days. There were no significant differences in clinical or audiological data between HBOT and NBOT groups. RESULTS: The average recovery of hearing both at high and speech frequencies was significantly better and tinnitus persisted less commonly after the HBOT than after the NBOT. Normal hearing at the end of the follow-up period was regained in 42 ears in the HBOT group and in 24 ears in the NBOT group (p<0.01).

Does the triad of fungi, bacteria and exposure to moisture have an impact on chronic hyperplastic sinusitis?

UNLABELLED: Here we evaluated a possible relationship between chronic hyperplastic sinusitis (CHS) and moisture exposure and secondly a seasonal variation of fungal and bacterial findings in the healthy nose. In 28 CHS patients sinus mucus was collected during endoscopic sinus surgery. Samples from the nasal cavities of 19 healthy volunteers were collected by nasal lavage (NAL) in January and in September. Bacterial culture and fungal staining and culture were carried out. Histological samples from the sinus mucosa were obtained. Patients' medical history and environmental factors were enquired. Mold odor or moisture problems in the home or work environment were reported by 46% of the CHS patients. Patients who reported moisture exposure did not differ significantly from those who had not been exposed with regards to microbiological findings, tissue eosinophilia, and earlier operations. Cladosporium (16%) and Alternaria (11%) were found in NAL fluid collected in the autumn from the control subjects. No fungi were isolated from samples taken during the winter. An association between CHS or fungal sinusitis and moisture damage was not apparent in the present study. The fungal findings in the nasal cavity reflect the environmental exposure. This should be taken into account when NAL is used for microbiological studies. PRACTICAL IMPLICATIONS: Living in a moldy house or working in a similar environment may increase the risk of respiratory symptoms and infections. However, our results suggest that chronic hyperplastic sinusitis and fungal sinusitis are not associated with moisture exposure. The nose is a good collector of particles in the air. Especially samples taken by the nasal lavage method reflect the environmental exposure. This should be taken into account when this method is used for microbiological studies.

Traditional tonsillectomy compared with bipolar radiofrequency thermal ablation tonsillectomy in adults: a pilot study.

OBJECTIVES: To assess the morbidity and efficacy of bipolar radiofrequency thermal ablation tonsillectomy and compare it with traditional cold dissection tonsillectomy with diathermy hemostasis. DESIGN: Prospective, randomized, single-blinded, controlled clinical study. SETTING: Helsinki University Central Hospital, Department of Otorhinolaryngology-Head & Neck Surgery, Helsinki, Finland. PATIENTS: Forty healthy volunteer patients aged 18 to 65 years admitted for elective tonsillectomy with recurrent or chronic tonsillitis, obstructive tonsillar hypertrophy, or history of quinsy. Two patients were excluded from the study and 1 patient cancelled the operation. INTERVENTIONS: Nineteen patients underwent a traditional cold dissection tonsillectomy with diathermy hemostasis, and 18 patients underwent a bipolar radiofrequency thermal ablation tonsillectomy. There was no intergroup difference in age, sex, weight, and indications for tonsillectomy. The subjects were not informed of the type of procedure until the telephone interview 3 weeks after the operation. MAIN OUTCOME MEASURES: Operating time and intraoperative blood loss; need for anesthetics during the operation; different recovery indicators in the recovery room (ie, duration and medications administered), surgical ward (ie, medications administered, use of corticosteroids, general condition, and status of the uvula on the first postoperative day), and in the 2 weeks following surgery (ie, visual analog scale scores on 6 symptoms, medications needed, the day patients returned to work, use of antibiotics, and retreatment acceptance); and complications and certain laboratory parameters. RESULTS: There was a statistically significant but clinically insignificant difference in operating time and intraoperative blood loss in favor of the traditional tonsillectomy group. The other outcome measures showed no statistically significant differences. CONCLUSION: Bipolar radiofrequency thermal ablation and traditional tonsillectomy were associated with similar postoperative morbidity.

Sleep-disordered breathing: radiofrequency thermal ablation is a promising new treatment possibility.

OBJECTIVES: The aim of this study was to assess the efficacy and morbidity of radiofrequency thermal ablation of the soft palate in subjects with sleep-disordered breathing. STUDY DESIGN: Prospective, nonrandomized study. Outpatient treatment and an extended follow-up time of 12 months. METHODS: Twenty-one healthy men who were 18 to 60 years of age (median age, 44 y) with sleep-disordered breathing were enrolled to the study. All the patients had habitual snoring for at least 1 year that was associated with excessive daytime sleepiness interfering with social or professional activities. Radiofrequency energy was delivered to the soft palate in two treatment sessions separated by 1 week at 460 +/- 1 kHz with an energy delivery of 600 and 300 J. Snoring Score, Epworth Sleepiness Scale, and cephalometric analysis were measured preoperatively and postoperatively. Certain inflammatory laboratory parameters and visual analogue scale scores of symptoms were measured related to the procedure. RESULTS: The changes in Snoring Score and Epworth Sleepiness Scale scores were statistically significant. The change in the length of the soft palate was statistically significant, whereas the change in palatal width was not. There were no notable changes in the laboratory parameters. The symptom visual analogue scores were low and transient, resolving within days. CONCLUSIONS: The radiofrequency thermal ablation of the soft palate in patients with sleep-disordered breathing seems to be effective. It is safe and associated with only a low morbidity. The promising results must be confirmed in a placebo-controlled study with a larger sample size and a long-term follow-up.

Cystic fibrosis gene mutations deltaF508 and 394delTT in patients with chronic sinusitis in Finland.

Previous studies have shown that cystic fibrosis (CF) gene mutations are linked to several severe chronic infections. Chronic sinusitis is one condition that may well be influenced by a mutation in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. We studied two prevalent CF mutations (AF508 and 394delTT) in a population with a low incidence of CF. The carrier frequency of the CF mutations in the Finnish population is approximately 1 in 80. We examined DNA specimens from 127 chronic sinusitis patients and found one patient who was heterozygous for 394delTT gene mutation. None of the DNA specimens had any AF508 mutation. This study shows that in a population with a low incidence of CF there was no abnormal carrier distribution of the two most common CF gene mutations in a group of chronic sinusitis patients. Routine screening of sinusitis patients for CF mutations provides no additional information on the etiology of chronic sinusitis.

Panfungal PCR and multiplex liquid hybridization for detection of fungi in tissue specimens.

A procedure based on panfungal PCR and multiplex liquid hybridization was developed for the detection of fungi in tissue specimens. The PCR amplified the fungal internal transcribed spacer (ITS) region (ITS1-5.8S rRNA-ITS2). After capture with specific probes, eight common fungal pathogens (Aspergillus flavus, Aspergillus fumigatus, Candida albicans, Candida krusei, Candida glabrata, Candida parapsilosis, Candida tropicalis, and Cryptococcus neoformans) were identified according to the size of the amplification product on an automated sequencer. The nonhybridized products were identified by sequencing. The performance of the procedure was examined with 12 deep-tissue specimens and 8 polypous tissue biopsies from the paranasal sinuses. A detection level of 0.1 to 1 pg of purified DNA (2 to 20 CFU) was achieved. Of the 20 specimens, PCR was positive for 19 (95%), of which 10 (53%) were hybridization positive. In comparison, 12 (60%) of the specimens were positive by direct microscopy, but only 7 (35%) of the specimens showed fungal growth. Sequencing of the nonhybridized amplification products identified an infecting agent in six specimens, and three specimens yielded only sequences of unknown fungal origin. The procedure provides a rapid (within 2 days) detection of common fungal pathogens in tissue specimens, and it is highly versatile for the identification of other fungal pathogens.

Effect of Alloiococcus otitidis and three pathogens of otitis media in production of interleukin-12 by human monocyte cell line.

Alloiococcus otitidis is detected in middle ear effusion of otitis media with effusion (OME). Only a limited number of studies are available concerning the immunological profile of A. otitidis. We have studied the ability of A. otitidis and three other representative pathogens of otitis media to stimulate the production of interleukin-12 (IL-12) from a monocytic cell line THP-1. Viable A. otitidis induced the production of IL-12 in THP-1 cells but IL-12 production was reduced if glutaraldehyde-fixed bacteria were used as stimulants. When viable bacteria were physically separated from THP-1 cells during the stimulation period, remarkable reductions of IL-12 secretion were shown after challenge with gram-positive bacteria A. otitidis and S. pneumoniae. When stimulated with soluble extracts of A. otitidis, THP-1 secreted IL-12 in a dose-dependent manner. The subfraction with a molecular mass over 100 kDa showed a strong ability to induce IL-12 production. Our results show that A. otitidis has immunostimulatory capacity with regard to IL-12 production. We also show that soluble antigen(s) of A. otitidis can modulate the immune response in OME.

Stimulation of adenoidal lymphocytes by Alloiococcus otitidis.

Otitis media with effusion (OME) is characterized by persistent effusion in the middle ear cavity and by chronic inflammation in the middle ear mucosa. Alloiococcus otitidis, a gram-positive aerobic bacterium, has been isolated in middle ear effusion, and by means of sensitive polymerase chain reaction detection assays it has been detected in as many as 20% of middle ear aspirates of patients with OME. Because A otitidis may freely interact with leukocytes in the middle ear effusion, it may potentially modulate the inflammatory reaction in OME. To study the nature of these interactions, we applied an in vitro assay in which killed A otitidis bacteria were incubated with peripheral blood and adenoidal mononuclear cells. The expression of the proliferation-associated surface marker CD69 was then measured in B lymphocytes and in CD4+ helper and CD8+ cytotoxic-suppressor T lymphocytes by means of multicolor flow cytometry. Alloiococcus otitidis induced the expression of CD69 in both peripheral blood and adenoidal T and B cells. Among the T cells, the cytotoxic-suppressor T lymphocytes were preferentially activated. It was also tested whether A otitidis would have an effect in another cytotoxic and immunoregulatory system, namely, the induction of natural killer cell activity in peripheral blood mononuclear cells. However, the effect was minimal compared with that of Salmonella minnesota or Staphylococcus aureus. The results show that A otitidis has a unique immunostimulatory capacity in vitro that is mainly confined to CD8+ T lymphocytes.

Apoptosis in auditory brainstem neurons after a severe noise trauma of the organ of Corti: intracochlear GDNF treatment reduces the number of apoptotic cells.

We have studied the morphological and cellular changes in the cochlear nucleus (CN) after cochlear nerve degeneration and whether these changes can be prevented by rescuing the primary cochlear neurons from degeneration with local glial cell line derived neurotrophic factor (GDNF) treatment. Degeneration of spiral ganglion neurons was seen to lead to a reduction of the volume of the anteroventral cochlear nucleus (AVCN); the size of the cell nuclei in the AVCN also was reduced. No differences were observed in cell density. After intrascalar GDNF treatment the volume of the AVCN was significantly larger when compared to the untreated side, and the size of the cell nuclei in the AVCN was significantly larger on the treated side. After degeneration of spiral ganglion neurons, an increased number of apoptotic cell nuclei were seen in ipsilateral CN and superior olivary complex. This increase was significantly smaller after intrascalar GDNF treatment. Degeneration of primary cochlear neurons seems to lead to an increase in the number of CN neurons undergoing apoptotic cell death. This can be prevented partially by rescuing primary cochlear neurons from degeneration with local GDNF treatment.

The relationship between audiogenic seizure (AGS) susceptibility and forebrain tone-responsiveness in genetically AGS-prone Wistar rats.

The present study characterized the intensity-response functions of extracellular field responsiveness of different cortical/subcortical structures of the forebrain following the free-field presentation of tone stimuli, within a population of genetically audiogenic seizure (AGS)-prone KM-Wistar rats. The neural response properties of each case were compared to its propensity to exhibit AGSs during the continuous tone stimulation (15 kHz, 90 s at max.). The amplitudes or slope components of the evoked responses and their peak latencies showed significant positive (amplitude and slope) and negative (peak latency) Bolzmann's sigmoid relationships with the onset-latency of AGS. These relationships, with areal differences in the slopes of saturation functions, applied for the three different data sets recorded simultaneously from the stratum radiatum dendritic layer of the hippocampal CA1 area, primary auditory cortex layers II-IV, and frontal cortex surface. In addition, the similar type of functions between the evoked response variables and AGS onset latency held when all the areas were considered together. These data suggest that the neural responsiveness to acoustic stimulation of the primary sensory, multimodal and association cortices of the forebrain may altogether contribute to the seizure initiation by that modality in the genetically AGS-prone rats. It has been previously shown that there exist abundant and dispersed auditory projections from these forebrain areas to the brain stem and spinal cord, structures that are generally considered to be the key predisposing factors in the generation of AGS. Hence, the types of correlation found reflect the subject-specific stage of forebrain responsiveness, being either related or unrelated to genetic AGS-specific changes, and possibly its triggering impact upon the lower brain AGS network. Accordingly, the mere comparison of forebrain response measures of these AGS-prone animals with those of the AGS-resistant ones could not reveal the result presented.

Interleukin-8 secretion of human epithelial and monocytic cell lines induced by middle ear pathogens.

Otitis media with effusion (OME) is one of the most common diseases in children. Alloiococcus otitidis, a new gram-positive bacterial species, was isolated from the middle ear fluid of children with OME; however, the pathogenic role of this bacteria is yet unknown. In this study, the ability of cultured epithelial cell lines (Hep-2 and Hela) and monocytic cell lines (THP-1 and U 937) to secrete chemokine interleukin-8 (IL-8) in response to the A. otitidis organism and three bacterial organisms mainly detected from middle ear fluid in OME, and bacterial cell components was investigated. When stimulated with four viable bacterial cells, epithelial cells and monocytes secreted IL-8 in a time-dependent manner. The monocytes produced significantly higher levels of IL-8 than the epithelial cells. Compared with that by viable bacterial cells, IL-8 secretion by stimulated epithelial cells and monocytes was reduced when the bacteria were heated and treated with glutaraldehyde. With bacterial stimulations, cell treatment of interferon-gamma caused monocytes to increase the induction of IL-8 production, however, the induction of monocyte differentiation caused monocytes to reduce the induction of IL-8 production. Furthermore, epithelial cells and monocytes stimulated by four viable bacterial organisms physically separated from cultured cells reduced the induction of IL-8 compared with directly stimulated cells, and monocytes stimulated with soluble extracts prepared from A. otitidis organisms produced IL-8 in a dose-dependent manner. These results suggest that part of the IL-8 stimulation of the A. otitidis organism may exist in a diffusable factor released by the bacteria or soluble components of the bacteria itself.

FGF/FGFR-2(IIIb) signaling is essential for inner ear morphogenesis.

Interactions between FGF10 and the IIIb isoform of FGFR-2 appear to be crucial for the induction and growth of several organs, particularly those that involve budding morphogenesis. We determined their expression patterns in the inner ear and analyzed the inner ear phenotype of mice specifically deleted for the IIIb isoform of FGFR-2. FGF10 and FGFR-2(IIIb) mRNAs showed distinct, largely nonoverlapping expression patterns in the undifferentiated otic epithelium. Subsequently, FGF10 mRNA became confined to the presumptive cochlear and vestibular sensory epithelia and to the neuronal precursors and neurons. FGFR-2(IIIb) mRNA was expressed in the nonsensory epithelium of the otocyst that gives rise to structures such as the endolymphatic and semicircular ducts. These data suggest that in contrast to mesenchymal-epithelial-based FGF10 signaling demonstrated for other organs, the inner ear seems to depend on paracrine signals that operate within the epithelium. Expression of FGF10 mRNA partly overlapped with FGF3 mRNA in the sensory regions, suggesting that they may form parallel signaling pathways within the otic epithelium. In addition, hindbrain-derived FGF3 might regulate otocyst morphogenesis through FGFR-2(IIIb). Targeted deletion of FGFR-2(IIIb) resulted in severe dysgenesis of the cochleovestibular membraneous labyrinth, caused by a failure in morphogenesis at the otocyst stage. In addition to the nonsensory epithelium, sensory patches and the cochleovestibular ganglion remained at a rudimentary stage. Our findings provide genetic evidence that signaling by FGFR-2(IIIb) is critical for the morphological development of the inner ear.

Puromycin-induced lipid peroxidation in the cochlea of ApoE knockout mice.

Puromycin-treated apolipoprotein E (ApoE)-deficient mice were used to study lipid peroxidation (LPO) in the cochlea. Puromycin causes accelerated peroxidation of lipids and induces both inner ear and renal lesions in experimental animals presenting with abnormally high serum cholesterol. To prevent LPO, we used probucol, an effective inhibitor of LPO, and, simultaneously, also a lipid-lowering drug. The mice were given a single injection of the aminonucleoside of puromycin (25 mg/100 g). Polyclonal malondialdehyde and 4-hydroxynonenal antibodies were used to localize the LPO products. LPO products were mainly found in the stria vascularis of puromycin-treated mice. No LPO products were observed in the hair cells. LPO product immunoreactivity was clearly diminished in the animal group treated with both puromycin and probucol. In the cochlea of the ApoE-deficient mouse, puromycin affects mainly the stria vascularis due to the accelerated peroxidation of structural lipids. Probucol treatment prevented the formation of LPO products.

Efficiency of hyperbaric oxygen therapy in experimental acute acoustic trauma from firearms.

In acute acoustic trauma (AAT), excessive noise exposure causes rupture of cell membranes and decreased cochlear blood flow. This leads to decreased oxygen tension in inner ear fluids and reduction of a variety of different oxygen-dependent cellular activities. Hyperbaric oxygen treatment (HBO) may help the cells suffering from hypoxia to survive. We exposed male Wistar rats to 60 impulses of 162-dB SPL from a 7.62-mm assault rifle equipped with a blank adaptor. After the exposure, 15 animals were given HBO treatment for 90 min daily for 10 consecutive days at 0.25 MPa. After a survival time of 4 weeks, auditory brainstem responses were measured and the left cochleae processed for light microscopy. The impulse noise caused permanent damage to the cochlea of all animals, with the most severe lesions in the lower middle coil, where a significantly smaller number of hair cells was missing in the HBO-treated group. The morphological damage was also reflected in function, as measured by auditory brainstem responses, which showed the greatest threshold shifts at 6.0, 8.0 and 10.0 kHz.

Intratympanic and systemic dexamethasone for Ménière's disease.

Intratympanic and systemic dexamethasone treatment of Ménière's disease (MD) was evaluated in a prospective study. Seventeen patients (6 men and 11 women) with MD (5 right-sided, 11 left-sided and 1 bilateral) were treated with three 0.2- to 0.4-ml injections of intratympanic dexamethasone hyaluronate (16 mg/ml) during a week and with an initial intramuscular dexamethasone injection of 15 mg. Most of patients were in stage 3, and the mean duration of MD was 5. 3 years. Pure-tone and speech audiometry and the symptom scale of the patients were followed up for 1 year after the treatment. Symptoms of aural fullness, hearing loss, tinnitus and vertigo did not improve significantly. However, sufficient control of vertigo was achieved in 76% of the patients. In conclusion, no definite treatment effect has yet been shown for intratympanic and systemic dexamethasone treatment. Therefore, the clinical use of dexamethasone in MD needs further investigation.

Lack of calbindin-D28k does not affect hearing level or survival of hair cells in acoustic trauma.

Calbindin is a cytosolic calcium-binding protein abundant in the hair cells of the inner ear and in distinct neurons of the auditory pathway. It is suggested to speed the return of potentially toxic calcium levels to normal. In this study, we show the basic hearing functions and the result of noise trauma from the calbindin null mutant mice generated by gene targeting. Auditory brainstem evoked response and distortion product otoacoustic emissions appear similar as in the control group. A moderate noise-induced trauma produced a similar loss of hair cells in calbindin null mutant mice than in wild-type controls. The result suggests that although calbindin is abundant in hair cells, it is not essential for the main hearing function and it does not provide physiological protection against a moderate noise-induced inner ear trauma in mice.

Rescue of hearing, auditory hair cells, and neurons by CEP-1347/KT7515, an inhibitor of c-Jun N-terminal kinase activation.

We have studied the mechanisms of auditory hair cell death after insults in vitro and in vivo. We show DNA fragmentation of hair cell nuclei after ototoxic drug and intense noise trauma. By using phospho-specific c-Jun-N-terminal kinase (JNK) and c-Jun antibodies in immunohistochemistry, we show that the JNK pathway, associated with stress, injury, and apoptosis, is activated in hair cells after trauma. CEP-1347, a derivative of the indolocarbazole K252a, is a small molecule that has been shown to attenuate neurodegeneration by blocking the activation of JNK (). Subcutaneously delivered CEP-1347 attenuated noise-induced hearing loss. The protective effect was demonstrated by functional tests, which showed less hearing threshold shift in CEP-1347-treated than in nontreated guinea pigs, and by morphometric methods showing less hair cell death in CEP-1347-treated cochleas. In organotypic cochlear cultures, CEP-1347 prevented neomycin-induced hair cell death. In addition to hair cells, CEP-1347 promoted survival of dissociated cochlear neurons. These results suggest that therapeutic intervention in the JNK signaling cascade, possibly by using CEP-1347, may offer opportunities to treat inner ear injuries.

Clinically applicable multiplex PCR for four middle ear pathogens.

The multiplex PCR method for the detection of Alloiococcus otitidis, Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae (P. H. Hendolin, A. Markkanen, J. Ylikoski, and J. J. Wahlfors, J. Clin. Microbiol. 35:2854-2858, 1997) in middle ear effusions (MEEs) was modified to be better suited for clinical use. To detect false-negative results, an internal amplification was added to the reaction, and to prevent carryover contamination, the dUTP-uracil-N-glycosidase system was incorporated into the procedure. Labor was minimized by using the heat-activatable AmpliTaq Gold polymerase in order to circumvent manual hot start and by detecting the amplification products on an automated sequencer. The performance of the improved protocol was verified with MEEs from patients with otitis media with effusion. In addition, a ligase detection reaction (LDR) was developed for confirmation of the PCR products. The modifications increased the reliability of the protocol and the hands-off time significantly. However, when two DNA extraction protocols were compared, gram-negative bacteria were detected more often in phenol-treated MEEs (94 versus 46%; P < 0.001), and gram-positive bacteria were detected more often in MEEs dissolved in sodium dodecyl sulfate-NaOH-chaotropic salt (83 versus 27%; P < 0.001). The LDR was found to be 100% specific. In all, the results demonstrate the feasibility of the rapid (7-h) multiplex PCR method for routine laboratory use.