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Accumulation of mitochondrial DNA mutations in the aging rat ear
Otolaryngology– Head and Neck Surgery Volume 129 Number 2
AM
Newborn and Childhood Hearing Loss: The Effect of Newborn Hearing Screening Scott Austin Schraff MD (presenter); Craig S Derkay MD Norfolk VA; Norfolk VA
Problem: To study the effect of newborn hearing screening by identifying and characterizing hearing loss in a consecutive cohort of newborns and children referred to our tertiary-care children’s hospital. Methods: The records of pediatric patient with the diagnosis of sensorineural hearing loss (SNHL), mixed hearing loss, and unspecified hearing loss from June 1, 1999, to December 1, 2002, were reviewed. Data collected included prenatal and postnatal risk factors for hearing loss, method of identification, diagnostic work-up, and hearing loss evaluation results. Results: 508 pediatric patient records with a hearing loss diagnosis were reviewed, of which 436 had all relevant data. 26 of 436 presented after failing universal newborn screening, and 20 of these failed additional testing and were diagnosed with hearing loss. These children had a median age of 4 months (interquartile range [IQR], 3-8 months) and 7 (35%) had no hearing loss risk factors. Of the additional 410 referred through traditional screening methods, 286 (70%) were eventually diagnosed with hearing loss. The median age of these children was 5 years (IQR, 2-9 years). The false-positive rate was 23% for UNHS and was 30% for traditional screening. In total, 306 (70%) of the 436 children referred for hearing evaluation actually had hearing loss. Conclusion: UNHS has markedly reduced the median age of diagnosis of those with newborn and childhood hearing loss and identified a significant percentage with no risk factors. Significance: With the institution of UNHS, we anticipate dramatic medical, social, and economic benefits for children born with hearing loss. Support: None reported.
11:20
AM
Evaluation of Relapses and Health-Related Quality of Life in BPPV Jose A Lopez-Escamez MD (presenter); Maria J Gamiz MD; Manuel Gomez-Finana MD; Antonio Fernandez-Perez MD; Isabel Sanchez-Canet MD El Ejido Spain; El Ejido Spain; El Ejido Spain; El Ejido Spain; El Ejido Spain
Problem: To evaluate the effect of relapses on healthrelated quality of life (HRQoL) in patients with benign paroxysmal positional vertigo (BPPV) treated by the particlerepositioning maneuver (PRM) using the MOS 36-Item Short
Form Health Survey (SF-36) and the Dizziness Handicap Inventory Short Form (DHI-S). Methods: Study design: Prospective, new cases of BPPV. Setting: Primary referral hospital. Patients: 40 individuals with BPPV were investigated. The diagnosis was based on the history of recurrent sudden crisis of vertigo and positionalinduced nystagmus during the Dix-Hallpike test (DHT). Interventions: All patients were treated by a single PRM and relapses were evaluated by DHT at 7, 30, 180, and 360 days post-treatment. A new PRM was performed if DHT was positive. Main outcome measures: Percentage of patients with negative DHT after treatment and scores obtained on SF-36 and DHI-S before and 30, 180, and 360 days post-treatment. Results: DHT was found to be negative in 76% (28/37) individuals at 30 days. Nine, seven, and four patients remained DH⫹ at 30, 180, and 360 days, respectively. The SF-36 and DHI-S had a good reliability (Cronbach’s alpha ⬎.7) in patients with BPPV. The average standardized score for each SF-36 scale was compared with the reference population normative data, showing differences with norms for all scales, except for vitality. After PRM, patients improved their scores with both instruments, indicating a restoration of HRQoL at 30 days. However, SF-36 standardized scores and DHI-S scores were not different between individuals with positive and negative DHT at 180 or 360 days, except for general health perception. Physical dimension scores improved from day 30 to 360. Conclusion: Patients with BPPV experienced a decrease in HRQoL which was restored by PRM. Relapses do not affect HRQoL at 180 or 360 days and physical and general health perception was maintained after PRM. Significance: PRM restores HRQoL in BPPV at long term. Support: Research Project 132/00 Consejeria de Salud, Junta de Andalucia, Spain; FIS PI021394 Project, Instituto de Saluc Carlos III, Spain 11:30
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Accumulation of Mitochondrial DNA Mutations in the Aging Rat Ear Hinrich Staecker MD PhD; Kim Baker MD (presenter) Baltimore MD; Baltimore MD
Problem: To determine the effect of aging on accumulation of mitochondrial DNA mutations and mitochondrial gene expression in the inner ear. Methods: Six- and twenty-month-old Fisher rats were obtained and hearing tested by auditory brainstem response (ABR). Rats were then divided into 2 groups. Group 1 rats were anesthetized and fixed via intracardiac perfusion with paraformaldehyde. The cochleae were removed and processed for histology. Hair cell counts, neuron counts, and measurements of the stria vascularis were made. Group 2 animals were anesthetized and decapitated. The cochleae
TUESDAY
11:00
Research Forum—Tuesday P103
Research Forum—Tuesday
were immediately removed. The cochleae were opened and the stria vascularis, the organ of Corti region, and the spiral ganglion removed by microdissection. The tissue then underwent mRNA and DNA extraction. Mitochondrial DNA deletions were assayed by PCR using rat primers and a Roche Lightcycler. Levels of mitochondrial gene expression were determined by semiquantitative RT-PCR. Results: Aged Fisher rats showed an average 25-dB hearing loss compared to young animals. There was a significant age-related accumulation of mitochondrial DNA deletions in the DNA from the spiral ganglion and the organ of Corti, as well as the stria vascularis. There was an age-related decrease in mitochondrial gene expression. Conclusion: There are age-related changes in mitochondria in the rat inner ear. The accumulation of large DNA deletions is demonstrated by this study, as are changes in overall mitochondrial gene expression. Data from aging in the central nervous system suggest that damage to mitochondrial DNA may be associated with a variety of neurodegenerative diseases. These data demonstrate that aging affects mitochondrial biology in the inner ear. Significance: Clinical correlates: Oxidative stress induced by otoxins and sound trauma may be a source of mitochondrial damage, and strategies to protect mitochondrial function may be beneficial in protecting inner ear function over time. Support: None reported. 11:40
while 30% presented worse results. In the group of cytalopran-treated patients 85.7% presented better performance in central auditory evaluation post-treatment while the remaining 14.3% presented no variation. Conclusion: This preliminary report presents a small number of patients that preclude statistical significance but indicate that the use of inhibitors of serotonin reuptake could be considered a therapeutic option in patients over 60 years old with auditory processing disorders. Significance: Possibility of improvement of auditory processing by increasing serotonin neurotransmitter. Support: Samples of cytalopran by Schering-Plough; placebo by Unipharmus Pharmacy
Room OCCC 203 •
Session Category: Session B: Head and Neck Surgery and Oncology Douglas Chepeha MD; Terry Day MD; Ted Teknos MD (moderators)
8:00
AM
CⴚMet Signaling in Head and Neck Squamous Cell Carcinoma Brian F Worden (presenter); Xinping Yang PhD; Bean Lee PhD; Carter Van Waes MD; Zhong Chen MD PhD Bethesda MD; Bethesda MD; Bethesda MD; Gaithersburg MD; Bethesda MD
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Serotonin Reuptake Inhibitors in Auditory Processing Disorders in Elderly Patients Oswaldo Laercio M Cruz MD; Cristiane Kasse MD (presenter); Maura Sanchez MA; Flavia A Barros MA; Flavio Barbosa MD Sao Paulo Brazil; Sao Paulo Brazil; Sao Paulo Brazil; Sao Paulo Brazil; Sao Paulo Brazil
Problem: Impairment of hearing cognition is commonly seen in hearing difficulties in the elderly. The decrease of neurotransmitters at brain synapses in more advanced age is one explanation for this fact, probably involving the serotonergic system. This prospective study analyzes the efficacy of a serotonin reuptake inhibitor (cytalopran) in improving auditory processing in patients over 60 years old. Methods: This is a preliminary report of a prospective, double-blind, placebo-controlled, randomized study. Initially, 28 subjects of both genders, aged 60 to 79 years, were analyzed. 14 patients received placebo for 2 months and 14 patients received 40 mg/d of cytalopran for 2 months. Audiometric and central auditory tests (Synthetic Sentence Identification with ipsilateral competitive message; Staggered Spondaic Word Test; Pitch Pattern Sequence Test) were performed before and after medication therapy. Results: In the placebo group 60% of patients presented a better performance in central auditory tests post-treatment
Problem: Constitutive and hepatocyte growth factor/scatter factor (HGF)–induced activation of c-Met is important in promoting tumor progression and metastasis in head and neck squamous cell carcinoma (HNSCC). We previously observed that increased serum levels of HGF in HNSCC patients were correlated with elevated serum levels of angiogenic factors VEGF and IL-8. However, the pathways downstream from c-Met responsible for promoting tumor progression and metastasis in HNSCC are poorly understood. Methods: cDNA microarrays were utilized to analyze gene expression profiles in ten UMSCC cell lines and normal human keratinocytes. Isolated RNA was used to generate Cy3- or Cy5-labeled cDNA by reverse transcription and the labeled cDNA products were hybridized to array slides. Fluorescent images were captured using a Genepix 4000, and data were analyzed by IP-lab software (Scanalytics/NHGRI/ NIH). ELISA-based DNA-binding assays (BD Biosystems) were utilized to investigate the activation of sixteen transcription factors involved in oncogenesis and inflammation. Nuclear extracts were isolated from UMSCC11A cells treated with recombinant human HGF, and DNA-binding assays were conducted according to manufacturer instructions. Results: Microarray results revealed the overexpression in HNSCC cells of numerous gene products known to be involved in signaling pathways downstream from c-Met includ-
TUESDAY
P104
Otolaryngology– Head and Neck Surgery August 2003
AM
Newborn and Childhood Hearing Loss: The Effect of Newborn Hearing Screening Scott Austin Schraff MD (presenter); Craig S Derkay MD Norfolk VA; Norfolk VA
Problem: To study the effect of newborn hearing screening by identifying and characterizing hearing loss in a consecutive cohort of newborns and children referred to our tertiary-care children’s hospital. Methods: The records of pediatric patient with the diagnosis of sensorineural hearing loss (SNHL), mixed hearing loss, and unspecified hearing loss from June 1, 1999, to December 1, 2002, were reviewed. Data collected included prenatal and postnatal risk factors for hearing loss, method of identification, diagnostic work-up, and hearing loss evaluation results. Results: 508 pediatric patient records with a hearing loss diagnosis were reviewed, of which 436 had all relevant data. 26 of 436 presented after failing universal newborn screening, and 20 of these failed additional testing and were diagnosed with hearing loss. These children had a median age of 4 months (interquartile range [IQR], 3-8 months) and 7 (35%) had no hearing loss risk factors. Of the additional 410 referred through traditional screening methods, 286 (70%) were eventually diagnosed with hearing loss. The median age of these children was 5 years (IQR, 2-9 years). The false-positive rate was 23% for UNHS and was 30% for traditional screening. In total, 306 (70%) of the 436 children referred for hearing evaluation actually had hearing loss. Conclusion: UNHS has markedly reduced the median age of diagnosis of those with newborn and childhood hearing loss and identified a significant percentage with no risk factors. Significance: With the institution of UNHS, we anticipate dramatic medical, social, and economic benefits for children born with hearing loss. Support: None reported.
11:20
AM
Evaluation of Relapses and Health-Related Quality of Life in BPPV Jose A Lopez-Escamez MD (presenter); Maria J Gamiz MD; Manuel Gomez-Finana MD; Antonio Fernandez-Perez MD; Isabel Sanchez-Canet MD El Ejido Spain; El Ejido Spain; El Ejido Spain; El Ejido Spain; El Ejido Spain
Problem: To evaluate the effect of relapses on healthrelated quality of life (HRQoL) in patients with benign paroxysmal positional vertigo (BPPV) treated by the particlerepositioning maneuver (PRM) using the MOS 36-Item Short
Form Health Survey (SF-36) and the Dizziness Handicap Inventory Short Form (DHI-S). Methods: Study design: Prospective, new cases of BPPV. Setting: Primary referral hospital. Patients: 40 individuals with BPPV were investigated. The diagnosis was based on the history of recurrent sudden crisis of vertigo and positionalinduced nystagmus during the Dix-Hallpike test (DHT). Interventions: All patients were treated by a single PRM and relapses were evaluated by DHT at 7, 30, 180, and 360 days post-treatment. A new PRM was performed if DHT was positive. Main outcome measures: Percentage of patients with negative DHT after treatment and scores obtained on SF-36 and DHI-S before and 30, 180, and 360 days post-treatment. Results: DHT was found to be negative in 76% (28/37) individuals at 30 days. Nine, seven, and four patients remained DH⫹ at 30, 180, and 360 days, respectively. The SF-36 and DHI-S had a good reliability (Cronbach’s alpha ⬎.7) in patients with BPPV. The average standardized score for each SF-36 scale was compared with the reference population normative data, showing differences with norms for all scales, except for vitality. After PRM, patients improved their scores with both instruments, indicating a restoration of HRQoL at 30 days. However, SF-36 standardized scores and DHI-S scores were not different between individuals with positive and negative DHT at 180 or 360 days, except for general health perception. Physical dimension scores improved from day 30 to 360. Conclusion: Patients with BPPV experienced a decrease in HRQoL which was restored by PRM. Relapses do not affect HRQoL at 180 or 360 days and physical and general health perception was maintained after PRM. Significance: PRM restores HRQoL in BPPV at long term. Support: Research Project 132/00 Consejeria de Salud, Junta de Andalucia, Spain; FIS PI021394 Project, Instituto de Saluc Carlos III, Spain 11:30
AM
Accumulation of Mitochondrial DNA Mutations in the Aging Rat Ear Hinrich Staecker MD PhD; Kim Baker MD (presenter) Baltimore MD; Baltimore MD
Problem: To determine the effect of aging on accumulation of mitochondrial DNA mutations and mitochondrial gene expression in the inner ear. Methods: Six- and twenty-month-old Fisher rats were obtained and hearing tested by auditory brainstem response (ABR). Rats were then divided into 2 groups. Group 1 rats were anesthetized and fixed via intracardiac perfusion with paraformaldehyde. The cochleae were removed and processed for histology. Hair cell counts, neuron counts, and measurements of the stria vascularis were made. Group 2 animals were anesthetized and decapitated. The cochleae
TUESDAY
11:00
Research Forum—Tuesday P103
Research Forum—Tuesday
were immediately removed. The cochleae were opened and the stria vascularis, the organ of Corti region, and the spiral ganglion removed by microdissection. The tissue then underwent mRNA and DNA extraction. Mitochondrial DNA deletions were assayed by PCR using rat primers and a Roche Lightcycler. Levels of mitochondrial gene expression were determined by semiquantitative RT-PCR. Results: Aged Fisher rats showed an average 25-dB hearing loss compared to young animals. There was a significant age-related accumulation of mitochondrial DNA deletions in the DNA from the spiral ganglion and the organ of Corti, as well as the stria vascularis. There was an age-related decrease in mitochondrial gene expression. Conclusion: There are age-related changes in mitochondria in the rat inner ear. The accumulation of large DNA deletions is demonstrated by this study, as are changes in overall mitochondrial gene expression. Data from aging in the central nervous system suggest that damage to mitochondrial DNA may be associated with a variety of neurodegenerative diseases. These data demonstrate that aging affects mitochondrial biology in the inner ear. Significance: Clinical correlates: Oxidative stress induced by otoxins and sound trauma may be a source of mitochondrial damage, and strategies to protect mitochondrial function may be beneficial in protecting inner ear function over time. Support: None reported. 11:40
while 30% presented worse results. In the group of cytalopran-treated patients 85.7% presented better performance in central auditory evaluation post-treatment while the remaining 14.3% presented no variation. Conclusion: This preliminary report presents a small number of patients that preclude statistical significance but indicate that the use of inhibitors of serotonin reuptake could be considered a therapeutic option in patients over 60 years old with auditory processing disorders. Significance: Possibility of improvement of auditory processing by increasing serotonin neurotransmitter. Support: Samples of cytalopran by Schering-Plough; placebo by Unipharmus Pharmacy
Room OCCC 203 •
Session Category: Session B: Head and Neck Surgery and Oncology Douglas Chepeha MD; Terry Day MD; Ted Teknos MD (moderators)
8:00
AM
CⴚMet Signaling in Head and Neck Squamous Cell Carcinoma Brian F Worden (presenter); Xinping Yang PhD; Bean Lee PhD; Carter Van Waes MD; Zhong Chen MD PhD Bethesda MD; Bethesda MD; Bethesda MD; Gaithersburg MD; Bethesda MD
AM
Serotonin Reuptake Inhibitors in Auditory Processing Disorders in Elderly Patients Oswaldo Laercio M Cruz MD; Cristiane Kasse MD (presenter); Maura Sanchez MA; Flavia A Barros MA; Flavio Barbosa MD Sao Paulo Brazil; Sao Paulo Brazil; Sao Paulo Brazil; Sao Paulo Brazil; Sao Paulo Brazil
Problem: Impairment of hearing cognition is commonly seen in hearing difficulties in the elderly. The decrease of neurotransmitters at brain synapses in more advanced age is one explanation for this fact, probably involving the serotonergic system. This prospective study analyzes the efficacy of a serotonin reuptake inhibitor (cytalopran) in improving auditory processing in patients over 60 years old. Methods: This is a preliminary report of a prospective, double-blind, placebo-controlled, randomized study. Initially, 28 subjects of both genders, aged 60 to 79 years, were analyzed. 14 patients received placebo for 2 months and 14 patients received 40 mg/d of cytalopran for 2 months. Audiometric and central auditory tests (Synthetic Sentence Identification with ipsilateral competitive message; Staggered Spondaic Word Test; Pitch Pattern Sequence Test) were performed before and after medication therapy. Results: In the placebo group 60% of patients presented a better performance in central auditory tests post-treatment
Problem: Constitutive and hepatocyte growth factor/scatter factor (HGF)–induced activation of c-Met is important in promoting tumor progression and metastasis in head and neck squamous cell carcinoma (HNSCC). We previously observed that increased serum levels of HGF in HNSCC patients were correlated with elevated serum levels of angiogenic factors VEGF and IL-8. However, the pathways downstream from c-Met responsible for promoting tumor progression and metastasis in HNSCC are poorly understood. Methods: cDNA microarrays were utilized to analyze gene expression profiles in ten UMSCC cell lines and normal human keratinocytes. Isolated RNA was used to generate Cy3- or Cy5-labeled cDNA by reverse transcription and the labeled cDNA products were hybridized to array slides. Fluorescent images were captured using a Genepix 4000, and data were analyzed by IP-lab software (Scanalytics/NHGRI/ NIH). ELISA-based DNA-binding assays (BD Biosystems) were utilized to investigate the activation of sixteen transcription factors involved in oncogenesis and inflammation. Nuclear extracts were isolated from UMSCC11A cells treated with recombinant human HGF, and DNA-binding assays were conducted according to manufacturer instructions. Results: Microarray results revealed the overexpression in HNSCC cells of numerous gene products known to be involved in signaling pathways downstream from c-Met includ-
TUESDAY
P104
Otolaryngology– Head and Neck Surgery August 2003