Protective effect of acetyl-l-carnitine against cisplatin ototoxicity: role of apoptosis-related genes and pro-inflammatory cytokines.

OBJECTIVES: Cisplatin is an anti-neoplastic agent treatment with which causes many side effects including ototoxicity. The aim of this study was to investigate whether acetyl-L-carnitine would have protective effects on cisplatin-induced ototoxicity in vitro, and if present, to reveal roles of apoptotic gene expressions and pro-inflammatory cytokines. MATERIALS AND METHODS: House Ear Institute-Organ of Corti 1 cell line was used for this study. Apoptotic genes were evaluated with an apoptosis PCR array and pro-inflammatory cytokine levels were measured using ELISA. RESULTS: Apoptotic cell death reduced by around 22% with acetyl-L-carnitine-cisplatin treatment compared to cisplatin alone. Genes displaying increase in expression of apoptosis, related to cisplatin treatment, were Casp8, Bcl10, Bcl2, Bcl2l1, Bcl2l2, Bid, Naip1, Bnip3l, Card6, Pak7, Cd40, Trp 53inp1, Cideb and Cd70. The acetyl-L-carnitine-cisplatin combination caused reduced expression of genes Casp8, Fas, Casp1, Tnfrsf11b, Tnfrsf10b induced by cisplatin. Acetyl-L-carnitine-cisplatin also caused reduced levels of IL-6, IL-1ß and TNF-α, pro-inflammatory cytokines, induced by cisplatin. CONCLUSION: Protective mechanisms of aceytl-L-carnitine against cisplatin induced apoptosis, mainly due to activation of anti-apoptotic Bcl family members' genes, and in an Akt-related gene expression dependent manner. This is the first study to indicate that acetyl-L-carnitine can be an effective agent against cisplatin ototoxicity in auditory cells, with induction of anti-apoptotic gene expression and attenuating levels of pro-inflammatory cytokines.

Results of multiple-frequency tympanometry measures in normal and otosclerotic middle ears.

Tympanometry is a non-invasive, quick, and inexpensive method for examining the middle-ear function. Its limited value in differentiating otosclerotic from normal middle ears caused researchers to develop new methods for evaluation of middle ears. Resonant frequency had been found to be higher in otosclerotic middle ears than normals. We conducted multiple-frequency tympanometry measurements in 25 surgically confirmed otosclerotic ears and 100 normal ears. Mean middle-ear resonant frequency for the otosclerotic group was found to be 1190 Hz and mean middle-ear resonant frequency of the control group was 934.6 Hz (p<0.001). With a cut off value of 1025 Hz (based on 95% confidence interval), sensitivity was 80% and specificity was 82%. The present findings confirm the advantage of the resonant frequency estimation over conventional tympanometry in detecting middle-ear status and mechanics in patients with otosclerosis. As a conclusion, detecting resonant frequency when evaluating patients for otosclerosis must be an essential part of examination. Nevertheless, further investigation is necessary for better diagnosis of otosclerosis preoperatively.

Long-term follow-up of otherwise healthy term infants with marked hyperbilirubinaemia: should the limits of exchange transfusion be changed in Turkey?

AIM: To evaluate prospectively non-haemolytic term infants with marked hyperbilirubinaemia treated by phototherapy only for evidence of bilirubin toxicity at 2-6 y of age, and to determine the suitability for Turkish children of the exchange transfusion limits recently reported by the American Academy of Pediatrics. METHODS: The study group included a total of 30 children, aged 2-6 y, who had developed marked hyperbilirubinaemia (20-24 mg dl(-1), 342-410 micromol l(-1)) during the newborn period (gestational age >37 wk, birthweight >2500 g) and were treated without exchange transfusion because intensive phototherapy, instituted during the preparations for exchange transfusion, was successful in decreasing their serum bilirubin levels. The control group consisted of 30 children of the same age group without clinical jaundice in the newborn period. Physical and neurological examinations, brainstem auditory-evoked potentials (BAEPs) and developmental tests for Turkish children were performed in both the study and control children. RESULTS: There was no difference between the groups with regard to mean BAEP latencies and developmental scores. None of the infants had hearing loss, developmental delay or abnormal neurological findings. CONCLUSION: The results suggest that successful intensive phototherapy without exchange transfusion in otherwise healthy term newborn infants with marked hyperbilirubinaemia (20-24 mg dl(-1), 342-410 micromol l(-1)) might not increase the risk of bilirubin brain injury and that the conventional limit of 20 mg dl(-1) (342 micromol l(-1)) could be changed to 22-24 mg dl(-1) (376-410 micromol l(-1)) for healthy term infants in Turkey. These limits, however, address only infants who do not have haemolytic disease, and the data are not sufficient to draw conclusions on the safety of even higher bilirubin levels (i.e. >24 mg dl(-1), 410 micromol l(-1)) in this population.

Long-term outcome of neonatal hyperbilirubinaemia: subjective and objective audiological measures.

Neonatal hyperbilirubinaemia is a common cause of early onset sensorineural hearing loss. There is no exact method to detect the extent of the neurotoxicity of bilirubin. On the other hand, the auditory pathway is known to be one of the most sensitive parts of the central nervous system (CNS) to this toxic agent. This prospective follow-up study was performed to evaluate and compare the factors related to the hearing of neonates with severe hyperbilirubinaemia and an age-matched control group. Both of these groups were tested using auditory brainstem response (ABR) as well as evoked otoacoustic emissions. Additionally, both of these groups of children were evaluated subjectively using an early speech-language-communication evaluation questionnaire. There was no significant difference in either objective (ABR and evoked otoacoustic emission) or subjective assessment (questionnaire) between the study and control groups. Furthermore, no correlation between serum total bilirubin levels and ABR latencies or thresholds was found within the study group.

TEOAE monitoring of Cisplatin induced ototoxicity in guinea pigs: the protective effect of vitamin B treatment.

OBJECTIVE: To evaluate Cisplatin (CP) induced ototoxicity and the effects of vitamin B treatment on ototoxicity in guinea pigs by using the Transient Evoked Otoacoustic Emission (TEOAE) technique. METHODS: Eleven guinea pigs were divided into two groups and they were tested by TEOAE before and after the experiment. A TEOAE response was regarded as positive when all of the following criteria were met: 1. The mean amplitude of the cochlear response in dB pe SPL should be greater than that of the noise in the external auditory canal; 2. The reproducibility rate of the response should be greater than 50%; 3. The stimulus stability rate should be greater than 65%; 4. The signal to noise ratio of the response in 1, 2, 3, 4 and 5 kHz band frequencies should be greater than 3 dB pe SPL in at least two bands. The first group included five animals that had only CP injections. Six animals in the second group received additional 0.2 ml/kg combined vitamin B preparations for 7 consecutive days. Thereafter, the right and left ears of all animals in both groups were tested by TEOAE. RESULTS: TEOAE responses recorded from 22 ears of 11 guinea pigs before drug administrations showed that the responses with maximum amplitude were originated from the mid-frequency region. Positive TEOAE responses were significantly reduced after CP administrations in both groups when compared with their respective pretreatment results (P<0.01). However, vitamin B injections, in addition to a single large dose of CP, resulted in significantly better TEOAE responses than those obtained after only CP injections (P<0.05). CONCLUSIONS: The routine use of TEOAE monitoring is recommended in clinical CP treatment protocols for the early detection and follow up of ototoxicity. Also, prospective clinical trials are needed in order to validate the protective effects of vitamin B treatment against ototoxicity.

Investigation of topical ciprofloxacin ototoxicity in guinea pigs.

Antibiotic eardrops mostly contain potentially ototoxic aminoglycosides. Ciprofloxacin is an alternative, and there is limited experience in its topical use. To investigate the topical ototoxicity of ciprofloxacin, 11 guinea pigs have been operated on. Transbullae silicone drug delivery tubes were placed to both ears of the animals. After the operation the guinea pigs were divided into two groups. The first group of animals received 0.2 ml of 4% gentamicin in one ear and 0.2 ml of 0.9% sodium chloride solution in the other. The second group received 0.2 ml of 0.2% ciprofloxacin in the test ear and 0.2 ml of 0.9% sodium chloride solution in the control ear. All drugs were given once a day on 7 consecutive days. Auditory brainstem response thresholds were recorded using click, 4 and 8 kHz logon stimuli before and after the operation, and after topical drug application. Results were statistically compared using Wilcoxon matched pairs signed-ranks test. Comparison of the thresholds before and after the operation, physiological saline application, as well as ciprofloxacin application yielded no statistically significant differences, whereas application of gentamicin resulted in total hearing loss. The results indicate that topical use of 0.2% ciprofloxacin is not ototoxic in guinea pigs.