AUDITORY TEMPORAL PROCESSING TESTS - NORMATIVE DATA FOR POLISH-SPEAKING ADULTS.

BACKGROUND: Several subjects exposed to neurotoxins in the workplace need to be assessed for central auditory deficit. Although central auditory processing tests are widely used in other countries, they have not been standardized for the Polish population. The aim of the study has been to evaluate the range of reference values for 3 temporal processing tests: the duration pattern test (DPT), the frequency pattern test (FPT) and the gaps in noise test (GIN). MATERIAL AND METHODS: The study included 76 normal hearing individuals (38 women, 38 men) at the age of 18 to 54 years old (mean ± standard deviation: 39.4±9.1). All study participants had no history of any chronic disease and underwent a standard ENT examination. RESULTS: The reference range for the DPT was established at 55.3% or more of correct answers, while for the FPT it stood at 56.7% or more of correct answers. The mean threshold for both cars in the GIN test was defined as 6 ms. In this study there were no significant associations between the DPT, FPT and GIN results and age or gender. Symmetry between the ears in the case of the DPT, FPT and GIN was found. CONCLUSIONS: Reference ranges obtained in this study for the DPT and FPT in the Polish population are lower than reference ranges previously published for other nations while the GIN test results correspond to those published in the related literature. Further investigations are needed to explain the discrepancies between normative values in Poland and other countries and adapt tests for occupational medicine purposes.

Genetic variants of CDH23 associated with noise-induced hearing loss.

OBJECTIVES: Noise-induced hearing loss (NIHL) is a complex disease resulting from the interaction between external and intrinsic/genetic factors. Based on mice studies, one of the most interesting candidate gene for NIHL susceptibility is CDH23-encoding cadherin 23, a component of the stereocilia tip links. The aim of this study was to analyze selected CDH23 single nucleotide polymorphisms (SNPs) and to evaluate their interaction with environmental and individual factors in respect to susceptibility for NIHL in humans. METHODS: A study group consisted of 314 worst-hearing and 313 best-hearing subjects exposed to occupational noise, selected out of 3,860 workers database. Five SNPs in CDH23 were genotyped using real-time PCR. Subsequently, the main effect of genotype and its interaction with selected environmental and individual factors were evaluated. RESULTS: The significant results within the main effect of genotype were obtained for the SNP rs3752752, localized in exon 21. The effect was observed in particular in the subgroup of young subjects and in those exposed to impulse noise; CC genotype was more frequent among susceptible subjects, whereas genotype CT appeared more often among resistant to noise subjects. The effect of this polymorphism was not modified by none of environmental/individual factors except for blood pressure; however, the latter one should be further investigated. Smoking was shown as an independent factor determining NIHL development. CONCLUSION: The results of this study confirm that CDH23 genetic variant may modify the susceptibility to NIHL development in humans, as it was earlier proven in mice. Because the differences between the 2 study groups were not necessarily related to susceptibility to noise but they also were prone to age-related cochlear changes, these results should be interpreted with caution until replication in another population.

Combined effect of X-ray radiation and static magnetic fields on reactive oxygen species in rat lymphocytes in vitro.

The aim of this study was to investigate the effect of static magnetic fields (SMF) on reactive oxygen species induced by X-ray radiation. The experiments were performed on lymphocytes from male albino Wistar rats. After exposure to 3 Gy X-ray radiation (with a dose rate of 560 mGy/min) the measurement of intracellular reactive oxygen species in lymphocytes, using a fluorescent probe, was done before exposure to the SMF, and after 15 min, 1 and 2 h of exposure to the SMF or a corresponding incubation time. For SMF exposure, 0 mT (50 µT magnetic field induction opposite to the geomagnetic field) and 5 mT fields were chosen. The trend of SMF effects for 0 mT was always opposite that of 5 mT. The first one decreased the rate of fluorescence change, while the latter one increased it.

Analysis of inner ear potassium recycling genes as potential factors associated with tinnitus.

UNLABELLED: Tinnitus is defined as a perception of sound in the absence of an external acoustic stimulus. Several factors are known to influence tinnitus, e.g. hearing loss, noise exposure, age, and hypertension. As only certain individuals develop tinnitus in the presence of the above risks and in approximately 50% of cases tinnitus is not attributed to any particular cause, the question arose whether this inter-individual susceptibility to tinnitus could be explained by the influence of genetic factors. OBJECTIVES: To test the hypothesis that genetic variability in genes of the potassium recycling pathway is associated with increased susceptibility to tinnitus. MATERIALS AND METHODS: The study group consisted of 626 subjects exposed to occupational noise (128 with tinnitus and 498 without tinnitus). 99 single nucleotide polymorphisms were investigated in 10 genes involved in the potassium recycling pathway in the inner ear, previously selected as putative noise-induced hearing loss (NIHL) candidate genes. RESULTS: Nominally significant associations were obtained for 2 variants in KCNE1 (potassium voltage-gated channel, Isk-related family, member 1) and SLC12A2 (solute carrier family 12, member 2) genes. The first gene contributed to tinnitus that developed independently of hearing loss, while the second one was associated with increased susceptibility to noise-induced hearing loss. CONCLUSIONS: Present findings lend support to the notion of potassium recycling pathway genes as possible risk modifiers of tinnitus in individuals with and without hearing loss. Due to the lack of replication in other independent populations these results should be seen as suggestive.

Static magnetic field affects oxidative stress in mouse cochlea.

OBJECTIVE: It has been shown that oxidative stress plays an important role in development of noise induced hearing loss. Since static magnetic fields (SMF) exposure may alter dynamics of oxidative processes in the tissue, the aim of the study was to assess the influence of SMF on noise-induced alteration in the cochlear level of reactive oxygen species (ROS) and hearing thresholds. MATERIALS AND METHODS: Auditory brainstem response (ABR), lipid peroxidation (LPO) levels, super-oxide dismutase (SOD) activity and catalase activity were assessed in the cochlea prior to, and at five time-points over two weeks following exposure of C57BL/6 mice to 8h, 119 dB SPL, 4 kHz octave band noise. RESULTS: The ABR indicated no permanent functional damage due to noise exposure either for the 4 kHz and 8 kHz SMF-exposed group or for animals not exposed to SMF. However, significant differences in LPO level, catalase and SOD activity between animals exposed to noise and SMF and those exposed to noise only were observed. CONCLUSIONS: The results suggest that SMF causes an increase in ROS level in the cochlea after noise exposure and, at the same time, it speeds up activation of antioxidative enzymes.

Analysis of gene polymorphisms associated with K ion circulation in the inner ear of patients susceptible and resistant to noise-induced hearing loss.

Noise-induced hearing loss (NIHL) is one of the leading occupational health risks in industrialized countries. It results from an interaction between environmental and genetic factors, however the nature of the genetic factors contributing to NIHL has not yet been clarified. Here, we investigated whether genetic variations in 10 genes putatively involved in the potassium recycling pathway in the inner ear may influence susceptibility to noise. 99 SNPs were genotyped in Polish noise-exposed workers, categorized into susceptible and resistant subjects. The most interesting results were obtained for KCNE1 and KCNQ4 as we replicated associations that were previously reported in a Swedish sample set, hence confirming that they are NIHL susceptibility genes. Additionally we report significant associations in GJB1, GJB2, GJB4, KCNJ10 and KCNQ1, however due to the lack of replication in the Swedish sample set, these results should be seen as suggestive.

Variations in HSP70 genes associated with noise-induced hearing loss in two independent populations.

Noise-induced hearing loss (NIHL) is one of the most important occupational health hazards. Millions of people worldwide are exposed daily to harmful levels of noise. NIHL is a complex disease resulting from an interaction between genetic and environmental factors. Although the environmental risk factors have been studied extensively, little is known about the genetic factors. Heat-shock proteins (HSPs) are induced after exposure to severe noise. When first induced by exposure to moderate sound levels, they can protect the ear from damage from excessive noise exposure. This protection is highly variable between individuals. An association of HSP70 genes with NIHL has been described by Yang et al (2006) in a Chinese sample set of noise-exposed workers. In this study, three polymorphisms (rs1043618, rs1061581 and rs2227956) in HSP70-1, HSP70-2 and HSP70-hom, respectively, were genotyped in 206 Swedish and 238 Polish DNA samples of noise-exposed subjects and analyzed. One SNP, rs2227956 in HSP70-hom, resulted in a significant association with NIHL in both sample sets. In addition, rs1043618 and rs1061581 were significant in the Swedish sample set. Analysis of the haplotypes composed of the three SNPs revealed significant associations between NIHL and haplotype GAC in both sample sets and with haplotype CGT in the Swedish sample set. In conclusion, this study replicated the association of HSP70 genes with NIHL in a second and third independent noise-exposed sample set, hereby adding to the evidence that HSP70 genes may be NIHL susceptibility genes.

The contribution of GJB2 (Connexin 26) 35delG to age-related hearing impairment and noise-induced hearing loss.

HYPOTHESIS: The common GJB2 (Connexin 26) 35delG mutation might contribute to the development of age-related hearing impairment (ARHI) and noise-induced hearing loss (NIHL). BACKGROUND: GJB2, a gene encoding a gap junction protein expressed in the inner ear, has been suggested to be involved in the potassium recycling pathway in the cochlea. GJB2 mutations account for a large number of individuals with nonsyndromic recessive hearing loss, with 35delG being the most frequent mutation in populations of European origin. Other genes involved in potassium homeostasis have been suggested to be associated with ARHI and NIHL, and distortion product otoacoustic emission distortions indicative of hearing loss alterations have been found in 35delG carriers. METHOD: We genotyped 35delG in two distinct sample sets: an ARHI sample set, composed of 2,311 Caucasian samples from nine different centers originating from seven different countries with an age range between 53 and 67 years, and an NIHL sample set consisting of 702 samples from the two extremes of a noise-exposed Polish sample. RESULTS: After statistical analysis, we were unable to detect an association between 35delG and ARHI, nor between 35delG and NIHL. CONCLUSION: Our findings indicate that there is no increased susceptibility in 35delG carriers for the development of ARHI or NIHL.

Association between variations in CAT and noise-induced hearing loss in two independent noise-exposed populations.

Noise-induced hearing loss (NIHL) is an important occupational hazard that results from an interaction between genetic and environmental factors. Although the environmental risk factors have been studied quite extensively, little is known about the genetic factors. On the basis of multiple studies, it was proposed that oxidative stress plays an important role in the development of NIHL. Here, we investigated whether variations (single nucleotide polymorphisms; SNPs) in the catalase gene (CAT), one of the genes involved in oxidative stress, influence noise susceptibility. Audiometric data from 1261 Swedish and 4500 Polish noise-exposed labourers were analysed. DNA samples were collected from the 10% most susceptible and the 10% most resistant individuals. Twelve SNPs were selected and genotyped. Subsequently, the interaction between noise exposure and genotypes and their effect on NIHL were analysed using logistic regression. Significant interactions were observed between noise exposure levels and genotypes of two SNPs for the Swedish population and of five SNPs for the Polish population. Two of these SNPs were significant in both populations. The interaction between predictor haplotypes and tagSNP haplotypes and noise exposure levels and their effect on NIHL were also analysed, resulting in several significant associations. In conclusion, this study identified significant associations between catalase SNPs and haplotypes and susceptibility to development of NIHL. These results indicate that catalase is a NIHL susceptibility gene, but that the effect of CAT polymorphisms can only be detected when noise exposure levels are taken into account.

Expression of bFGF and NGF and their receptors in chick's auditory organ following overexposure to noise.

Growth factors are known to activate signaling cascades for DNA replication; they participate in the regulation of cell differentiation and are required as positive signals for cell survival. Thus, many of them may be regarded as potential candidates stimulating regeneration processes in the inner ear. We analyzed the expression of basic fibroblast growth factor (bFGF) and nerve growth factor (NGF) and their receptor (bFGFR and NGFR)-like immunoreactivity in chick basilar papillae, along with bFGF and NGF mRNA expression. The evaluation was made 1 and 5 days after exposure to wide-band noise with two increasing levels of acoustic energy. For both factors, the immunoreactivity was shown predominantly in the middle part of basilar papilla, in noise-exposed, but not control birds. It was localized in the cytoplasm of hair cells, nuclei of supporting cells and cytoplasm of ganglion cells. Strong immunoreactivity of bFGFR and NGFR was found both in control and noise-exposed animals, with the cell localization similar to that of growth factors. The increase in mRNA expression for bFGF and NGF was found in noise-exposed animals only after lower exposure to noise, on day 5 after exposure (p<0.01). A lack of increased expression after higher exposure could be excused by larger damage of hair cells followed by the increase of mRNA for beta-actin to which the results were referred. The results suggest bFGF and NGF involvement in postinjury regeneration of the basilar papilla.

The effect of weak 50 Hz magnetic fields on the number of free oxygen radicals in rat lymphocytes in vitro.

The aim of the work was verification of the hypothesis that weak power frequency (50 Hz) magnetic fields (MF) affected the number of free oxygen radicals in living biological cells and that these changes could be qualitatively explained by the radical pair mechanism. The experiments were performed on rat lymphocytes. One-hour exposure to 50 Hz MF at 20, 40, or 200 microT flux densities was performed inside a pair of Helmholtz coils with axis along or crosswise to the Earth's static MF. Iron ions (FeCl2) were used as a stimulator of the oxidation processes. Oxygen radicals were measured by fluorimetry using a DCF-DA fluorescent probe. Only in the lymphocytes exposed at 40 microT MF directed along the Earth's static MF there was a decrease of fluorescence in relation to non-exposed samples. Our observation seems to confirm the hypothesis that low level power frequency MF affects oxidative processes which occur in living biological cells and that this effect can be explained by the radical pair mechanism.

Effects of in vitro exposure to power frequency magnetic fields on UV-induced DNA damage of rat lymphocytes.

The mechanisms of biological effects of 50/60 Hz (power frequency) magnetic fields (MF) are still poorly understood. There are a number of studies indicating that MF affect biochemical processes in which free radicals are involved, such as the biological objects' response to ultraviolet radiation (UVA). Therefore, the present study was aimed to assess the effect of 50 Hz MFs on the oxidative deterioration of DNA in rat lymphocytes irradiated in vitro by UVA. UVA radiation (150 J/m2) was applied for 5 min for all groups and 50 Hz MF (40 microT rms) exposure was applied for some of the groups for 5 or 60 min. The level of DNA damage was assessed using the alkaline comet assay, the fluorescence microscope, and image analysis. It has been found that the 1 h exposure to MF caused an evident increase in all parameters consistent with damaged DNA. This suggest that MF affects the radical pairs generated during the oxidative or enzymatic processes of DNA repair.

Acute exposure to 930 MHz CW electromagnetic radiation in vitro affects reactive oxygen species level in rat lymphocytes treated by iron ions.

The aim of this study was to test the hypothesis that the 930 MHz continuous wave (CW) electromagnetic field, which is the carrier of signals emitted by cellular phones, affects the reactive oxygen species (ROS) level in living cells. Rat lymphocytes were used in the experiments. A portion of the lymphocytes was treated with iron ions to induce oxidative processes. Exposures to electromagnetic radiation (power density 5 W/m2, theoretical calculated SAR = 1.5 W/kg) were performed within a GTEM cell. Intracellular ROS were measured by the fluorescent probe dichlorofluorescin diacetate (DCF-DA). The results show that acute (5 and 15 min) exposure does not affect the number of produced ROS. If, however, FeCl2 with final concentration 10 microg/ml was added to the lymphocyte suspensions to stimulate ROS production, after both durations of exposure, the magnitude of fluorescence (ROS level during the experiment) was significantly greater in the exposed lymphocytes. The character of the changes in the number of free radicals observed in our experiments was qualitatively compatible with the theoretical prediction from the model of electromagnetic radiation effect on radical pairs.

[Protective effect of melatonin and vitamin E against prooxidative action of iron ions and static magnetic field]. / Ochronny wplyw melatoniny i witaminy E w prooksydacyjnym dzialaniu jonów zelaza oraz stalego pola magnetycznego.

The purpose of this study was to examine the effect of melatonin and vitamin E (trolox) on the level of lipid peroxidation in rat blood lymphocytes after in vitro (3 h) exposure to iron ions and/or 7mT static magnetic field (SMF). The lipid peroxidation process was chosen as a marker of free radical mechanism of SMF in cells. The cells were supplemented with (0.5 mM) melatonin or (0.1 mM) vitamin E (trolox) in preincubation. During SMF exposure in Helmholtz coils some samples were treated with ferrous chloride (10 mg/ml or 20 mg/ml), while the rest served as controls. There is a significant increase in the amount of lipid peroxidation end-products (4-HNE + MDA) in rat lymphocytes after simultaneous exposure to 7 mT SMF and iron ions (versus control samples and those exposed to SMF alone). Instead, when the cells were treated with melatonin or trolox and then exposed to iron ions and 7 mT SMF, the level of lipid peroxidation was significantly reduced. The results also indicated that melatonin is less effective than vitamin E (trolox) in inhibiting lipid peroxidation under the experimental conditions used.

Effect of 7 mT static magnetic field and iron ions on rat lymphocytes: apoptosis, necrosis and free radical processes.

Simultaneous exposure of rat lymphocytes to 7 mT static magnetic field (SMF) and iron ions caused an increase in the number of cells with DNA damage. The mechanism by which MF induces DNA damage and the possible cytotoxic consequences are not known. However, we suppose that free radicals are involved. Potentially, the deterioration of DNA molecules by simultaneous exposure to 7 mT SMF and iron ions may lead to cell death: apoptosis or necrosis. The possible prooxidative properties of these two agents may result in an induction of the lipid peroxidation process as a marker of free radical mechanism in the cells. Experiments were performed on rat blood lymphocytes incubated for 3 h in Helmholtz coils at SMF of flux density 7 mT. During SMF exposure, some samples were treated with ferrous chloride (10 microg/ml), the rest serving as controls. We used the dye exclusion method with the DNA-fluorochromes: ethidium bromide and acridine orange. No significant differences were observed between unexposed lymphocytes incubated with medium alone and lymphocytes exposed to 7 mT SMF. Three-hour incubation with FeCl(2) (10 microg/ml) did not affect cell viability. However, when lymphocytes were exposed to 7 mT SMF and simultaneously treated with FeCl(2), there was a significant increase in the percentage of apoptotic and necrotic cells accompanied by significant alterations in cell viability. As compared to lipid peroxidation, there is a significant increase in the amount of lipid peroxidation end products MDA+4 HNE in rat lymphocytes after simultaneous exposure to 7 mT SMF and FeCl(2) (vs. to the control samples and those exposed to SMF alone). This suggests that 7 mT static magnetic field in the presence of Fe(2+) ions can increase the concentration of oxygen free radicals and thus may lead to cell death.