Identification of a Circadian Clock in the Inferior Colliculus and Its Dysregulation by Noise Exposure.

UNLABELLED: Circadian rhythms regulate bodily functions within 24 h and long-term disruptions in these rhythms can cause various diseases. Recently, the peripheral auditory organ, the cochlea, has been shown to contain a self-sustained circadian clock that regulates differential sensitivity to noise exposure throughout the day. Animals exposed to noise during the night are more vulnerable than when exposed during the day. However, whether other structures throughout the auditory pathway also possess a circadian clock remains unknown. Here, we focus on the inferior colliculus (IC), which plays an important role in noise-induced pathologies such as tinnitus, hyperacusis, and audiogenic seizures. Using PER2::LUC transgenic mice and real-time bioluminescence recordings, we revealed circadian oscillations of Period 2 protein in IC explants for up to 1 week. Clock genes (Cry1, Bmal1, Per1, Per2, Rev-erbα, and Dbp) displayed circadian molecular oscillations in the IC. Averaged expression levels of early-induced genes and clock genes during 24 h revealed differential responses to day or night noise exposure. Rev-erbα and Dbp genes were affected only by day noise exposure, whereas Per1 and Per2 were affected only by night noise exposure. However, the expression of Bdnf was affected by both day and night noise exposure, suggesting that plastic changes are unlikely to be involved in the differences in day or night noise sensitivity in the IC. These novel findings highlight the importance of circadian responses in the IC and emphasize the importance of circadian mechanisms for understanding central auditory function and disorders. SIGNIFICANCE STATEMENT: Recent findings identified the presence of a circadian clock in the inner ear. Here, we present novel findings that neurons in the inferior colliculus (IC), a central auditory relay structure involved in sound processing, express a circadian clock as evidenced at both the mRNA and protein levels. Using a reporter mouse that expresses a luciferase protein coupled to the core clock protein PERIOD2 (PER2::LUC), we could observe spontaneous circadian oscillations in culture. Furthermore, we reveal that the mRNA profile of clock-related genes in the IC is altered differentially by day or night noise exposure. The identification of a clock in the IC is relevant for understanding the mechanisms underlying dysfunctions of the IC such as tinnitus, hyperacusis, or audiogenic seizures.

Role of cysteinyl leukotriene signaling in a mouse model of noise-induced cochlear injury.

Noise-induced hearing loss is one of the most common types of sensorineural hearing loss. In this study, we examined the expression and localization of leukotriene receptors and their respective changes in the cochlea after hazardous noise exposure. We found that the expression of cysteinyl leukotriene type 1 receptor (CysLTR1) was increased until 3 d after noise exposure and enhanced CysLTR1 expression was mainly observed in the spiral ligament and the organ of Corti. Expression of 5-lipoxygenase was increased similar to that of CysLTR1, and there was an accompanying elevation of CysLT concentration. Posttreatment with leukotriene receptor antagonist (LTRA), montelukast, for 4 consecutive days after noise exposure significantly decreased the permanent threshold shift and also reduced the hair cell death in the cochlea. Using RNA-sequencing, we found that the expression of matrix metalloproteinase-3 (MMP-3) was up-regulated after noise exposure, and it was significantly inhibited by montelukast. Posttreatment with a MMP-3 inhibitor also protected the hair cells and reduced the permanent threshold shift. These findings suggest that acoustic injury up-regulated CysLT signaling in the cochlea and cochlear injury could be attenuated by LTRA through regulation of MMP-3 expression. This study provides mechanistic insights into the role of CysLTs signaling in noise-induced hearing loss and the therapeutic benefit of LTRA.

Effects of the optimal flexor/extensor ratio on G-tolerance.

[Purpose] The aim of this study was to examine the flexor/extensor ratio of the knee joints and compare it with the results of Korean Air Force students in G-tolerance test. [Subjects and Methods] The body composition of Korea Air Force students (n=77) was measured by an impedance method. A muscular function test was performed using a Humac Norm (USA) at angular speeds of 60°/sec and 240°/sec and an isokinetic muscular function test was also conducted. [Results] In the failed C and passing groups, muscle mass and fat percentages were significantly higher than those of students in the failed A group. The BMI of the failed C and passing groups were significantly higher than that of the failed A group. The group that passed had a significantly higher value of left knee 60°/sec flexion peak torque than the failed B group. Moreover, the total work of left knee extension of the failed C group and the passing group was significantly higher than that of the failed A group. The C group and the passing group had significantly higher values of the trunk 60°/sec flexor/extensor ratio than the failed A group, and the total work flexor/extensor ratio of the passing group was significantly higher than that of the failed A group. [Conclusions] Based on these results, balance the right and left knee flexor/extensor ratio, and a high flexor/extensor ratio of the trunk are required to endure a high G-tolerance test (+6G/30 sec). Moreover, an improvement in the maximum muscular strength is necessary to endure a situation of rapidly increasing acceleration in the early stage.

Salmonellosis outbreaks linked to eggs at 2 gimbap restaurants in Korea.

OBJECTIVES: Salmonellosis outbreaks occurred at 2 restaurants 2 days apart, and an epidemiological investigation was conducted to determine whether the outbreaks were connected. METHODS: Case studies were conducted for both outbreaks. Stool samples were collected from individuals, and food samples were collected from the restaurants. Pulsed-field gel electrophoresis (PFGE) and whole-genome sequencing analyses were performed on outbreak-related Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) isolates. Traceback investigations were also conducted for the ingredients from gimbap restaurants A and B. RESULTS: In total, 106 people from gimbap restaurant A and 5 from gimbap restaurant B met the case definition. Salmonella Enteritidis was detected in samples from 2 food handlers, 22 patients, and 1 food (iceberg lettuce) at gimbap restaurant A and from 1 patient at gimbap restaurant B. According to PFGE, all isolates were identified as SEGX01.089. The molecular typing of all isolates showed the same pattern, and the genetic distance was close according to phylogenetic analysis. Eggs were the only food ingredient that was supplied to both gimbap restaurants. CONCLUSIONS: The outbreaks were caused by Salmonella Enteritidis, and the source of infections was suspected to be contaminated eggs. To prevent foodborne outbreaks of Salmonella, restaurants should heat eggs sufficiently, and egg farms need to establish management systems that prevent Salmonella infections.

Circadian Regulation of Cochlear Sensitivity to Noise by Circulating Glucocorticoids.

The cochlea possesses a robust circadian clock machinery that regulates auditory function. How the cochlear clock is influenced by the circadian system remains unknown. Here, we show that cochlear rhythms are system driven and require local Bmal1 as well as central input from the suprachiasmatic nuclei (SCN). SCN ablations disrupted the circadian expression of the core clock genes in the cochlea. Because the circadian secretion of glucocorticoids (GCs) is controlled by the SCN and GCs are known to modulate auditory function, we assessed their influence on circadian gene expression. Removal of circulating GCs by adrenalectomy (ADX) did not have a major impact on core clock gene expression in the cochlea. Rather it abolished the transcription of clock-controlled genes involved in inflammation. ADX abolished the known differential auditory sensitivity to day and night noise trauma and prevented the induction of GABA-ergic and glutamate receptors mRNA transcripts. However, these improvements were unrelated to changes at the synaptic level, suggesting other cochlear functions may be involved. Due to this circadian regulation of noise sensitivity by GCs, we evaluated the actions of the synthetic glucocorticoid dexamethasone (DEX) at different times of the day. DEX was effective in protecting from acute noise trauma only when administered during daytime, when circulating glucocorticoids are low, indicating that chronopharmacological approaches are important for obtaining optimal treatment strategies for hearing loss. GCs appear as a major regulator of the differential sensitivity to day or night noise trauma, a mechanism likely involving the circadian control of inflammatory responses.

Circadian regulation of auditory function.

The circadian system integrates environmental cues to regulate physiological functions in a temporal fashion. The suprachiasmatic nucleus, located in the hypothalamus, is the master clock that synchronizes central and peripheral organ clocks to orchestrate physiological functions. Recently, molecular clock machinery has been identified in the cochlea unravelling the potential involvement in the circadian regulation of auditory functions. Here, we present background information on the circadian system and review the recent findings that introduce circadian rhythms to the auditory field. Understanding the mechanisms by which circadian rhythms regulate auditory function will provide fundamental knowledge on the signalling networks that control vulnerability and resilience to auditory insults.

Differential Phase Arrangement of Cellular Clocks along the Tonotopic Axis of the Mouse Cochlea Ex Vivo.

Topological distributions of individual cellular clocks have not been demonstrated in peripheral organs. The cochlea displays circadian patterns of core clock gene expression [1, 2]. PER2 protein is expressed in the hair cells and spiral ganglion neurons of the cochlea in the spiral ganglion neurons [1]. To investigate the topological organization of cellular oscillators in the cochlea, we recorded circadian rhythms from mouse cochlear explants using highly sensitive real-time tracking of PER2::LUC bioluminescence. Here, we show cell-autonomous and self-sustained oscillations originating from hair cells and spiral ganglion neurons. Multi-phased cellular clocks were arranged along the length of the cochlea with oscillations initiating at the apex (low-frequency region) and traveling toward the base (high-frequency region). Phase differences of 3 hr were found between cellular oscillators in the apical and middle regions and from isolated individual cochlear regions, indicating that cellular networks organize the rhythms along the tonotopic axis. This is the first demonstration of a spatiotemporal arrangement of circadian clocks at the cellular level in a peripheral organ. Cochlear rhythms were disrupted in the presence of either voltage-gated potassium channel blocker (TEA) or extracellular calcium chelator (BAPTA), demonstrating that multiple types of ion channels contribute to the maintenance of coherent rhythms. In contrast, preventing action potentials with tetrodotoxin (TTX) or interfering with cell-to-cell communication the broad-spectrum gap junction blocker (CBX [carbenoxolone]) had no influence on cochlear rhythms. These findings highlight a dynamic regulation and longitudinal distribution of cellular clocks in the cochlea.

The effects of isometric exercise types on pain and muscle activity in patients with low back pain.

The purpose of the present study is to evaluate the effects of isometric exercise types on low back pain (LBP) patients. Isometric exercise types were mat exercise and I-Zer exercise. Subjects were divided into 3 groups: LBP control group, LBP mat exercise group, and LBP I-Zer exercise group in 23-25 aged men. Visual analogue scale (VAS) and electromyography (EMG) were used to evaluate the degree of pain and the muscle activity in LBP patients. Root mean square (RMS), median frequency (MDF), and mean frequency (MNF) were checked by EMG power spectrum analysis on longissimus thoracic (LT), iliocostalis lumborum (IL), mulitifidus (M), and rectus abdominis (RA). LBP mat exercise program and LBP I-Zer exercise program were conducted 5 sets once time, 3 times per week during 6 weeks. The two-way ANOVA with repeated measure was used to check the pain degree and muscle activity. The present results showed that muscle activity in the LBP I-Zer exercise group was increased compared to the LBP mat exercise group and LBP control group (P<0.05). LBP I-Zer exercise group and LBP mat exercise group showed increased mean frequency in LT, IL, M, and RA muscles than the LBP control group. Therefore, LBP patients performed isometric exercise may have positive effect to reduce pain degree and to increase muscle activity. Especially, LBP I-Zer exercise type showed more effectiveness in reducing pain degree and enhancing muscle activity.

The effect of exercise on expression of myokine and angiogenesis mRNA in skeletal muscle of high fat diet induced obese rat.

PURPOSE: The purpose of this study was to investigate the effect of regular treadmill exercise on the mRNA expressions of myokines and angiogenesis factors in the skeletal muscle of obese rats. METHODS: Thirty two male Sprague-Dawley rats (4weeks old) were divided into the CO (control) and HF (high fat diet) groups. Obesity was induced in the HF group by consumption of 45% high-fat diet for 15 weeks. These groups were further subdivided into training groups (COT and HFT); the training groups conducted moderate intensity treadmill training for 8 weeks. Soleus muscles were excised and analyzed by real-time quantitative PCR. RESULTS: mRNA expression of myokines, such as PGC-1α, IL-6, and IL-15, in the COT and HFT groups (which conducted regular exercise), were higher as compared with the CO and HF groups (p < 0.05). Also, the levels in the HF group were significantly lower when compared with CO group (p < 0.05). Expression of angiogenesis mRNA, namely mTOR, VEGF, and FLT1, were significantly lower in the HF group, as compared to the CO group (p < 0.05). In addition, COT group had a higher expression of mTORC1, mTORC2, VEGF and FLT mRNA, than the CO group (p < 0.05); the HFT group also had higher expressions of mTOR, VEGF and FLT1 mRNA than the HF group (p < 0.05). CONCLUSION: These results indicate that mRNA expression of myokines was increased through the activity of muscle contraction, and it also promoted the mRNA expression of angiogenesis due to activation of mTOR. Thus, we conclude that not only under normal health conditions, but in obesity and excess nutritional circumstances also, regular exercise seems to act positively on the glycemic control and insulin sensitivity through the angiogenesis signaling pathway.

The phosphorylation of STAT6 during ischemic reperfusion in rat cerebral cortex.

For many years, brain ischemia has been known to be a leading cause of adult neurological disorder. In particular, many reports have shown that hyperexcitability of neurons and inflammatory response of the glia induced by ischemic reperfusion (I/R) determine the fate of cells in the ischemic core and the penumbra region. Although there are many reports on the activation and roles of signal transducer and activator of transcription (STAT) proteins (STAT1, STAT3, and STAT5) during hyperexcitation in the neuron and inflammation occurring following I/R, the temporal and spatial activation of STAT6 protein in the ischemic cortex still remain elusive. In this study, using a transient rat middle cerebral artery occlusion model, we primarily investigated the time-course expression of the phosphorylated STAT6 (pSTAT6) in the ischemic core region following I/R, which was compared with that of pSTAT3. We found that pSTAT6 significantly decreases at 1 and 12 h following I/R, whereas pSTAT3 markedly increases at each follow-up time point. In addition, the level of pSTAT6 is reduced in the ischemic core in comparison with the penumbra region at 12 h following I/R. However, there is no significant difference in pSTAT3 expression between the ischemic core and the penumbra. Taken together, our data suggest that pSTAT6 and pSTAT3 are modulated differently following I/R during ischemic stroke.

Tympanic membrane regeneration using a water-soluble chitosan patch.

Chronic otitis media or tympanic membrane (TM) perforation is one of the most common otologic diseases. Surgical tympanoplasty remains the best treatment option despite the fact that paper patches are frequently used. Although paper patches are not biocompatible or effective, tympanoplasty is an expensive, complex surgery. Tissue engineering techniques offer a new treatment strategy for TM regeneration. In this study, novel tissue-engineered artificial eardrums were fabricated from water-soluble chitosan, which is known to be a good wound-healing biomaterial. The characteristics, cytotoxicity, and healing effects of several water-soluble chitosan patches (WSCPs) made using various concentrations of water-soluble chitosan and glycerol were investigated. The optimal WSCP was fabricated with 3% water-soluble chitosan and 3% glycerol, and it had a thickness of about 35 mum, a tensile strength of 7 MPa, a percent elongation of 101%, a hydrophilic surface, and no cytotoxicity. In vivo studies showed that the WSCPs were more effective than spontaneous healing for the repair of traumatic TM perforations. The healed TMs to which WSCPs were applied had a much higher density of collagen fibers and a better lamina propria layer structure than spontaneously healed TMs.

Development of water-insoluble chitosan patch scaffold to repair traumatic tympanic membrane perforations.

Perforated tympanic membranes (TM) and otitis media can be managed with a paper patch or tympanoplasty. However, a paper patch is not biocompatible and tympanoplasty requires complex aseptic surgical procedures. A novel biocompatible patch with a water-insoluble chitosan as the main component was prepared. Optimal mechanical characteristics of a water-insoluble chitosan patch scaffold (CPS) was approximately 40 microm in thickness, 7 MPa in tensile strength, and 107% in percent elongation, even though the characteristics varied significantly depending on the concentrations of chitosan and glycerol. SEM of the CPSs showed a very smooth surface as compared with that of the paper patches. These CPSs showed no cytotoxicity and had a stimulating effect on the proliferation of TM cells in in vitro study. In in vivo study, 4 (21.1%) and 17 (89.5%) TMs out of 19 adult rats with CPSs showed no perforations at 1 and 2 weeks, respectively. However, left control TMs showed healing of 0 (0%) at 1 week and 18 (94.7%) at 2 weeks. TEM findings of regenerated eardrums using CPSs showed thinner, smoother, and more compact tissues than spontaneously healed eardrums. A CPS was more effective than spontaneous healing to repair traumatic TM perforations.

Effect of septoplasty on inferior turbinate hypertrophy.

OBJECTIVE: To measure the effect of septoplasty on the volume of inferior turbinate in patients with a deviated nasal septum. DESIGN: In this retrospective analysis, patients who underwent septoplasty without turbinate surgery from May 1, 2003, through April 30, 2006, were studied. The thicknesses and cross-sectional areas of mucosa and conchal bones were measured with computed tomography before the operations and at least 1 year after the operations. SETTING: University hospital. PATIENTS: A total of 20 patients who presented with a chief concern of nasal obstruction. MAIN OUTCOME MEASURES: The thicknesses of the medial mucosa, bone, and lateral mucosa and the cross-sectional area of turbinate before and after septoplasty were compared using the Wilcoxon signed rank test. P < .05 was considered statistically significant. RESULTS: The medial mucosa and cross-sectional area of the inferior turbinate on the concave side of the septum were significantly decreased by septoplasty (both, P = .01), and the medial mucosa and cross-sectional area of the inferior turbinate on the convex side of the septum were significantly increased by septoplasty (P = .01). The thicknesses and cross-sectional areas of the conchal bone on the concave and convex sides of the septum were not affected by septoplasty. CONCLUSION: After septoplasty, inferior turbinate hypertrophy, especially in the medial mucosa, may reverse.