Improvements in blood IGF-1 and skeletal age following adenotonsillectomy for growth delay in children with obstructive sleep apnea.

OBJECTIVE: In children with obstructive sleep apnea (OSA) who underwent adenotonsillectomy (AT), we measured body height and weight using standard deviation (SD) scores, insulin-like growth factor 1 (IGF-1), and skeletal age using carpal radiography. We then compared these values before and after surgery with the aim of investigating postoperative improvements in growth hormone (GH) deficiency. METHODS: Subjects comprised 35 children between 2 and 9 years of age (21 boys, 14 girls; mean age, 5.85 ± 1.75 years). Respiratory event index (REI), 3 % oxygen desaturation index (3 % ODI), height SD score, body mass index (BMI) percentile, blood IGF-1 level, and skeletal age from carpal radiographs were measured before surgery and both 3 and 12 months after surgery, and compared. RESULTS: Height SD score improved significantly from preoperatively (-0.44 ± 1.13) to both 3 months postoperatively (-0.22 ± 1.14) and 12 months postoperatively (-0.13 ± 0.94). However, no significant improvement in height SD score was seen from 3 months to 12 months after AT. BMI percentile improved significantly from preoperatively (35.6 ± 26.7) to both 3 months postoperatively (44.7 ± 26.5) and 12 months postoperatively (49.1 ± 22.15), with significant improvement also seen from 3 months to 12 months after AT. SD score for IGF-1 showed significant improvement from before (-0.57 ± 1.00) to 12 months after surgery (-0.12 ± 0.89). No significant improvement in the difference between skeletal and chronological ages was seen from before to after surgery, but the number of patients for whom skeletal age normalized from before to after surgery increased significantly (74.3 % vs. 94.3 %), and the number with advanced or delayed skeletal age decreased significantly (25.7 % vs. 5.7 %) CONCLUSION: Early improvements can be obtained with surgical treatment in children with OSA who have short height and poor weight gain due to GH deficiency.

Anti-Inflammatory Effects of EM900 on Cultured Human Nasal Epithelial Cells.

OBJECTIVES: Macrolide therapy is an important conservative therapy for chronic rhinosinusitis, especially in Japan. The mechanism underlying this therapy involves anti-inflammatory and not antimicrobial activity. However, the administration of long-term low-dose macrolides (LTLMs) causes an increase in the number of antibiotic-resistant bacteria. EM900 is a derivative of erythromycin (EM), with anti-inflammatory but not antibacterial effects. It does not induce macrolide-resistant bacteria as shown by LTLM. In the present study, we analyzed the inhibitory effects of EM900 in comparison with those of clarithromycin (CAM) on inflammatory cytokine production in human nasal epithelial cells (HNEpCs). METHODS: After HNEpCs were cultured for 4 days, CAM or EM900 was added into the culture, followed by stimulation with tumor necrosis factor (TNF)-α. Interleukin (IL)-8 and vascular endothelial growth factor (VEGF) levels were measured using real-time polymerase chain reaction (RT-PCR) and an enzyme-linked immunosorbent assay (ELISA). RESULTS: Both the ELISA and RT-PCR showed that EM900 and CAM significantly inhibited IL-8 production in HNEpCs. In contrast, EM900 and CAM did not suppress the increased VEGF production when HNEpCs were stimulated with TNF-α. CONCLUSION: EM900 showed an anti-inflammatory effect, such as that of CAM, due to the inhibitory effect on IL-8 production in HNEpCs.

Inhibitory Effects of S-carboxymethylcystein on Goblet Cell Proliferation in Cultured Epithelium.

BACKGROUND: The influence of S-carboxymethylcystein (S-CMC) on the proliferation ability of goblet cells in nasal polyp epithelium in response to inflammatory stimulation was examined. METHODS: The subjects were patients with chronic paranasal sinusitis. An epithelial cell culture system was established using nasal polyp mucosa excised during endoscopic paranasal sinus surgery. The samples were divided into 4 groups (group a: control group, group b: 10 ng/mL tumor necrosis factor-α (TNF-α) treatment group, group c: 10-7 M S-CMC and 10 ng/mL TNF-α treatment group, group d: 10-5 M S-CMC and 10 ng/mL TNF-α treatment group). The total number of epithelial cells and number of goblet cells were measured under a microscope, and the ratio of goblet cells to the total number of epithelial cells was calculated. RESULTS: In group b, 10 ng/mL of TNF-α significantly increased the number of goblet cells compared with group a, suggesting involvement of TNF-α in goblet cell proliferation. In addition, the number of goblet cells significantly decreased in group d compared with that in group b, and it also decreased in group c compared with that in group b, although the difference was not significant, and the decrease was smaller than that in group d, suggesting that S-CMC inhibited goblet cell proliferation in a concentration-dependent manner. CONCLUSION: TNF-α promoted goblet cell proliferation in nasal polyps, suggesting its influence on nasal polyp formation. As S-CMC inhibited inflammatory stimulation-induced goblet cell proliferation in nasal polyp epithelium, it may be useful for the treatment of sinusitis.

Relationship between swallowing function and breathing/phonation.

OBJECTIVE: Clarification of the association between the swallowing function and respiratory and phonatory functions. METHODS: The subjects were 30 patients with a chief complaint of swallowing disorder with clear consciousness capable of retaining a sitting position. Patients with organic and functional diseases of the larynx were excluded. Twenty-two and eight patients were male and female, respectively, and the mean age was 77.0±14.6years old. The chest expansion score was measured as an index of the respiratory function, and the maximum phonation time (MPT) was measured as an index of the phonatory function. The presence or absence of aspiration was judged using videoendoscopic swallowing study (VESS) and videofluoroscopic swallow studies (VFSS). The patients were divided into those with and without aspiration, and the chest expansion score and MPT were compared. In addition, the distance of laryngeal elevation was measured in the lateral view of VFSS, and its correlations with the chest expansion score and MPT were closely analyzed. To evaluate reliability of the test, the distance of laryngeal elevation and videoendoscopic score were compared between the presence and absence of aspiration. RESULTS: The distance of laryngeal elevation was significantly shortened and the videoendoscopic score was significantly higher in the group with aspiration, as previously reported. On comparison of the chest expansion score between the groups with and without aspiration, no significant difference was noted at the axillary or xiphoid process level, and shortening was significant only at the 10th rib level in the group with aspiration. On comparison of MPT, it was significantly shortened in the group with aspiration. In addition, a significant positive correlation with the distance of laryngeal elevation was noted in both chest expansion score and MPT. CONCLUSION: It was suggested that declines of the respiratory and phonatory functions are risk factors of aspiration through limiting laryngeal elevation, and the chest expansion score at the 10th rib level and MPT are useful for screening of aspiration.

Expression Profiling of MicroRNAs in the Inner Ear of Elderly People by Real-Time PCR Quantification.

The molecular mechanisms underlying age-related hearing loss are unknown, and currently, there is no treatment for this condition. Recent studies have shown that microRNAs (miRNAs) and age-related diseases are intimately linked, suggesting that some miRNAs may present attractive therapeutic targets. In this study, we obtained 8 human temporal bones from 8 elderly subjects at brain autopsy in order to investigate the expression profile of miRNAs in the inner ear with miRNA arrays. A mean of 478 different miRNAs were expressed in the samples, of which 348 were commonly expressed in all 8 samples. Of these, levels of 16 miRNAs significantly differed between young elderly and old elderly subjects. miRNAs, which play important roles in inner ear development, were detected in all samples, i.e., in both young and old elderly subjects, whether with or without hearing loss. Our results suggest that these miRNAs play important roles not only in development, but also in the maintenance of inner ear homeostasis.

[EFFECT OF PROPHYLACTIC TREATMENT BY LEUKOTORIENE RECEPTOR ANTAGONIST ON JAPANESE CEDAR POLLINOSIS OF PRE- AND POST-SCHOOL CHILDREN JAPANESE CEDAR POLLINOSIS].

UNLABELLED: Backgroud: It has already been reported that the prophylactic treatment by leukotoriene receptor antagonists is more effective on reducing symptoms of Japanese cedar pollinosis than the authentic treatment after the pollen dispersal. However, the treatment above has never evaluated in children cases around school age in ENT out-patient clinic. This study about the prophylactic treatment was planned to focus on the effect in the generation of pre- and post-elementary school entrance. METHODS: Children of pre- and post-elementary school entrance were enrolled for this study. This study was achieved in seasons of Japanese cedar pollinosis both in 2013 and 2014, and was designed as the comparison of clinical symptoms and quality of life in between two such groups as one group with the prophylactic treatment and another with the authentic treatment. RESULTS: Efficacy of prophylactic treatment by leukotoriene receptor antagonists was elucidated as follows; quality of sleep was significantly better both in 2013 and 2014, and more kinds of clinical symptoms or quality of life impairments were significantly more suppressed than in the group with the authentic treatment in 2014 when less pollen was dispersed. CONCLUSION: Even in the children of pre- and post-elementary school entrance, the prophylactic treatment by leukotoriene receptor antagonists is more effective on reducing symptoms of Japanese cedar pollinosis than the authentic treatment.

Treatment of nostril and nasal stenosis due to facial burn using a self-expandable metallic esophageal stent.

For the treatment of nasal and nostril stenosis caused by facial burn, it is necessary to perform rhinoplasty and nasal vestibuloplasty using various flaps, perform cicatrectomy of the nostrils with a rhinosurgical procedure, and prevent restenosis of the nostrils and nasal cavity for a certain period by methods such as placement of a nasal retainer or transnasal airway and gauze packing of the nasal cavity. With all methods, postoperative placement of a retainer or nasal treatment is necessary for the prevention of postoperative restenosis, and the patient's cooperation is essential. In a patient who did not cooperate in postoperative treatments due to autism and had recurrences of nasal and nostril stenosis after conventional surgical treatments, adequate patency of the nasal cavity and nostrils could be maintained with minimal postoperative treatment by placing a self-expandable metallic esophageal stent.

Cochlin expression in the rat perilymph during postnatal development.

CONCLUSIONS: The changes in the cochlin isoforms in the perilymph may provide important insights to the understanding of cochlin function and the pathogenesis of related inner ear diseases. OBJECTIVES: Cochlin is involved in various pathologies of the inner ear. Altered levels of cochlin isoforms in developing inner ear tissue were reported previously. The purpose of this study was to elucidate the cochlin isoform expression in the perilymph of rats during postnatal development in relation to Coch gene mRNA expression. METHODS: We studied the cochlin isoforms in the rat perilymph during postnatal development by Western blot analysis. Real-time PCR was also performed to elucidate the expression level of Coch mRNA in the developing inner ear of rats. RESULTS: Western blot analysis showed that the expression of p63s in the perilymph was highest on the 12th day after birth (DAB12), the earliest age at which we could identify the perilymphatic space microscopically, and it decreased gradually as the cochlea developed. On the other hand, the expression of Cochlin-tomoprotein (CTP)was lowest on DAB12 and increased gradually up to DAB24. COCH mRNA was detected from DAB3 and gradually increased to DAB15, and then gradually decreased to DAB70.

Cochlin-tomoprotein (CTP) detection test identifies traumatic perilymphatic fistula due to penetrating middle ear injury.

CONCLUSIONS: The cochlin-tomoprotein (CTP) detection test can be used to make a definite, objective diagnosis of traumatic perilymphatic fistula (PLF), and therefore offers valuable information on patient selection for surgical treatment. OBJECTIVES: Penetrating middle ear injury can cause traumatic PLF, which is a surgically treatable otologic emergency. Recently, we have reported on CTP, a novel perilymph-specific protein. The purpose of this study was to determine if the CTP detection test is useful for the diagnosis of traumatic PLF. METHODS: This was a prospective study of CTP detection in penetrating middle ear injury cases with tympanic membrane perforation and hearing loss. RESULTS: A total of seven individuals were included in this study. CTP was detected in three of four cases with posterosuperior quadrant perforation of the tympanic membrane. In one of these three cases, even though the high resolution CT scan was not suggestive of PLF and the perilymph leakage could not be visualized intraoperatively, the CTP detection test was able to detect PLF. In two cases, the preoperative positive test results enabled us to make a diagnosis of PLF and a decision for surgical treatment. CTP was not detected in the cases with anterior or inferior tympanic membrane perforation.

Molecular cloning of the Coch gene of guinea pig inner ear and its expression analysis in cultured fibrocytes of the spiral ligament.

CONCLUSIONS: We have cloned guinea pig Coch cDNA and the sequence information will be useful for future molecular study combined with physiological experiments. Proper Coch gene expression appears to be dependent on the unique extracellular micro-environment of the inner ear in vivo. These results provide insight into the Coch gene expression and its regulation. OBJECTIVE: To characterize the guinea pig Coch gene, we performed molecular cloning and expression analysis in the inner ear and cultured fibrocytes of the spiral ligament. METHODS: The Coch cDNA was isolated using RACE. Cochlin isofoms were studied by Western blot using three different types of mammalian inner ear. The cochlear fibrocytes were cultured and characterized by immunostaining. Coch gene expression in the fibrocytes was investigated and the influence of cytokine stimulation was evaluated. RESULTS: The full-length 1991 bp Coch cDNA that encodes a 553 amino acid protein was isolated. The sequence had significant homology with other mammals, and the sizes of the Cochlin isoforms were identical. In the cultured fibrocytes, Coch mRNA was expressed in a very small amount and the isoform production was different, compared with the results in vivo. Cytokine stimulation did not alter the level of mRNA expression or isoform formation.

Expression of cochlin mRNA splice variants in the inner ear.

Proteomic analysis of inner ear proteins revealed unique properties of cochlin, encoded by the COCH gene. We detected 3 cochlin isoforms, p63s, p44s and p40s, in the inner ear tissue and a short 16-kDa isoform, cochlin-tomoprotein (CTP), in the perilymph. The role of the cochlin isoforms has not been elucidated. To improve our understanding of the mechanism of cochlin isoform expression, we investigated rat cochlin mRNA expression in the inner ear and other organs. We performed RNA-ligation-mediated amplification of cDNA ends (RLM-RACE) using RNA isolated from the inner ear and spleen of rats, which are known to express abundant cochlin mRNA. We also examined the expression profile of full-length cochlin mRNA by nested RT-PCR in the cerebrum, cerebellum/brain stem, eye, inner ear, thyroid gland, thymus gland, lung, heart, liver, spleen, adrenal gland, kidney and blood. We verified CTP expression in rat perilymph by Western blot. By RLM-RACE, alternately spliced variants of cochlin mRNA with 3 different lengths were detected (2442, 2008 and 724 bp). The two longer mRNAs encode full-length cochlin with different polyadenylation signals in the 3'-untranslated region, which are expressed both in the ear and spleen. The short variant encodes the limulus factor C, cochlin, late gestation lung protein (LCCL) domain and the N-terminal sequence of the von Willebrand factor A (vWFA1) domain, and this variant was detected only in the ear. All 3 variants have the same transcriptional start site. By RT-PCR, we found that full-length cochlin was expressed in all organs examined, with a splice variant in the heart. By Western blot, we detected short isoforms (11-17 kDa) in the perilymph. Cochlin isoform formation is regulated, at least in part, by alternative splicing at the transcriptional level. The short mRNA was detected only in the inner ear, and this variant may provide a clue to understanding the formation and function of cochlin isoforms.

Cochlin-tomoprotein: a novel perilymph-specific protein and a potential marker for the diagnosis of perilymphatic fistula.

BACKGROUND: Perilymphatic fistula (PLF) is an abnormal connection between the inner and middle ear. A procedure for obtaining definite proof of a PLF remains elusive, and methods of diagnosis remain controversial. To date, there is no clinically relevant biochemical marker for perilymph leakage. Using proteomic analysis of inner ear proteins, we have previously found unique properties of cochlin, encoded by the COCH gene. We detected 3 cochlin isoforms (p63s, p44s and p40s) in the inner ear tissue and a short 16-kDa isoform of cochlin-tomoprotein (CTP) in the perilymph. Since cochlin was found to be highly specific to the inner ear, we speculated that CTP might also be specific to the perilymph. The aim of this study was to determine whether CTP, a novel perilymph-specific protein, could be used as a marker for the diagnosis of PLF. METHODS: By Western blotting, we investigated the specificity of CTP expression in a range of body fluids that included perilymph, serum, saliva and cerebrospinal fluid. To elucidate the detection limit of CTP, serially diluted recombinant human (rh)CTP as well as human perilymph was tested. RESULTS: CTP was selectively expressed in all 20 perilymph samples tested, but not in 77 samples of the other body fluids. The detection limit of rhCTP was 0.27 ng or 0.022 microl of perilymph per well on Western blot analysis. CONCLUSION: The results strongly suggest that CTP can be a specific marker of perilymph leakage. Moreover, CTP has the potential to be a biochemical marker that allows a definitive diagnosis of the etiology of PLF-related hearing loss and vestibular disorders.