Reduction of DNA Folding by Ionic Liquids and Its Effects on the Analysis of DNA-Protein Interaction Using Solid-State Nanopore.

DNA folding is not desirable for solid-state nanopore techniques when analyzing the interaction of a biomolecule with its specific binding sites on DNA since the signal derived from the binding site could be buried by a large signal from the folding of DNA nearby. To resolve the problems associated with DNA folding, ionic liquids (ILs), which are known to interact with DNA through charge-charge and hydrophobic interactions are employed. 1-n-butyl-3-methylimidazolium chloride (C4 mim) is found to be the most effective in lowering the incident of DNA folding during its translocation through solid-state nanopores (4-5 nm diameter). The rate of folding signals from the translocation of DNA-C4 mim is decreased by half in comparison to that from the control bare DNA. The conformational changes of DNA upon complexation with C4 mim are further examined using atomic force microscopy, showing that the entanglement of DNA which is common in bare DNA is not observed when treated with C4 mim. The stretching effect of C4 mim on DNA strands improves the detection accuracy of nanopore for identifying the location of zinc finger protein bound to its specific binding site in DNA by lowering the incident of DNA folding.

Analysis of Effect of Eggshell Membrane Patching for Moderate-to-Large Traumatic Tympanic Membrane Perforation.

BACKGROUND AND OBJECTIVES: To evaluate the effect of eggshell membrane (ESM) patching for tympanic membrane (TM) perforation, and to investigate correlations between healing time and age, gender, patching time, perforation size, and perforation location. SUBJECTS AND METHODS: One hundred and seventy-five patients diagnosed with traumatic TM perforation at the Department of Otorhinolaryngology-Head and Neck Surgery, Korea University Ansan Hospital, between January 2008 and October 2013. Patients were divided into two groups, according to perforation edge approximation or ESM patching treatment. Healing time was compared between the group that received perforation edge approximation and the group that received ESM patching. Perforation grade, age, onset, and location were also compared between the two groups. RESULTS: ESM patching significantly improved healing time compared to spontaneous healing, especially in patients with moderate or large traumatic TM perforations (≥grade II). However, patient age, gender, perforation location, and especially timing of procedure, did not significantly affect healing time. CONCLUSIONS: ESM patching can be a good treatment choice to promote tympanic membrane healing in large traumatic TM perforations.

Short-Term Outcomes of Acute Low-Tone Sensorineural Hearing Loss According to Treatment Modality.

BACKGROUND AND OBJECTIVES: We compared improvements in hearing thresholds in acute low-tone sensorineural hearing loss (ALHL) patients after two different treatments: steroid alone and steroid and diuretic combined. We analyzed how the duration between the onset of symptoms and the initiation of treatment affected hearing loss improvement and investigated the relation between presence of vertigo in ALHL patients and ALHL progression to Ménière's disease (MD). SUBJECTS AND METHODS: We retrospectively analyzed the medical records of 47 ALHL patients aged 21 to 76 years. Patients received either orally administered steroid alone (n=12) or steroid and diuretic combined (n=35). We compared improvements in the two groups' hearing thresholds at three lower frequencies (125, 250, and 500 Hz) after participants had received one month of each respective treatment. RESULTS: Our two treatments did not show any statistical difference in hearing loss improvement after one month. Forty percent of ALHL patients with vertigo developed MD, which was a significantly higher rate than the 12.5% of ALHL patients without vertigo who developed MD. The shorter duration between the onset of symptoms and the initiation of treatment significantly increased improvement in the sum of lower frequency hearing threshold after one month. CONCLUSIONS: The current study suggests that steroid and diuretic administered together and steroid alone similarly improve the hearing threshold in ALHL patients after one month. We concluded that patients should initiate ALHL treatment as soon as they experience symptoms. ALHL patients should also be notified of their higher risk of developing MD.

Protective effects of caffeic acid phenethyl ester (CAPE) against neomycin-induced hair cell damage in zebrafish.

OBJECTIVE: Caffeic acid phenethyl ester (CAPE) is known to reduce the generation of oxygen-derived free radicals, which is a major mechanism of aminoglycoside-induced ototoxicity. The objective of the present study was to evaluate the effects of CAPE on neomycin-induced ototoxicity in zebrafish (Brn3c: EGFP). METHODS: Five-day post-fertilization zebrafish larvae (n=10) were exposed to 125 µM neomycin and one of the following CAPE concentrations for 1h: 50, 100, 250, 500, or 1000 µM. Ultrastructural changes were evaluated using scanning electron microscopy (SEM). The terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick-end labeling (TUNEL) assay and 2-[4-(dimethylamino)styryl]-N-ethylpyridiniumiodide (DASPEI) assay were performed for evaluation of apoptosis and mitochondrial damage. RESULTS: CAPE decreased neomycin-induced hair cell loss in the neuromasts (500 µM CAPE: 12.7 ± 1.1 cells, 125 µM neomycin only: 6.3 ± 1.1 cells; n = 10, P < 0.05). In the ultrastructural analysis, structures of mitochondria and hair cells were preserved when exposed to 125 µM neomycin and 500 µM CAPE. CAPE decreased apoptosis and mitochondrial damage. CONCLUSION: In the present study, CAPE attenuated neomycin-induced hair cell damage in zebrafish. The results of the current study suggest that neomycin induces apoptosis, and the apoptotic cell death can be prevented by treatment with CAPE in zebrafish.

Increased expression of YKL-40 in mild and moderate/severe persistent allergic rhinitis and its possible contribution to remodeling of nasal mucosa.

BACKGROUND: Prominent expression of YKL-40 has been associated with pathological conditions characterized by tissue remodeling. We determined the expression level and distribution pattern of YKL-40 in allergic nasal mucosa and evaluated the effect of YKL-40 on the proliferation and migration of fibroblasts, the production of the mediators related to tissue remodeling, and collagen production. Additionally, the cytokine-driven regulation of YKL-40 expression was evaluated in cultured epithelial cells. METHODS: The expression of YKL-40 in normal, mild, and moderate/severe allergic nasal mucosa was evaluated using real-time polymerase chain reaction (PCR), Western blot, and immunohistochemistry. Fibroblast migration was observed using a scratch wound method, and proliferation was determined by the MTT methods. Expression of proteoglycans, transforming growth factor (TGF) beta1, MMP2, MMP9, TIMP1, TIMP2, and collagen concentration were analyzed in fibroblasts treated with YKL-40. The expression levels of YKL-40 in cultured epithelial cells were examined after stimulation with mediators including Th2 cytokines, interferon (IFN)gamma, and TNF-alpha with real-time PCR and ELISA. RESULTS: The expression of YKL-40 was up-regulated in allergic rhinitis and distributed in superficial epithelium, submucosal glands, and vascular endothelium, in addition to infiltrating cells. TGF-beta1, TIMP1, MMP9, and biglycan were up-regulated in fibroblasts on stimulation with YKL-40, accompanying increased proliferation and migration, and collagen production. IL-13, IFN-gamma, and TNF-alpha induced the increased production of YKL-40 in cultured epithelial cells. CONCLUSION: YKL-40 is up-regulated in mild and moderate/severe persistent allergic rhinitis, and its expression can be regulated differentially by different cytokines, possibly contributing to the remodeling of nasal mucosa in allergic rhinitis.

Optimal level of continuous positive airway pressure: auto-adjusting titration versus titration with a predictive equation.

OBJECTIVES: The aims of the present study were twofold. We sought to compare two methods of titrating the level of continuous positive airway pressure (CPAP) - auto-adjusting titration and titration using a predictive equation - with full-night manual titration used as the benchmark. We also investigated the reliability of the two methods in patients with obstructive sleep apnea syndrome (OSAS). METHODS: Twenty consecutive adult patients with OSAS who had successful, full-night manual and auto-adjusting CPAP titration participated in this study. The titration pressure level was calculated with a previously developed predictive equation based on body mass index and apnea-hypopnea index. RESULTS: The mean titration pressure levels obtained with the manual, auto-adjusting, and predictive equation methods were 9.0 +/- 3.6, 9.4 +/- 3.0, and 8.1 +/- 1.6 cm H2O,respectively. There was a significant difference in the concordance within the range of +/- 2 cm H2O (p = 0.019) between both the auto-adjusting titration and the titration using the predictive equation compared to the full-night manual titration. However, there was no significant difference in the concordance within the range of +/- 1 cm H2O (p > 0.999). CONCLUSIONS: When compared to full-night manual titration as the standard method, auto-adjusting titration appears to be more reliable than using a predictive equation for determining the optimal CPAP level in patients with OSAS.

Chronic rhinosinusitis with polyps and without polyps is associated with increased expression of suppressors of cytokine signaling 1 and 3.

BACKGROUND: Chronic rhinosinusitis with nasal polyps (CRSwNP) or without nasal polyps (CRSsNP) is associated with expression of various cytokines. Suppressors of cytokine signaling (SOCS) regulate cytokine activity in a variety of cells, modulating inflammatory responses. OBJECTIVE: We analyzed the expression and distribution pattern of SOCS1 and SOCS3 in CRSwNP and CRSsNP, and their cytokine-driven expression regulation in sinus mucosa. In addition, the expression levels of various cytokines were evaluated in CRSwNP and CRSsNP. METHODS: The expression levels of SOCS1 and SOCS3 in CRSwNP and CRSsNP and in control samples were assessed by using real-time PCR, Western blot, and immunohistochemistry. Nasal epithelial cell culture was used to elucidate the effect of IL-4, IL-5, IL-6, IL-10, IL-13, IFN-γ, TNF-α, and TGF-ß1 on SOCS1 and SOCS3 expression in sinus mucosa. The expression levels of these cytokines were also evaluated in normal and inflammatory sinus mucosa by using real-time PCR and Western blot. RESULTS: The expression levels of SOCS1 and SOCS3 were increased in CRS, irrespective of the presence of nasal polyp, and they were distributed in superficial epithelium, submucosal glands, and vascular endothelium in sinus mucosa. SOCS1 was induced by IL-4, IL-13, IFN-γ, and TNF-α, while SOCS3 expression was upregulated by IL-6, IL-13, IFN-γ, and TNF-α. IL-4 and IL-13 levels were increased in CRSwNP, while IL-4, IFN-γ, TNF-α, and TGF-ß1 levels were increased in CRSsNP. CONCLUSION: SOCS1 and SOCS3 are increased in CRS, irrespective of nasal polyp presence. This may be a response to elevated levels of various cytokines increasingly expressed in inflammatory sinus mucosa.

Expression levels of endogenous hydrogen sulfide are altered in patients with allergic rhinitis.

OBJECTIVES/HYPOTHESIS: Hydrogen sulfide (H(2) S), the third endogenous gaseous transmitter, may be a crucial mediator in airway hyper-responsiveness and airway inflammation, including asthma. To elucidate the role of H(2) S in allergic rhinitis, the present study was undertaken to determine the level of expression of H(2) S in healthy nasal mucosa and mild and moderate/severe persistent allergic nasal mucosa as well as peripheral blood obtained from each patient. The expression and distribution pattern of the H(2) S-synthesizing enzymes cystathione γ-lyase (CSE) and cystathione ß-synthase (CBS) were investigated in healthy and allergic nasal mucosa. STUDY DESIGN: Controlled, prospective study. METHODS: The concentration of H(2) S in nasal mucosa and plasma was determined by zinc trap spectrophotometry. The expression levels and patterns of distribution of CSE and CBS mRNA and proteins were evaluated using real time polymerase chain reaction, Western blot, and immunohistochemistry. RESULTS: The levels of expression of H(2) S in nasal mucosa and plasma were increased in patients with mild and moderate/severe persistent allergic rhinitis compared with healthy controls. CSE was localized in vascular endothelium and surrounding muscles, and submucosal glands, whereas CBS was exclusively distributed in the superficial epithelium and submucosal glands. Their expression levels were increased in mild and moderate/severe persistent allergic rhinitis. CONCLUSIONS: The current findings indicate that, in parallel with increased expression levels of CSE and CBS, H(2) S is upregulated in nasal mucosa and plasma of allergic patients. Based on localization of CSE and CBS, H(2) S may play multiple functions in human nasal mucosa, contributing to the development of allergic symptoms such as rhinorrhea, sneezing, and nasal stuffiness.

Tuberculous otitis media with facial paralysis combined with labyrinthitis.

Tuberculosis otitis media is a very rare cause of otorrhea, so that it is infrequently considered in differential diagnosis because clinical symptoms are nonspecific, and standard microbiological and histological tests for tuberculosis often give false-negative results. We present a rare case presenting as a rapidly progressive facial paralysis with severe dizziness and hearing loss on the ipsilateral side that was managed with facial nerve decompression and anti-tuberculosis therapy. The objective of this article is to create an awareness of ear tuberculosis, and to consider tuberculosis in the differential diagnosis of chronic otitis media with complications.