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Background and Objectives Surgery for bilateral chronic otitis media (COM) is usually performed sequentially, not simultaneously. The main reason is to prevent iatrogenic bilateral conductive hearing loss during recovery period. However, with asymmetric hearing loss, the difference in patient inconvenience between sequential and simultaneous surgery may be the same. This study evaluates the efficacy of simultaneous COM surgery in patients with asymmetric hearing.Subjects Materials and Method From 2012 to 2018, 9 patients underwent simultaneous bilateral COM surgery. The period of patients’ hospital stay, the success rate of tympanoplasty, tolerability for operation, and hearing thresholds were analyzed statistically.Results For the 8 same-day surgery patients, the mean hospital period was 4.4±0.7 days, which was much shorter than that of sequential surgery (8 days). Tympanoplasty was successful in all patients without complications. In the better side, the preoperative/postoperative mean air conduction thresholds (AC) were 62.97±12.89 dB/47.81±19.07 dB (p=0.017), the bone conduction thresholds (BC) were 46.72±10.31 dB/37.66±16.99 dB (p=0.161) and the air-bone gaps (ABG) were 16.25±8.81 dB/10.16±7.78 dB (p=0.176). In the worse side, the preoperative/ postoperative mean AC were 86.56±18.22 dB/72.18±29.43 dB (p=0.035), BC were 53.28± 11.10 dB/48.13±18.41 dB (p=0.173), and ABG were 33.28±11.22 dB/24.06±14.80 dB (p=0.500). In both ear, the postoperative AC and BC were better than or equivocal to those of the preoperative value, and the result was similar with each of the unilateral ear audiological results.Conclusion Despite the fact that there is no complete consensus to date, simultaneous bilateral COM surgery can be an option when patients have asymmetric hearing loss. Simultaneous bilateral COM surgery could save time, cost, and lead to similar results with sequential surgery, so it could be a considerable surgical option for patients with bilateral COM.

Objectives. To investigate the neurocognition of aged patients with chronic tinnitus and reveal the possible association between tinnitus severity and cognitive function, with attention to mild cognitive impairment (MCI).Methods. Fifty-eight elderly patients (≥65 years old) with chronic tinnitus (≥6 months) were prospectively enrolled in this study. All patients assessed the neurocognitive batteries including the Korean version of the patient health questionnaire-9 (K-PHQ-9), the Lawton instrumental activities of daily living scale (K-IADL), and the Montreal cognitive assessment (MoCA-K). After initial evaluation to exclude moderate or severe cognitive impairment by a psychiatrist, the patients were classified into two groups: MCI and non-MCI, according to the MoCA-K scores (cutoff value, 22/23). All patients underwent audiological examinations including psychoacoustic tests of tinnitus.Results. Of 58 patients, 10 (17.2%) met the MCI criteria. The tinnitus handicap inventory (THI) score in the MCI group was significantly higher than that in the non-MCI group. Based on multivariate regression analysis, a significant association between tinnitus severity and MoCA-K score was also detected. Specifically, bothersome tinnitus (THI score ≥30) was closely linked to the presence of MCI. Meanwhile, the impact of MCI on both K-PHQ-9 and K-IADL scores was not evident in patients with chronic tinnitus.Conclusion. Tinnitus severity appears to be a potential independent determinant for predicting the MCI, suggesting the underlying mechanism between chronic tinnitus and cognitive deficit. Given that MCI highly links to dementia, the evaluation of cognitive functions in aged patients with chronic tinnitus need to be considered at the initial assessment of tinnitus.

Objectives Sodium salicylate (SS) is well known for its ototoxic properties that induce functional and morphological changes in the cochlea and brain. Ginkgo biloba extract (GBE) has been widely used for treatment of various neurodegenerative diseases; however, its effects on salicylate-induced ototoxicity remain unclear. Herein, we examined the effects of EGb 761 (EGb), a standard form of GBE, on the plasticity of the N-methyl-D-aspartate receptor subunit 2B (GluN2B) in the inferior colliculus (IC) following SS administration. Methods Seven-week-old Sprague Dawley rats (n=24) were randomly allocated to control, SS, EGb, and EGb+SS groups. The SS group received a single intraperitoneal SS injection (350 mg/kg), the EGb group received EGb orally for 5 consecutive days (40 mg/kg), and the EGb+SS group received EGb for 5 consecutive days, followed by an SS injection. The auditory brainstem responses (ABRs) were assessed at baseline and 2 hours after SS administration. GluN2B expression was examined by Western blot and immunohistochemistry. Results There were no significant differences in ABR threshold shifts among the groups. The expression of the GluN2B protein normalized by which of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was significantly lower in the EGb+SS group, as compared to the SS group (P=0.012). Weak and diffused GluN2B immunoreactivity was detected in the IC neural cells of the EGb+SS group, while those of the SS group exhibited strong and diffused GluN2B positivity. Conclusion EGb may play a role in regulating the GluN2B expression in the IC of salicylate-induced ototoxicity model.

Background: Korean ginseng has been widely evaluated to treat human diseases; however, most studies on Korean ginseng have focused on its root. In this study, polysaccharides [acidic-polysaccharide-linked glycopeptide (APGP) extracted with 90% ethanol and hot water] were prepared from Korean ginseng berries, and their effect on immunosenescence was explored. Methods: The effect of APGP on thymic involution was evaluated by measuring the size of thymi dissected from aged mice. The effect of APGP on populations of immune cells, including natural killer (NK) cells, dendritic cells, age-correlated CD11c-positive B cells, and several subtypes of T cells [CD4-positive, CD8-positive, and regulatory (Treg) T cells] in the thymi and spleens of aged mice was analyzed by fluorescence-activated cell sorting analysis. Serum levels of interleukin (IL)-2 and IL-6 were evaluated by enzyme-linked immunosorbent assay analysis. Profiles of APGP components were evaluated by high-performance liquid chromatography (HPLC) analysis. Results: APGP suppressed thymic involution by increasing the weight and areas of thymi in aged mice. APGP increased the population of NK cells, but showed no effect on the population of dendritic cells in the thymi and spleens of aged mice. APGP decreased the population of age-correlated CD11c-positive B cells in the spleens of aged mice. APGP showed no effect on the populations of CD4- and CD8-positive T cells in the thymi of aged mice, whereas it increased the population of Treg cells in the spleens of aged mice. APGP further decreased the reduced serum levels of IL-2 in aged mice, but serum levels of IL-6 were not statistically changed by APGP in aged mice. Finally, HPLC analysis showed that APGP had one major peak at 15 min (a main type of polysaccharide) and a long tail up to 35 min (a mixture of a variety of types of polysaccharides). Conclusion: These results suggested that APGP exerted an anti-immunosenescent effect by suppressing thymic involution and modulating several types of immune cells. Keywords: APGP, ginseng berry, immunosenescence, NK cell, Treg cell

Since gaze estimation plays a crucial role in recognizing human intentions, it has been researched for a long time, and its accuracy is ever increasing. However, due to the wide variation in eye shapes and focusing abilities between the individuals, accuracies of most algorithms vary depending on each person in the test group, especially when the initial calibration is not well performed. To alleviate the user-dependency, we attempt to derive features that are general for most people and use them as the input to a deep network instead of using the images as the input. Specifically, we use the pupil shape as the core feature because it is directly related to the 3D eyeball rotation, and thus the gaze direction. While existing deep learning methods learn the gaze point by extracting various features from the image, we focus on the mapping function from the eyeball rotation to the gaze point by using the pupil shape as the input. It is shown that the accuracy of gaze point estimation also becomes robust for the uncalibrated points by following the characteristics of the mapping function. Also, our gaze network learns the gaze difference to facilitate the re-calibration process to fix the calibration-drift problem that typically occurs with glass-type or head-mount devices.

Importance The lateral pharyngeal wall is recognized as an important site of upper airway collapse during sleep in patients with obstructive sleep apnea (OSA), and expansion sphincter pharyngoplasty (ESP) may have promising clinical utility in patients with OSA and lateral pharyngeal wall collapse. Objectives To evaluate the therapeutic outcomes of ESP in conjunction with other surgical procedures and to investigate indications for ESP in patients with OSA. Design, Setting, and Participants Cohort study of 63 patients with OSA diagnosed with lateral pharyngeal collapse under drug-induced sleep endoscopy who underwent ESP combined with tonsillectomy, uvuloplasty, or nasal surgery at Seoul National University Hospital in Seoul, Korea, between March 1, 2015, and December 1, 2016. Main Outcomes and Measures The primary outcome measure was the change in the apnea-hypopnea index (AHI) after surgery (AHI represents the number of apnea-hypopnea events per hour). Other outcome measures were differences in the surgical response rates, lowest oxygen saturation, subjective visual analog scale scores for snoring and apnea, and Epworth Sleepiness Scale score. Results Fifty of the 63 patients (79%) were male; the mean age was 42.1 (range, 20-54) years, and the mean body mass index (calculated as weight in kilograms divided by height in meters squared) was 27.6 (range, 19.0-32.1). Expansion sphincter pharyngoplasty was performed in patients with OSA with an AHI greater than 15 events per hour, more than 75% retropalatal circumferential narrowing when awake, and narrowed oropharynx due to bulky soft tissue around the lateral pharyngeal wall. In 42 of the 63 patients (67%), ESP was objectively successful in correcting lateral pharyngeal collapse; there was a significant reduction in mean AHI from 35.5 to 17.3 (mean difference, 18.1; 95% CI, 16.3-20.0) and improvement of the lowest mean (SD) oxygen saturation measurement from 78.2% (21.3%) to 86.4% (10.6%) (mean difference, 8.60%; 95% CI, 6.60%-10.60%) 6 months after the operation. The rate of postoperative complications, including pain and bleeding, was minimal after ESP, and a few patients reported an abnormal sensation around the soft palate and swallowing difficulty after ESP. Conclusions and Relevance Expansion sphincter pharyngoplasty appears to be a promising surgical technique to reduce lateral pharyngeal collapse in patients with moderate or severe OSA. Clinical data suggest that both severe palatal circumferential narrowing and bulky lateral pharyngeal tissue are favorable surgical indications for ESP in patients with OSA.

Purpose The previously reported Japanese clinical scoring study (JESREC) suggests that chronic rhinosinusitis (CRS) can be divided into 4 subtypes according to the degree of eosinophilic CRS (ECRS) and offers the information regarding the prognosis of CRS to clinicians. However, this scoring system has not yet been validated by an immunological study and needs to provide treatment guidelines based on underlying immunologic profiles. We investigated the immunologic profile of each CRS subgroup according to the JESREC classification and suggest its clinical application. Methods A total of 140 CRS patients and 20 control subjects were enrolled. All patients were classified into 4 groups according to the JESREC (non-, mild, moderate and severe ECRS). Nasal tissues were analyzed for mRNA expression of major cytokines (IL-5, IL-10, IL-13, IL-17A, IL-22, IL-23p19, IFN-γ, periostin, thymic stromal lymphopoietin [TSLP] and ST2), major chemokines (CCL11, CCL24, CXCL1 and CXCL2), transcription factors (T-bet, GATA3, RORC and FOXP3) and COL1A1 for type I collagen. Protein levels of 3 major cytokines (IL-5, IL-17A and IFN-γ) were also measured by multiplex immunoassay. Principal component analysis (PCA) was conducted to investigate the overall profile of multiple mediators. Results The moderate/severe ECRS showed up-regulation of type 2-related mediators (IL-5, IL-13, periostin, TSLP and ST-2), whereas INF-γ (type 1 cytokine) and CXCL1 (neutrophil chemokine) expressions were increased in non-/mild ECRS compared with moderate/severe ECRS. The JESREC classification reflected an immunological endotype. In PCA data, PCA1 indicates a relative type 2 profile, whereas PCA2 represents a type 1/type 17-related profile. In this analysis, mild ECRS was indistinguishable from non-ECRS, whereas moderate to severe ECRS showed a distinct distribution compared with non-ECRS. The JESREC classification could be divided into 2 categories, non-/mild vs. moderate/severe ECRS based on underlying immunological analyses. Conclusions The CRS clinical scoring system from the JESREC study reflects an inflammatory endotype. However, the immunologic profile of mild ECRS was similar to that of non-ECRS. Therefore, we propose type 2-targeted medical treatment for moderate to severe ECRS and type 1/type 17-targeted for non-ECRS and mild ECRS as the first treatment option.

Finding the location of a pupil center is important for the human-computer interaction especially for the user interface in AR/VR devices. In this paper, we propose an indirect use of the convolutional neural network (CNN) for the task, which first segments the pupil region by a CNN, and then finds the center of mass of the region. For this, we create a dataset by labeling the pupil area on 111,581 images from 29 IR video sequences. We also label the pupil region of widely used datasets to test and validate our method on a variety of inputs. Experiments show that the proposed method provides better accuracies than the conventional ones, showing robustness to the noise.

Currently virus-like particles (VLPs) are receiving much attention as platforms for next generation vaccines. However, chromatography-based methods for purifying VLPs remain challenging. Unlike traditional methods using density gradient for purifying VLPs, there have been few advances in explaining how assembled particles can be obtained by chromatography. Nervous necrosis virus (NNV) infects over 30 species of fish and leads to large economic losses in the farmed fish industry. Previously we developed a heparin chromatography-based method for purifying red-spotted grouper NNV (RGNNV) VLPs. However it is unclear how the assembled RGNNV VLPs are obtained by this method. It is known that assembly of NNV capsid proteins depends on calcium ions. In the present study, we found that the yield of purified RGNNV capsid protein in heparin chromatography was enhanced when calcium ions were present during binding. Also, it appears that the capsid protein of RGNNV undergoes partial disassembly and reassembly during sample preparation prior to heparin chromatography and the protein finally undergoes assembly during the chromatography. Therefore, our results indicated that heparin-binding affinity of RGNNV capsid protein is linked to its ability for VLP formation. The assembly of RGNNV capsid proteins recombinantly produced is a good model for explaining VLP formation during chromatography-based purification processes.

Objectives Recently, the endoscope has been increasingly introduced for middle-ear surgery. To evaluate the postoperative outcomes of endoscopic ear surgery (EES) in pediatric patients, we did a qualitative analysis with a systematic review and quantitative analysis with meta-analysis of available literature. Methods Studies reporting the comparative surgical outcomes of EES in pediatric patients were systematically reviewed by searching the MEDLINE, PubMed, and Embase databases from database inception through 2017. The selected articles included clinical studies conducted with at least 30 subjects and at least one postoperative parameter, including residual or recurrent cholesteatoma and graft success in tympanoplasty. Two investigators independently reviewed all studies and extracted the data using a standardized form. A meta-analysis was performed using a random-effects model and qualitative review was performed on the smaller studies. Results Ten studies were identified as appropriate for quantitative meta-analysis and 19 studies for qualitative analysis. In the meta-analysis, residual or recurrence rate of cholesteatoma was significantly lower in the EES group than in the microscopic ear surgery (MES) group (odds ratio [OR]: 0.56, 95% confidence interval [CI]: 0.38-0.84, P = .005). The graft success rate of tympanoplasty was not statistically different between EES and MES groups (OR: 0.72, 95% CI: 0.41-1.26, P = .249). In the qualitative analysis, most of the studies reported similar audiological outcomes after tympanoplasty and success rate of cholesteatoma removal between the two groups. Conclusions It appears that EES reduces the risk of residual cholesteatoma in children and that the success of perforation closure is equivalent to MES. Laryngoscope, 129:1444-1452, 2019.

Virus-like particles (VLPs) are multi protein complexes mimicking the structural properties of the native virus. The development of freeze-dried formulations of such complex protein structures remains a challenge. Red-spotted grouper nervous necrosis virus (RGNNV) causes mass mortality in fish culture, and RGNNV VLPs have been suggested to be promising vaccine candidates. In the present study, the stability of RGNNV VLPs in the liquid state was investigated over a 4-week period, along with the influence of freeze-drying on VLP stability. RGNNV VLPs were completely degraded after one week at 37 °C followed by 3 weeks at ambient temperature, and they were partially degraded after 4 weeks at 4 °C. Therefore, the inherent stability of RGNNV VLP in an aqueous milieu is insufficient for long-term storage. When RGNNV VLPs were freeze-dried in the presence or absence of sugar stabilizers, sorbitol was found to improve VLP stability whereas mannitol reduced it. VLP preparations freeze-dried with sorbitol or without stabilizer were as immunogenic as control (non-freeze dried) VLPs, whereas VLPs freeze-dried in mannitol were less immunogenic. These results indicate that freeze-dried RGNNV VLPs have potential as vaccines.

To investigate the predictive factors of polyp regression after intravitreal ranibizumab injections for polypoidal choroidal vasculopathy (PCV). Sixty-nine eyes (69 patients) with treatment-naive PCV received an initial series of 3-monthly intravitreal ranibizumab injections, followed by as-needed injections. Polyp regression was determined after 6 months from baseline by means of indocyanine angiography and correlated with clinical characteristics at baseline. After treatment, 26.1 % (18 out of 69 eyes) achieved complete polyp regression and 73.9 % (51 out of 69 eyes) showed persistent polyps. The mean logarithm of the minimum angle of resolution (logMAR) of best-corrected visual acuity (BCVA) was significantly improved in both groups (P = 0.02). No statistically significant difference in BCVA improvement was observed between the groups. However, the proportion of subjects with BCVA improvement by more than three lines was significantly higher in the polyp regression group (P = 0.02). Univariate analysis of the baseline characteristics identified the number of polyps (P = 0.02), the total polyp area (P = 0.009) and the polyp pattern (P = 0.01) as predictive factors for polyp regression. Multivariate logistic regression identified the total polyp area (P = 0.01) as an independent predictor for polyp regression after intravitreal ranibizumab injections. The regression of polypoidal lesions in PCV patients after intravitreal ranibizumab injections was associated with a smaller total polyp area. This result could be helpful in predicting polypoidal changes after ranibizumab treatment and in deciding the best treatment strategy for PCV patients.

Nervous necrosis viruses (NNV) cause serious economic losses in marine fish cultivation. The red-spotted grouper NNV (RGNNV) is the most common species of NNV worldwide. There have been many efforts to develop prophylactic NNV vaccines, and various types of vaccine candidate have been suggested. However, most were designed as injectable vaccines, which are not suitable for large-scale vaccination and cause too much stress to the fish. Oral vaccination through voluntary feeding is an ideal way to provide protective immunity to fish. In the present study, recombinant Saccharomyces cerevisiae producing RGNNV capsid protein was used as oral vaccine. The recombinant yeast was prepared in freeze-dried form after disruption. Convict groupers were divided into three groups, control, and oral and parenteral vaccination groups, each consisting of 700 fishes. The control group received no treatment, the parenteral group received one intraperitoneal injection of RGNNV virus-like particles, and the oral vaccination group consumed feed containing the lysed recombinant yeast; voluntary intake was allowed four times at one-week intervals. Both vaccination groups produced serum RGNNV neutralizing antibody titers of >103 (log 2, 9.96), sustained for at least 95days post-immunization. In addition, in response to challenge with RGNNV both groups suffered significantly reduced mortality and had reduced brain RGNNV titers. These results indicate that recombinant yeast-based oral fish vaccines have great potential for large-scale vaccination.

Philasterides dicentrarchi is a causative agent of scuticociliatosis in olive flounder Paralichthys olivaceus, aquaculture in Korea. In this study, a cDNA encoding a cathepsin L-like cysteine protease (PdCtL) of P. dicentrarchi (synonym Miamiensis avidus) was identified. To express the PdCtL recombinant protein in a heterologous system, 10 codons were redesigned to conform to the standard eukaryotic genetic code using polymerase chain reaction (PCR)-based site-directed mutagenesis. The recombinant P. dicentrarchi procathepsin L (proPdCtL) was expressed at high levels in E. coli Rosetta (DE3) pLysS with a pPET21a vector, and successfully refolded, purified, and activated into a functional and enzymatically active form. The optimal pH for protease activity was 5. Similar to other cysteine proteases, enzyme activity was inhibited by E64 and leupeptin. Immunogenicity of recombinant PdCtL was assessed by enzyme-linked immunosorbent assay, western blot, and specific anti-recombinant PdCtL antibodies were detected. Our results suggest that the biochemical characteristics of the recombinant ciliate proPdCtL protein are similar to those of the cathepsin L-like cysteine protease, that the PCR-based site-direct mutated ciliate gene was successfully expressed in a biochemically active form, and that the recombinant PdCtL acted as a specific epitope in olive flounder.

This paper proposes a gaze estimation algorithm using 3-D eyeball model and eyelid shape. The gaze estimation suffers from differences of eye shapes and individual behaviors, and requires user-specific gaze calibration. The proposed method exploits the usual 3-D eyeball model and shapes of the eyelid to estimate gaze without user-specific calibration and learning. Since the gaze is closely related to the 3-D rotation of eyeball, this paper first derives the relation between 2-D pupil location extracted in the eye image and 3-D rotation of eyeball. This paper also models the shapes of the eyelid to adjust gaze based on the observation that the shapes of the eyelid are deformed with respect to the gaze. This paper models the curvature of eyelid curve to compensate for the gaze. According to the various experiments, the proposed method shows good results in gaze estimation. The proposed method does not need user-specific calibration or gaze learning since the general 3-D eyeball and eyelid models are exploited in the localized eye region. Therefore, it is expected that the proposed gaze estimation algorithm is suitable for various applications such as VR/AR devices, driver gaze tracking, gaze-based interfaces, and so on.

Purpose Endotype in chronic rhinosinusitis (CRS) has been established in the last decade. However, the exact immunologic profile of CRS still has controversy because it has a considerable immunologic heterogeneity. Therefore, we investigated various inflammatory mediators according to different nasal tissues in chronic rhinosinusitis and compared them within the same subject. Methods We collected uncinate process mucosa (UP) and nasal polyp (NP) tissues from controls, CRS without NP (CRSsNP) and CRS with NP (CRSwNP). Expression levels of 28 inflammatory mediators including T helper (Th) 1, Th2, Th17, proinflammatory cytokines and remodeling markers were determined by multiplex immunoassay and were analyzed using paired tests as well as principal component analysis (PCA) to investigate endotype in each subtype of CRS. Results Signature inflammatory mediators are interleukin (IL)-5, C-C motif chemokine ligand (CCL)-24, monocyte chemoattractant protein (MCP)-4, and vascular cell adhesion molecule (VCAM)-1 in eosinophilic NP, whereas IL-17A, IL-1β, and matrix metallopeptidase (MMP)-9 were detected as signature inflammatory markers in non-eosinophilic NP. Despite differences in inflammatory cytokine profile between eosinophilic and non-eosinophilic NP, the common upregulation of IL-5, CCL-11, IL-23, IL-2Rα, VCAM-1, MMP-3 and MMP-9 were shown in NP compared to UP within the same subject. In the PCA, we observed that Th2 immune response was helpful in discriminating between nasal tissues in subtypes of CRS and that there was a partial overlap between non-eosinophilic NP and eosinophilic NP in terms of Th2 mediators. Conclusions Commonly upregulated mediators in NP were Th2-associated, compared with UP regardless of CRS subtypes, whereas signature markers were distinct in each NP subtype. These findings imply that Th2 inflammatory responses may play a role in the development of NP regardless of CRSwNP subtypes.

The principal objective of this study was to investigate the incidence, risk assessment, antibiotic resistance, and genotyping of Vibrio parahaemolyticus in Korean seafood. The incidence of V. parahaemolyticus in seafood obtained from several fish markets in Korea was investigated from May to December of 2009, except between July and September. Two selective mediums (TCBS [thiosulfate, citrate, bile salts, and sucrose] agar and CHROMagar™ Vibrio) were used, and the V. parahaemolyticus strains were identified via polymerase chain reaction (PCR) amplification (Vp. flaE, tl, and toxR). 16S rRNA gene sequencing and their virulence were analyzed via the detection of tdh, trh, ORF8, toxRS/old, and toxRS/new genes. We collected 24 strains of V. parahaemolyticus: 19 seafood isolates, three environmental isolates, and two clinical (human) isolates. Among these strains, two tdh+ strains, two ORF8+ strains, 16 toxRS/old+ strains, and one toxRS/new+ strain were isolated. Twenty-two commercial antibiotics were used to assess the antibiotic susceptibility of isolates, and all the strains evidenced resistance to more than four antibiotics. The strains harboring antibiotic-resistant genes such as TetA (25%) and strB (4.16%) were detected via PCR. Repetitive extragenic palindromic sequence (REP)-PCR analysis revealed differences in the V. parahaemolyticus strains from other species and intraspecific strains.

In October 2010, cuticular deformities with abnormal growth suppression were detected among cultured Litopenaeus vannamei shrimp in one farm located on the Taean peninsula along the western coast of Korea. We detected infectious hypodermal and haematopoietic necrosis virus (IHHNV) from the moribund L. vannamei shrimp and its pond water via PCR assays. The IHHNV strain isolated from the shrimps was designated K1 and the strain isolated from the pond water was designated K2, and the capsid genes were sequenced for genetic comparison and phylogenetic analysis with others obtained from different geographical regions. The results demonstrated that the Korean strains were similar to infectious IHHNV and were closely related to the Type 1 strains. This is the first report, to the best of our knowledge, of the detection of IHHNV from L. vannamei in Korean cultured shrimp.

We evaluated the protective efficacy of a combined vaccine containing formalin-inactivated Edwardsiella tarda, Streptococcus iniae, and S. parauberis using olive flounder Paralichthys olivaceus. The fish were injected intraperitoneally with 0.1 mL of the combined vaccine. The fish were then challenged by intraperitoneal injection of the three bacterial strains. Mortality was significantly lower in the vaccinated groups compared with the control groups 3 wk post vaccination. Furthermore, there was a significant increase in antibody titer up to 16 wk after vaccination. Our results suggest that the combined vaccine successfully induced humoral and protective immunity against the three pathogens in olive flounder.

A pandemic influenza A (H1N1) virus strain was isolated from a pig farm in Korea in December 2009. The strain was propagated in and isolated from both the Madin-Darby canine kidney cell line and embryonated eggs. The partial and complete sequences of the strain were identical to those of A/California/04/2009, with >99% sequence similarity in the HA, NA, M, NS, NP, PA, PB1, and PB2 genes. The isolated strain was inactivated and used to prepare a swine influenza vaccine. This trial vaccine, containing the new isolate that has high sequence similarity with the pandemic influenza A (H1N1) virus, resulted in seroconversion in Guinea pigs and piglets. This strain could therefore be a potential vaccine candidate for swine influenza control in commercial farms.

Antimicrobial resistance patterns in a collection of 33 motile Aeromonas species were described in this study. Quinolone has been frequently employed for treatment of Aeromonas -related diseases, and prolonged use of antimicrobial compounds has led to development of resistant strains. In a sample of diseased fish and environmental water, we evaluated nalidixic acid ( n = 19) and ciprofloxacin ( n = 4) resistance via minimum inhibitory concentration (MIC) assays and the genetic basis was also investigated. Among the isolated Aeromonas spp., 17 strains encoded for chromosomal mutations of quinolone resistance-determining regions (QRDRs) in gyrA , 11 strains encoded for mutations of QRDRs in parC , 1 strain harbored plasmid-mediated quinolone resistance (PMQR) qnrS1- like gene and 4 strains harbored the PMQR qnrS2 gene. In particular, the new variant ( qnrS1 -like) differed from qnrS1 by 6 amino acid substitutions at positions 5 (Asn 5 →Arg 5 ), 120 (Ser 120 →Thr 120 ), 148 (Asn 148 →His 148 ), 206 (Leu 206 →Glu 206 ), 207 (Ile 207 → Leu 207 ), and 216 (Tyr 216 →Phe 216 ), and the gene was designated qnrS5 . These resistant strains may function as reservoirs of quinolone resistance.

The discovery of germ cell-specific general transcription factor and coactivator variants has suggested that reproductive tissues control gene expression somewhat differently than somatic tissues. One of these factors, ALF (TFIIAtau), was first described as a testis-specific counterpart of the large (alpha/beta) subunit of TFIIA. Here we characterize endogenous ALF and TFIIA activities in the African clawed frog Xenopus laevis. ALF is present in both testis and ovary in this organism, and it completely replaces TFIIA in immature oocytes. When oocytes undergo progesterone-induced maturation, ALF activity disappears, and TFIIA activity is restored. Reactivation occurs through the translational up-regulation of two maternal TFIIAalpha/beta mRNAs and involves polyadenylation of a conserved 3'-untranslated region module. The effects of ALF overexpression and ALF immunodepletion on a thymidine kinase promoter construct demonstrate that this factor serves as an active replacement for TFIIA. In contrast, overexpression of TFIIA inhibits transcription, indicating that the somatic factor fails to function properly in the context of the oocyte transcription machinery. Overall, the results show that the translationally regulated reciprocal expression of ALF and TFIIA allows for the production of an active TFIIA-like general transcription factor throughout oogenesis.

Received November 10, 2012 Revised February 19, 2013 Accepted February 19, 2013 Address for correspondence Chang Ki Yeo, MD Department of Otorhinolaryngology, School of Medicine, Keimyung University, 56 Dalseong-ro, Jung-gu, Daegu 700-712, Korea Tel +82-53-250-7715 Fax +82-53-256-0325 E-mail ckyeo@dsmc.or.kr Ultrasonograms and thyroglobulin measurements have high sensitivity for evaluation of recurrent thyroid cancer and they allow early detection of recurrent or residual thyroid tumors. Patients treated for thyroid cancer can show recurrent or residual tumors in 5-40% of cases. Radioiodine therapy and surgical excision are effective for recurrent or residual thyroid tumors, but up to 30% of tumors will not reveal iodine uptake, and need further observation or surgical excision. Clinically, patients choose surgical excision rather than observation. During reoperation, the risk of surgical complications is increased because of fibrosis, neovascularization, and modified anatomical relationships. In the past, surgeons have occasionally used methylene blue during reoperation but it tends to spread into the surrounding tissues. Recently, charcoal injection has been used for recurrent thyroid microcarcinoma. The authors successfully used charcoal injection for the removal of tumor in two cases of recurrent microcarcinoma of the thyroid bed after total thyroidectomy. Korean J Otorhinolaryngol-Head Neck Surg 2013;56:522-4 Key WordsZZCharcoal ᆞThyroid cancer ᆞThyroidectomy.

Background and ObjectivesZZThe evaluation of vestibular function is very important for patients with sudden sensorineural hearing loss (SNHL) as it is a well-known prognostic factor. The purpose of this study was to evaluate subclinical vestibular dysfunction (SVD) and its association with prognosis in sudden SNHL patients with and without dizziness using the vestibular function tests. Subjects and MethodZZSeventy eight patients who had been diagnosed with unilateral sudden SNHL were examined by dividing them into three groups according to their clinical findings such as vestibular function and subjective symptoms: 1) normal, 2) SVD and 3) dizziness. Vestibular dysfunction and hearing recovery rates were also assessed. ResultsZZPatients with SVD accounted for 62.5% of patients even though they did not complain subjective dizziness. The extent of vestibular dysfunction in the dizziness group was greater than that in the SVD group (p=0.05). The hearing recovery rates for normal, SVD and dizziness group were 72% (13/18), 57% (17/30) and 48% (12/25), respectively. The incidence and extent of vestibular dysfunction was greater in patients with severe hearing loss than in those with moderate to severe loss. A significant correlation was noted between hearing recovery rate and vestibular evoked myogenic potential (VEMP) testing. Normal and abnormal VEMP data were associated with good and poor recovery (p=0.001). ConclusionZZThe evaluation of vestibular function is important in sudden SNHL patients with dizziness; however, SVD also explains a significant proportion of sudden SNHL in the absence of dizziness. Notably, VEMP is useful tools for evaluating prognosis of sudden SNHL. � Korean J Otorhinolaryngol-Head Neck Surg 2013;56:700-5

A series of alkyl 15,35,55,75,95,115-hexa-tert-butyl-1,3,5,7,9,11-hexa[1,3]benzenacyclododecaphane esters have been prepared that selectively bind to alkylammonium cations. Ester ligands showed extraction efficiency toward alkylamines in the sequence of ammonium Pr > Pri > Bu > Bui≃ Bus > tert-butylammonium guests. Single and competitive transport experiments of butylamines also confirmed the binding properties obtained with extraction experiments and the transport selectivity for Bu/But pair exceeds 25. Conformational properties, such as Tc and ΔGc‡, of ester ligands were investigated by means of temperature-dependent 1H NMR spectroscopy. Conformational freedom was generally found to decrease as the size of substituent of ester side chain increased. 1H NMR complexation studies of ethyl ester 2 with caesium tetraphenylborate and butylammonium picrate guests suggest that the conformational reorganization into a cone conformation has provoked complex formation.