Evaluation of safety and anti-obesity effects of DWP16001 in naturally obese dogs.

BACKGROUND: The aim of this study was to investigate the anti-obesity effects of DWP16001, a sodium-glucose cotransporter-2 (SGLT2 inhibitor), in naturally obese dogs. A total of 20 dogs were divided into four equal groups: one obese control (OC group), and three treated groups; DWP0.2 group, DWP0.5 group, and DWP1 group. OC group fed with food for maintenance and treated groups were fed with food for maintenance with 0.2 mg/kg DWP16001, 0.5 mg/kg DWP16001 and 1 mg/kg DWP16001, respectively. The food for maintenance was provided to dogs as 2 RER (Resting energy requirement) in kcal and DWP16001-supplemented food was administered once a day for 8 weeks. RESULTS: Body condition score, body weight, and fat thickness were significantly reduced (P < 0.05) in the DWP0.2 group compared with the OC group, respectively without affecting the food consumption. At the 10th week the food consumption rate was 101.35 ± 2.56, 166.59 ± 4.72, 98.47 ± 1.44 and 123.15 ± 2.45% compared with initial food consumption rate. Body fat percentage, chest and waist circumference, blood glucose, and insulin were reduced compared to OC group but not significantly different from those of the OC group during experimental period. Serum alanine aminotransferase, alkaline phosphatase, creatine phosphokinase, and creatinine were significantly reduced in DWP0.2 group on 8 weeks. Serum cholesterol and triglycerides were reduced but not significantly. No specific adverse effects were observed throughout the experiment, and hematological parameters were unchanged. The results indicate that DWP16001 was not harmful to the dogs in our study and might have anti-obesity effects in naturally obese dogs. CONCLUSIONS: The above results and discussion suggest that DWP16001 is safe and might have anti-obesity effects in naturally obese dogs.

Macrophage Depletion in CCR2-/- Mice Delays Bacterial Clearance and Enhances Neutrophil Infiltration in an Acute Otitis Media Model.

BACKGROUND: Otitis media (OM) is a common and potentially serious disease of childhood. Although OM is multifactorial on origin, bacterial infection is a unifying component. Many studies have established a critical role for innate immunity in bacterial clearance and OM resolution. A key component of innate immunity is the recruitment of immune and inflammatory cells, including macrophages. METHODS: To explore the role of macrophages in OM, we evaluated the expression of genes related to macrophage function during a complete episode of acute OM in the mouse caused by middle ear (ME) inoculation with Haemophilus influenzae. We also combined CCR2 deficiency with chlodronate liposome toxicity to deplete macrophages during OM. RESULTS: Macrophage genes were robustly regulated during OM. Moreover, macrophage depletion enhanced and prolonged the infiltration of neutrophils into the infected ME and increased the persistence of bacterial infection. CONCLUSIONS: The results illustrate the critical role played by macrophages in OM resolution.

Postoperative results of ventilation tube insertion: a retrospective multicenter study for suggestion of grading system of otitis media with effusion.

BACKGROUND: In otitis media with effusion (OME), it is important to know when to surgically intervene and when careful monitoring is more appropriate. This study aimed to visualize and classify the clinical manifestations of OME and the correlation between the new grading system and postoperative results after ventilation tube insertion (VTI). METHODS: We classified the collective 1,012 ears from 506 patients into six groups: grade 0 (no effusion), grade I (scant effusion, but abnormal), grade II (effusion less than half of the tympanic cavity), grade III (effusion over half of the tympanic cavity, with air bubbles), grade IV (complete effusion), and grade V (retracted tympanic membrane or hemotympanum without air bubbles). RESULTS: The mean age at VTI was 5.2 (±2.9) years and mean duration between diagnosis and operation was 4.1 (±1.8) months. Between the grades, the nature of the middle ear effusion was also significantly different (p < 0.001). The duration of ventilation tube retention after VTI was significantly different when compared between two groups: grade I-IV and grade V (p = 0.019). Our results showed that the recurrence rate, as well as rate of revision VTI, increased as the grade increased (p < 0.001). CONCLUSIONS: The new grading system of OME using endoscopic otoscope evaluation had a significant correlation with the age at VTI, the nature of middle ear effusion, the recurrence rate of OME, and the rate of revision VTI.

Lack of the hyaluronan receptor CD44 affects the course of bacterial otitis media and reduces leukocyte recruitment to the middle ear.

BACKGROUND: CD44 is a multifunctional molecule that plays major roles in both leukocyte recruitment and tissue proliferation. Since mucosal hyperplasia and leukocyte infiltration of the middle ear cavity are major features of otitis media, we evaluated the role of CD44 in the pathophysiology and course of this disease in a mouse model of middle ear infection. Expression of genes related to CD44 function were evaluated using gene arrays in wild-type mice. The middle ears of mice deficient in CD44 were inoculated with non-typeable Haemophilus influenzae. Histopathology and bacterial clearance were compared to that seen in wild-type controls. RESULTS: We observed strong up-regulation of CD44 and of genes related to its role in leukocyte extravasation into the middle ear, during the course of acute otitis media. Mice deficient in CD44 exhibited reduced early mucosal hyperplasia and leukocyte recruitment, followed by delayed resolution of infection and persistent inflammation. CONCLUSIONS: CD44 plays an important role in OM pathogenesis by altering the mucosal growth and neutrophil enlistment. Targeted therapies based on CD44 could be useful adjuncts to the treatment of middle ear infections.

Modulation of the intestinal microbiota of dogs by kefir as a functional dairy product.

Kefir is a traditional dairy product with multiple probiotic characteristics derived from its associated microorganisms, including more than 50 species of lactic acid bacteria and yeast. For centuries, many people have produced kefir for human consumption; its consumption and potential role as a probiotic supplement in companion animals have never been tested. The present study explored the potential application of kefir as a probiotic supplement for dogs. Kefir was orally administered to healthy adult dogs (n = 6) for 2 wk. On d 0 and 14 (before and after kefir consumption, respectively), gut microbiota was analyzed comprehensively using quantitative PCR and 16S rDNA amplicon-based community analysis using fresh fecal samples. The 16S rDNA amplicon-based community analysis showed that the relative abundance of the phylum Fusobacteria was significantly decreased after kefir consumption. Furthermore, the relative abundance of the families Prevotellaceae, Selenomonadaceae, and Sutterellaceae increased significantly, whereas that of the families Clostridiaceae, Fusobacteriaceae, and Ruminococcaceae decreased significantly. The quantitative PCR assay showed that kefir consumption significantly increased the population of lactic acid bacteria and the lactic acid bacteria:Enterobacteriaceae ratio and significantly decreased the Firmicutes:Bacteroidetes ratio. In summary, 2-wk kefir administration successfully modified the gut microbiota without causing any clinically evident adverse effects. Therefore, kefir could be further developed as a novel probiotic food supplement for dogs to improve the quality of life of dogs.

Idiopathic sudden sensorineural hearing loss in dialysis patients.

Although sudden sensorineural hearing loss (SSNHL) affects chronic kidney disease (CKD) patients more frequently than non-CKD patients, few reports have described SSNHL in dialysis patients. We aimed to review the characteristics of SSNHL in chronic dialysis patients and evaluate treatment responses to steroid therapy. We retrospectively reviewed the records of dialysis patients diagnosed with idiopathic SSNHL at Asan Medical Center between January 2000 and December 2014. Pure tone and speech audiometry analyzes were performed before and 2 weeks and 2 months after treatment onset to evaluate outcomes. Twenty-two patients (11 men, 11 women; mean age: 49.9 ± 11.7 years) were included; 16 (72%) and 6 (28%) had undergone hemodialysis and peritoneal dialysis, respectively, for a median of 49.2 ± 41.4 (1-144) months. End-stage renal disease was most frequently caused by diabetic nephropathy (11 cases), chronic glomerulonephritis (1 case) and unknown factors (7 cases). Common accompanying symptoms included tinnitus (68.2%), ear fullness (45.5%) and vertigo (27.3%). The mean pure tone audiometry threshold at the initial presentation was 82.6 ± 22.4 dB. At 2 months post-steroid treatment, 4 (18.2%), 4 (18.2%) and 6 (27.3%) patients exhibited a complete, partial, or slight recovery, respectively; 8 patients (36.3%) showed no improvement. Although we could not identify the specific cause of SSNHL in this population, our relatively large case series elucidates the precise clinical features of SSNHL in this population and demonstrates the outcomes of steroid treatment.

Comparison Of Isokinetic Muscle Strength And Clinical Outcomebetween Isolated And Combined PCL Reconstruction.

The purposes of this study was to 1) compare the functional result and muscle strength between patients who underwent reconstruction by isolated posterior cruciate ligament (PCL) and combined PCLposeterolateral corner sling (PLCS) reconstruction. Nineteen (Group I: isolated PCL reconstruction) and 30 (Group II: combined PCL-PLCS reconstruction) were compared. The clinical results and isokinetic muscle strength were compared between groups. The clinical results were comparable between groups and posterior stability was not different between the two groups, either. The isokinetic strength test result was not significantly different between the two groups, although absolute values of the flexion strength in Group I was higher than those in Group II. The HQ ratio of the affected side in Group I was similar to that of the contralateral side. Isokinetic muscle strength, especially flexion, was not fully recovered as compared with the contralateral side in both groups, although clinical and stability results were improved. However, the isolated PCL reconstruction group that was managed with the accelerated rehabilitation protocol showed an HQ ratio similar to that of the contralateral side. Therefore, an earlier active strengthening program would be also necessary in PCL-PLCS reconstructed groups, and justification for prevention of early weight bearing in PCL-PLCS reconstruction should be re-evaluated because weight bearing is helpful for muscle strengthening.

Simultaneous versus Sequential Intratympanic Steroid Treatment for Severe-to-Profound Sudden Sensorineural Hearing Loss.

OBJECTIVES: Severe-to-profound sudden sensorineural hearing loss (SSNHL) has a poor prognosis. We aimed to compare the efficacy of simultaneous and sequential oral and intratympanic steroids for this condition. METHODS: Fifty patients with severe-to-profound SSNHL (>70 dB HL) were included from 7 centers. The simultaneous group (27 patients) received oral and intratympanic steroid injections for 2 weeks. The sequential group (23 patients) was treated with oral steroids for 2 weeks and intratympanic steroids for the subsequent 2 weeks. Pure-tone averages (PTA) and word discrimination scores (WDS) were compared before treatment and 2 weeks and 1 and 2 months after treatment. Treatment outcomes according to the modified American Academy of Otolaryngology-Head and Neck Surgery (AAO-HNS) criteria were also analyzed. RESULTS: The improvement in PTA and WDS at the 2-week follow-up was 23 ± 21 dB HL and 20 ± 39% in the simultaneous group and 31 ± 29 dB HL and 37 ± 42% in the sequential group; this was not statistically significant. Complete or partial recovery at the 2-week follow-up was observed in 26% of the simultaneous group and 30% of the sequential group; this was also not significant. The improvement in PTA and WDS at the 2-month follow-up was 40 ± 20 dB HL and 37 ± 35% in the simultaneous group and 41 ± 25 dB HL and 48 ± 41% in the sequential group; this was not statistically significant. Complete or partial recovery at the 2-month follow-up was observed in 33% of the simultaneous group and 35% of the sequential group; this was also not significant. Seven patients in the sequential group did not need intratympanic steroid injections for sufficient improvement after oral steroids alone. CONCLUSIONS: Simultaneous oral/intratympanic steroid treatment yielded a recovery similar to that produced by sequential treatment. Because the addition of intratympanic steroids can be decided upon based on the improvement after an oral steroid, the sequential regimen can be recommended to avoid unnecessary intratympanic injections.

Novel stilbene-based Fischer base analog of leuco-TAM - (2E,2'Z)-{2-(4-(E)-styrylphenyl)propane-1,3-diylidene}bis(1,3,3-trimethylindoline) - derivatives: synthesis and structural consideration by 1D NMR and 2D NMR spectroscopy.

We report the synthesis of a series of novel stilbene-based (St) Fischer base analogs of leuco-triarylmethane (LTAM) dyes by treating Fischer base with (E)-4-styrylbenzaldehyde derivatives. All St-LTAM molecules examined herein are characterized by 1D and 2D NMR. They were found to exhibit ZE configuration and isomerize to their diastereomers EE and ZZ in 2-3 h. They exhibit type I behavior of diastereomeric isomerization.

A multicenter randomized double-blind study: comparison of the Epley, Semont, and sham maneuvers for the treatment of posterior canal benign paroxysmal positional vertigo.

We evaluated the short-term efficacy of Epley, Semont, and sham maneuvers for resolving posterior canal benign paroxysmal positional vertigo (BPPV) in a prospective multicenter randomized double-blind controlled study. Subjects were randomly divided into three groups: Epley (36 patients), Semont (32 patients), and sham (Epley maneuver for the unaffected side, 31 patients). Out of 14 institutes which participated in this study, 5 institutes had previous experience of the Epley but not the Semont maneuver and the other 9 had previous experience of both maneuvers. Each maneuver was repeated twice if there was still positional vertigo or nystagmus on day 0, and the presence of nystagmus and vertigo on positional testing were evaluated immediately, 1 day, and 1 week after treatment. After the first maneuver, the Epley group showed a significantly higher resolution rate of positional nystagmus than the Semont or sham groups (63.9, 37.5, and 38.7%, respectively). After the second maneuver, the resolution rate (83.3%) of the Epley group was significantly higher than that (51.6%) of the sham group. At 1 day and 1 week after treatment, the resolution rate of the Epley group was significantly higher than those of the other groups. Similar results were seen for the resolution of positional vertigo. The Epley maneuver showed persistent resolution rates of positional vertigo and nystagmus without a fatigue phenomenon. The Epley maneuver was significantly more effective per maneuver than Semont or sham maneuvers for the short-term treatment of posterior canal BPPV. The Semont maneuver showed a higher success rate than the sham maneuver, but it was not significantly different.

Vulnerability to acoustic trauma in the normal hearing ear with contralateral hearing loss.

OBJECTIVES: We undertook an animal study to investigate the functional and histological changes that occur in the normal hearing ear of following acoustic trauma. METHODS: As an animal model of unilateral hearing loss, the right ears of CBA mice were deafened by cochlear destruction at 6 weeks of age (SSD group). The control groups included mice that underwent a sham surgery, and mice that were exposed to noise binaurally and monaurally (by plugging the right ear completely). At 10 weeks of age, all mice were exposed to acoustic trauma (110 dB sound pressure level for 1 hour) that induced a transient threshold shift (TTS). Changes in the hearing thresholds of the left ear were assessed over the next 4 weeks by measuring the auditory brainstem responses (ABRs) and distortion product otoacoustic emissions (DPOAEs). RESULTS: Following the noise exposure, the SSD group showed a permanent threshold shift (PTS) of about 10 dB, whereas the other groups showed full recovery from the TTS. The threshold of the DPOAEs of the left ears were increased after noise exposure but returned to normal in all groups, with no significant differences among the groups. Histological evaluation showed no apparent cellular loss or apoptosis in the left ears of all groups, including the SSD group. CONCLUSIONS: These results suggest that normal hearing ears are more vulnerable to acoustic trauma following contralateral unilateral cochlear ablation. This increased vulnerability may be due to damaged neural structures.

Results of anterior facial nerve rerouting procedures for removing skull base tumors.

OBJECTIVES: The purpose of this study was to estimate the rates of functional recovery of the facial nerve and of total tumor resection in patients who undergo short anterior rerouting and long anterior rerouting of the facial nerve in removal of skull base tumors. METHODS: We retrospectively collected data on 37 patients with skull base tumors who underwent facial nerve rerouting during the procedure for tumor removal. Information on the rerouting technique, the completeness of tumor resection, and changes in facial nerve function were obtained from the medical records. Rerouting techniques were classified as short anterior rerouting or long anterior rerouting. RESULTS: Ten of 16 patients (62.5%) in the group with short anterior rerouting showed postoperative facial palsy, and all completely recovered within 1 year. In the group with long anterior rerouting, 18 of 21 patients (85.7%) showed postoperative facial palsy, and recovery to a preoperative level of facial function was found in 10 patients at 1 year of follow-up. Total tumor resection was possible in 94% and 81% of patients with short rerouting and long rerouting, respectively. The mean operation time was not significantly related to the postoperative recovery of facial function. CONCLUSIONS: Short rerouting techniques, when appropriately chosen on the basis of tumor and patient characteristics, offer excellent preservation of facial function and tumor resection, comparable to those of long rerouting techniques.

Effect of the sodium-glucose cotransporter-2 inhibitor, DWP16001, as an add-on therapy to insulin for diabetic dogs: A pilot study.

BACKGROUND: Sodium-glucose cotransporter-2 (SGLT2) inhibitors are a novel class of anti-hyperglycaemic agents. OBJECTIVE: This study aimed to evaluate the safety and the adjuvant glycaemic control effect of an SGLT2 inhibitor, DWP16001, in diabetic dogs receiving insulin treatment. METHODS: Nineteen diabetic dogs receiving insulin treatment (NPH, porcine lente and glargine insulin) were divided into two groups according to dosing frequency: DWP TOD group (n = 10) and DWP SID group (n = 9). In the DWP TOD group, 0.025 mg/kg of DWP16001 was administered once every 3 days, whereas, in the DWP SID group, 0.025 mg/kg of DWP16001 was administered once a day. Food intake was maintained during the trial period. Hypoglycaemia, ketoacidosis or unexpected life-threatening reactions were assessed as adverse effects before and after DWP16001 administration. We compared insulin requirement reduction and blood glucose level control between two groups. RESULTS: No specific adverse effects were observed during the clinical trial, and haematological parameter remained unchanged. Moreover, the fasting glucose levels and daily insulin dose in the DWP TOD group were lower than the pre-administration values, but not significantly different for 8 weeks. Systolic blood pressure, fructosamine and insulin dose decreased significantly in the DWP SID group compared to the DWP TOD group at 8 weeks (p < 0.05) without affecting food consumption. Among these patients, 10 patients were monitored while receiving DWP16001 for 12 months (DWP TOD group n = 5, DWP SID group n = 5). The fasting glucose and fructosamine levels and daily insulin dose were reduced in both groups at 12 months compared with those before receiving DWP16001. CONCLUSION: When DWP16001, an SGLT2 inhibitor, was supplied to dogs with type 1 diabetes, no adverse effects were observed, and it was confirmed that the administered insulin dose can be reduced in controlling blood glucose.

Expression of matrix metalloproteinase 2 and 9 and tissue inhibitor of metalloproteinase 1 in nonrecurrent vs recurrent nasal polyps.

BACKGROUND: Matrix metalloproteinases (MMPs) 2 and 9 are a group of Zn2+-dependent endopeptidases to remodel the extracellular matrix. The association of these 2 endopeptidases with the development of morphologic changes in nasal polyps was previously described. OBJECTIVE: To determine whether MMP-2, MMP-9, and tissue inhibitor of MMP-1 (TIMP-1) play a role in the recurrence of nasal polyps. METHODS: To compare MMP-2, MMP-9, and TIMP-1 expression in recurrent vs nonrecurrent polyps, nasal polyps (NPs) and recurrent nasal polyps (RNPs) were obtained from 15 NP patients with chronic rhinosinusitis (CRS) undergoing endoscopic sinus surgery (ESS) and 15 RNP patients with CRS undergoing revision ESS. Fifteen specimens of inferior turbinate mucosa from patients undergoing nasal septal surgery were used as control. Through real-time polymerase chain reaction and immunohistochemistry, MMP-2, MMP-9, and TIMP-1 expressions were measured among controls, NP patients, and RNP patients. RESULTS: Expression of MMP-9 messenger RNA in the NP patients was significantly higher than in the controls. In addition, expression of MMP-9 messenger RNA in the RNP patents was significantly increased compared with NP patients. With immunohistochemistry, a more increased expression of MMP-9 was observed in NP patients than controls. Expression of MMP-9 in RNP patients was also significantly high compared with NP patients, particularly in stroma. CONCLUSION: Expression of MMP-9 is increased in NP patients, and it is so more in the mucosa of RNP patients. Increased expression of MMP-9 is particularly found in the stoma of RNP patients, and it may contribute to the recurrence of NP.

Hierarchical Heterogeneous NiFe Layered Double Hydroxides for Efficient Solar-Powered Water Oxidation.

Highly active, stable, and low-cost oxygen evolution reaction (OER) electrocatalysts are urgently needed for the realization of large-scale industrial hydrogen production via water electrolysis. Layered double hydroxides (LDHs) stand out as one of the most promising nonprecious electrocatalysts worth pursuing. Here, a hierarchical heterogeneous Ni2+Fe3+@Ni2+Fe2+ LDH was successfully synthesized via a sequential electrodeposition technique using separate electrolytes containing iron precursors with different valence states (Fe2+, Fe3+). The underlying highly crystalline Ni2+Fe2+ LDH nanosheet array provides a large surface for the catalytically more active Ni2+Fe3+ LDH overlayer with low crystallinity. The resulting Ni2+Fe3+@Ni2+Fe2+ LDH demonstrates excellent OER activity with overpotentials of 218 and 265 mV to reach current densities of 10 and 100 mA cm-2, respectively, as well as good long-term stability for 30 h even at a high current density of 500 mA cm-2. In an overall water splitting, an electrolyzer using an electrocatalyst of Sn4P3/CoP2 as a cathode requires only a cell voltage of 1.55 V at 10 mA cm-2. Furthermore, the solar-powered overall water splitting system consisting of our electrolyzer and a perovskite/Si tandem solar cell exhibits a high solar-to-hydrogen conversion efficiency of 15.3%.

Congenital cholesteatoma: clinical features and growth patterns.

OBJECTIVE: The clinical features and patterns of growth, relative to age, were examined in patients with congenital cholesteatoma to investigate disease progression and site of origin. PATIENTS AND METHODS: We retrospectively reviewed 72 children younger than 15 years with intraoperatively confirmed congenital cholesteatoma diagnosed using the inclusion criteria of Levenson et al. Patient demographics, history, and otoscopic and operative findings were evaluated. Correlation between mass volumetric and operation age was analyzed in patients with closed-type masses. Parameters of disease extent of middle ear quadrant, ossicular erosion, attic involvement, invasion of the mastoid cavity, and type of mass were assessed relative to age at operation. RESULTS: Mean patient age was 63.1 months (range, 20-179 months), with 51 patients (70.8%) being asymptomatic and diagnosed incidentally. Nineteen patients (26.4%) had closed-type cystic masses, and 53 (73.6%) had open-type lesions. The volume of closed-type masses was linearly correlated with age at operation. Logistic regression showed that increased age at operation was associated with a lower proportion of anterosuperior quadrant lesions, resulting in uncertainty about the site of origin. The possibility of open-type masses also increased according to age at operation. CONCLUSION: Congenital cholesteatoma shows growth and extension over time. Early detection and intervention are necessary to avoid advanced disease.

Distinctive clinical characteristics and suicidal tendencies of patients with anxious depression.

This study evaluated clinical characteristics and suicidality of patients with anxious depression in a large cohort of samples. Data were collected from 1003 patients who were depressed. A total of 461 patients were diagnosed with anxious depression and 542 were diagnosed with nonanxious depression. After adjusting for the severity of depression, those in the anxious depression group had significantly younger onset age, had been suffering from depression for a longer period, were more likely to experience a recurrence, and obtained lower scores on a scale assessing quality of life. The anxious depression group was characterized by a significantly higher proportion of individuals reporting significant suicidal ideation and previous suicide attempts, and those in this group tended to obtain higher scores on the Scale for Suicide Ideation. The present findings that were drawn from detailed evaluation of suicidality strongly support previous results assessed only with the help of clinical reports. More attention should be paid to assess suicide risk in these patients.

Recurrent bilateral branch retinal artery occlusion with hearing loss and encephalopathy: the first case report of Susac syndrome in Korea.

We report the first case of Susac syndrome in Koreans, in a 23-yr-old female patient who presented with sudden visual loss and associated neurological symptoms. Ophthalmic examination and fluorescein angiography showed multiple areas of branch retinal artery occlusion, which tended to recur in both eyes. Magnetic resonance imaging showed dot-like, diffusion-restricted lesions in the corpus callosum and left fornix, and audiometry showed low-frequency sensory hearing loss, compatible with Susac syndrome. She received immunosuppressive therapy with oral steroid and azathioprine. Three months later all the symptoms disappeared but obstructive vasculitis have been relapsing. This patient demonstrated the entire clinical triad of Susac syndrome, which tends to occur in young females. Although this disorder has rarely been reported in Asian populations, a high index of suspicion is warranted for early diagnosis and timely treatment.

Smad, PI3K/Akt, and Wnt-dependent signaling pathways are involved in BMP-4-induced ESC self-renewal.

It is known that bone morphogenetic protein 4 (BMP-4) has a diverse effect on ESCs. However, its precise mechanism in mouse ESCs is not fully understood. We evaluated the effect of BMP-4 on ESC proliferation and its related signal cascades in this study. BMP-4 significantly increased the level of [(3)H]-thymidine incorporation in time- (> or =8 hours) and dose- (> or =10 ng/ml) dependent manners. Additionally, BMP-4 increased cyclin D1 and decreased p27(kip1) expression values in a time-dependent manner. The increases in BMP-4-induced [(3)H]-thymidine incorporation and cyclin D1 expression were inhibited by the BMP-4 receptor antagonist noggin. BMP-4 increased Wnt1 expression. Wnt1 expression was attenuated by Smad4 small interfering RNA (siRNA), and BMP-4-induced cyclin D1 expression was inhibited by Smad4 and Wnt1 siRNAs. BMP-4 also activated beta-catenin, which was blocked by Smad4 and Wnt1 siRNAs. In addition, BMP-4 induced Akt phosphorylation. BMP-4-induced beta-catenin activation and cyclin D1 expression were attenuated by phosphatidyl inositol 3-kinase (PI3K) siRNA and Akt inhibitor. Additionally, downregulation of Smad4, Wnt1, and PI3K expression by siRNA decreased the levels of pluripotency marker mRNAs of ESCs, including Oct4, Sox2, and FoxD3. Our results suggested that BMP-4-induced [(3)H]-thymidine incorporation was significantly attenuated by Smad4, Wnt1, and PI3K knockdown. In conclusion, BMP-4 contributed to the maintenance of cell proliferation and the pluripotent state by Smad, PI3K/Akt, and Wnt1/beta-catenin in mouse ESCs.

Intratympanic steroid injection for sudden sensorineural hearing loss: Impact of injection interval on therapeutic efficacy.

OBJECTIVE: To compare the effect of injection time intervals of intratympanic (IT) dexamethasone (DEX) in patients with idiopathic sudden sensorineural hearing loss (ISSNHL). METHODS: Seventy-five adults with ISSNHL were grouped into four groups according to the IT DEX interval. In addition to concurrent oral steroid medication for two weeks, patients received IT DEX injections every 1, 2, 3, and four days, respectively. (Group 1, Group 2, Group 3, and Group 4). We evaluated the treatment outcomes according to modified criteria from "Clinical Practice Guideline: Sudden Hearing Loss" of the American Academy of Otolaryngology-Head and Neck Surgery (AAOHNS) to justify treatment success. RESULTS: There were no significant differences in demographic and baseline audiometric data. The mean of pure tone audiometry (PTA) and speech discrimination score (SDSs) were significantly improved after oral steroid and IT DEX treatment in all four groups. Group 1 showed significantly higher improvement than Group 4 in PTA after treatment. There was a significantly higher complete recovery (CR) rate in Group 1 than Group 4. CONCLUSION: We found a statistically significant difference in the complete hearing recovery rate and audiometric results (PTA) between the group with a daily interval of injections and the group with a four-day time interval. Therefore, daily time intervals in intratympanic steroid injection may be considered as an option for better improvement of hearing in patients with ISSNHL.