Photobiomodulation with near-infrared laser for tinnitus management: preliminary animal experiments and randomized clinical trials.

This study investigates the effectiveness of photobiomodulation therapy (PBMT) in treating chronic high-frequency tinnitus with the TINI device, a near-infrared (830 nm) laser. The study includes preliminary animal experiments with 28 mice and a randomized controlled trial with 56 participants to examine the functional and molecular changes in the auditory system that PBMT may cause. The animal model used sodium salicylate to induce tinnitus, followed by PBMT, which showed promising reductions in the behavioral evidence of tinnitus and a reversal of tinnitus-associated upregulation of vesicular glutamate transporters 2 expression in the ipsilateral dorsal cochlear nucleus (p < 0.05). In the clinical trial, participants with chronic high-frequency tinnitus received trans-tympanic application of the TINI device. The results did not show a significant difference in tinnitus score at the final time point when compared to the sham group. However, questionnaires revealed significant improvements in tinnitus symptoms and psychological outcomes following treatment with the TINI device compared to before treatment (p < 0.05). These findings suggest that while PBMT has potential benefits, its clinical effectiveness may be unclear due to its complex nature and interaction with other conditions. Further research is required to optimize treatment parameters and gain a complete understanding of the therapeutic potential of PBMT in managing tinnitus.

Diagnostic Roles of α-Methylacyl-CoA Racemase (AMACR) Immunohistochemistry in Gastric Dysplasia and Adenocarcinoma.

Background and Objectives: This study aimed to elucidate the diagnostic role of α-Methylacyl-CoA racemase (AMACR) immunohistochemistry in gastric dysplasia and adenocarcinoma. Materials and Methods: Immunohistochemistry for AMACR was performed on 39 gastric dysplasia and 40 gastric adenocarcinoma cases. The expression patterns of AMACR were investigated and divided into luminal and cytoplasmic expression patterns in the gastric lesions. In addition, correlations between AMACR expression and patient age, sex, and tumor size were evaluated. Results: AMACR was expressed in 26 of 39 cases of gastric dysplasia (66.7%) and 17 of 40 cases of gastric adenocarcinomas (42.5%). The AMACR expression rates in high- and low-grade dysplasia were 80.0% and 52.6%, respectively. A detailed analysis of the expression patterns revealed that the luminal expression pattern was significantly higher in low-grade dysplasia than in high-grade dysplasia and gastric adenocarcinoma (p < 0.001). The cytoplasmic expression pattern, without luminal expression, was predominant in high-grade dysplasia and gastric adenocarcinoma. In addition, the rates of loss of expression in the overall area were 15.1 ± 23.9%, 49.0 ± 29.9%, and 59.0 ± 32.2% in low-grade dysplasia, high-grade dysplasia, and gastric adenocarcinoma, respectively. The negative rate of low-grade dysplasia was significantly lower than that of high-grade dysplasia and gastric adenocarcinoma (p < 0.001 and p < 0.001, respectively). Conclusions: AMACR is a useful diagnostic marker for differentiating low-grade dysplasia from high-grade dysplasia and gastric adenocarcinoma. Luminal or cytoplasmic expression patterns and the extent of loss of expression are important for differentiation.

The prognostic value of a combined immune score in tumor and immune cells assessed by immunohistochemistry in triple-negative breast cancer.

BACKGROUND: This study aimed to develop a novel combined immune score (CIS)-based model assessing prognosis in triple-negative breast cancer (TNBC). METHODS: The expression of eight immune markers (PD-1, PD-L1, PD-L2, IDO, TIM3, OX40, OX40L, and H7-H2) was assessed with immunohistochemistry on the tumor cells (TCs) and immune cells (ICs) of 227 TNBC cases, respectively, and subsequently associated with selected clinicopathological parameters and survival. Data retrieved from The Cancer Genome Atlas (TCGA) were further examined to validate our findings. RESULTS: All immune markers were often expressed in TCs and ICs, except for PD-1 which was not expressed in TCs. In ICs, the expression of all immune markers was positively correlated between one another, except between PD-L1 and OX40, also TIM3 and OX40. In ICs, PD-1, PD-L1, and OX40L positive expression was associated with a longer progression-free survival (PFS; p = 0.040, p = 0.020, and p = 0.020, respectively). In TCs, OX40 positive expression was associated with a shorter PFS (p = 0.025). Subsequently, the TNBC patients were classified into high and low combined immune score groups (CIS-H and CIS-L), based on the expression levels of a selection of biomarkers in TCs (TCIS-H or TCIS-L) and ICs (ICIS-H or ICIS-L). The TCIS-H group was significantly associated with a longer PFS (p < 0.001). Furthermore, the ICIS-H group was additionally associated with a longer PFS (p < 0.001) and overall survival (OS; p = 0.001), at significant levels. In the multivariate analysis, both TCIS-H and ICIS-H groups were identified as independent predictors of favorable PFS (p = 0.012 and p = 0.001, respectively). ICIS-H was also shown to be an independent predictor of favorable OS (p = 0.003). The analysis of the mRNA expression data from TCGA also validated our findings regarding TNBC. CONCLUSION: Our novel TCIS and ICIS exhibited a significant prognostic value in TNBC. Additional research would be needed to strengthen our findings and identify the most efficient prognostic and predictive biomarkers for TNBC patients.

Transition-Metal-Free Upscaling of 2,5-Furandicarboxylic Acid Synthesis and Investigation of the Reaction Mechanism.

An environmentally friendly oxidation system has proposed for the practical and scalable production of value-added 2,5-furandicarboxylic acid from 1 kg of 5-hydroxymethylfurfural. The system is composed of a simple base, oxygen, and a green solvent, thereby providing a sustainable and economical approach to organic synthesis. To gain insight into the mechanism of this oxidation process, NMR spectroscopic analysis and kinetic study are used for the mechanistic investigation of this environmentally friendly oxidation process.

Prevalence, Trend, and Risk Factors for Early Chronic Obstructive Pulmonary Disease: An Analysis of the Nationwide Population-Based Survey from 2010 to 2019 in South Korea.

This study aimed to evaluate the prevalence, trends, and risk factors of early chronic obstructive pulmonary disease (COPD) by using a nationally representative sample. The datasets of the Korea National Health and Nutrition Examination Survey 2010-2019 were used, where 80,860 individuals were identified; of these, 9,045 participants aged 40-49 years who underwent spirometry with no missing data were analyzed. Early COPD was defined as forced expiratory volume in 1 s /forced vital capacity ratio < the lower limit of normal (2.5th percentile) in individuals aged <50 years without a history of asthma, inhaler therapy, or persistent respiratory symptoms. The prevalence and trend of early COPD were estimated according to features such as smoking status and pack-years. Joinpoint regression analysis was used to analyze the significant annual change in the trend according to sex, smoking status, and pack-years. A complex sample multivariable-adjusted regression model was used to identify factors affecting early COPD. The estimated population size during 2010-2019 was 82,326,178. Early COPD was present in 4.5% of patients (6.5% of men and 2.3% of women). It was present in 7.7% of current smokers, followed by former and never smokers. Among smokers with ≥ 10 pack-years, early COPD was present in 8.2%, whereas it was present in 2.6% of smokers with < 10 pack-years. Joinpoint regression analyses found a recent decrease in the trend of prevalence in males who were former and current smokers. The multivariable-adjusted logistic regression model showed that being male, lower educational level, smoking status, and pack-years were factors that affected the presence of early COPD. Continued surveillance of this pre-disease condition is required, and further research are warrant.

USP13 regulates HMGB1 stability and secretion through its deubiquitinase activity.

BACKGROUND: High mobility group box 1 (HMGB1) is a damage-associated molecular pattern (DAMP) molecule that plays a central role in innate immunity. HMGB1 acts as a late mediator of inflammation when actively secreted in response to inflammatory stimuli. Several post-translational modifications (PTMs), including acetylation, phosphorylation, and oxidation, are involved in HMGB1 secretion. However, the E3 ligases of HMGB1 and the mechanism by which DUBs regulate HMGB1 deubiquitination are not well known. METHODS: LC-MS/MS, proximity ligation assay, immunoprecipitation were used to identify ubiquitin-specific protease 13 (USP13) as a binding partner of HMGB1 and to investigate ubiquitination of HMGB1. USP13 domain mutant was constructed for domain study and Spautin-1 was treated for inhibition of USP13. Confocal microscopy image showed localization of HMGB1 by USP13 overexpression. The data were analyzed using one-way analysis of variance with Tukey's honestly significant difference post-hoc test for multiple comparisons or a two-tailed Student's t-test. RESULTS: We identified ubiquitin-specific protease 13 (USP13) as a novel binding partner of HMGB1 and demonstrated that USP13 plays a role in stabilizing HMGB1 from ubiquitin-mediated degradation. USP13 overexpression increased nucleocytoplasmic translocation of HMGB1 and promoted its secretion, which was inhibited by treatment with Spautin-1, a selective inhibitor of USP13. CONCLUSION: Taken together, we suggest that USP13 is a novel deubiquitinase of HMGB1 that regulates the stability and secretion of HMGB1.

Inner Ear Symptoms Are Prevalent in Patients with High Head Abbreviated Injury Scale Scores after Blunt Head Trauma.

INTRODUCTION: The purpose of this article was to determine the prevalence of inner ear symptoms in patients with blunt head trauma and to explore whether the severity of head trauma was associated with the incidence of such symptoms. METHODS: We performed a retrospective review of 56 patients admitted with blunt head trauma who underwent audiovestibular evaluation within 1 month after injury. Two scales were used to measure the severity of trauma; these were the Glasgow Coma Scale (GCS) and the Head Abbreviated Injury Scale (H-AIS). Patients with sensorineural-type hearing loss, or dizziness with nystagmus, were considered to have inner ear symptoms. RESULTS: About half of all patients (45%) with blunt head trauma showed trauma-related inner ear symptoms. Patients with inner ear symptoms were significantly more likely to have H-AIS scores ≥4 than those without inner ear symptoms (p = 0.004), even without concomitant temporal bone fracture (p > 0.05). Also, patients with inner ear symptoms required a statistically significantly longer time (measured from admission) before undergoing their ontological evaluations than did those without such symptoms (p = 0.002), possibly due to prolonged bed rest and use of sedatives. CONCLUSION: Thus, detailed history-taking and early evaluation using trauma scales are essential for all patients suffering from severe head trauma. It may be necessary to initiate early treatment of traumatic inner ear diseases.

An Exploration Into Patients' Experiences That Make Them Feel Safe During Hospitalization: A Qualitative Study.

BACKGROUND: Patient safety is a crucial indicator of health care quality. It is necessary to check the subjective perception of patient safety from the patient's point of view as a consumer of healthcare services. To identify patients' experiences of safety and the themes that constitute the patients' feeling of safety during hospitalization. METHODS: A qualitative study, comprising five focus group discussions (seven people each), was conducted in South Korea between May and July 2018. Patients who were hospitalized for at least three days within one year were included. Researchers analyzed the transcribed script, and a content analysis was performed to describe patients' hospitalized experiences of safety. RESULTS: A total of 35 patients with an average age of 45.4 years participated in the study, and had experience of hospitalization for up to 32 days. The findings revealed four core themes and 14 sub-themes. Patients wanted to take initiative in controlling his/her reception of information and wanted healthcare providers to make the patient feel safe. Patients felt safe when hospitals provided unstinted and generous support. Also, public sentiment about national healthcare and safety made an effect on patient safety sentiment. CONCLUSION: Patients felt safe during hospitalization not only because of the explanation, attitude, and professionalism of the healthcare providers but also because of the support, system, and procedure of the medical institution. Healthcare providers and medical institutions should strive to narrow the gap in patient safety awareness factors through activities with patients. Furthermore, the government and society should make an effort to create a safe medical environment and social atmosphere.

Understanding Physicians' and Nurses' Adaption of National-Leading Patient Safety Culture Policy: A Qualitative Study in Tertiary and General Hospitals in Korea.

BACKGROUND: In Korea, the safety culture is led by national policy. How the policy ensures a patient safety culture needs to be investigated. This study aimed to examine the way in which physicians and nurses regard, understand, or interpret the patient safety-related policy in the hospital setting. METHODS: In this qualitative study, we conducted four focus group interviews (FGIs) with 25 physicians and nurses from tertiary and general hospitals in South Korea. FGIs data were analyzed using thematic analysis, which was conducted in an inductive and interpretative way. RESULTS: Three themes were identified. The healthcare providers recognized its benefits in the forms of knowledge, information and training at least although the policy implemented by the law forcibly and temporarily. The second theme was about the interaction of the policy and the Korean context of healthcare, which makes a "turning point" in the safety culture. The final theme was about some strains and conflicts resulting from patient safety policy. CONCLUSION: To provide a patient safety culture, it is necessary to develop a plan to improve the voluntary participation of healthcare professionals and their commitment to safety. Hospitals should provide more resources and support for healthcare professionals.

Management of facial nerve schwannoma: when is the timing for surgery.

PURPOSE: Although the estimated prevalence is extremely low, facial nerve schwannoma (FNS) is the most common primary tumor of the facial nerve (FN). In the present study, the outcome of surgical management in 18 patients with FNS was analyzed and an appropriate time for surgery was proposed. MATERIALS AND METHODS: A total of 18 patients with FNS who underwent surgical management by a single surgeon from 1999 to 2018 were retrospectively analyzed. RESULTS: Among the 18 patients, five had no facial paralysis before surgery. Near-total removal was performed in three cases, and two cases were managed with decompression. In 13 cases with various degree of preoperative facial palsy, nerve continuity was lost during surgery. FN was reconstructed using cable graft in ten cases, direct anastomosis in one case, and facial-hypoglossal nerve transfer in one case. Facial reanimation surgery without FN reconstruction was performed in one case due to a long-standing facial paralysis before surgery. Preoperative House-Brackmann (H-B) grade in all patients was significantly worse as tumor size increased. The correlation was not observed between the duration and severity of preoperative facial palsy. Analysis of 12 patients who underwent FN reconstruction revealed that all patients with good preoperative facial function (H-B grade II-III) recovered to H-B grade III after surgery (7/7, 100%). However, patients with poor preoperative facial function (H-B grade IV or worse) had only a 40% (2/5) chance of improving to grade III after surgery. Preoperative tumor size and duration of facial palsy did not affect postoperative final facial function. CONCLUSION: We suggest that H-B grade III facial palsy is the best time for surgical intervention, regardless of the tumor size or duration of facial palsy.

The collagen structure of C1q induces wound healing by engaging discoidin domain receptor 2.

BACKGROUND: C1q has been reported to reveal complement-independent roles in immune and non-immune cells. C1q binds to its specific receptors to regulate distinct functions that rely on the environment and cell types. Discoidin domain receptor 2 (DDR2) is activated by collagen and functions in wound healing by controlling matrix metalloproteinase (MMP) expression. Since C1q exhibits a collagen-like structure, we hypothesized that C1q might engage DDR2 to regulate wound healing and extracellular matrix (ECM) remodeling. METHODS: Cell-based assay, proximity ligation assay, ELISA, and surface plasmon analysis were utilized to investigate DDR2 and C1q binding. We also investigate the C1q-mediated in vitro wound healing ability using the human fibrosarcoma cell line, HT1080. RESULTS: C1q induced the phosphorylation of DDR2, p38 kinase, and ERK1/2. C1q and DDR2 binding improved cell migration and induced MMP2 and MMP9 expression. DDR2-specific shRNA reduced C1q-mediated cell migration for wound healing. CONCLUSIONS: C1q is a new DDR2 ligand that promotes wound healing. These findings have therapeutic implications in wound healing-related diseases.

Genetic diagnosis of infantile-onset epilepsy in the clinic: Application of whole-exome sequencing following epilepsy gene panel testing.

This study aimed to evaluate the clinical utility of whole-exome sequencing in a group of infantile-onset epilepsy patients who tested negative for epilepsy using a gene panel test. Whole-exome sequencing was performed on 59 patients who tested negative on customized epilepsy gene panel testing. We identified eight pathogenic or likely pathogenic sequence variants in eight different genes (FARS2, YWHAG, KCNC1, DYRK1A, SMC1A, PIGA, OGT, and FGF12), one pathogenic structural variant (8.6 Mb-sized deletion on chromosome X [140 994 419-149 630 805]), and three putative low-frequency mosaic variants from three different genes (GABBR2, MTOR, and CUX1). Subsequent whole-exome sequencing revealed an additional 8% of diagnostic yield with genetic confirmation of epilepsy in 55.4% (62/112) of our cohort. Three genes (YWHAG, KCNC1, and FGF12) were identified as epilepsy-causing genes after the original gene panel was designed. The others were initially linked with mitochondrial encephalopathy or different neurodevelopmental disorders, although an epilepsy phenotype was listed as one of the clinical features. Application of whole-exome sequencing following epilepsy gene panel testing provided 8% of additional diagnostic yield in an infantile-onset epilepsy cohort. Whole-exome sequencing could provide an opportunity to reanalyze newly recognized epilepsy-linked genes without updating the gene panel design.

Wnt Modulation Enhances Otic Differentiation by Facilitating the Enucleation Process but Develops Unnecessary Cardiac Structures.

Otic organoids have the potential to resolve current challenges in hearing loss research. The reproduction of the delicate and complex structure of the mammalian cochlea using organoids requires high efficiency and specificity. Recent attempts to strengthen otic organoids have focused on the effects of the Wnt signaling pathway on stem cell differentiation. One important aspect of this is the evaluation of undesirable effects of differentiation after Wnt activation. In the present study, we differentiated mouse embryonic stem cell embryoid bodies (EB) into otic organoids and observed two morphologies with different cell fates. EBs that underwent a core ejection process, or 'enucleation,' were similar to previously reported inner ear organoids. Meanwhile, EBs that retained their core demonstrated features characteristic of neural organoids. The application of a Wnt agonist during the maturation phase increased enucleation, as well as otic organoid formation, in turn leading to sensory hair cell-like cell generation. However, with a longer incubation period, Wnt activation also led to EBs with 'beating' organoids that exhibited spontaneous movement. This observation emphasizes the necessity of optimizing Wnt enhancement for the differentiation of specific cells, such as those found in the inner ear.

Inflachromene inhibits autophagy through modulation of Beclin 1 activity.

Autophagy is a central intracellular catabolic mechanism that mediates the degradation of cytoplasmic proteins and organelles, and regulation of autophagy is essential for homeostasis. HMGB1 is an important sepsis mediator when secreted and also functions as an inducer of autophagy by binding to Beclin 1. In this study, we studied the effect of inflachromene (ICM), a novel HMGB1 secretion inhibitor, on autophagy. ICM inhibited autophagy by inhibiting nucleocytoplasmic translocation of HMGB1 and by increasing Beclin 1 ubiquitylation for degradation by enhancing the interaction between Beclin 1 and E3 ubiquitin ligase RNF216. These data suggest that ICM could be used as a potential autophagy suppressor.

Importance of early diagnosis in LMNA-related muscular dystrophy for cardiac surveillance.

INTRODUCTION: The identification of LMNA-related muscular dystrophy is important because it poses life-threatening cardiac complications. However, diagnosis of LMNA-related muscular dystrophy based on clinical features is challenging. METHODS: We reviewed the clinical phenotypes of 14 children with LMNA variants, focusing on the cardiac function and genotypes. RESULTS: Most patients presented with motor developmental delay or gait abnormalities. Eight (57%) patients had prominent neck extensor weakness or contractures. All patients showed ankle contractures at an early stage. Regular cardiac surveillance allowed for the detection of dysrhythmias in 57% of patients at a mean age of 14 years (range, 5-26). All patients had missense variants; however, there were no clear phenotype-genotype correlations. DISCUSSION: Early diagnosis of LMNA-related muscular dystrophy provides an opportunity for cardiac surveillance, potentially leading to the prevention of cardiac mortality in children.

Clinical Implications of Poloxamer 407 as Packing Material in an Animal Model.

BACKGROUND: Endoscopic ear surgery has recently increased, but it is still inconvenient and time-consuming to place packing material in the middle ear with one hand. Poloxamer 407 (P407) is a thermo-reversible gel that can be easily administered with one hand into the middle ear cavity in liquid form. Upon warming to body temperature, the gel form of P407 can support the graft in the target position and is known to prevent postsurgical tissue adhesion. OBJECTIVES: We aim to investigate the feasibility of P407 as packing material in an animal model. Male Hartley guinea pigs (350 and 400 g) were utilized in this study. METHOD: The animals were randomly divided into 3 groups according to the packing material: the control group, the P407 group, and the gelatin group. To assess the role of packing material on bacterial colonization, left ears were inoculated with Streptococcus pneumoniae through the tympanic membrane using a 0° endoscope. Five days after inoculation, the middle ear cavity was packed through a transbullar approach using 18% P407 or gelatin in both ears. In the control group, no ear pack was inserted. The tympanic membrane was examined every week using a 0° 1.9-mm endoscope until 6 weeks. Half of the animals in each group were sacrificed 6 weeks after placement of the packing materials. RESULTS: Compared with the absorbable gelatin sponge, the P407 group showed little inflammation or fibrosis in the tympanic membrane and middle ear mucosa regardless of bacterial inoculation. The gelatin group showed severe otorrhea or perforation until 2 weeks in the right ear (2 of 4) and the left ear (1 of 4). Even though the endoscopic findings were similar between both packing groups at 6 weeks, histological analysis showed persistent packing material, inflammatory cells, and fibrosis in the gelatin group compared to the P407 group. CONCLUSIONS: This study suggested that P407 is feasible as a packing material to handle with one hand and to prevent adhesion, especially in infected middle ear mucosa. Although there is a lack of data on how well P407 supports grafts, we suggest that P407 could be a candidate for packing material in endoscopic ear surgery.

Experiences and responses of second victims of patient safety incidents in Korea: a qualitative study.

BACKGROUND: Healthcare professionals who experience trauma due to patient safety incidents can be considered second victims, and they also suffer from various difficulties. In order to support second victims, it is necessary to determine the circumstances of the incidents in question, along with the symptoms that the victims are experiencing and the support they require. A qualitative study on healthcare professionals of various occupations, such as physicians and nurses working in Korea, was conducted, and the experiences and response methods and processes of second victims were examined. METHODS: In-depth interviews were conducted with 16 healthcare professionals (six physicians, eight nurses, and two pharmacists) who had experienced a patient safety incident. All interviews were recorded and transcribed, and the data analysis was conducted in accordance with Strauss and Corbin's grounded theory. Both open coding and axial coding were performed. Consolidated criteria for reporting qualitative research (COREQ) were applied in this study. RESULTS: The results of the open coding demonstrated that the experiences of second victims can be categorized into "the reactions of the first victim and surrounding people after the incident," "Influence of factors aside from the incident," "the initial complex responses of the participants to the incident," "open discussion of the incident," "the culture in medical institutions regarding early-stage incident response," "the coping responses of the participants after incidents," and "living with the incident." Then, the seven categories in the open coding stage were rearranged according to the paradigm model, and the reaction process of the second victims was analyzed through process analysis, being divided into the "entanglement stage," "agitating stage," "struggling stage," "managing stage," and "indurating stage." CONCLUSIONS: This research is significant because it provides a comprehensive understanding of second victims' experiences in the eastern region of Korea, by obtaining data using a qualitative research method. The findings of the study also highlight the five stages of the second victim response process, and can be used to design a specialized second victim support program in Korea.

CaBP1 regulates Cav1 L-type Ca2+ channels and their coupling to neurite growth and gene transcription in mouse spiral ganglion neurons.

CaBP1 is a Ca2+ binding protein that is widely expressed in neurons in the brain, retina, and cochlea. In heterologous expression systems, CaBP1 interacts with and regulates voltage-gated Cav Ca2+ channels but whether this is the case in neurons is unknown. Here, we investigated the cellular functions of CaBP1 in cochlear spiral ganglion neurons (SGNs), which express high levels of CaBP1. Consistent with the role of CaBP1 as a suppressor of Ca2+-dependent inactivation (CDI) of Cav1 (L-type) channels, Cav1 currents underwent greater CDI in SGNs from mice lacking CaBP1 (C-KO) than in wild-type (WT) SGNs. The coupling of Cav1 channels to downstream signaling pathways was also disrupted in C-KO SGNs. Activity-dependent repression of neurite growth was significantly blunted and unresponsive to Cav1 antagonists in C-KO SGNs in contrast to WT SGNs. Moreover, Cav1-mediated Ca2+ signals and phosphorylation of cAMP-response element binding protein were reduced in C-KO SGNs compared to WT SGNs. Our findings establish a role for CaBP1 as an essential regulator of Cav1 channels in SGNs and their coupling to downstream pathways controlling activity-dependent transcription and neurite growth.

A preliminary study on the efficacy and safety of low level light therapy in the management of cochlear tinnitus: A single blind randomized clinical trial.

OBJECTIVES: To evaluate the efficacy and safety of low-level light therapy (LLLT) using new irradiation parameters for chronic unilateral tinnitus with cochlear dysfunction. DESIGN: A single-blind, randomized clinical trial. SETTING: Tertiary-care hospital center. PARTICIPANTS: Participants who had a history of chronic unilateral tinnitus (≥ 3 months) and pure-tone thresholds greater than 15dB (averaged for 3k, 4k, and 6k Hz). MAIN OUTCOME MEASURES: Numerical rating scales (NRS) measuring loudness, duration, and annoyance, the tinnitus handicap inventory (THI), and psychoacoustical matches of tinnitus loudness and minimum masking levels (MML). RESULTS: Thirty-eight participants were received either a 100-mW diode laser at 830-nm (TINI group; n=19) or placebo (sham group; n=19) irradiation through the tympanic membrane. No adverse events were reported during 2 weeks of 10-interventions (20 minutes/day, five days/week). The NRS measuring duration of tinnitus and psychoacoustical matches of tinnitus loudness significantly decreased over times in the TINI group (p<0.05). However, post-hoc analysis revealed that there was no significant decrease of tinnitus among different time points (baseline, during LLLT, immediately after LLLT, and two weeks after LLLT). There was no placebo effect in the Sham group. Participants who improved the duration by at least one point or improved the loudness matches by more than 5 dB SL two weeks after LLLT tended to have worse pure-tone thresholds. It may suggest that further study is needed in patients with worse pure-tone thresholds to evaluate the therapeutic efficacy of LLLT. CONCLUSION: Although this preliminary result is insufficient to support the therapeutic efficacy of new laser device for chronic tinnitus, further study is needed in a large number of selected patients.

N-linked glycosylation plays a crucial role in the secretion of HMGB1.

HMGB1 protein is a delayed mediator of sepsis that is secreted to the extracellular milieu in response to various stimulants, inducing a pro-inflammatory response. HMGB1 is devoid of an endoplasmic reticulum (ER)-targeting signal peptide; hence, the mechanism of extracellular secretion is not completely understood, although HMGB1 is secreted after being subjected to post-translational modifications. Here, we identified the role of N-glycosylation of HMGB1 in extracellular secretion. We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. Inhibition of N-glycosylation with tunicamycin resulted in a molecular shift of HMGB1 as assessed by gel electrophoresis. Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. These mutant proteins had reduced secretion even after acetylation, phosphorylation, oxidation and exposure to pro-inflammatory stimuli. Taken together, we propose that HMGB1 is N-glycosylated, and that this is important for its DNA interaction and is a prerequisite for its nucleocytoplasmic transport and extracellular secretion.