Antibodies and B cells recognising citrullinated proteins display a broad cross-reactivity towards other post-translational modifications.

OBJECTIVE: Autoantibodies against antigens carrying distinct post-translational modifications (PTMs), such as citrulline, homocitrulline or acetyllysine, are hallmarks of rheumatoid arthritis (RA). The relation between these anti-modified protein antibody (AMPA)-classes is poorly understood as is the ability of different PTM-antigens to activate B-cell receptors (BCRs) directed against citrullinated proteins (CP). Insights into the nature of PTMs able to activate such B cells are pivotal to understand the 'evolution' of the autoimmune response conceivable underlying the disease. Here, we investigated the cross-reactivity of monoclonal AMPA and the ability of different types of PTM-antigens to activate CP-reactive BCRs. METHODS: BCR sequences from B cells isolated using citrullinated or acetylated antigens were used to produce monoclonal antibodies (mAb) followed by a detailed analysis of their cross-reactivity towards PTM-antigens. Ramos B-cell transfectants expressing CP-reactive IgG BCRs were generated and their activation on stimulation with PTM-antigens investigated. RESULTS: Most mAbs were highly cross-reactive towards multiple PTMs, while no reactivity was observed to the unmodified controls. B cells carrying CP-reactive BCRs showed activation on stimulation with various types of PTM-antigens. CONCLUSIONS: Our study illustrates that AMPA exhibit a high cross-reactivity towards at least two PTMs indicating that their recognition pattern is not confined to one type of modification. Furthermore, our data show that CP-reactive B cells are not only activated by citrullinated, but also by carbamylated and/or acetylated antigens. These data are vital for the understanding of the breach of B-cell tolerance against PTM-antigens and the possible contribution of these antigens to RA-pathogenesis.

[Autoantibodies and the autoreactive immune response : There is more to ACPA than ACPA]. / Autoantikörper und die autoreaktive Immunantwort : ACPA sind mehr als nur ACPA.

The immune response against posttranslationally modified (PTM) antigens, in particular the generation of anti-citrullinated protein antibodies (ACPA), is a very specific hallmark of rheumatoid arthritis. The factors that initiate this immune response and the triggers that stimulate the transition from asymptomatic autoimmunity to autoimmune disease are so far unknown. Genetic risk factors and the maturation of the ACPA response prior to the onset of arthritis indicate an important role for helper T cells in this process. Antigens that trigger this process, however, remain to be defined. Notably, recent data demonstrate that ACPA do not only recognize citrullinated protein antigens. Other posttranslational protein modifications such as homocitrulline and acetyllysine are also recognized. This cross-reactivity towards different PTM antigens was found for various monoclonal ACPA and broadens the spectrum of antigens that can stimulate and activate ACPA-expressing B cells. Also, it suggests that such B cells could receive help from autoreactive but also from non-autoreactive T cells. This review summarizes these recent findings and provides insight into their potential relevance for the disease rheumatoid arthritis.

Effects of type II collagen epitope carbamylation and citrullination in human leucocyte antigen (HLA)-DR4(+) monozygotic twins discordant for rheumatoid arthritis.

The aim of this study is to investigate the effect of the native, citrullinated or carbamylated type II human collagen T cell- and B cell-epitopes on the adaptive immune response in rheumatoid arthritis (RA). Peripheral blood T and B cells obtained from a human leucocyte D4-related (antigen DR4(-) HLA-DR4)(+) woman with early RA, her healthy monozygotic twin and an unrelated HLA-DR3(+) woman with early RA were analysed for activation (CD154/CD69), apoptosis (annexin/7-aminoactinomycin), cytokine production [interferon (IFN)γ/interleukin (IL)-17/IL-4/IL-10/IL-6] and functional phenotype (CD45Ra/CCR7) after stimulation with the collagen native T cell epitope (T261-273), the K264 carbamylated T cell epitope (carT261-273), the native B cell epitope (B359-369) or the R360 citrullinated B cell epitope (citB359-369), and the combinations of these. The T cell memory compartment was activated by T cell epitopes in both discordant DR4(+) twins, but not in the DR3(+) RA. The collagen-specific activation of CD4(+) T cells was induced with both the native and carbamylated T cell epitopes only in the RA twin. Both T cell epitopes also induced IL-17 production in the RA twin, but a greater IL-4 and IL-10 response in the healthy twin. The citrullinated B cell epitope, particularly when combined with the carbamylated T cell epitope, induced B cell activation and an increased IL-6/IL-10 ratio in the RA twin compared to a greater IL-10 production in the healthy twin. Our data suggest that circulating collagen-specific T and B cells are found in HLA-DR4(+) subjects, but only RA activated cells express co-stimulatory molecules and produce proinflammatory cytokines. Carbamylation and citrullination further modulate the activation and cytokine polarization of T and B cells.

Ethyl-Zinc(II)-Cation Equivalents: Synthesis and Hydroamination Catalysis.

Ion-like ethylzinc(II) compounds with weakly coordinating aluminates [Al(OR(F))4](-) and [(R(F)O)3Al-F-Al(OR(F))3](-) (R(F)=C(CF3)3) were synthesized in a one-pot reaction and fully characterized by single-crystal X-ray diffraction, NMR and vibrational spectroscopy, and by quantum chemical calculations. The catalytic activity of ion-like Et-Zn[Al(OR(F))4] in intermolecular hydroamination and in the unusual double hydroamination of anilines and alkynes was investigated. Favorable performance was also found in comparison to the Et2Zn/[PhNMe2H](+)[B(C6F5)4](-) system generated in situ at lower catalyst loadings of 2.5 mol %.

[Biomarkers and personalized medicine]. / Biomarker und personalisierte Medizin.

Biomarkers form the basis for patient stratification and the development of individualized treatment strategies. As the understanding of the pathophysiological processes underlying rheumatic diseases increases, novel biomarkers will become available and established markers can be used more efficiently. Autoantibodies in rheumatoid arthritis (RA), for example, define a subgroup of patients with a specific risk profile and response to therapy. For this reason they have been added to the classification criteria for RA and are part of current treatment guidelines. In addition, novel markers are being evaluated and validated. For the concept of personalized medicine this indicates that the use of future therapeutic substances will be more frequently coupled to the detection of specific biomarkers. While this will decrease the number of patients who become eligible for certain treatments, it will increase efficacy and safety for patients and potentially the cost-effectiveness.

Structure-activity relationships of actively FhuE transported rifabutin derivatives with potent activity against Acinetobacter baumannii.

Hospital-acquired infections are on the rise and represent both, a clinical and financial burden. With resistance emerging and an ever-dwindling armamentarium at hand, infections caused by Acinetobacter baumannii are particularly problematic, since these bacteria have a high level of resistance and resilience to traditional and even last-resort antibiotics. The antibiotic rifabutin was recently found to show potent in vitro and in vivo activity against extensively drug resistant A. baumannii. Building on this discovery, we report on the synthesis and activity of rifabutin analogs, with a focus on N-functionalization of the piperidine ring. The antimicrobial testing uncovered structure activity relationships (SAR) for A. baumannii that were not reflected in Staphylococcus aureus. The cellular activity did not correlate with cell-free transcription inhibition, but with bacterial intracellular compound accumulation. Mass spectrometry-based accumulation studies confirmed the involvement of the siderophore receptor FhuE in active compound translocation at low concentrations, and they showed a strong impact of the culture medium on the accumulation of rifabutin. Overall, the study underlines the structural feature required for strong accumulation of rifabutin in A. baumannii and identifies analogs as or more potent than rifabutin against A. baumannii.

Toward a data-driven evaluation of the 2010 American College of Rheumatology/European League Against Rheumatism criteria for rheumatoid arthritis: is it sensible to look at levels of rheumatoid factor?

OBJECTIVE: Recently, new classification criteria for rheumatoid arthritis (RA) have been devised by methodology that used first a quantitative approach (data from databases), then a qualitative approach (consensus; based on paper patients), and finally a common sense-based approach (evaluation of the former phases). Now the individual items that make up these criteria are being evaluated. This study was undertaken to analyze the item "autoantibodies," in particular rheumatoid factor (RF) level. METHODS: Three separate cohorts comprising a total of 972 patients with undifferentiated arthritis were studied for RA development (according to the 1987 American College of Rheumatology criteria) and arthritis persistence. The positive predictive value (PPV), negative predictive value (NPV), and likelihood ratios (LRs) were compared between different levels of RF and the presence of anti-citrullinated protein antibody (ACPA). A similar comparison was made in 686 RA patients for the rate of joint destruction and achievement of sustained disease-modifying antirheumatic drug-free remission during 7 years of followup. The variation in RF levels obtained by different measurement methods in the same RF-positive sera was explored. RESULTS: Compared to high RF levels, presence of ACPA had a better balance between positive LR and negative LR and between PPV and NPV for RA development. The additive value of ACPA assessment after testing for RF level was higher than vice versa. The association between high RF level and RA severity was not as strong as that between ACPA antibodies and RA severity. The RF level obtained by different methods in the same patients' sera varied considerably. CONCLUSION: Our findings indicate that determination of RF level is subject to large variation; high RF level has limited additive prognostic value compared to ACPA positivity. Thus, omitting RF level and using RF presence, ACPA presence, and ACPA level may improve the 2010 criteria for RA.

Anti-citrullinated protein antibodies have a low avidity compared with antibodies against recall antigens.

OBJECTIVES: Anti-citrullinated protein antibodies (ACPA) are highly specific for rheumatoid arthritis (RA) and have been implicated in disease pathogenesis. Recent ongoing evidence indicates that the ACPA response broadens before precipitation of full-blown RA, as indicated by a more extensive isotype usage and an increased citrullinated epitope recognition profile. Nonetheless, the evolution of the ACPA response is still poorly understood and might intrinsically differ from the protective responses against pathogens. METHODS: The avidity and the avidity maturation of ACPA in relation to the avidity of antibodies against recall antigens were analysed. RESULTS: The avidity of ACPA was significantly lower than the avidity of antibodies to the recall antigens tetanus toxoid and diptheria toxoid. Moreover, ACPA did not show avidity maturation during longitudinal follow-up and ACPA avidity was also relatively low in patients who displayed extensive isotype switching. CONCLUSIONS: These observations indicate that the natural evolution of ACPA differs from the development of antibodies against recall antigens. These data also indicate that ACPA avidity maturation and isotype switching are disconnected, whereby extensive isotype switching occurs in the setting of restricted avidity maturation. Intrinsic differences between the RA-specific autoantibody system and protective antibody responses against pathogens could be of relevance for designing novel B cell-targeted therapies for RA.

The next generation of current measurement for ionization chambers.

Re-entrant ionization chambers (ICs) are essential to radionuclide metrology and nuclear medicine for maintaining standards and measuring half-lives. The requirements of top-level metrology demand that systems must be precise and stable to 0.1 % over many years, and linear from 10-14 A to 10-8 A. Thus, laboratories depend on bespoke current measurement systems and often rely on sealed sources to generate reference currents. To maintain and improve present capabilities, metrologists need to overcome two looming challenges: ageing electronics and decreasing availability of sealed sources. Possible solutions using Ultrastable Low-Noise Current Amplifiers (ULCAs), resistive-feedback electrometers, and (quantum) single-electron pumps are reviewed. Broader discussions of IC design and methodology are discussed. ULCAs show promise and resistive-feedback systems which take advantage of standard resistor calibrations offer an alternative.

Effects of rectilinear acceleration and optokinetic and caloric stimulations in space.

During the flight of Spacelab 1 the crew performed a number of experiments to explore changes in vestibular function and visual-vestibular interactions on exposure to microgravity. Measurements were made on the threshold for detection of linear oscillation, vestibulo-ocular reflexes elicited by angular and linear movements, oculomotor and posture responses to optokinetic stimulations, and responses to caloric stimulation. Tests were also conducted on the ground, during the 4 months before and on days 1 to 6 after flight. The most significant result was that caloric mystagmus of the same direction as on the earth could also be evoked in the weightless environment.

The improvement of middle ear ventilation by laser ablation of the epipharyngeal eustachian tube: a prospective study.

A long-lasting dysfunction of the eustachian tube seems to be the etiologic origin for development of chronic otitis media (COM) with mesotympanic perforation, otitis media with effusion (OME), and chronic atelectasis of the middle ear. Surgical interventions in the middle ear generally treat the sequelae of the tube dysfunction but not the dysfunction itself. This prospective clinical study investigated how far fiber-guided laser ablation of the posterior half of the epipharyngeal tubal ostium led to better middle ear ventilation in the otologic disease patterns mentioned below. There were 38 adult patients included in the analysis. The patients in one group had a perforated tympanic membrane [COM before primary tympanoplasty (n = 14) or revision tympanoplasty (n = 5)]; the patients in a second group had an intact eardrum [OME resistant to therapy (n = 3), with an atelectasis of the middle ear (n = 2) or problems of pressure equalization with fast changes in ambient pressure (diving, flying) (n = 14)]. Laser ablation of the posterior half of the epipharyngeal tubal ostium was performed, generally with local anesthesia, if tubal function testing was pathologic (Valsalva maneuver, passive tube opening, tympanogram). In patients with COM the procedure was performed 8 weeks before the middle ear surgery. All patients were checked 8 weeks postoperatively and in the course of the following year. The intervention seemed to have had a positive effect on tube function in 68.4% of patients operated on (P = 0.001). In 26 of the 38 patients that had undergone operation, an improvement the results of tubal function tests could be seen in the postoperative follow-up. In the COM group the Valsalva maneuver improved in 14 of 19 patients (73.6%) (P = 0.001), and the passive tube opening improved in nine of 18 patients (50%). In the group with an intact eardrum the Valsalva maneuver improved in 13 of 18 patients (72.2%) (P = 0.001). The resulting condition remained stable after 1 year. None of the patients showed any complications as a result of the therapy. Minimally invasive shaping of the distal eustachian tube under topical anesthesia can be recommended for patients with the above-mentioned diagnoses who have pathologic middle ear ventilation. Especially prior to tympanoplasties, and especially in otologic revision procedures, where middle ear ventilation is a prerequisite for successful otologic surgery, the function of the eustachian tube can be optimized in 70% of the patients, particularly if there are pathological findings (tubal tonsil, narrow orifice of the tubal ostium, adenoids). The placement of permanent ear tubes in adults with recurrent OME can also be avoided by the procedure described. The resultant conditions remained stable for the next year. Patients with tympanic ventilation problems due to rapid pressure changes (flying, diving) can also benefit from this procedure.

[Hyperkeratotic inverted seborrheic keratosis of the outer auditory canal : a first description]. / Hyperkeratotische invertierte seborrhoische Keratose des äusseren Gehörgangs : Eine Erstbeschreibung.

Seborrheic keratosis is one of the most common benign neoplasms seen at the trunk, extremities, head and neck of older individuals which rarely occurs in the auditory canal. Even rarer is the occurrence of an inverted or hyperkeratotic variant in this localization. We present the clinical picture of a 74-year-old man with a recurrent hyperkeratotic, partially inverted seborrheic keratosis of the right external auditory canal. This disease is sometimes difficult to diagnose, particulary to mark down malignant lesions. An uncritical diagnosis must be avoided, since these lesions and especially the irritated subtype might be falsely assessed as basal cell carcinomas which they may resemble histologically.

Noise-optimized ultrastable low-noise current amplifier.

We have developed a noise-optimized ultrastable low-noise current amplifier (ULCA) aimed at reducing the uncertainty at low currents. It involves a thin-film resistor network with 6.75 GΩ at the high-ohmic path which reduces the noise level to 1.6 fA/Hz. Noise investigations as well as short-term and long-term stability studies were carried out. The stability of the input current gain was measured using a cryogenic current comparator at ±6.1 nA. Methods for investigating the measurement accuracy at low input currents of about 100 pA at a level of below one part in 107 are introduced and experimentally verified. The performance of the noise-optimized ULCA is compared with that of the standard variant introduced in 2014. It is shown that the reduced noise floor is achieved without impairing the stability of the transresistance.

Synaptoporin, a novel putative channel protein of synaptic vesicles.

By homology screening of a rat brain library, we have isolated cDNAs that encode a novel member of the synaptophysin/connexin channel protein superfamily. The deduced protein, named synaptoporin, displays 58% amino acid identity to synaptophysin, with highly conserved transmembrane segments, but a divergent cytoplasmic tail. Northern blot analysis and PCR amplification of RNA from different rat tissues indicate expression of synaptoporin transcripts in the CNS. Antibodies against a synthetic peptide or a fusion construct encompassing the cytoplasmic tail region of synaptoporin detect a polypeptide of 37 kd that copurifies with small synaptic vesicles. Our data suggest the existence of a family of vesicular channel proteins whose members may be differently distributed among synaptic vesicle subpopulations.

[Adenoid cystic carcinoma of the oropharynx. Brachytherapy for the treatment of local recurrences]. / Adenoidzystisches Karzinom des Oropharynx. Brachytherapie zur Behandlung eines lokalen Rezidivs.

PURPOSE: Adenoid cystic carcinomas are rare tumours. Regional lymphatic spread is rather uncommon. Distant metastases occur in approximately half of all patients. High local failure rates of up to 50% after surgery have been reported. Even after adjuvant radiation, the results remain poor for locally advanced tumours after incomplete resection. We report on a safe and effective salvage option in the treatment of adenoid cystic carcinoma. MATERIAL AND METHODS: A 44 year old woman with the diagnosis of adenoid cystic carcinoma of the oropharynx and simultaneous pulmonary metastases received a mixed beam photon radiotherapy and a neutron boost. Three years later, the patient developed a dysphagia grade 2, dysarthria and pain. A local recurrence in the base of tongue and floor of mouth was histologically confirmed. RESULTS: Because of the local symptoms, we offered the patient Ir-192-HDR brachytherapy with palliative intent. Under general anaesthesia, eight needles were implanted into the palpable tumour. On the basis of a pretreatment MRI, a single dose of 5 Gy encompassing the palpable tumour was given to a total dose of 15 Gy. The patient remains free from local symptoms at 8 month after brachytherapy. CONCLUSION: HDR-Ir brachytherapy is an effective method for the treatment of local recurrences of an adenoid cystic carcinoma, even after preradiotherapy. Excellent palliative effects can be achieved without increased toxicity after previous radiotherapy.

[Cervicoproprioceptive provocation of horizontal and vertical nystagmus in test subjects]. / Zervikopropriozeptive Provokation von horizontalen und vertikalen Nystagmen bei Probanden.

BACKGROUND: The cervico-ocular reflex (COR) has been confirmed in numerous animal experiments. On the other hand, its clinical assessment is disputed. Anteflexion and retroflexion of the head are among the main movements of the atlanto-occipital joint. We investigated whether these head movements produce neck proprioceptive stimulation in the vestibular system. SUBJECTS AND METHOD: We investigated 50 students under the experimental conditions of strictly cervical provocation. The trunk was rotated, anteflexed, and retroflexed into its end positions for 60 s under video-oculographic control without changing the head-space relationship. During the cervical provocation, horizontal, vertical, or rotational nystagmus within a time window of 5-30 s after the start of the provocation was detected as a target parameter. RESULTS: The results showed that, compared to the baseline, there was a significant increase in vertical and horizontal nystagmus after the cervical provocation. The horizontal nystagmus reaction, rather than the vertical, was increased significantly by the maximum trunk rotation. In addition, we found a significant amount of vertical rather than horizontal nystagmus under trunk flexion provocation. CONCLUSION: Under cervical provocation of test subjects, we were able to elicit horizontal and vertical nystagmus (upbeat nystagmus) via cervico-ocular roots. Due to a neck proprioceptive activation of the vestibular system we interpret our result as a "cervico-tonic provocation nystagmus".

[Topical immunomodulation. A milestone for the treatment of therapy-resistant noninfectious chronic external otitis?]. / Topische Immunmodulation. Ein Meilenstein in der Behandlung der therapieresistenten nichtinfektiösen chronischen Otitis externa?

BACKGROUND: Recurrent exacerbation of chronic external otitis represents a special challenge for the attending physician. The goal of our study was to evaluate the effectiveness of novel topical immunomodulators acting through an anti-inflammatory, nonsteroidal mechanism. PATIENTS AND METHODS: In a prospective study, in 33 patients an ear wick containing tacrolimus ointment (Protopic 0.1%) was inserted every 2-3 days. Altogether, the wick was changed three times. Therapeutic outcomes were assessed by reexaminations, video-otoscopy, and a standardized findings sheet. RESULTS: Twenty-eight patients showed significant improvement of clinical symptoms, with 13 of them showing complete healing (follow-up 10-22 months). Relapses (15 cases) were associated with significantly extended symptom-free intervals and reduced numbers of further recurrent episodes. CONCLUSIONS: Because of the safe and successful anti-inflammatory effects, topical immunomodulators represent a new alternative in chronic inflammatory stages of otherwise therapy-resistant external otitis.

Spatial organisation of microbiota in quiescent adenoiditis and tonsillitis.

BACKGROUND: The reasons for recurrent adenotonsillitis are poorly understood. METHODS: The in situ composition of microbiota of nasal (5 children, 25 adults) and of hypertrophied adenoid and tonsillar tissue (50 children, 20 adults) was investigated using a broad range of fluorescent oligonucleotide probes targeted to bacterial rRNA. None of the patients had clinical signs of infection at the time of surgery. RESULTS: Multiple foci of ongoing purulent infections were found within hypertrophied adenoid and tonsillar tissue in 83% of patients, including islands and lawns of bacteria adherent to the epithelium, with concomitant marked inflammatory response, fissures filled with bacteria and pus, and diffuse infiltration of the tonsils by bacteria, microabscesses, and macrophages containing phagocytosed microorganisms. Haemophilusinfluenzae mainly diffusely infiltrated the tissue, Streptococcus and Bacteroides were typically found in fissures, and Fusobacteria,Pseudomonas and Burkholderia were exclusively located within adherent bacterial layers and infiltrates. The microbiota were always polymicrobial. CONCLUSIONS: Purulent processes persist during asymptomatic periods of adenotonsillitis. Most bacteria involved in this process are covered by a thick inflammatory infiltrate, are deeply invading, or are located within macrophages. The distribution of the bacteria within tonsils may be responsible for the failure of antibiotic treatment.

Snoring noise pollution--the need for objective quantification of annoyance, regulatory guidelines and mandatory therapy for snoring.

Habitual snoring without episodes of apnea or hypoventilation and without respiratory related arousals is considered to be annoying and without any need for treatment. However, studies seem to suggest an enormous psychosocial impact of annoyance for the bed partner. Apart from subjective questionnaires there still exists no generally accepted mode of measurement that can describe snoring objectively. We therefore adapted methods developed for environmental medicine and established a new snore score using psycho-acoustic parameters. For quantification of snoring noise we conducted nocturnal measurements in 19 habitual snorers. Free-field snore sounds were acquired with two low-cost non-contact microphones and transferred to a PC (sampling frequency 11 kHz). The data were recorded, analysed and stored automatically using a MATLAB script. Following the analysis of sound characteristics and levels, the score was computed from relevant parameters containing the rating level (L(R)), maximum level, two percentile levels for frequent maxima (L(5)S; L(1)) and snoring time. The determined values substantially exceeded the prescribed limits defined by WHO noise guidelines, and mainly affected the equivalent continuous sound exposure level, rating level and the immission standard values of brief noise peaks, whose maximum was exceeded by up to 32 dB(A). The Berlin snore score illustrated the objective acoustic annoyance on a scale from 0 to 100. It allows inter-individual comparison and objectifies the need for therapy. The clinical applicability of evaluating the reduction of snoring after surgical therapy is discussed exemplarily. The presented measuring method was found to be suitable for quantifying snoring noise and can be easily integrated into existing polysomnographic applications. In the case of habitual snoring with objective evidence of psychosocially disturbing acoustic annoyance, health fund providers should assume the costs of mandatory medical therapy.

Intratympanic treatment of acute acoustic trauma with a cell-permeable JNK ligand: a prospective randomized phase I/II study.

OBJECTIVES: Intratympanic administration of a cell-permeable JNK ligand has been shown to prevent hearing loss after acute acoustic trauma in animal models. CONCLUSIONS: Functional and morphological analysis of the treated ears revealed that AM-111 had an excellent otoprotective effect, even when administered hours after the noise exposure. Blocking the signal pathway with D-JNKI-1 is therefore a promising way to protect the morphological integrity and physiological function of the inner ear in various conditions involving acute sensorineural hearing loss. SUBJECTS AND METHODS: For the first application of AM-111 in humans, we organized a clinical phase I/II trial in patients with acute acoustic trauma after exposure to firecrackers in Berlin and Munich on New Year's Eve 2005/2006. We randomly selected 11 patients for intratympanic treatment with AM-111 at a concentration of 0.4 mg/ml or 2 mg/ml within 24 h after noise exposure. Pure tone audiometry and otoacoustic emissions were assessed before treatment and on days 3 and 30 thereafter. RESULTS: Based on clinical experience and on a calculation using an empirically derived exponential hearing recovery function AM-111 seems to have had a therapeutic effect. A total of 13 adverse events were reported in 5 study participants. None of the adverse events were serious or severe.