Stem cells have been touted as a source of potential replacement neurons for inner ear degeneration for almost two decades now; yet to date, there are few studies describing the use of human pluripotent stem cells (hPSCs) for this purpose. If stem cell therapies are to be used clinically, it is critical to validate the usefulness of hPSC lines in vitro and in vivo. Here, we present the first quantitative evidence that differentiated hPSC-derived neurons that innervate both the inner ear hair cells and cochlear nucleus neurons in coculture, with significantly more new synaptic contacts formed on target cell types. Nascent contacts between stem cells and hair cells were immunopositive for both synapsin I and VGLUT1, closely resembling expression of these puncta in endogenous postnatal auditory neurons and control cocultures. When hPSCs were cocultured with cochlear nucleus brainstem slice, significantly greater numbers of VGLUT1 puncta were observed in comparison to slice alone. New VGLUT1 puncta in cocultures with cochlear nucleus slice were not significantly different in size, only in quantity. This experimentation describes new coculture models for assessing auditory regeneration using well-characterised hPSC-derived neurons and highlights useful methods to quantify the extent of innervation on different cell types in the inner ear and brainstem.

Accurate perception of voice pitch plays a vital role in speech understanding, especially for tonal languages such as Mandarin. Lexical tones are primarily distinguished by the fundamental frequency (F0) contour of the acoustic waveform. It has been shown that the auditory system could extract the F0 from the resolved and unresolved harmonics, and the tone identification performance of resolved harmonics was better than unresolved harmonics. To evaluate the neural response to the resolved and unresolved components of Mandarin tones in quiet and in speech-shaped noise, we recorded the frequency-following response. In this study, four types of stimuli were used: speech with either only-resolved harmonics or only-unresolved harmonics, both in quiet and in speech-shaped noise. Frequency-following responses (FFRs) were recorded to alternating-polarity stimuli and were added or subtracted to enhance the neural response to the envelope (FFRENV) or fine structure (FFRTFS), respectively. The neural representation of the F0 strength reflected by the FFRENV was evaluated by the peak autocorrelation value in the temporal domain and the peak phase-locking value (PLV) at F0 in the spectral domain. Both evaluation methods showed that the FFRENV F0 strength in quiet was significantly stronger than in noise for speech including unresolved harmonics, but not for speech including resolved harmonics. The neural representation of the temporal fine structure reflected by the FFRTFS was assessed by the PLV at the harmonic near to F1 (4th of F0). The PLV at harmonic near to F1 (4th of F0) of FFRTFS to resolved harmonics was significantly larger than to unresolved harmonics. Spearman's correlation showed that the FFRENV F0 strength to unresolved harmonics was correlated with tone identification performance in noise (0 dB SNR). These results showed that the FFRENV F0 strength to speech sounds with resolved harmonics was not affected by noise. In contrast, the response to speech sounds with unresolved harmonics, which were significantly smaller in noise compared to quiet. Our results suggest that coding resolved harmonics was more important than coding envelope for tone identification performance in noise.

Introduction: Tremor of the upper limbs is a disabling symptom that is present during several neurological disorders and is currently without treatment. Functional MRI (fMRI) is an essential tool to investigate the pathophysiology of tremor and aid the development of treatment options. However, no adequately or standardized protocols for fMRI exists at present. Here we present a novel, online available fMRI task that could be used to assess the in vivo pathology of tremor. Objective: This study aims to validate the tremor-evoking potential of the fMRI task in a small group of tremor patients outside the scanner and assess the reproducibility of the fMRI task related activation in healthy controls. Methods: Twelve HCs were scanned at two time points (baseline and after 6-weeks). There were two runs of multi-band fMRI and the tasks included a "brick-breaker" joystick game. The game consisted of three conditions designed to control for most of the activation related to performing the task by contrasting the conditions: WATCH (look at the game without moving joystick), MOVE (rhythmic left/right movement of joystick without game), and PLAY (playing the game). Task fMRI was analyzed using FSL FEAT to determine clusters of activation during the different conditions. Maximum activation within the clusters was used to assess the ability to control for task related activation and reproducibility. Four tremor patients have been included to test ecological and construct validity of the joystick task by assessing tremor frequencies captured by the joystick. Results: In HCs the game activated areas corresponding to motor, attention and visual areas. Most areas of activation by our game showed moderate to good reproducibility (intraclass correlation coefficient (ICC) 0.531-0.906) with only inferior parietal lobe activation showing poor reproducibility (ICC 0.446). Furthermore, the joystick captured significantly more tremulous movement in tremor patients compared to HCs (p = 0.01) during PLAY, but not during MOVE. Conclusion: Validation of our novel task confirmed tremor-evoking potential and reproducibility analyses yielded acceptable results to continue further investigations into the pathophysiology of tremor. The use of this technique in studies with tremor patient will no doubt provide significant insights into the treatment options.

Patients with intractable tremor not alleviated by pharmaceutical therapies can be successfully treated using Deep Brain Stimulation of subcortical areas such as the Subthalamic Nucleus (STN). Current DBS devices produce constant-current or constant-voltage biphasic pulse trains and offer a range of pulse rate and pulse duration options as well as different current or voltage levels. The clinical method of setting the parameters by observation is non-ideal, firstly because of the number of possible parameter combinations, and a current lack of knowledge of how the parameters interact with each other, and secondly because subjective clinical observation is prone to observer error and bias. In this study, we aimed to develop an objective method of recording the severity of tremor and to use it to explore the effect on tremor of stimulus parameters. Six patients with Essential Tremor who had been fitted with a DBS device in the Posterior Subthalamic Area (PSA) participated in the study. Tremor was measured by position sensors attached to the arms and wrists while the patient was holding both arms stretched out in front of them and when performing a finger-nose pointing task. Clinical rating of tremor was also performed. Results of the experiments showed that the optimal stimulus parameters were subject specific and the effects of each parameter were non-monotonic, often with a very specific range providing therapeutic benefit (for example, Fig. 1). The objective measures were more sensitive than clinical judgement and show that an objective fitting method could improve benefits in individual patients.

Deep Brain Stimulation (DBS) is rapidly emerging as a safe and effective treatment option for mitigating the effects of tremor. Despite the relative success of DBS for treating tremor, a common and typically unquantified adverse effect of treatment is dysarthria (slurred speech). Current assessment protocols are driven by the qualitative judgements of treating clinicians and lack the sensitivity and objectivity required to make reliable decisions about treatment optimisation. Therefore we aimed to pilot a speech evaluation procedure that would form the basis of an objective clinical DBS optimisation tool for use in patients with tremor. Six patients diagnosed with essential tremor receiving treatment via deep brain stimulation of the posterior sub-thalamic nucleus were recruited. Electrical stimulation parameters (i.e., pulse rate, pulse duration, and current amplitude) were systematically adjusted and speech samples recorded to identify the patient-specific settings required for optimal therapeutic benefit (reduced tremor) with minimal adverse effects (dysarthria). Altered speech production between stimulation parameters was quantified via acoustic analysis. Measures of timing (e.g., speech rate), intonation (e.g., pitch variation) and quality (e.g., noise-to-harmonics ratio) reflected increasing/decreasing levels of dysarthria (see associated figure). Via this protocol we aim to understand the inter-relationship between the effects of the parameters as well as to develop a real-time objective system for surgeons to optimise these parameters for each patient. A secondary outcome is to increase our understanding of how electrical parameter settings are related to movement and speech, and how the optimal parameters are related to the nature of the individual’s pathology.

BACKGROUND: Tremor is a debilitating symptom of Multiple Sclerosis (MS). Little is known about its pathophysiology and treatments are limited. Clinical trials investigating new interventions often rely on subjective clinical rating scales to provide supporting evidence of efficacy. NEW METHOD: We present a novel instrument (TREMBAL) which uses electromagnetic motion capture technology to quantify MS tremor. We aim to validate TREMBAL by comparison to clinical ratings using regression modelling with 310 samples of tremor captured from 13 MS participants who performed five different hand exercises during several follow-up visits. Minimum detectable change (MDC) and test-retest reliability were calculated and comparisons were made between MS tremor and data from 12 healthy volunteers. RESULTS: Velocity of the index finger was most congruent with clinical observation. Regression modelling combining different features, sensor configurations, and labelling exercises did not improve results. TREMBAL MDC was 84% of its initial measurement compared to 91% for the clinical rating. Intra-class correlations for test-retest reliability were 0.781 for TREMBAL and 0.703 for clinical ratings. Tremor was lower (p =  0.002) in healthy subjects. COMPARISON WITH EXISTING METHODS: Subjective scales have low sensitivity, suffer from ceiling effects, and mitigation against inter-rater variability is challenging. Inertial sensors are ubiquitous, however, their output is nonlinearly related to tremor frequency, compensation is required for gravitational artefacts, and their raw data cannot be intuitively comprehended. CONCLUSIONS: TREMBAL, compared with clinical ratings, gave measures in agreement with clinical observation, had marginally lower MDC, and similar test-retest reliability.

IMPORTANCE: Multiple sclerosis produces neurological impairments that are variable in duration, severity and quality. Speech is frequently impaired, resulting in decreased communication skills and quality of life. Advancements in technology now makes it possible to use quantitative acoustic assessment of speech as biomarkers of disease progression. OBSERVATIONS: Four domains of speech have been identified: articulation (slow articulation and imprecise consonants), voice (pitch and loudness instability), respiration (decreased phonatory time and expiratory pressure) and prosody (longer and frequent pauses, deficient loudness control). Studies also explored I) predictive models for diagnosis of MS and of ataxia using speech variables, II) the relationship of dysarthria with cognition and III) very few studies correlated neuroimaging with dysarthria. We could not identify longitudinal studies of speech or dysarthria in Multiple Sclerosis. CONCLUSION AND RELEVANCE: Refinement of objective measures of speech has enhanced our understanding of Multiple Sclerosis-related deficits in cross-sectional analysis while both integrative and longitudinal studies are identified as major gaps. This review highlights the potential for using quantitative acoustic assessments as clinical endpoints for diagnosing, monitoring progression and treatment in disease modifying trials.

According to 2010 estimates from The National Institute on Deafness and other Communication Disorders, approximately 17% (36 million) American adults have reported some degree of hearing loss. Currently, the only clinical treatment available for those with severe-to-profound hearing loss is a cochlear implant, which is designed to electrically stimulate the auditory nerve in the absence of hair cells. Whilst the cochlear implant has been revolutionary in terms of providing hearing to the severe-to-profoundly deaf, there are variations in cochlear implant performance which may be related to the degree of degeneration of auditory neurons following hearing loss. Hence, numerous experimental studies have focused on enhancing the efficacy of cochlear implants by using neurotrophins to preserve the auditory neurons, and more recently, attempting to replace these dying cells with new neurons derived from stem cells. As a result, several groups are now investigating the potential for both embryonic and adult stem cells to replace the degenerating sensory elements in the deaf cochlea. Recent advances in our knowledge of stem cells and the development of induced pluripotency by Takahashi and Yamanaka in 2006, have opened a new realm of science focused on the use of induced pluripotent stem (iPS) cells for therapeutic purposes. This review will provide a broad overview of the potential benefits and challenges of using iPS cells in combination with a cochlear implant for the treatment of hearing loss, including differentiation of iPS cells into an auditory neural lineage and clinically relevant transplantation approaches.

An experiment was conducted to investigate the feasibility of using functional near-infrared spectroscopy (fNIRS) to image cortical activity in the language areas of cochlear implant (CI) users and to explore the association between the activity and their speech understanding ability. Using fNIRS, 15 experienced CI users and 14 normal-hearing participants were imaged while presented with either visual speech or auditory speech. Brain activation was measured from the prefrontal, temporal, and parietal lobe in both hemispheres, including the language-associated regions. In response to visual speech, the activation levels of CI users in an a priori region of interest (ROI)-the left superior temporal gyrus or sulcus-were negatively correlated with auditory speech understanding. This result suggests that increased cross-modal activity in the auditory cortex is predictive of poor auditory speech understanding. In another two ROIs, in which CI users showed significantly different mean activation levels in response to auditory speech compared with normal-hearing listeners, activation levels were significantly negatively correlated with CI users' auditory speech understanding. These ROIs were located in the right anterior temporal lobe (including a portion of prefrontal lobe) and the left middle superior temporal lobe. In conclusion, fNIRS successfully revealed activation patterns in CI users associated with their auditory speech understanding.

Wireless power and data transfer to medical implants is a research area where improvements in current state-of-the-art technologies are needed owing to the continuing efforts for miniaturization. At present, lithographical patterning of evaporated metals is widely used for miniature coil fabrication. This method produces coils that are limited to low micron or nanometer thicknesses leading to high impedance values and thus limiting their potential quality. In the present work we describe a novel technique, whereby trenches were milled into a diamond substrate and filled with silver active braze alloy, enabling the manufacture of small, high cross-section, low impedance microcoils capable of transferring up to 10 mW of power up to a distance of 6 mm. As a substitute for a metallic braze line used for hermetic sealing, a continuous metal loop when placed parallel and close to the coil surface reduced power transfer efficiency by 43%, but not significantly, when placed perpendicular to the microcoil surface. Encapsulation of the coil by growth of a further layer of diamond reduced the quality factor by an average of 38%, which can be largely avoided by prior oxygen plasma treatment. Furthermore, an accelerated ageing test after encapsulation showed that these coils are long lasting. Our results thus collectively highlight the feasibility of fabricating a high-cross section, biocompatible and long lasting miniaturized microcoil that could be used in either a neural recording or neuromuscular stimulation device.

Supraparticles (SPs) composed of smaller colloidal particles provide a platform for the long-term, controlled release of therapeutics in biomedical applications. However, current synthesis methods used to achieve high drug loading and those involving biocompatible materials are often tedious and low throughput, thereby limiting the translation of SPs to diverse applications. Herein, we present a simple, effective, and automatable alginate-mediated electrospray technique for the assembly of robust spherical silica SPs (Si-SPs) for long-term (>4 months) drug delivery. The Si-SPs are composed of either porous or nonporous primary Si particles within a decomposable alginate matrix. The size and shape of the Si-SPs can be tailored by controlling the concentrations of alginate and silica primary particles used and key electrospraying parameters, such as flow rate, voltage, and collector distance. Furthermore, the performance (including drug loading kinetics, loading capacity, loading efficiency, and drug release) of the Si-SPs can be tuned by changing the porosity of the primary particles and through the retention or removal (via calcination) of the alginate matrix. The structure and morphology of the Si-SPs were characterized by electron microscopy, dynamic light scattering, N2 adsorption-desorption analysis, and X-ray photoelectron spectroscopy. The cytotoxicity and degradability of the Si-SPs were also examined. Drug loading kinetics and loading capacity for six different types of Si-SPs, using a model protein drug (fluorescently labeled lysozyme), demonstrate that Si-SPs prepared from primary silica particles with large pores can load significant amounts of lysozyme (∼10 μg per SP) and exhibit sustained, long-term release of more than 150 days. Our experiments show that Si-SPs can be produced through a gel-mediated electrospray technique that is robust and automatable (important for clinical translation and commercialization) and that they present a promising platform for long-term drug delivery.