Predicting most comfortable listening levels of ESRT from ECAP thresholds in paediatric cochlear implant users.

Studies have reported a varied correlation strength between the electrically evoked compound action potential (ECAP) and electrically evoked stapedial reflex thresholds (ESRT) in cochlear implant recipients. However, there is a lack of information on the relationship between the two measures in paediatric cochlear implant users. This study was aimed to compare the ESRT and ECAP measures and determine where ECAP thresholds fall within the dynamic range of ESRT-based Maps in paediatric cochlear implant users.

A new objective method to estimate the charge integration efficiency in cochlear implant users.

Objective: The present study aimed to objectively assess the charge integration efficiency (CIE) of the auditory nerve using electrically-evoked stapedial reflex threshold (eSRT) measurements in paediatric cochlear implant users.

Design: The eSRT was estimated in two ways: by keeping pulse width constant while increasing pulse amplitude and vice versa. The electrical dynamic range (EDR) obtained for eSRT was measured with increasing pulse amplitude (EDR) and pulse width (EDR) by calculating the difference in charge units between eSRT and behavioural thresholds; further, CIE was estimated.

Effect of Age on Distortion Product Otoacoustic Emissions at Extended High Frequencies.

The inner ear is most susceptible to the aging effects. Distortion product otoacoustic emissions (DPOAEs) are a good indicator for interpreting the age effects but are usually recorded at up to 8000 Hz frequencies in routine audiologic testing. The present study was designed to assess and compare the DPOAEs at conventional frequencies and at extended high frequencies (EHFs) across different age groups.

Electrically evoked late latency response using single electrode stimulation and its relation to speech perception among paediatric cochlear implant users.

Introduction: Aided auditory late latency response (LLR) serves as an objective tool for evaluating auditory cortical maturation following cochlear implantation in children. While aided LLR is commonly measured using sound-field acoustic stimulation, recording electrically evoked LLR (eLLR) offer distinct advantages, such as improved stimulus control and the capability for single electrode stimulation. Hence, the study aimed to compare eLLR responses with single electrode stimulation in the apical, middle, and basal regions and to evaluate their relationship with speech perception in paediatric cochlear implant (CI) recipients.

Functional redundancy and formin-independent localization of tropomyosin isoforms in .

Tropomyosin is an actin binding protein which protects actin filaments from cofilin-mediated disassembly. Distinct tropomyosin isoforms have long been hypothesized to differentially sort to subcellular actin networks and impart distinct functionalities. Nevertheless, a mechanistic understanding of the interplay between Tpm isoforms and their functional contributions to actin dynamics has been lacking.

IntAct: A nondisruptive internal tagging strategy to study the organization and function of actin isoforms.

Mammals have 6 highly conserved actin isoforms with nonredundant biological functions. The molecular basis of isoform specificity, however, remains elusive due to a lack of tools. Here, we describe the development of IntAct, an internal tagging strategy to study actin isoforms in fixed and living cells.

A monomeric StayGold fluorescent protein.

StayGold is an exceptionally bright and stable fluorescent protein that is highly resistant to photobleaching. Despite favorable fluorescence properties, use of StayGold as a fluorescent tag is limited because it forms a natural dimer. Here we report the 1.

Evaluation of the Electrically-Evoked Stapedial Reflex Threshold in Pediatric Cochlear Implant Users with High-Frequency Probe Tones.

 Measurement of the electrically-evoked stapedial reflex threshold (ESRT) is an objective tool used to set the comfort levels in pediatric cochlear implant (PCI) users. The levels of ESRT have a strong correlation with comfort levels. However, the clinical utility of ESRT is limited because the ESRT response is not observed in all cochlear implant users.

ALIBY: ALFA Nanobody-Based Toolkit for Imaging and Biochemistry in Yeast.

Specialized epitope tags continue to be integral components of various biochemical and cell biological applications such as fluorescence microscopy, immunoblotting, immunoprecipitation, and protein purification. However, until recently, no single tag could offer this complete set of functionalities on its own. Here, we present a plasmid-based toolkit named ALIBY (FA toolkit for maging and iochemistry in east) that provides a universal workflow to adopt the versatile ALFA tag/ALFA system within the well-established model organism Saccharomyces cerevisiae.

mNG-tagged fusion proteins and nanobodies to visualize tropomyosins in yeast and mammalian cells.

Tropomyosins are structurally conserved α-helical coiled-coil proteins that bind along the length of filamentous actin (F-actin) in fungi and animals. Tropomyosins play essential roles in the stability of actin filaments and in regulating myosin II contractility. Despite the crucial role of tropomyosin in actin cytoskeletal regulation, in vivo investigations of tropomyosin are limited, mainly due to the suboptimal live-cell imaging tools currently available.

Asgard archaea shed light on the evolutionary origins of the eukaryotic ubiquitin-ESCRT machinery.

The ESCRT machinery, comprising of multiple proteins and subcomplexes, is crucial for membrane remodelling in eukaryotic cells, in processes that include ubiquitin-mediated multivesicular body formation, membrane repair, cytokinetic abscission, and virus exit from host cells. This ESCRT system appears to have simpler, ancient origins, since many archaeal species possess homologues of ESCRT-III and Vps4, the components that execute the final membrane scission reaction, where they have been shown to play roles in cytokinesis, extracellular vesicle formation and viral egress. Remarkably, metagenome assemblies of Asgard archaea, the closest known living relatives of eukaryotes, were recently shown to encode homologues of the entire cascade involved in ubiquitin-mediated membrane remodelling, including ubiquitin itself, components of the ESCRT-I and ESCRT-II subcomplexes, and ESCRT-III and Vps4.

Calponin-homology domain mediated bending of membrane-associated actin filaments.

Actin filaments are central to numerous biological processes in all domains of life. Driven by the interplay with molecular motors, actin binding and actin modulating proteins, the actin cytoskeleton exhibits a variety of geometries. This includes structures with a curved geometry such as axon-stabilizing actin rings, actin cages around mitochondria and the cytokinetic actomyosin ring, which are generally assumed to be formed by short linear filaments held together by actin cross-linkers.

An organelle-tethering mechanism couples flagellation to cell division in bacteria.

In some free-living and pathogenic bacteria, problems in the synthesis and assembly of early flagellar components can cause cell-division defects. However, the mechanism that couples cell division with the flagellar biogenesis has remained elusive. Herein, we discover the regulator MadA that controls transcription of flagellar and cell-division genes in Caulobacter crescentus.

Collective dynein transport of the nucleus by pulling on astral microtubules during Saccharomyces cerevisiae mitosis.

Positioning the nucleus at the bud neck during Saccharomyces cerevisiae mitosis involves pulling forces of cytoplasmic dynein localized in the daughter cell. Although genetic analysis has revealed a complex network positioning the nucleus, quantification of the forces acting on the nucleus and the number of dyneins driving the process has remained difficult. To better understand the collective forces involved in nuclear positioning, we compare a model of dyneins-driven microtubule (MT) pulling, MT pushing, and cytoplasmic drag to experiments.

Time-varying mobility and turnover of actomyosin ring components during cytokinesis in .

Cytokinesis in many eukaryotes is dependent on a contractile actomyosin ring (AMR), composed of F-actin, myosin II, and other actin and myosin II regulators. Through fluorescence recovery after photobleaching experiments, many components of the AMR have been shown to be mobile and to undergo constant exchange with the cytosolic pools. However, how the mobility of its components changes at distinct stages of mitosis and cytokinesis has not been addressed.

Phosphoregulation of tropomyosin-actin interaction revealed using a genetic code expansion strategy.

Tropomyosins are coiled-coil proteins that regulate the stability and / or function of actin cytoskeleton in muscle and non-muscle cells through direct binding of actin filaments. Recently, using the fission yeast, we discovered a new mechanism by which phosphorylation of serine 125 of tropomyosin (Cdc8), reduced its affinity for actin filaments thereby providing access for the actin severing protein Adf1/Cofilin to actin filaments causing instability of actin filaments. Here we use a genetic code expansion strategy to directly examine this conclusion.

Genetic suppression of defective profilin by attenuated Myosin II reveals a potential role for Myosin II in actin dynamics in vivo in fission yeast.

The actin cytoskeleton plays a variety of roles in eukaryotic cell physiology, ranging from cell polarity and migration to cytokinesis. Key to the function of the actin cytoskeleton is the mechanisms that control its assembly, stability, and turnover. Through genetic analyses in , we found that -S1 (-G515D), a Myosin II mutant allele, was capable of rescuing lethality caused by partial defects in actin nucleation/stability caused, for example, through compromised function of the actin-binding protein Cdc3-profilin.

Phosphoregulation of tropomyosin is crucial for actin cable turnover and division site placement.

Tropomyosin is a coiled-coil actin binding protein key to the stability of actin filaments. In muscle cells, tropomyosin is subject to calcium regulation, but its regulation in nonmuscle cells is not understood. Here, we provide evidence that the fission yeast tropomyosin, Cdc8, is regulated by phosphorylation of a serine residue.

Measuring Binaural Temporal-Fine-Structure Sensitivity in Hearing-Impaired Listeners, Using the TFS-AF Test.

Background: Various methods have been used to measure temporal-fine-structure (TFS) sensitivity in hearing-impaired (HI) listeners. A new method called TFS-adaptive frequency (TFS-AF) test, tracks the highest frequency up to which a person can detect a given interaural phase difference (IPD) in bursts of pure tones. So far, the test was only administered to listeners with normal hearing (NH) or impaired low-frequency hearing.

Rapid production of pure recombinant actin isoforms in .

Actins are major eukaryotic cytoskeletal proteins, and they are involved in many important cell functions, including cell division, cell polarity, wound healing and muscle contraction. Despite obvious drawbacks, muscle actin, which is easily purified, is used extensively for biochemical studies of the non-muscle actin cytoskeleton. Here, we report a rapid and cost-effective method to purify heterologous actins expressed in the yeast Actin is expressed as a fusion with the actin-binding protein thymosin β4 and purified by means of an affinity tag introduced in the fusion.

Steric hindrance in the upper 50 kDa domain of the motor Myo2p leads to cytokinesis defects in fission yeast.

Cytokinesis in many eukaryotes requires a contractile actomyosin ring that is placed at the division site. In fission yeast, which is an attractive organism for the study of cytokinesis, actomyosin ring assembly and contraction requires the myosin II heavy chain Myo2p. Although -E1, a temperature-sensitive mutant defective in the upper 50 kDa domain of Myo2p, has been studied extensively, the molecular basis of the cytokinesis defect is not understood.

Actin turnover maintains actin filament homeostasis during cytokinetic ring contraction.

Cytokinesis in many eukaryotes involves a tension-generating actomyosin-based contractile ring. Many components of actomyosin rings turn over during contraction, although the significance of this turnover has remained enigmatic. Here, using , we investigate the role of turnover of actin and myosin II in its contraction.

Motor Activity Dependent and Independent Functions of Myosin II Contribute to Actomyosin Ring Assembly and Contraction in Schizosaccharomyces pombe.

Cytokinesis depends on a contractile actomyosin ring in many eukaryotes [1-3]. Myosin II is a key component of the actomyosin ring, although whether it functions as a motor or as an actin cross-linker to exert its essential role is disputed [1, 4, 5]. In Schizosaccharomyces pombe, the myo2-E1 mutation affects the upper 50 kDa sub-domain of the myosin II heavy chain, and cells carrying this lethal mutation are defective in actomyosin ring assembly at the non-permissive temperature [6, 7].

Myo2p is the major motor involved in actomyosin ring contraction in fission yeast.

Cytokinesis in many eukaryotes requires an actomyosin-based contractile ring [1]. In fission yeast, cytokinesis involves the type II myosins Myo2p and Myp2p and the type V myosin Myo51p [2]. A recent study by Laplante et al.