CUL3-KBTBD6/KBTBD7 ubiquitin ligase cooperates with GABARAP proteins to spatially restrict TIAM1-RAC1 signaling.

The small Rho GTPase RAC1 is an essential regulator of cellular signaling that controls actin rearrangements and cell motility. Here, we identify a novel CUL3 RING ubiquitin ligase complex, containing the substrate adaptors KBTBD6 and KBTBD7, that mediates ubiquitylation and proteasomal degradation of TIAM1, a RAC1-specific GEF. Increasing the abundance of TIAM1 by depletion of KBTBD6 and/or KBTBD7 leads to elevated RAC1 activity, changes in actin morphology, loss of focal adhesions, reduced proliferation, and enhanced invasion. KBTBD6 and KBTBD7 employ ATG8 family-interacting motifs to bind preferentially to GABARAP proteins. Surprisingly, ubiquitylation and degradation of TIAM1 by CUL3(KBTBD6/KBTBD7) depends on its binding to GABARAP proteins. Our study reveals that recruitment of CUL3(KBTBD6/KBTBD7) to GABARAP-containing vesicles regulates the abundance of membrane-associated TIAM1 and subsequently spatially restricted RAC1 signaling. Besides their role in autophagy and trafficking, we uncovered a previously unknown function of GABARAP proteins as membrane-localized signaling scaffolds.

TECPR2 Cooperates with LC3C to Regulate COPII-Dependent ER Export.

Hereditary spastic paraplegias (HSPs) are a diverse group of neurodegenerative diseases that are characterized by axonopathy of the corticospinal motor neurons. A mutation in the gene encoding for Tectonin ß-propeller containing protein 2 (TECPR2) causes HSP that is complicated by neurological symptoms. While TECPR2 is a human ATG8 binding protein and positive regulator of autophagy, the exact function of TECPR2 is unknown. Here, we show that TECPR2 associates with several trafficking components, among them the COPII coat protein SEC24D. TECPR2 is required for stabilization of SEC24D protein levels, maintenance of functional ER exit sites (ERES), and efficient ER export in a manner dependent on binding to lipidated LC3C. TECPR2-deficient HSP patient cells display alterations in SEC24D abundance and ER export efficiency. Additionally, TECPR2 and LC3C are required for autophagosome formation, possibly through maintaining functional ERES. Collectively, these results reveal that TECPR2 functions as molecular scaffold linking early secretion pathway and autophagy.

Telehealth Management of Dysphagia in Adults: A Survey of Speech Language Pathologists' Experiences and Perceptions.

The goal of this study was to explore telehealth use for dysphagia management in response to COVID-19 to understand variables associated with clinician confidence and perceived effectiveness of this service delivery model and determine clinician-perceived benefits and challenges of managing dysphagia via telehealth. Speech-language pathologists (SLPs, n = 235) completed a web-based survey, providing information on demographics, telehealth use during the pandemic, and perspectives on current and future tele-management of dysphagia. Analyses included descriptive statistics to examine usage patterns; logistic regression to determine which variables were associated with telehealth use, clinician confidence, and perceived-effectiveness; and conventional content analysis to analyze responses to open-ended questions. Results revealed a sharp increase in the tele-management of dysphagia during the pandemic. Years of experience with dysphagia management (p = .031) and pre-pandemic use of telehealth (p < .001) were significantly associated with current use patterns. Working in the outpatient setting was associated with greater clinician confidence (p = .003) and perceived effectiveness (p = .007), and use of guidelines (p = .042) was also associated with greater clinician confidence. Key challenges identified included inadequate technological infrastructure, inadequate patient digital literacy, and reimbursement restrictions. Key benefits were treatment continuity, improving access to care, and time savings. The majority (67%) of respondents reported that they would use telehealth in the future. These findings demonstrate SLPs' abilities and desire to expand their practice patterns to include telehealth for dysphagia management. Therefore, clinician training and more research on best practices for assessment and treatment of dysphagia via telehealth is warranted to refine models of care for dysphagia tele-management.

Oxygen-dependent asparagine hydroxylation of the ubiquitin-associated (UBA) domain in Cezanne regulates ubiquitin binding.

Deubiquitinases (DUBs) are vital for the regulation of ubiquitin signals, and both catalytic activity of and target recruitment by DUBs need to be tightly controlled. Here, we identify asparagine hydroxylation as a novel posttranslational modification involved in the regulation of Cezanne (also known as OTU domain-containing protein 7B (OTUD7B)), a DUB that controls key cellular functions and signaling pathways. We demonstrate that Cezanne is a substrate for factor inhibiting HIF1 (FIH1)- and oxygen-dependent asparagine hydroxylation. We found that FIH1 modifies Asn35 within the uncharacterized N-terminal ubiquitin-associated (UBA)-like domain of Cezanne (UBACez), which lacks conserved UBA domain properties. We show that UBACez binds Lys11-, Lys48-, Lys63-, and Met1-linked ubiquitin chains in vitro, establishing UBACez as a functional ubiquitin-binding domain. Our findings also reveal that the interaction of UBACez with ubiquitin is mediated via a noncanonical surface and that hydroxylation of Asn35 inhibits ubiquitin binding. Recently, it has been suggested that Cezanne recruitment to specific target proteins depends on UBACez Our results indicate that UBACez can indeed fulfill this role as regulatory domain by binding various ubiquitin chain types. They also uncover that this interaction with ubiquitin, and thus with modified substrates, can be modulated by oxygen-dependent asparagine hydroxylation, suggesting that Cezanne is regulated by oxygen levels.

Fluorescence-based ATG8 sensors monitor localization and function of LC3/GABARAP proteins.

Autophagy is a cellular surveillance pathway that balances metabolic and energy resources and transports specific cargos, including damaged mitochondria, other broken organelles, or pathogens for degradation to the lysosome. Central components of autophagosomal biogenesis are six members of the LC3 and GABARAP family of ubiquitin-like proteins (mATG8s). We used phage display to isolate peptides that possess bona fide LIR (LC3-interacting region) properties and are selective for individual mATG8 isoforms. Sensitivity of the developed sensors was optimized by multiplication, charge distribution, and fusion with a membrane recruitment (FYVE) or an oligomerization (PB1) domain. We demonstrate the use of the engineered peptides as intracellular sensors that recognize specifically GABARAP, GABL1, GABL2, and LC3C, as well as a bispecific sensor for LC3A and LC3B. By using an LC3C-specific sensor, we were able to monitor recruitment of endogenous LC3C to Salmonella during xenophagy, as well as to mitochondria during mitophagy. The sensors are general tools to monitor the fate of mATG8s and will be valuable in decoding the biological functions of the individual LC3/GABARAPs.

Multi-task prioritization during the performance of a postural-manual and communication task.

Individuals are often required to complete two tasks simultaneously, such as walking while talking. Although the influence of performing a cognitive task during upright standing has been studied, less is known regarding how individuals prioritize the performance of multiple goal-directed tasks. The purpose of this study was to examine how young adults prioritize the performance of two goal-directed tasks while standing: generating speech (communicating) while performing a postural-manual task. Fifteen young adult female participants completed four conditions of the postural-manual task, quietly and while talking; a control condition of speech while standing was also completed. Task accuracy was defined as holding a block within an opening without hitting the perimeter and speaking clearly. Participants generally prioritized accurate communication over accurate completion of the postural-manual task, as evidenced by modifying their manual and postural strategies and decreasing manual accuracy.

Sentence production in Parkinson's disease.

While growing evidence reports changes in language use in non-demented individuals with Parkinson's disease (PD), the presence and nature of the deficits remain largely unclear. Researchers have proposed that dysfunctioning fronto-basal ganglia circuit results in impaired grammatical processes, predicting qualitatively similar language impairments between individuals with PD and agrammatic Broca's aphasia, whereas others suggest that PD is not associated with language-specific grammatical impairment. In addition, there is a paucity of research examining syntactic production in PD at the sentence-level. This study examined sentence production of individuals with PD, healthy older adults, and individuals with agrammatic Broca's aphasia. In Experiment 1, using a Cinderella story-telling task, proportion of grammatical sentences, number of embedded clauses and production of verb arguments in sentences were examined. In Experiment 2, a structured sentence elicitation task was used in which syntactic complexity of sentences (canonical vs. non-canonical word order) was systematically manipulated while minimizing demands for non-syntactic processing. Only the participants with agrammatic Broca's aphasia showed significantly impaired syntactic production in both experiments. Participants with PD did not show impaired syntactic production in either task, despite impairments in lexical retrieval, repetition of words and sentences, and speech production. These findings suggest that impaired syntactic processing may not be a core deficit underlying the changes in language use in non-demented PD. Changes in language use in PD are qualitatively different from language deficits in aphasia.

Structural and Functional Analysis of a Novel Interaction Motif within UFM1-activating Enzyme 5 (UBA5) Required for Binding to Ubiquitin-like Proteins and Ufmylation.

The covalent conjugation of ubiquitin-fold modifier 1 (UFM1) to proteins generates a signal that regulates transcription, response to cell stress, and differentiation. Ufmylation is initiated by ubiquitin-like modifier activating enzyme 5 (UBA5), which activates and transfers UFM1 to ubiquitin-fold modifier-conjugating enzyme 1 (UFC1). The details of the interaction between UFM1 and UBA5 required for UFM1 activation and its downstream transfer are however unclear. In this study, we described and characterized a combined linear LC3-interacting region/UFM1-interacting motif (LIR/UFIM) within the C terminus of UBA5. This single motif ensures that UBA5 binds both UFM1 and light chain 3/γ-aminobutyric acid receptor-associated proteins (LC3/GABARAP), two ubiquitin (Ub)-like proteins. We demonstrated that LIR/UFIM is required for the full biological activity of UBA5 and for the effective transfer of UFM1 onto UFC1 and a downstream protein substrate both in vitro and in cells. Taken together, our study provides important structural and functional insights into the interaction between UBA5 and Ub-like modifiers, improving the understanding of the biology of the ufmylation pathway.

The efficacy of the Microsoft KinectTM to assess human bimanual coordination.

The Microsoft Kinect has been used in studies examining posture and gait. Despite the advantages of portability and low cost, this device has not been used to assess interlimb coordination. Fundamental insights into movement control, variability, health, and functional status can be gained by examining coordination patterns. In this study, we investigated the efficacy of the Microsoft Kinect to capture bimanual coordination relative to a research-grade motion capture system. Twenty-four healthy adults performed coordinated hand movements in two patterns (in-phase and antiphase) at eight movement frequencies (1.00-3.33 Hz). Continuous relative phase (CRP) and discrete relative phase (DRP) were used to quantify the means (mCRP and mDRP) and variability (sdCRP and sdDRP) of coordination patterns. Between-device agreement was assessed using Bland-Altman bias with 95 % limits of agreement, concordance correlation coefficients (absolute agreement), and Pearson correlation coefficients (relative agreement). Modest-to-excellent relative and absolute agreements were found for mCRP in all conditions. However, mDRP showed poor agreement for the in-phase pattern at low frequencies, due to large between-device differences in a subset of participants. By contrast, poor absolute agreement was observed for both sdCRP and sdDRP, while relative agreement ranged from poor to excellent. Overall, the Kinect captures the macroscopic patterns of bimanual coordination better than coordination variability.

Nanogold-Decorated Silica Monoliths as Highly Efficient Solid-Phase Adsorbent for Ultratrace Mercury Analysis in Natural Waters.

We propose a novel analytical method for mercury (Hg) trace determination based on direct Hg preconcentration from aqueous solution onto a gold nanoparticle-decorated silica monolith (AuNP@SiO2). Detection of Hg is performed after thermal desorption by means of atomic fluorescence spectrometry. This new methodology benefits from reagent-free, time- and cost-saving procedure, due to most efficient solid-phase adsorbent and results in high sensitive quantification. The excellent analytical performance of the whole procedure is demonstrated by a limit of detection as low as 1.31 ng L(-1) for only one-min accumulation duration. A good reproducibility with standard deviations ≤5.4% is given. The feasibility of the approach in natural waters was confirmed by a recovery experiment in spiked seawater with a recovery rate of 101%. Moreover, the presented method was validated through reference analysis of a submarine groundwater discharge sample by cold vapor-atomic fluorescence spectrometry resulting in a very good agreement of the found values. Hence the novel method is a very promising new tool for low-level Hg monitoring in natural waters providing easy-handling on-site preconcentration, reagent-free stabilization as well as reagent-free, highly sensitive detection.

An Examination of Cognitive Heterogeneity in Parkinson Disease: The Dual-Syndrome Hypothesis.

PURPOSE: Cognitive impairment is one of the most debilitating nonmotor symptoms in Parkinson disease (PD), and its presentation is heterogeneous. One proposed model to explain cognitive variability in PD is the dual-syndrome hypothesis. This hypothesis delineates two cognitive profiles, a "fronto-striatal" profile and a "posterior cortical" profile according to symptom presentation, associated motor phenotype, and risk for dementia. The current study examined the dual-syndrome hypothesis in individuals with idiopathic PD to evaluate the existence of these profiles, determine the association with the motor phenotype (tremor dominant vs. postural instability/gait disorder), and assess the relative risk for dementia. METHOD: A retrospective examination was conducted using data from the Parkinson's Progression Markers Initiative database at baseline (within 2 years of diagnosis) and 5 years after baseline. Descriptive categorizations, cluster analyses, generalized linear mixed models, and logistic regressions were used to address the research questions. RESULTS: There was emerging evidence of cognitive profiles; however, these were not fully supported by cluster analyses. Baseline cognitive profile was associated with later motor phenotype, and as predicted, dementia risk was greatest in persons with baseline posterior cortical impairments. CONCLUSION: The current results provide mixed support for the dual-syndrome hypothesis, with some evidence that the posterior cortical cognitive profile is associated with postural instability and gait disorder as well as greater dementia risk.

A Preliminary Study of Voluntary Cough Motor Performance and Learning With Skill Training and Biofeedback.

PURPOSE: Sensorimotor cough skill training (CST) has been shown to improve cough strength, as well as facilitate changes during training (i.e., motor performance) and generalization to untrained tasks (i.e., motor learning). However, there is a gap in our understanding of the effects of voluntary CST (without sensory stimuli) on motor performance and learning. Furthermore, the contribution of physiologic factors, such as lung volume, a driver of cough strength in healthy adults, and treatment-specific factors, such as biofeedback, remains unexamined. METHOD: Twenty individuals with Parkinson's disease (PD) completed pre- and post-CST single voluntary, sequential voluntary, and reflex cough testing. Participants were randomized to biofeedback or no biofeedback groups. They completed one CST session involving 25 trials of voluntary coughs, with the treatment target set 25% above baseline peak flow. Participants were instructed to "cough hard" to exceed the target. In the biofeedback group, participants received direct visualization of the target line in real time. RESULTS: Cough peak flow showed positive improvements in motor performance (ß = .02; 95% credible interval [CI]: 0.01, 0.03) and learning (ß = .26; 95% CI: 0.03, 0.47). Changes in lung volume from pre- to post-CST did not predict treatment response. No differences in treatment response were detected between the biofeedback groups. CONCLUSIONS: A single session of voluntary CST improved voluntary cough motor performance and learning. Although lung volume increased during CST, changes to lung volume did not predict treatment response. These findings demonstrate the potential of voluntary CST to improve motor performance and motor learning among individuals with PD and cough dysfunction. SUPPLEMENTAL MATERIAL AND OPEN SCIENCE FORM: https://doi.org/10.23641/asha.25447444.

Behavioral Management of Respiratory/Phonatory Dysfunction for Dysarthria Associated With Neurodegenerative Disease: A Systematic Review.

PURPOSE: This systematic review represents an update to previous reviews of the literature addressing behavioral management of respiratory/phonatory dysfunction in individuals with dysarthria due to neurodegenerative disease. METHOD: Multiple electronic database searches and hand searches of prominent speech-language pathology journals were conducted in accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses standards. RESULTS: The search yielded 1,525 articles, from which 88 met inclusion criteria and were reviewed by two blinded co-investigators. A large range of therapeutic approaches have been added to the evidence base since the last review, including expiratory muscle strength training, singing, and computer- and device-driven programs, as well as a variety of treatment modalities, including teletherapy. Evidence for treatment in several different population groups-including cerebellar ataxia, myotonic dystrophy, autosomal recessive spastic ataxia of Charlevoix-Saguenay, Huntington's disease, multiple system atrophy, and Lewy body dementia-were added to the current review. Synthesis of evidence quality provided strong evidence in support of only one behavioral intervention: Lee Silverman Voice Treatment Program (LSVT LOUD) in people with Parkinson's disease. No other treatment approach or population included in this review demonstrated more than limited evidence, reflecting that these approaches/populations require urgent further examination. CONCLUSION: Suggestions about where future research efforts could be significantly strengthened and how clinicians can apply research findings to their practice are provided. SUPPLEMENTAL MATERIAL: https://doi.org/10.23641/asha.24964473.

Task-dependent postural control throughout the lifespan.

Routine activities performed while standing and walking require the ability to appropriately and continuously modulate postural movements as a function of a concurrent task. Changes in task-dependent postural control contribute to the emergence, maturation, and decline of complex motor skills and stability throughout the lifespan.

The Effects of Speech Task on Lexical Stress in Parkinson's Disease.

PURPOSE: Hypokinetic dysarthria associated with Parkinson's disease (PD) is characterized by dysprosody, yet the literature is mixed with respect to how dysprosody affects the ability to mark lexical stress, possibly due to differences in speech tasks used to assess lexical stress. The purpose of this study was to compare how people with and without PD modulate acoustic dimensions of lexical stress-fundamental frequency, intensity, and duration-to mark lexical stress across three different speech tasks. METHOD: Twelve individuals with mild-to-moderate idiopathic PD and 12 age- and sex-matched older adult controls completed three speech tasks: picture description, word production in isolation, and word production in lists. Outcome measures were the fundamental frequency, intensity, and duration of the vocalic segments of two trochees (initial stress) and two iambs (final stress) spoken in all three tasks. RESULTS: There were very few group differences. Both groups marked trochees by modulating intensity and fundamental frequency and iambs by modulating duration. Task had a significant impact on the stress patterns used by both groups. Stress patterns were most differentiated in words produced in isolation and least differentiated in lists of words. CONCLUSIONS: People with PD did not demonstrate impairments in the production of lexical stress, suggesting that dysprosody associated with PD does not impact all types of prosody in the same way. However, there were reduced distinctions in stress marking that were more apparent in trochees than iambs. In addition, the task used to assess prosody has a significant effect on all acoustic measures. Future research should focus on the use of connected speech tasks to obtain more generalizable measures of prosody in PD.

Ultra-trace determination of mercury in river waters after online UV digestion of humic matter.

A fully automated online ultraviolet (UV) digestion method for subsequent mercury (Hg) quantification in humic matter containing river waters is reported. The new developed flow injection analysis system (FIAS) consists basically of a UV lamp, a meander-form quartz glass reaction tube for online irradiation of the sample, and a nano-gold collector for preconcentration of dissolved mercury species. The FIAS is coupled to an atomic fluorescence spectrometer (AFS) for Hg detection. The optimized procedure allows accurate mercury quantification in water samples with up to 15 mg CL(-1) as dissolved organic carbon by addition of only 1% (v/v) of hydrogen peroxide solution and online UV irradiation for 6 min. Addition of strong oxidants and any other reagents is avoided due to the use of the catalytic active nano-gold collector. Here, preconcentration of Hg species, release of mercury as Hg(0), and AFS measurement are performed without addition of any reagents. Hence, the proposed approach offers significant advantages over existing methods. Analytical figures of merit showed the good performance of the developed method: The limit of quantification was found to be as low as 0.14 ng Hg L(-1). The linear working range is from 0.1 to 200 ng Hg L(-1) and relative standard deviation is <6.0% (n = 9). The system was successfully validated by comparison of the mercury concentrations found in model and real water samples obtained by the reference method EPA 1631 and the proposed method. Furthermore, application to six real river waters confirmed the feasibility of the proposed approach.

Perception of Physical Demand, Mental Demand, and Performance: A Comparison of Two Voice Interventions for Parkinson's Disease.

PURPOSE: The purpose of the study was to examine the effect of two voice intervention approaches for hypophonia secondary to Parkinson's disease (PD) on self-reported measures of physical demand, mental demand, and vocal performance. METHOD: Thirty-four persons with hypophonia secondary to PD were assigned to one of three groups: Lee Silverman Voice Treatment (LSVT) LOUD (n = 12), SpeechVive (n = 12), and nontreatment clinical control (n = 10). The LSVT LOUD and the SpeechVive participants received 8 weeks of voice intervention following the standardized protocol previously described for each approach. To confirm the effectiveness of each voice intervention, sound pressure level (dB SPL) data were analyzed for the experimental and control participants for a monologue sample obtained pretreatment, midtreatment, and posttreatment. During the voice intervention period, the LSVT LOUD and the SpeechVive participants were instructed to complete a modified version of the National Aeronautics and Space Administration Task Load Index rating scale to indicate the mental and physical demand required to complete the intervention activities, and to indicate how well they performed in completing the assigned vocal tasks. RESULTS: The LSVT LOUD and the SpeechVive participants demonstrated a significant posttreatment increase in SPL (dB), in comparison to the clinical controls, thus confirming a positive intervention effect. The LSVT LOUD participants reported significantly higher ratings of physical and mental demand over the course of treatment, in comparison to the SpeechVive participants. CONCLUSION: Consideration of the mental and physical demand associated with two voice intervention approaches, commonly used for PD, may help to foster improved therapeutic compliance and treatment outcomes.

Longitudinal Effects of Parkinson's Disease on Speech Breathing During an Extemporaneous Connected Speech Task.

PURPOSE: A critical component to the development of any type of intervention to improve speech production in individuals with Parkinson's disease (PD) is a complete understanding of the speech impairments present at each stage of the disease and how these impairments change with disease progression. The purpose of this longitudinal study was to examine the impact of disease on speech production and speech breathing during an extemporaneous speech task in individuals with PD over the course of approximately 3.5 years. METHOD: Eight individuals with PD and eight age- and sex-matched control participants produced an extemporaneous connected speech task on two occasions (Time 1 and Time 2) an average of 3 years 7 months apart. Dependent variables included sound pressure level; utterance length; speech rate; lung volume initiation, termination, and excursion; and percent vital capacity per syllable. RESULTS: From Time 1 to Time 2, individuals with PD demonstrated decreased utterance length and lung volume initiation, termination, and excursion and increased speech rate. Control participants demonstrated decreased utterance length and lung volume termination and increased lung volume excursion and percent vital capacity per syllable from Time 1 to Time 2. CONCLUSIONS: Changes in speech production and speech breathing variables experienced by individuals with PD over the course of several years are related to their disease process and not typical aging. Changes to speech breathing highlight the need to provide intervention focused on increasing efficient respiratory patterning for speech production.

The effects of vocal exertion on lung volume measurements and acoustics in speakers reporting high and low vocal fatigue.

PURPOSE: Vocal exertion is common and often results in reduced respiratory and laryngeal efficiency. It is unknown, however, whether the respiratory kinematic and acoustic adjustments employed during vocal exertion differ between speakers reporting vocal fatigue and those who do not. This study compared respiratory kinematics and acoustic measures in individuals reporting low and high levels of vocal fatigue during a vocal exertion task. METHODS: Individuals reporting low (N = 20) and high (N = 10) vocal fatigue participated in a repeated measures design study over 2 days. On each day, participants completed a 10-minute vocal exertion task consisting of repeated, loud vowel productions at elevated F0 sustained for maximum phonation time. Respiratory kinematic and acoustic measures were analyzed on the 1st vowel production (T0), and the vowels produced 2 minutes (T2), 5 minutes (T5), 7 minutes (T7), and 10 minutes (T10) into the vocal exertion task. Vowel durations were also measured at each time point. RESULTS: No differences in respiratory kinematics were observed between low and high vocal fatigue groups at T0. As the vocal exertion task progressed (T2-T10), individuals reporting high vocal fatigue initiated phonation at lower lung volumes while individuals with low vocal fatigue initiated phonation at higher lung volumes. As the exertion task progressed, total lung volume excursion decreased in both groups. Differences in acoustic measures were observed, as individuals reporting high vocal fatigue produced softer, shorter vowels from T0 through T10. CONCLUSIONS: Individuals reporting high vocal fatigue employed less efficient respiratory strategies during periods of increased vocal demand when compared with individuals reporting low vocal fatigue. Individuals reporting high vocal fatigue had shorter maximum phonation time on loud vowels. Further study should examine the potential screening value of loud maximum phonation time, as well as the clinical implications of the observed respiratory patterns for managing vocal fatigue.