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Despite extensive structure-function analyses, the molecular mechanisms of normal and pathological tau action remain poorly understood. How does the C-terminal microtubule-binding region regulate microtubule dynamics and bundling? In what biophysical form does tau transfer trans-synaptically from one neuron to another, promoting neurodegeneration and dementia? Previous biochemical/biophysical work led to the hypothesis that tau can dimerize via electrostatic interactions between two N-terminal 'projection domains' aligned in an anti-parallel fashion, generating a multivalent complex capable of interacting with multiple tubulin subunits. We sought to test this dimerization model directly. Native gel analyses of full-length tau and deletion constructs demonstrate that the N-terminal region leads to multiple bands, consistent with oligomerization. Ferguson analyses of native gels indicate that an N-terminal fragment (tau(45-230) ) assembles into heptamers/octamers. Ferguson analyses of denaturing gels demonstrates that tau(45-230) can dimerize even in sodium dodecyl sulfate. Atomic force microscopy reveals multiple levels of oligomerization by both full-length tau and tau(45-230) . Finally, ion mobility-mass spectrometric analyses of tau(106-144) , a small peptide containing the core of the hypothesized dimerization region, also demonstrate oligomerization. Thus, multiple independent strategies demonstrate that the N-terminal region of tau can mediate higher order oligomerization, which may have important implications for both normal and pathological tau action. The microtubule-associated protein tau is essential for neuronal development and maintenance, but is also central to Alzheimer's and related dementias. Unfortunately, the molecular mechanisms underlying normal and pathological tau action remain poorly understood. Here, we demonstrate that tau can homo-oligomerize, providing novel mechanistic models for normal tau action (promoting microtubule growth and bundling, suppressing microtubule shortening) and pathological tau action (poisoning of oligomeric complexes).

Microtubules (MTs) are hollow cytoskeletal filaments assembled from αβ-tubulin heterodimers. Tau, an unstructured protein found in neuronal axons, binds to MTs and regulates their dynamics. Aberrant Tau behavior is associated with neurodegenerative dementias, including Alzheimer's. Here, we report on a direct force measurement between paclitaxel-stabilized MTs coated with distinct Tau isoforms by synchrotron small-angle X-ray scattering (SAXS) of MT-Tau mixtures under osmotic pressure (P). In going from bare MTs to MTs with Tau coverage near the physiological submonolayer regime (Tau/tubulin-dimer molar ratio; ΦTau = 1/10), isoforms with longer N-terminal tails (NTTs) sterically stabilized MTs, preventing bundling up to PB ∼ 10,000-20,000 Pa, an order of magnitude larger than bare MTs. Tau with short NTTs showed little additional effect in suppressing the bundling pressure (PB ∼ 1,000-2,000 Pa) over the same range. Remarkably, the abrupt increase in PB observed for longer isoforms suggests a mushroom to brush transition occurring at 1/13 < ΦTau < 1/10, which corresponds to MT-bound Tau with NTTs that are considerably more extended than SAXS data for Tau in solution indicate. Modeling of Tau-mediated MT-MT interactions supports the hypothesis that longer NTTs transition to a polyelectrolyte brush at higher coverages. Higher pressures resulted in isoform-independent irreversible bundling because the polyampholytic nature of Tau leads to short-range attractions. These findings suggest an isoform-dependent biological role for regulation by Tau, with longer isoforms conferring MT steric stabilization against aggregation either with other biomacromolecules or into tight bundles, preventing loss of function in the crowded axon environment.

Chlamydia trachomatis is a major bacterial pathogen throughout the world. Although antibiotic therapy can be implemented in the case of early detection, a majority of the infections are asymptomatic, requiring the development of preventive measures. Efforts have focused on the production of a vaccine using the C. trachomatis major outer membrane protein (MOMP). MOMP is purified in its native (n) trimeric form using the zwitterionic detergent Z3–14, but its stability in detergent solutions is limited. Amphipols (APols) are synthetic polymers that can stabilize membrane proteins (MPs) in detergent-free aqueous solutions. Preservation of protein structure and optimization of exposure of the most effective antigenic regions can avoid vaccination with misfolded, poorly protective protein. Previously, we showed that APols maintain nMOMP secondary structure and that nMOMP/APol vaccine formulations elicit better protection than formulations using either recombinant or nMOMP solubilized in Z3–14. To achieve a greater understanding of the structural behavior and stability of nMOMP in APols, we have used several spectroscopic techniques to characterize its secondary structure (circular dichroism), tertiary and quaternary structures (immunochemistry and gel electrophoresis) and aggregation state (light scattering) as a function of temperature and time. We have also recorded NMR spectra of 15N-labeled nMOMP and find that the exposed loops are detectable in APols but not in detergent. Our analyses show that APols protect nMOMP much better than Z3–14 against denaturation due to continuous heating, repeated freeze/thaw cycles, or extended storage at room temperature. These results indicate that APols can help improve MP-based vaccine formulations.

Presents schemes for classifying the properties of matter and the conditions which may affect these properties. (WB)

Introduction: Voice disorders are a common communication disorder in children. Behavioral voice therapy is recommended by both Otolaryngologists and Speech-Language Pathologists as a first-line approach for treatment of benign vocal fold lesions that affect children in large numbers. However, the role of cognitive mechanisms critical to voice therapy have not yet been explored., Objective: This proof-of-concept study aims to provide preliminary data on the potential relation between cognitive abilities and behavioral results of voice therapy for children with benign phonotraumatic vocal fold lesions., Methods: Six children (4;05 -9;02 years) diagnosed with vocal fold nodules completed a battery of cognitive tests from the NEuroPSYchological (NEPSY-II) Assessment and a standard course of "Adventures in Voice" therapy. Recordings pre and post intervention were analyzed acoustically using the Cepstral Spectral Index of Dysphonia (CSID) and perceptually using visual analog scales (VAS) for severity and resonance separately. Raw and age-corrected scaled scores from the NEPSY-II were then examined for their possible relation to voice outcomes., Results: Multiple cognitive functions correlated with voice outcomes. Raw score measures for Design Fluency, Inhibition, Fingertip Tapping, and Narrative Memory correlated favorably with all voice outcome measures. Age correlated with all NEPSY-II raw scores and perceptual voice outcome measures. Scaled scores for Auditory Attention, Design Fluency, Fingertip Tapping (Dominant hand), and Narrative Memory correlated with all voice outcome measures., Conclusion: Results suggest that there is merit to further investigation of the relation between cognitive skills and their development and voice treatment outcomes in children with benign phonotraumatic lesions. Future studies with larger samples will build on present findings., (Copyright © 2021 The Voice Foundation. Published by Elsevier Inc. All rights reserved.)

Objectives: The primary aim was to compare two methods for perceptual evaluation of voice - paired comparison (PC) and visual analog scale (VAS) ratings. Secondary aims were to assess the correspondence between two dimensions of voice- overall severity of voice quality and resonant voice, and to investigate the influence of rater experience on perceptual rating scores and rating confidence scores., Study Design: Experimental design., Methods: Voice samples from six children (pre and post therapy) were rated by 15 Speech-Language Pathologists specialized in voice. Raters completed four tasks corresponding to the two rating methods and voice qualities: PC-severity, PC-resonance, VAS-severity, and VAS-resonance. For PC tasks, raters chose the better of two voice samples (better voice quality or better resonance, depending on the task) and indicated the degree of confidence in each choice. Rating and confidence score were combined to produce a number on a 1-10 scale (PC-confidence adjusted). VAS ratings involved rating voices on a scale for degree of severity and resonance, respectively., Results: PC-confidence adjusted and VAS ratings were moderately correlated for overall severity and also vocal resonance. VAS ratings were normally distributed and had greater rater consistency than PC-confidence adjusted ratings. VAS scores reliably predicted binary PC choices (choice of voice sample only). Overall severity and vocal resonance were weakly correlated and rater experience was not linearly related to rating scores or confidence., Conclusions: Results suggest that the VAS rating method holds advantages over PC, including normally distributed ratings, superior consistency of ratings, and the ability to provide more finely grained detail regarding the auditory perception of voice. Overall severity and vocal resonance were not redundant in the current data set, suggesting that resonant voice and overall severity are not isomorphic. Finally, the number of years of clinical experience was not linearly related to perceptual ratings or rating confidence., (Copyright © 2023 The Voice Foundation. Published by Elsevier Inc. All rights reserved.)

Purpose: The study's primary aim was to investigate developmental changes in the perception of vocal loudness and voice quality in children 3-6 years of age. A second aim was to evaluate a testing procedure-the intermodal preferential looking paradigm (IPLP)-for the study of voice perception in young children., Method: Participants were categorized in two age groups: 3- to 4-year-olds and 5- to 6-year-olds. Children were tested remotely via a Zoom appointment and completed two perceptual tasks: (a) voice discrimination and (b) voice identification. Each task consisted of two tests: a vocal loudness test and a voice quality test., Results: Children in the 5- to 6-year-old group were significantly more accurate than children in the 3- to 4-year-old group in discriminating and identifying differences between voices for both loudness and voice quality. The IPLP, used in the identification task, was found to successfully detect differences between the age groups for overall accuracy and for most of the sublevels of vocal loudness and voice quality., Conclusions: Results suggest that children's ability to discriminate and identify differences in vocal loudness and voice quality improves with age. Findings also support the use of the IPLP as a useful tool to study voice perception in young children.

Purpose: The purpose of this study was to establish the frequency response of a selection of low-cost headset microphones that could be given to subjects for remote voice recordings and to examine the effect of microphone type and frequency response on key acoustic measures related to voice quality obtained from speech and vowel samples., Method: The frequency responses of three low-cost headset microphones were evaluated using pink noise generated via a head-and-torso model. Each of the headset microphones was then used to record a series of speech and vowel samples prerecorded from 24 speakers who represented a diversity of sex, age, fundamental frequency ( F ), and voice quality types. Recordings were later analyzed for the following measures: smoothed cepstral peak prominence (CPP; dB), low versus high spectral ratio (L/H ratio; dB), CPP o ), and voice quality types. Recordings were later analyzed for the following measures: smoothed cepstral peak prominence (CPP; dB), low versus high spectral ratio (L/H ratio; dB), CPP F o (Hz), and cepstral spectral index of dysphonia (CSID)., Results: The frequency response of the microphones under test was observed to have nonsignificant effects on measures of the CPP and CPP F , significant effects on the CSID in speech contexts, and strong and significant effects on the measure of spectral tilt (L/H ratio). However, the correlations between the various headset microphones and a reference precision microphone were excellent ( o , significant effects on the CSID in speech contexts, and strong and significant effects on the measure of spectral tilt (L/H ratio). However, the correlations between the various headset microphones and a reference precision microphone were excellent ( r s > .90)., Conclusions: The headset microphones under test all showed the capability to track a wide range of diversity in the voice signal. Though the use of higher quality microphones that have demonstrated specifications is recommended for typical research and clinical purposes, low-cost electret microphones may be used to provide valid measures of voice, specifically when the same microphone and signal chain is used for the evaluation of pre- versus posttreatment change or intergroup comparisons.

Purpose This systematic review aims to identify, classify, and evaluate existing information regarding treatment for benign vocal fold lesions in children and to identify gaps and limitations that may limit effective pediatric voice treatment. Method A literature search was performed using electronic databases (PubMed and Google Scholar) as well as reference lists from previous reviews, studies, and books. Included in the present review are studies that described behavioral treatment for children with benign vocal fold lesions presumed to be phonotraumatic (vocal fold nodules and edema). Results Twenty-one studies were eligible for inclusion in the review. Eight different research designs were used, and three intervention types were identified: direct voice intervention (voice training), indirect treatment (vocal hygiene or counseling), and comparative studies that contrasted different treatment methods. The most commonly used treatment method was eclectic direct intervention, which focused on vocal exercises or voicing patterns. Postintervention improvement was reported in all studies. In general, findings suggested an advantage of direct over indirect intervention and of longer treatment duration over short-term approaches. Conclusions The findings suggest that behavioral voice therapy may be generally effective in treating children with vocal fold nodules. Several limitations emerged in the corpus of studies reviewed including heterogeneity of research methods, missing information about outcome measures, and inappropriate statistical analyses. Thus, a need exists for further well-designed controlled studies to enhance the body of knowledge about developmental factors affecting vocal treatment outcomes, in particular, vocal fold structure as well as cognitive and linguistic development.

Chlamydia trachomatis is a major bacterial pathogen throughout the world. Although antibiotic therapy can be implemented in the case of early detection, a majority of the infections are asymptomatic, requiring the development of preventive measures. Efforts have focused on the production of a vaccine using the C. trachomatis major outer membrane protein (MOMP). MOMP is purified in its native (n) trimeric form using the zwitterionic detergent Z3-14, but its stability in detergent solutions is limited. Amphipols (APols) are synthetic polymers that can stabilize membrane proteins (MPs) in detergent-free aqueous solutions. Preservation of protein structure and optimization of exposure of the most effective antigenic regions can avoid vaccination with misfolded, poorly protective protein. Previously, we showed that APols maintain nMOMP secondary structure and that nMOMP/APol vaccine formulations elicit better protection than formulations using either recombinant or nMOMP solubilized in Z3-14. To achieve a greater understanding of the structural behavior and stability of nMOMP in APols, we have used several spectroscopic techniques to characterize its secondary structure (circular dichroism), tertiary and quaternary structures (immunochemistry and gel electrophoresis) and aggregation state (light scattering) as a function of temperature and time. We have also recorded NMR spectra of (15)N-labeled nMOMP and find that the exposed loops are detectable in APols but not in detergent. Our analyses show that APols protect nMOMP much better than Z3-14 against denaturation due to continuous heating, repeated freeze/thaw cycles, or extended storage at room temperature. These results indicate that APols can help improve MP-based vaccine formulations.

Objective: To evaluate the safety and effectiveness of tinzaparin for the prevention and treatment of venous thromboembolism (VTE)., Data Sources: A MEDLINE and PubMed database search (1980-December 2002) was conducted. Only articles written in English were reviewed., Study Selection and Data Extraction: Articles reporting the safety, efficacy, and cost-effectiveness of tinzaparin in humans were evaluated. Emphasis was placed on randomized, controlled trials., Data Synthesis: Tinzaparin sodium is a low-molecular-weight heparin (LMWH) that exerts its anticoagulant effect through inhibition of factors Xa and IIa and release of tissue factor pathway inhibitor from the vascular epithelium. Tinzaparin is indicated for treatment of acute symptomatic deep-vein thrombosis (DVT), with or without pulmonary embolism. Clinical studies suggest that tinzaparin is also effective for VTE prophylaxis, as well as other indications. Once-daily subcutaneous tinzaparin is equally or more effective than intravenous unfractionated heparin (UFH) for prevention and treatment of VTE, at least as safe as UFH for bleeding complications, and requires little or no monitoring. No dose "cap" is required for obese patients, and no initial dosing adjustments are necessary in elderly and/or renally impaired patients, although some monitoring is recommended. The few comparative data available suggest that tinzaparin efficacy may be comparable to that of other LMWHs; more comparative studies are needed. Pharmacoeconomic studies indicate a favorable cost-benefit ratio., Conclusions: Tinzaparin is safe and effective for prevention and treatment of DVT. Consistent once-daily dosing may facilitate self-administration of tinzaparin in the outpatient setting.

Handedness was examined in relation to sex, race, age, education, occupation, marital status, and religious preferences for two random samples of adults drawn from an urban population. There were statistically significant differences on each variable for the total of 2083 respondents as well as for many subgroups based on combinations of sex, race, and age. The results are compared to previous findings.