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Purpose: The purpose of this investigation was to examine single-word speech intelligibility outcomes following sound production treatment in a group of 22 speakers with chronic acquired apraxia of speech (AOS) and aphasia. Also, the stability of repeated posttreatment intelligibility measures was examined for two scoring methods., Method: The Assessment of Intelligibility of Dysarthric Speech was administered twice to each participant at pretreatment and twice at 8 weeks posttreatment. The test-retest reliability of the pretreatment samples was evaluated in a prior study; repeated samples were found to be stable over sampling times. For the current study, the three expert listeners who had rated the pretreatment samples scored the posttreatment samples using transcription and multiple-choice scoring formats. An additional expert listener, blinded to sampling time, scored pre- and posttreatment samples., Results: The posttreatment samples were found to be stable over sampling times for the group. Posttreatment intelligibility scores were statistically significantly higher than the pretreatment scores for both scoring methods (i.e., increases of 9%-10%). Scores derived from the two scoring methods were strongly, positively correlated, with multiple-choice scores being significantly higher than transcription. The scoring methods did not differ significantly in the amount of change found from pre- to posttreatment. There were no statistically significant correlations between AOS severity and changes in pre- to posttreatment intelligibility scores. Performance for most participants was similar to group performance., Conclusions: Stability of posttreatment intelligibility samples supports use of intelligibility scores as outcome measures. This group of speakers demonstrated statistically significant increases in single-word speech intelligibility following sound production treatment., Supplemental Material: https://doi.org/10.23641/asha.26972425.

Purpose: Neurogenic speech and language disorders-such as acquired apraxia of speech (AOS) and aphasia with phonemic paraphasia (APP)-are often misdiagnosed due to similarities in clinical presentation. Word syllable duration (WSD)-a measure of average syllable length in multisyllabic words-serves as a proxy for speech rate, which is an important and arguably more objective clinical characteristic of AOS and APP. This study reports stability of WSD over time for speakers with AOS (and aphasia)., Method: Twenty-nine participants with AOS and aphasia (11 women and 18 men, M age = 53.5 years, SD = 13.3) repeated 30 multisyllabic words (of three-, four-, and five-syllable lengths) on three occasions across 4 weeks. WSDs were calculated for each word and then averaged across each list (i.e., word length), as well as across combined lists (i.e., all 30 words) to yield four WSDs for each participant at each time point. Stability over time was calculated using Friedman's test for the group and using Spearman's rho for the individual level. Effects of time and word length were examined using robust mixed-effects linear regression., Results: Friedman's tests and correlations indicated no significant difference in WSDs across sampling occasions for each word length separately or combined. WSD correlated positively with AOS severity and negatively with intelligibility but was not correlated with aphasia severity. Regression analyses confirmed WSD to be stable over time, while WSD calculated from only five tokens (i.e., WSD-5) was less stable over time., Conclusions: Results indicate that WSD can be a stable measure over time, at the individual and group level, providing support for its use in diagnosis and/or as an outcome measure, both clinically and for research. In general, WSD outperformed WSD-5, suggesting that it may be better to calculate WSD from more than five tokens. Stability of WSD in other populations and suitability for differential diagnosis need to be determined. Currently, differentiating disorders by speaking rate, alone, is not recommended., Supplemental Material: https://doi.org/10.23641/asha.25438735.

Background: Every year, many thousands of travellers return to the United Kingdom (UK) from visits to other countries and some will become unwell due to infections acquired abroad. Many imported infections have similar clinical presentations, such as fever and myalgia, so diagnostic testing is an important tool to improve patient management and outcomes. The aim of this study was to examine the demographics, travel history, presenting symptoms and diagnostic outcomes of referrals to the UK's specialist diagnostic Rare & Imported Pathogens Laboratory (RIPL) for the period 2015-2020., Methods: Anonymised clinical and laboratory data were extracted from RIPL's Laboratory Information Management System and cleaned prior to descriptive analysis of the data. Travel history data were mapped to one of eight world regions, whereas symptom data were categorised into presenting syndromes. Diagnostic data were categorised as either positive, equivocal or negative., Results: During the period 2015-2020, RIPL received 73 951 samples from 53 432 patients suspected of having infections that are rare in the UK. The most common age group for unwell returning travellers was 30-39 years and the most commonly reported travel destination was Southern and SE Asia. Dengue virus was the most diagnosed infection overall, followed by chikungunya, Zika, leptospirosis and spotted fever group Rickettsia. Dengue virus was among the top three most frequent diagnoses for all world regions except Europe and represented 62.5% of all confirmed/probable diagnoses., Conclusions: None of the top five infections diagnosed by RIPL in travellers are vaccine-preventable, therefore understanding traveller demographics, destination-specific risk factors and encouraging preventative behaviours is the best available strategy to reduce the number of returning travellers who become infected. Prompt referral of acute samples with a detailed travel history, including purpose of travel and activities undertaken as well as dates and destinations can be a valuable tool in designing public health interventions and diagnostic algorithms., (© International Society of Travel Medicine 2023. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)

Clinical phonetic transcription is regarded as a highly specialised skill requiring hours of practice for mastery. Although this skill is a critical part of students' clinical preparation to become speech-language pathologists, students often report feeling unprepared to apply the skill in clinical practice. Previous studies suggest that increased opportunities for practice and timely feedback on transcriptions are needed in order to develop skill confidence. However, providing more opportunities for practice can be impeded by the limited resources to manage the grading of additional assignments. The purpose of this study is to show the implementation of a web-based learning management system (LMS) designed in our labs for phonetics instruction. The Automated Phonetic Transcription Grading Tool (APTgt LMS) was developed to provide a platform for assignment delivery and automated grading of transcription assignments. The APTgt LMS has three embedded IPA keyboards (basic, advanced, and full IPA) and an automated edit distance algorithm modified by phonetic alignment principles, which allows for individualised scoring and visual course-level feedback in an interactive online environment. For pilot testing, student confidence was queried before and after practice opportunities using APTgt. A concurrent mixed methods research design was used to analyse four Likert scale and three open-ended questions. Student confidence in transcribing disordered speech was found to significantly increase (p <0.001) following additional practice. Students reported concerns related to accurate transcription of disordered speech and that additional practice is still needed. Tools like APTgt can aid in facilitating student learning and increasing student confidence in applied transcription.

Arthrogryposis multiplex congenita (AMC) [also known as multiple joints contracture or Fetal Akinesia Deformation Sequence (FADS)] is etiologically a heterogeneous condition with an estimated incidence of approximately 1 in 3000 live births and much higher incidence when prenatally diagnosed cases are included. The condition can be acquired or secondary to fetal exposures and can also be caused by a variety of single-gene disorders affecting the brain, spinal cord, peripheral nerves, neuromuscular junction, muscle, and a variety of disorders affecting the connective tissues (Niles et al., Prenatal Diagnosis, 2019; 39:720-731). The introduction of next-generation gene sequencing uncovered many genes and causative variants of AMC but also identified genes that cause both dominant and recessive inherited conditions with the variability of clinical manifestations depending on the genes and variants. Molecular diagnosis in these cases is not only important for prognostication but also for the determination of recurrence risk and for providing reproductive options including preimplantation and prenatal diagnosis. TTN, the largest known gene in the human genome, has been known to be associated with autosomal dominant dilated cardiomyopathy. However, homozygote and compound heterozygote pathogenic variants with recessive inheritance have rarely been reported. We report the effect of recessive variants located within the fetal IC and/or N2BA isoforms in association with severe FADS in three families. All parents were healthy obligate carriers and none of them had cardiac or skeletal muscle abnormalities. This report solidifies FADS as an alternative phenotypic presentation associated with homozygote/compound heterozygous pathogenic variants in the TTN., (© 2022 Wiley Periodicals LLC.)

Many computerized tools for comparing phonetic transcriptions have been proposed and shared in the past; however, previous tools are relatively difficult to access and incorporate into clinical and research practice, or require users to learn additional phonetic symbol systems. The purpose of this project was to develop and test a readily available web-based application for quantitatively comparing phonetic transcriptions that are input using International Phonetic Alphabet (IPA) symbols. A web-based computer application was developed to allow for IPA phonetic transcription comparison. A point-and-click keyboard was developed to provide support for character input of the full IPA, as well as most symbols in the extIPA set. The application compares phonetic transcriptions using a modified edit distance algorithm following phonologically informed alignment. Visualizations of the algorithm's optimal alignment and scoring operations are shown for each comparison input by a user. The application, named the Automated Phonetic Transcription Comparison Tool (APTct), was thoroughly tested for accurate implementation of the algorithm principles. Validity tests through two sample use cases were also performed by comparison of hand calculations to APTct calculations. In function testing, the APTct showed excellent agreement with an expert's hand scoring using the same algorithm principles. In a validity test, only minor differences between the APTct and hand calculations were observed, primarily due to errors inputting the transcriptions into the APTct. The web-based APTct is a validated and versatile tool for quantitatively comparing even complex IPA phonetic transcriptions. It is freely available to clinicians and researchers, who may find it useful for a variety of potential scenarios. We invite researchers, clinicians, and other individuals to use the tool in their clinical and research work. The permalink for the website is as follows: https://aptct.auburn.edu.

Portland cement based grouts used for radioactive waste immobilization contain high replacement levels of supplementary cementitious materials, including blast-furnace slag and fly ash. The minerals formed upon hydration of these cements may have capacity for binding actinide elements present in radioactive waste. In this work, the minerals ettringite (Ca 6 Al 2 (SO 4 ) 3 (OH) 12 ·26H 2 O) and hydrotalcite (Mg 6 Al 2 (OH) 16 CO 3 ·4H 2 O) were selected to investigate the importance of minor cement hydrate phases in sequestering and immobilizing U VI from radioactive waste streams. U L III -edge X-ray absorption spectroscopy (XAS) was used to probe the U VI coordination environment in contact with these minerals. For the first time, solid-state 27 Al magic angle spinning nuclear magnetic resonance (MAS-NMR) spectroscopy was applied to probe the Al coordination environment in these U VI -contacted minerals and make inferences on the U VI coordination, in conjunction with the X-ray spectroscopy analyses. The U L III -edge XAS analysis of the U VI -contacted ettringite phases found them to be similar (>∼70%) to the uranyl oxyhydroxides present in a mixed becquerelite/metaschoepite mineral. Fitting of the EXAFS region, in combination with 27 Al NMR analysis, indicated that a disordered Ca- or Al-bearing U VI secondary phase also formed. For the U VI -contacted hydrotalcite phases, the XAS and 27 Al NMR data were interpreted as being similar to uranyl carbonate, that was likely Mg-containing., (open access.)

High-field asymmetric waveform ion mobility spectrometry (FAIMS) has gained popularity in the proteomics field for its capability to improve mass spectrometry sensitivity and to decrease peptide co-fragmentation. The recent implementation of FAIMS on Tribrid Orbitrap instruments enhanced proteome coverage and increased the precision of quantitative measurements. However, the FAIMS interface has not been available on older generation Orbitrap mass spectrometers such as the Q-Exactive. Here, we report the integration of the FAIMS Pro device with embedded electrical and gas connections to a Q-Exactive HF mass spectrometer. Proteomic experiments performed on HeLa tryptic digests with the modified mass spectrometer improved signal to noise and reduced interfering ions, resulting in an increase of 42% in peptide identification. FAIMS was also combined with segmented ion fractionation where 100 m / z windows were obtained in turn to further increase the depth of proteome analysis by reducing the proportion of chimeric MS/MS spectra from 50 to 27%. We also demonstrate the application of FAIMS to improve quantitative measurements when using isobaric peptide labeling. FAIMS experiments performed on a two-proteome model revealed that FAIMS Pro provided a 65% improvement in quantification accuracy compared to conventional LC-MS/MS experiments.

A digital colour image may be composed of hundreds of thousands of pixels, every pixel exhibiting a single colour. Each colour can be described as a combination of red, green and blue (RGB) components, of discrete values between 0-255. The RGB data contained within the pixels of an image could, therefore, be used to quantitatively establish the colour of nuclear material powders from digital images, particularly for use in nuclear forensics applications, where there is a need for consistent, objective analysis. This paper sets out a standard method for the photography and analysis of digital images of uranium oxide powder, for the objective quantification of colour by mean RGB values. Eight heat treated (up to 550°C) powder samples of studtite ([(UO 2 )(O 2 )(H 2 O) 2 ]·2H 2 O) were photographed at room temperature and analysed by the RGB method. Hue, saturation and value of the coloured samples were obtained alongside mean RGB values, both of which were used to successfully determine the heating temperatures of unknown specimens of studtite., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Crown Copyright © 2020. Published by Elsevier B.V. All rights reserved.)

Materials from GeoMelt® In-Container Vitrification (ICV)™ of simulant UK nuclear wastes were characterised to understand the partitioning of elements, including inactive surrogates for radionuclide species of interest, within the heterogeneous products. Aqueous durability analysis was performed to assess the potential disposability of the resulting wasteforms. The vitrification trial aimed to immobilise a variety of simulant legacy waste streams representative of decommissioning operations in the UK, including plutonium contaminated material, Magnox sludges and ion-exchange materials, which were vitrified upon the addition of glass forming additives. Two trials with different wastes were characterised, with the resultant vitreous wasteforms comprising olivine and pyroxene crystalline minerals within glassy matrices. Plutonium surrogate elements were immobilised within the glassy fraction rather than partitioning into crystalline phases. All vitrified products exhibited comparable or improved durability to existing UK high level waste vitrified nuclear wasteforms over a 28 day period., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

Technetium-99 immobilization in low-temperature nuclear waste forms often relies on additives that reduce environmentally mobile pertechnetate (TcO 4 - ) to insoluble Tc(IV) species. However, this is a short-lived solution unless reducing conditions are maintained over the hazardous life cycle of radioactive wastes (some ∼10,000 years). Considering recent experimental observations, this work explores how rapid formation of ettringite [Ca 6 Al 2 (SO 4 ) 3 (OH) 12 ·26(H 2 O)], a common mineral formed in cementitious waste forms, may be used to directly immobilize TcO 4 - . Results from ab initio molecular dynamics (AIMD) simulations and solid-phase characterization techniques, including synchrotron X-ray absorption, fluorescence, and diffraction methods, support successful incorporation of TcO 4 - into the ettringite crystal structure via sulfate substitution when synthesized by aqueous precipitation methods. One sulfate and one water are replaced with one TcO 4 - and one OH - during substitution, where Ca 2+ -coordinated water near the substitution site is deprotonated to form OH - for charge compensation upon TcO 4 - substitution. Furthermore, AIMD calculations support favorable TcO 4 - substitution at the SO 4 2- site in ettringite rather than gypsum (CaSO 4 ·2H 2 O, formed as a secondary mineral phase) by at least 0.76 eV at 298 K. These results are the first of their kind to suggest that ettringite may contribute to TcO 4 - immobilization and the overall lifetime performance of cementitious waste forms.

One challenge faced by teachers of phonetics, speech science, and clinical speech disorders courses is providing meaningful instruction that closes the theory to practice gap. One barrier to providing this type of deep learning experience is the lack of publicly available examples of speech recordings that illustrate comparisons between typical and disordered speech production across a broad range of disorder populations. Data of this type exist, but are typically collected for specific research projects under narrowly written IRB protocols that do not allow for release of even de-identified speech recordings to other investigators or teachers. As a partial corrective to this problem, we have developed an approved publicly available database of speech recordings that provides illustrative examples of adult and child speech production from individuals with and without speech disorders. The recorded speech materials were designed to illustrate important clinical concepts, and the recordings were collected under controlled conditions using high-quality equipment. The ultimate goal of creating this corpus is to improve practitioners' and scientists' understanding of the scientific bases of knowledge in our profession and improve our ability to develop clinical scientists and young researchers in the field.

We introduce a graphene-based nanofluidic cell that facilitates in situ imaging of liquid samples via transmission electron microscopy. The cell combines the benefits of graphene liquid cells-namely, high resolution, reduced charging effects, and excellent sample stability-with the ability to introduce reactants and control fluid concentrations as provided by conventional silicon-nitride-windowed flow cells. The graphene flow cell offers significantly less window bowing compared to existing commercial holders. We demonstrate the performance of the flow cell by imaging gold nanoparticle dynamics and uranyl acetate crystallization. Our results confirm the utility of graphene flow cells in obtaining the high spatial and temporal resolution required for probing the complex dynamics of nanoparticles and nucleation pathways in aqueous solutions.

The synthesis of a palladium-containing iodovanadinite derivative, hypothetically "PdPb 9 (VO 4 ) 6 I 2 ", was attempted using PdI 2 as a source of iodine in searching for a novel waste form for radioiodine. Stoichiometric amounts of Pb 3 (VO 4 ) 2 and PdI 2 were batched and reacted at elevated temperatures in sealed vessels. Batched material was also subjected to high-energy ball-milling (HEBM) in order to reduce reaction time and the potential for iodine volatilization during subsequent reaction at 200-500 °C. The resulting products were characterized using X-ray diffraction, scanning electron microscopy, energy-dispersive X-ray analysis, IR spectroscopy, thermal analysis and Pd K XANES. Results showed that PdI 2 can function as a sacrificial iodine source for the formation of iodovanadinite, prototypically Pb 10 (VO 4 ) 6 I 2 , however, the incorporation of Pd into this phase was not definitively observed. The sacrificial reaction mechanism involved the decomposition of PdI 2 to Pd metal and nascent I 2 , with the latter incorporated into the iodovanadinite Pb 10 (VO 4 ) 6 I 2 phase. In comparison to processing using standard solid state reaction techniques, the use of HEBM prior to high temperature reaction generates a more homogeneous end-product with better iodine retention for this system. Overall, the key novelty and importance of this work is in demonstrating a method for direct immobilisation of undissolved PdI 2 from nuclear fuel reprocessing, in a composite wasteform in which I-129 is immobilised within a durable iodovandinite ceramic, encapsulating Pd metal., Competing Interests: There are no conflicts of interest to declare., (This journal is © The Royal Society of Chemistry.)

The rise of sample multiplexing in quantitative proteomics for the dissection of complex phenotypic comparisons has been advanced by the development of ever more sensitive and robust instrumentation. Here, we evaluated the utility of the Orbitrap Eclipse Tribrid mass spectrometer (advanced quadrupole filter, optimized FTMS scan overhead) and new instrument control software features (Precursor Fit filtering, TurboTMT and Real-time Peptide Search filtering). Multidimensional comparisons of these novel features increased total peptide identifications by 20% for SPS-MS 3 methods and 14% for HRMS 2 methods. Importantly Real-time Peptide Search filtering enabled a ∼2× throughput improvement for quantification. Across the board, these sensitivity increases were attained without sacrificing quantitative accuracy. New hardware and software features enable more efficient characterization in pursuit of comparative whole proteome insights.

Purpose Verbs with low concreteness are frequent in discourse samples but rarely targeted in aphasia treatments for verbs. These verbs are an important part of functional communication, and recent studies have called for more research regarding aphasia and treatment stimuli with low concreteness. The aim of this study was to pilot the use of verbs with low concreteness in a novel sentence production intervention with persons with aphasia. Method The study took the form of a single-case experimental design with multiple baselines across behaviors and across participants. Three persons with chronic nonfluent aphasia and apraxia of speech participated in the study. Each participant received treatment designed to increase the semantic networks of verbs with high frequency and low concreteness. Sentence production was closely examined over the course of treatment for treated and untreated verbs of varying concreteness levels. Additional measures of language and cognitive functioning were also taken before and after treatment. Results Results indicated improved sentence production with target verbs attributable to the treatment in the 1st phase of 2 phases for 2 of the 3 participants. The increases corresponded with the application of treatment, despite the difference in number of baseline sessions for the participants. Where there were treatment effects, there was also considerable generalization to untreated sets of items during the 1st treatment phase. Word retrieval also improved for 2 participants. Conclusions The results suggest that the novel treatment may improve sentence production and word retrieval in persons with aphasia, even when using target verbs with low concreteness ratings. Future research is warranted into the use of low concreteness verbs. Supplemental Material https://doi.org/10.23641/asha.10870958.

Background: Acquired apraxia of speech (AOS) involves speech-production deficits on both the segmental and suprasegmental levels. Recent research has identified a non-linear interaction between the metrical structure of bisyllabic words and word-production accuracy in German speakers with AOS, with trochaic words (strong-weak stress) being resistant to errors compared with iambic words (weak-strong)., Aims: To replicate previous findings in English speakers with AOS, to measure the test-retest reliability of the effect, and to examine the potential impact of different methods of word scoring., Methods & Procedures: Speech samples were collected from 27 speakers with AOS and aphasia. Participants were at least 12 months post-stroke or penetrating brain injury, and represented a large range of AOS and aphasia severities. Productions were elicited via verbal model. Sampling was conducted on three separate occasions: the initial data-collection session and then repeated samplings at 1- and 4-week intervals. Bisyllabic words with a CVCVC segmental structure were selected. The list was divided into sublists representing differing lexical stress patterns: A list of 42 trochees, and one of 37 iambs. All speech samples were phonetically transcribed and then aligned with canonical transcriptions via an edit distance algorithm that followed transcription alignment principles. Phonetic-level errors (distortions) were penalized less severely than phonemic-level errors. Per cent consonants correct and whole-word accuracy were also examined. Trochee and iamb lists were analysed separately., Outcomes & Results: Paired samples t-tests indicated that the modified edit distance was significantly lower for the trochee lists than for the iamb lists. There was a lack of a significant effect of time on the absolute difference between modified edit distance for both lists. Intraclass coefficients suggested the list and procedures used are appropriate as an outcome measure for group research., Conclusions & Implications: The results suggest that in English, as in German, the trochaic structure is more resistant to segmental errors in persons with AOS and aphasia, providing replication of the findings of Aichert et al. in 2016. Further, this effect is stable over repeated sampling occasions. Implications for clinical management of AOS include possible ways to scaffold item difficulty and potentially improve stimulus generalization., (Published 2019. This article is a U.S. Government work and is in the public domain in the USA.)

Purpose This investigation was designed to provide interrater reliability data for the Apraxia of Speech Rating Scale 3.0 (ASRS 3.0; Strand, Duffy, Clark, & Josephs, 2014 ). Importantly, ratings were completed by investigators who were not involved with the ASRS development. Another aim was to evaluate the relationship of the ASRS 3.0 total score to word intelligibility. Method Two investigators independently completed ASRS 3.0 ratings for 28 participants with chronic apraxia of speech and aphasia. Intelligibility scores were obtained for all participants. Results Consistency of ratings for each feature and total score of the ASRS 3.0 was measured using intraclass correlation coefficients. Twelve of 13 intraclass correlation coefficients for feature ratings reached significance; clinical meaningfulness ranged from poor to excellent. Interrater reliability for the total scores was excellent. Similarly, absolute difference of ratings was minimal for the total scores, but varied across the 13 feature ratings. Correlations between the intelligibility scores and ASRS 3.0 total score were moderate to strong. Conclusion The total ASRS 3.0 score may be viewed as a reliable indicator of prevalence of apraxia of speech features. Although there was good to acceptable correspondence in ratings of the majority of the individual features, additional operationalization of rating procedures may be needed to improve interrater reliability for a few features.

The speciation of Ti in trinitite, the explosive melt glass derived from the Trinity Test of 16 th of July 1945, was investigated by X-ray absorption spectroscopy (XAS). Ti K-edge XANES showed that Ti was present in the Ti(iv) oxidation state for all samples. Fitting of pre-edge features by Gaussian functions and comparison with standards of known Ti coordination revealed significant variation in Ti coordination environment between samples. The variation of Ti coordination may be attributed to several factors including specific local chemistry and thermal histories of samples, in keeping with the highly heterogeneous microstructure of trinitite and the arkosic sand source material., Competing Interests: There are no conflicts of interest to declare., (This journal is © The Royal Society of Chemistry.)

Multiplexed, isobaric tagging methods are powerful techniques to increase throughput, precision, and accuracy in quantitative proteomics. The dynamic range and accuracy of quantitation, however, can be limited by coisolation of tag-containing peptides that release reporter ions and conflate quantitative measurements across precursors. Methods to alleviate these effects often lead to the loss of protein and peptide identifications through online or offline filtering of interference containing spectra. To alleviate this effect, high-Field Asymmetric-waveform Ion Mobility Spectroscopy (FAIMS) has been proposed as a method to reduce precursor coisolation and improve the accuracy and dynamic range of multiplex quantitation. Here we tested the use of FAIMS to improve quantitative accuracy using previously established TMT-based interference standards (triple-knockout [TKO] and Human-Yeast Proteomics Resource [HYPER]). We observed that FAIMS robustly improved the quantitative accuracy of both high-resolution MS 2 (HRMS 2 ) and synchronous precursor selection MS 3 (SPS-MS 3 )-based methods without sacrificing protein identifications. We further optimized and characterized the main factors that enable robust use of FAIMS for multiplexed quantitation. We highlight these factors and provide method recommendations to take advantage of FAIMS technology to improve isobaric-tag-quantification moving forward.

The depth of proteomic analyses is often limited by the overwhelming proportion of confounding background ions that compromise the identification and quantification of low abundance peptides. To alleviate these limitations, we present a new high field asymmetric waveform ion mobility spectrometry (FAIMS) interface that can be coupled to the Orbitrap Tribrid mass spectrometers. The interface provides several advantages over previous generations of FAIMS devices, including ease of operation, robustness, and high ion transmission. Replicate LC-FAIMS-MS/MS analyses ( n = 100) of HEK293 protein digests showed stable ion current over extended time periods with uniform peptide identification on more than 10,000 distinct peptides. For complex tryptic digest analyses, the coupling of FAIMS to LC-MS/MS enabled a 30% gain in unique peptide identification compared with non-FAIMS experiments. Improvement in sensitivity facilitated the identification of low abundance peptides, and extended the limit of detection by almost an order of magnitude. The reduction in chimeric MS/MS spectra using FAIMS also improved the precision and the number of quantifiable peptides when using isobaric labeling with tandem mass tag (TMT) 10-plex reagent. We compared quantitative proteomic measurements for LC-MS/MS analyses performed using synchronous precursor selection (SPS) and LC-FAIMS-MS/MS to profile the temporal changes in protein abundance of HEK293 cells following heat shock for periods up to 9 h. FAIMS provided 2.5-fold increase in the number of quantifiable peptides compared with non-FAIMS experiments (30,848 peptides from 2,646 proteins for FAIMS versus 12,400 peptides from 1,229 proteins with SPS). Altogether, the enhancement in ion transmission and duty cycle of the new FAIMS interface extended the depth and comprehensiveness of proteomic analyses and improved the precision of quantitative measurements., (© 2018 Pfammatter et al.)

Liquid chromatography (LC) prefractionation is often implemented to increase proteomic coverage; however, while effective, this approach is laborious, requires considerable sample amount, and can be cumbersome. We describe how interfacing a recently described high-field asymmetric waveform ion mobility spectrometry (FAIMS) device between a nanoelectrospray ionization (nanoESI) emitter and an Orbitrap hybrid mass spectrometer (MS) enables the collection of single-shot proteomic data with comparable depth to that of conventional two-dimensional LC approaches. This next generation FAIMS device incorporates improved ion sampling at the ESI-FAIMS interface, increased electric field strength, and a helium-free ion transport gas. With fast internal compensation voltage (CV) stepping (25 ms/transition), multiple unique gas-phase fractions may be analyzed simultaneously over the course of an MS analysis. We have comprehensively demonstrated how this device performs for bottom-up proteomics experiments as well as characterized the effects of peptide charge state, mass loading, analysis time, and additional variables. We also offer recommendations for the number of CVs and which CVs to use for different lengths of experiments. Internal CV stepping experiments increase protein identifications from a single-shot experiment to >8000, from over 100 000 peptide identifications in as little as 5 h. In single-shot 4 h label-free quantitation (LFQ) experiments of a human cell line, we quantified 7818 proteins with FAIMS using intra-analysis CV switching compared to 6809 without FAIMS. Single-shot FAIMS results also compare favorably with LC fractionation experiments. A 6 h single-shot FAIMS experiment generates 8007 protein identifications, while four fractions analyzed for 1.5 h each produce 7776 protein identifications.

A suite of uranium brannerites for the disposal of MOX residues, formulated (U 0.9 Ce 0.1 ) 1- x M x Ti 2 O 6 (M = Ca 2+ and/or Gd 3+ ), were prepared using a mixed oxide route under oxidising, inert and reducing atmospheres (air, argon and H 2 /N 2 ). Gd 3+ was added to act as a neutron absorber in the final Pu bearing wasteform and Ce added to function as a structural analogue for Pu. X-ray powder diffraction of the synthesised specimens found that phase distribution was strongly affected by the processing atmosphere and Gd content. In all cases prototypical brannerite was formed, accompanied by different secondary phases dependent on processing atmosphere. Microstructural analysis (SEM) of the sintered samples confirmed the results of the X-ray powder diffraction. Bulk XANES found that Ti remained in the Ti 4+ oxidation state whereas Ce was uniformly reduced to the Ce 3+ oxidation state regardless of processing conditions or stoichiometry. Micro-focus XANES was used to determine U oxidation in the brannerite phase and showed that U oxidised to higher U oxidation states to charge compensate. It was concluded that the charge balance mechanism was a combination of U oxidation and A-site vacancies., Competing Interests: There are no conflicts of interest to declare., (This journal is © The Royal Society of Chemistry.)

Background: This study investigates maternal and perinatal outcomes for women with low-risk pregnancies laboring in free-standing birth centers compared with laboring in a hospital maternity unit in a large New Zealand health district., Methods: The study used observational data from 47 381 births to women with low-risk pregnancies in South Auckland maternity facilities 2003-2010. Adjusted odds ratios with 95% confidence intervals were calculated for instrumental delivery, cesarean section, blood transfusion, neonatal unit admission, and perinatal mortality., Results: Labor in birth centers was associated with significantly lower rates of instrumental delivery, cesarean section and blood transfusion compared with labor in hospital. Neonatal unit admission rates were lower for infants of nulliparous women laboring in birth centers. Intrapartum and neonatal mortality rates for birth centers were low and were not significantly different from the hospital population. Transfers to hospital for labor and postnatal complications occurred in 39% of nulliparous and 9% of multiparous labors. Risk factors identified for transfer were nulliparity, advanced maternal age, and prolonged pregnancy ≥41 weeks' gestation., Conclusions: Labor in South Auckland free-standing birth centers was associated with significantly lower maternal intervention and complication rates than labor in the hospital maternity unit and was not associated with increased perinatal morbidity., (© 2017 Wiley Periodicals, Inc.)

Purpose: The purpose of this article is to quantify and describe stuttering-like disfluencies in speakers with acquired apraxia of speech (AOS), utilizing the Lidcombe Behavioural Data Language (LBDL). Additional purposes include measuring test-retest reliability and examining the effect of speech sample type on disfluency rates., Method: Two types of speech samples were elicited from 20 persons with AOS and aphasia: repetition of mono- and multisyllabic words from a protocol for assessing AOS (Duffy, 2013), and connected speech tasks (Nicholas & Brookshire, 1993). Sampling was repeated at 1 and 4 weeks following initial sampling. Stuttering-like disfluencies were coded using the LBDL, which is a taxonomy that focuses on motoric aspects of stuttering., Results: Disfluency rates ranged from 0% to 13.1% for the connected speech task and from 0% to 17% for the word repetition task. There was no significant effect of speech sampling time on disfluency rate in the connected speech task, but there was a significant effect of time for the word repetition task. There was no significant effect of speech sample type., Conclusions: Speakers demonstrated both major types of stuttering-like disfluencies as categorized by the LBDL (fixed postures and repeated movements). Connected speech samples yielded more reliable tallies over repeated measurements. Suggestions are made for modifying the LBDL for use in AOS in order to further add to systematic descriptions of motoric disfluencies in this disorder.

Purpose: The purpose of this investigation was to compare the effects of schedule of practice (i.e., blocked vs. random) on outcomes of Sound Production Treatment (SPT; Wambaugh, Kalinyak-Fliszar, West, & Doyle, 1998) for speakers with chronic acquired apraxia of speech and aphasia., Method: A combination of group and single-case experimental designs was used. Twenty participants each received SPT administered with randomized stimuli presentation (SPT-R) and SPT applied with blocked stimuli presentation (SPT-B). Treatment effects were examined with respect to accuracy of articulation as measured in treated and untreated experimental words produced during probes., Results: All participants demonstrated improved articulation of treated items with both practice schedules. Effect sizes were calculated to estimate magnitude of change for treated and untreated items by treatment condition. No significant differences were found for SPT-R and SPT-B relative to effect size. Percent change over the highest baseline performance was also calculated to provide a clinically relevant indication of improvement. Change scores associated with SPT-R were significantly higher than those for SPT-B for treated items but not untreated items., Conclusion: SPT can result in improved articulation regardless of schedule of practice. However, SPT-R may result in greater gains for treated items., Supplemental Materials: https://doi.org/10.23641/asha.5116831.

Purpose: Awareness of errors has been considered a clinical feature of acquired apraxia of speech (AOS). However, there is limited research examining error awareness in speakers with AOS. The purpose of this investigation was to examine awareness of errors and explore the relationship between awareness of errors and treatment outcomes in speakers with AOS., Method: Twenty speakers with AOS and aphasia produced mono- and multisyllabic words in a repetition task. Following each production, speakers were asked to judge the accuracy of their production (i.e., correct or incorrect). Then, speakers received Sound Production Treatment., Results: Judgment accuracy of productions for the group ranged from 20% to 96%. There was a weak relationship between judgment accuracy and probe performance at posttreatment (r = .47) and a moderate relationship between judgment accuracy and probe performance at follow-up (r = .53)., Conclusion: Findings indicate that speakers with AOS varied in their ability to judge the accuracy of their productions. For some speakers, the ability to judge the accuracy of their productions did not coincide with their production accuracy of treatment stimuli at posttreatment and at follow-up. Further research is needed to explore the relationship between error awareness and treatment outcomes.

The estrogen-synthesizing enzyme aromatase is abundant at the synapse in the zebra finch hippocampus (HP), and its inhibition impairs spatial memory function. To more fully test the role of local estradiol (E2) synthesis in memory, the HP of adult male zebra finches was exposed to either control pellets or those containing the aromatase inhibitor 1,4,6-androstatriene-3,17-dione (ATD), ATD and E2, ATD and the G protein-coupled estrogen receptor (GPER) agonist G1, or the antagonist G15 alone. Birds were tested for spatial memory acquisition and performance, and HP levels of the postsynaptic protein PSD95 were measured. ATD-treated birds took longer to reach criterion than control birds, whereas acquisition in ATD+E2 and ATD+G1 birds was indistinguishable from control and ATD treatments. Interestingly, all G15 birds failed to acquire the task. Following a retention interval, ATD birds took the longest to reach the (formerly) baited cup and made the most mistakes. ATD+E2 animals displayed the lowest retention latencies and made fewer mistakes than ATD-treated birds, and ATD+G1 birds did not significantly differ from controls in retention latencies. The amount of PSD95 in the HP was lowest in ATD-treated animals compared with birds with silicone-only-implanted craniotomies, ATD+E2, and ATD+G1 birds, who did not differ in this expression. Thus, spatial memory acquisition and performance appear aromatase and E2 dependent, an effect more reliably revealed after consolidation and/or recall compared to acquisition. E2 may exert this effect via GPERs, resulting in an increase in PSD95 levels that may modify receptor activity or intracellular signaling pathways to increase synaptic strength., (Copyright © 2017 Endocrine Society.)

We describe ProteomeTools, a project building molecular and digital tools from the human proteome to facilitate biomedical research. Here we report the generation and multimodal liquid chromatography-tandem mass spectrometry analysis of >330,000 synthetic tryptic peptides representing essentially all canonical human gene products, and we exemplify the utility of these data in several applications. The resource (available at http://www.proteometools.org) will be extended to >1 million peptides, and all data will be shared with the community via ProteomicsDB and ProteomeXchange.

Purpose: The ability to recognize one's own speech errors has long been considered a clinical feature of acquired apraxia of speech (AOS) despite limited empirical data supporting this notion. This study was designed to (a) investigate the ability of speakers with AOS to self-judge the accuracy of their own word productions and (b) examine the test-retest stability of a measure to quantify the self-judgments of speakers with AOS., Method: Twenty-four speakers with AOS and aphasia repeated mono- and multisyllabic words. After each word, they indicated whether their production was correct or incorrect. This procedure was repeated 1 week later to examine performance stability., Results: Percentage of incorrect word productions was stable for the group across times. Accuracy of judgments ranged from 64% to 100% at Time 1 and from 56% to 100% at Time 2. Inaccurate judgments of error productions (false positives) occurred much more frequently than inaccurate judgments of correct productions (false negatives)., Conclusions: Error production was remarkably stable in our participants. As a group, the participants failed to detect almost one third of words produced erroneously. However, accuracy and stability of judgments over sampling times varied across participants. Findings suggest that error awareness might be a worthwhile target for treatment in some individuals with AOS.

Cardiac lipomas are rare and usually present as benign, encapsulated masses outside the heart; however, they can also be found within the atria. No single theory-including molecular genetic mutation-adequately explains why this occurs. Extensive career experience and broadened knowledge in embryology and cardiac physiology have helped us to develop a hypothesis based on invagination of extracardiac tumors. This report describes a vexing case of a giant right atrial lipoma, from 1985, in which the diagnosis was made incidentally during management of a patient's acute limb ischemia. In addition, we discuss the imaging and treatment of cardiac lipoma.

Acute kidney injury (AKI) is common among emergency department patients admitted to hospital. There is evidence of inadequate management of the condition leading to adverse outcomes. We present an illustrative case of AKI complicating a gastrointestinal disorder in an older adult. We discuss the clinical presentation, assessment and management of AKI with reference to recent consensus guidelines on classification and treatment., (Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/)

The release of radionuclides from spent fuel in a geological disposal facility is controlled by the surface mediated dissolution of UO2 in groundwater. In this study we investigate the influence of reactive surface sites on the dissolution of a synthesized CeO2 analogue for UO2 fuel. Dissolution was performed on the following: CeO2 annealed at high temperature, which eliminated intrinsic surface defects (point defects and dislocations); CeO2-x annealed in inert and reducing atmospheres to induce oxygen vacancy defects and on crushed CeO2 particles of different size fractions. BET surface area measurements were used as an indicator of reactive surface site concentration. Cerium stoichiometry, determined using X-ray Photoelectron Spectroscopy (XPS) and supported by X-ray Diffraction (XRD) analysis, was used to determine oxygen vacancy concentration. Upon dissolution in nitric acid medium at 90 °C, a quantifiable relationship was established between the concentration of high energy surface sites and CeO2 dissolution rate; the greater the proportion of intrinsic defects and oxygen vacancies, the higher the dissolution rate. Dissolution of oxygen vacancy-containing CeO2-x gave rise to rates that were an order of magnitude greater than for CeO2 with fewer oxygen vacancies. While enhanced solubility of Ce(3+) influenced the dissolution, it was shown that replacement of vacancy sites by oxygen significantly affected the dissolution mechanism due to changes in the lattice volume and strain upon dissolution and concurrent grain boundary decohesion. These results highlight the significant influence of defect sites and grain boundaries on the dissolution kinetics of UO2 fuel analogues and reduce uncertainty in the long term performance of spent fuel in geological disposal.

We describe a new method to accomplish multiplexed, absolute protein quantification in a targeted fashion. The approach draws upon the recently developed neutron encoding (NeuCode) metabolic labeling strategy and parallel reaction monitoring (PRM). Since PRM scanning relies upon high-resolution tandem mass spectra for targeted protein quantification, incorporation of multiple NeuCode labeled peptides permits high levels of multiplexing that can be accessed from high-resolution tandem mass spectra. Here we demonstrate this approach in cultured cells by monitoring a viral infection and the corresponding viral protein production over many infection time points in a single experiment. In this context the NeuCode PRM combination affords up to 30 channels of quantitative information in a single MS experiment.

Purpose: The purpose of this study was to examine divided attention over a large age range by looking at the effects of 3 nonspeech tasks on concurrent speech motor performance. The nonspeech tasks were designed to facilitate measurement of bidirectional interference, allowing examination of their sensitivity to speech activity. A cross-sectional design was selected to explore possible changes in divided-attention effects associated with age., Method: Sixty healthy participants were separated into 3 groups of 20: younger (20s), middle-aged (40s), and older (60s) adults. Each participant completed a speech task (sentence repetitions) once in isolation and once concurrently with each of 3 nonspeech tasks: a semantic-decision linguistic task, a quantitative-comparison cognitive task, and a manual motor task. The nonspeech tasks were also performed in isolation., Results: Data from speech kinematics and nonspeech task performance indicated significant task-specific divided attention interference, with divided attention affecting speech and nonspeech measures in the linguistic and cognitive conditions and affecting speech measures in the manual motor condition. There was also a significant age effect for utterance duration., Conclusions: The results increase what is known about bidirectional interference between speech and other concurrent tasks as well as age effects on speech motor control.

Purpose: This investigation was designed to provide a quantification and synthesis of a series of single-case experimental design investigations into the effects of sound production treatment, an articulatory-kinematic treatment for acquired apraxia of speech (AOS). The main purpose was to perform a meta-analysis of aggregated sound production treatment data in order to provide benchmarks to serve as indicators of magnitude of change. Additional analyses explored various factors influencing effect sizes and level of performance., Method: Effect sizes were calculated for treated and untreated items for 24 participants across 10 investigations. Benchmarks were calculated as the quartiles of the distributions of the effect sizes. Correlational analyses were performed to examine (a) end-of-treatment performance relative to follow-up performance, (b) response of trained items relative to untrained items, and (c) effect size relative to participant variables., Results: Effect sizes were predominantly large and positive; benchmarks for treated items were larger than those for untreated items. End of treatment and follow-up performance were positively correlated, and response generalization was positively correlated with AOS severity., Conclusion: These benchmarks may assist in evaluating the effects of interventions for individuals with AOS utilizing similar outcome measures in both clinical and research settings.

The field of proteomics almost uniformly relies on peptide cation analysis, leading to an underrepresentation of acidic portions of proteomes, including relevant acidic posttranslational modifications. Despite the many benefits negative mode proteomics can offer, peptide anion analysis remains in its infancy due mainly to challenges with high-pH reversed-phase separations and a lack of robust fragmentation methods suitable for peptide anion characterization. Here, we report the first implementation of activated ion negative electron transfer dissociation (AI-NETD) on the chromatographic timescale, generating 7,601 unique peptide identifications from Saccharomyces cerevisiae in single-shot nLC-MS/MS analyses of tryptic peptides-a greater than 5-fold increase over previous results with NETD alone. These improvements translate to identification of 1,106 proteins, making this work the first negative mode study to identify more than 1,000 proteins in any system. We then compare the performance of AI-NETD for analysis of peptides generated by five proteases (trypsin, LysC, GluC, chymotrypsin, and AspN) for negative mode analyses, identifying as many as 5,356 peptides (1,045 proteins) with LysC and 4,213 peptides (857 proteins) with GluC in yeast-characterizing 1,359 proteins in total. Finally, we present the first deep-sequencing approach for negative mode proteomics, leveraging offline low-pH reversed-phase fractionation prior to online high-pH separations and peptide fragmentation with AI-NETD. With this platform, we identified 3,467 proteins in yeast with trypsin alone and characterized a total of 3,730 proteins using multiple proteases, or nearly 83% of the expressed yeast proteome. This work represents the most extensive negative mode proteomics study to date, establishing AI-NETD as a robust tool for large-scale peptide anion characterization and making the negative mode approach a more viable platform for future proteomic studies., (© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.)

Gas chromatography/mass spectrometry (GC/MS) has long been considered one of the premiere analytical tools for small molecule analysis. Recently, a number of GC/MS systems equipped with high-resolution mass analyzers have been introduced. These systems provide analysts with a new dimension of information, accurate mass measurement to the third or fourth decimal place; however, existing data processing tools do not capitalize on this information. Beyond that, GC/MS spectral reference libraries, which have been curated over the last several decades, contain almost exclusively unit resolution MS spectra making integration of accurate mass data dubious. Here we present an informatic approach, called high-resolution filtering (HRF), which bridges this gap. During HRF, high-resolution mass spectra are assigned putative identifications through traditional spectral matching at unit resolution. Once candidate identities have been assigned, all unique combinations of atoms from these candidate precursors are generated and matched to m/z peaks using narrow mass tolerances. The total amount of measured signal that is annotated is used as a metric of plausibility for the presumed identification. Here we demonstrate that the HRF approach is both feasible and highly specific toward correct identifications.

This article is part of a Special Issue "Estradiol and cognition". In addition to their well-studied and crucial effects on brain development and aging, an increasing number of investigations across vertebrate species indicate that estrogens like 17β-estradiol (E2) have pronounced and rapid effects on cognitive function. The incidence and regulation of the E2-synthesizing enzyme aromatase at the synapse in regions of the brain responsible for learning, memory, social communication and other complex cognitive processes suggest that local E2 production and action affect the acute and chronic activity of individual neurons and circuits. Songbirds in particular are excellent models for the study of this "synaptocrine" hormone provision given that aromatase is abundantly expressed in neuronal soma, dendrites, and at the synapse across many brain regions in both sexes. Additionally, songbirds readily acquire and recall memories in laboratory settings, and their stereotyped behaviors may be manipulated and measured with relative ease. This leads to a rather unparalleled advantage in the use of these animals in studies of the role of neural aromatization in cognition. In this review we describe the results of a number of experiments in songbird species with a focus on the influence of synaptic E2 provision on two cognitive processes: auditory discrimination reliant on the caudomedial nidopallium (NCM), a telencephalic region likely homologous to the auditory cortex in mammals, and spatial memory dependent on the hippocampus. Data from these studies are providing evidence that the local and acute provision of E2 modulates the hormonal, electrical, and cognitive outputs of the vertebrate brain and aids in memory acquisition, retention, and perhaps the confluence of memory systems., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Recent advances in chromatography and mass spectrometry (MS) have made rapid and deep proteomic profiling possible. To maximize the performance of the recently produced Orbitrap hybrid mass spectrometer, we have developed a protocol that combines improved sample preparation (including optimized cellular lysis by extensive bead beating) and chromatographic conditions (specifically, 30-cm capillary columns packed with 1.7-μm bridged ethylene hybrid material) and the manufacture of a column heater (to accommodate flow rates of 350-375 nl/min) that increases the number of proteins identified across a single liquid chromatography-tandem MS (LC-MS/MS) separation, thereby reducing the need for extensive sample fractionation. This strategy allowed the identification of up to 4,002 proteins (at a 1% false discovery rate (FDR)) in yeast (Saccharomyces cerevisiae strain BY4741) over 70 min of LC-MS/MS analysis. Quintuplicate analysis of technical replicates reveals 83% overlap at the protein level, thus demonstrating the reproducibility of this procedure. This protocol, which includes cell lysis, overnight tryptic digestion, sample analysis and database searching, takes ∼24 h to complete. Aspects of this protocol, including chromatographic separation and instrument parameters, can be adapted for the optimal analysis of other organisms.

We describe a synthesis strategy for the preparation of lysine isotopologues that differ in mass by as little as 6 mDa. We demonstrate that incorporation of these molecules into the proteomes of actively growing cells does not affect cellular proliferation, and we discuss how to use the embedded mass signatures (neutron encoding (NeuCode)) for multiplexed proteome quantification by means of high-resolution mass spectrometry. NeuCode SILAC amalgamates the quantitative accuracy of SILAC with the multiplexing of isobaric tags and, in doing so, offers up new opportunities for biological investigation. We applied NeuCode SILAC to examine the relationship between transcript and protein levels in yeast cells responding to environmental stress. Finally, we monitored the time-resolved responses of five signaling mutants in a single 18-plex experiment., (© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.)

In the safety case for the geological disposal of nuclear waste, the release of radioactivity from the repository is controlled by the dissolution of the spent fuel in groundwater. There remain several uncertainties associated with understanding spent fuel dissolution, including the contribution of energetically reactive surface sites to the dissolution rate. In this study, we investigate how surface features influence the dissolution rate of synthetic CeO2 and ThO2, spent nuclear fuel analogues that approximate as closely as possible the microstructure characteristics of fuel-grade UO2 but are not sensitive to changes in oxidation state of the cation. The morphology of grain boundaries (natural features) and surface facets (specimen preparation-induced features) was investigated during dissolution. The effects of surface polishing on dissolution rate were also investigated. We show that preferential dissolution occurs at grain boundaries, resulting in grain boundary decohesion and enhanced dissolution rates. A strong crystallographic control was exerted, with high misorientation angle grain boundaries retreating more rapidly than those with low misorientation angles, which may be due to the accommodation of defects in the grain boundary structure. The data from these simplified analogue systems support the hypothesis that grain boundaries play a role in the so-called "instant release fraction" of spent fuel, and should be carefully considered, in conjunction with other chemical effects, in safety performance assessements for the geological disposal of spent fuel. Surface facets formed during the sample annealing process also exhibited a strong crystallographic control and were found to dissolve rapidly on initial contact with dissolution medium. Defects and strain induced during sample polishing caused an overestimation of the dissolution rate, by up to 3 orders of magnitude.

We have developed a multiplexed quantitative analysis method for carboxylic acids by liquid chromatography high resolution mass spectrometry. The method employs neutron encoded (NeuCode) methylamine labels ((13)C or (15)N enriched) that are affixed to carboxylic acid functional groups to enable duplex quantitation via mass defect measurement. This work presents the first application of NeuCode quantitation to small molecules. We have applied this technique to detect adulteration of olive oil by quantitative analysis of fatty acid methyl amide derivatives, and the quantitative accuracy of the NeuCode analysis was validated by GC/MS. Currently, the method enables duplex quantitation and is expandable to at least 6-plex analysis.

A mass spectrometry (MS) method is described here that can reproducibly identify hundreds of peptides across multiple experiments. The method uses intelligent data acquisition to precisely target peptides while simultaneously identifying thousands of other, nontargeted peptides in a single nano-LC-MS/MS experiment. We introduce an online peptide elution order alignment algorithm that targets peptides based on their relative elution order, eliminating the need for retention-time-based scheduling. We have applied this method to target 500 mouse peptides across six technical replicate nano-LC-MS/MS experiments and were able to identify 440 of these in all six, compared with only 256 peptides using data-dependent acquisition (DDA). A total of 3757 other peptides were also identified within the same experiment, illustrating that this hybrid method does not eliminate the novel discovery advantages of DDA. The method was also tested on a set of mice in biological quadruplicate and increased the number of identified target peptides in all four mice by over 80% (826 vs 459) compared with the standard DDA method. We envision real-time data analysis as a powerful tool to improve the quality and reproducibility of proteomic data sets.

The ability to acquire highly accurate quantitative data is an increasingly important part of any proteomics experiment, whether shotgun or top-down approaches are used. We recently developed a quantitation strategy for peptides based on neutron encoding, or NeuCode SILAC, which uses closely spaced heavy isotope-labeled amino acids and high-resolution mass spectrometry to provide quantitative data. We reasoned that the strategy would also be applicable to intact proteins and could enable robust, multiplexed quantitation for top-down experiments. We used yeast lysate labeled with either (13)C6(15)N2-lysine or (2)H8-lysine, isotopologues of lysine that are spaced 36 mDa apart. Proteins having such close spacing cannot be distinguished during a medium resolution scan, but upon acquiring a high-resolution scan, the two forms of the protein with each amino acid are resolved and the quantitative information revealed. An additional benefit NeuCode SILAC provides for top down is that the spacing of the isotope peaks indicates the number of lysines present in the protein, information that aids in identification. We used NeuCode SILAC to quantify several hundred isotope distributions, manually identify and quantify proteins from 1:1, 3:1, and 5:1 mixed ratios, and demonstrate MS(2)-based quantitation using ETD.