Validity and reliability of a shoe-embedded sensor module for measuring foot progression angle during over-ground walking

Wearable systems are becoming increasingly popular for gait assessment outside of laboratory settings. A single shoe-embedded sensor module can measure the foot progression angle (FPA) during walking. The FPA has important clinical utility, particularly in populations with knee osteoarthritis, as it is a target for biomechanical treatments. However, the validity and the day-to-day reliability of FPA measurement using wearable systems during over-ground walking has yet to be established. Two gait analysis sessions on 20 healthy adults were conducted. During both sessions, participants performed natural over-ground walking in a motion capture laboratory and on a 100 m linear section of outdoor athletics track. FPA was measured in the laboratory via marker trajectory data, while the sensor module measured FPA during the outdoor track walking. Validity was examined by comparing the laboratory- and sensor-measured average FPA. Day-to-day reliability was examined by comparing the sensor-measured FPA between the first and second gait analysis sessions. Average absolute error between motion capture and sensor measured FPA were 1.7° and 2.1° at session 1 and 2, respectively. A Bland and Altman plot indicated no systematic bias, with 95% limit of agreement widths of 4.2° – 5.1°. Intraclass correlation coefficient (ICC_2k) analysis resulted in good to excellent validity (ICC = 0.89 – 0.91) and reliability (ICC = 0.95). Overall, the shoe-embedded sensor module is a valid and reliable method of measuring FPA during over-ground walking without the need for laboratory equipment.     

Center of pressure and center of mass behavior during gait initiation on inclined surfaces: A statistical parametric mapping analysis

This study analyzed gait initiation (GI) on inclined surfaces with 68 young adult subjects of both sexes. Ground reaction forces and moments were collected using two AMTI force platforms, of which one was in a horizontal position and the other was inclined by 8% in relation to the horizontal plane. Departing from a standing position, each participant executed three trials in the following conditions: horizontal position (HOR), inclined position at ankle dorsi-flexion (UP), and inclined position at ankle plantar-flexion (DOWN). Statistical parametric mapping analysis was performed over the entire center of pressure (COP) and center of mass (COM) time series. COP excursion did not show significant differences in the medial-lateral (ML) direction in both inclined conditions, but it was greater in the anterior-posterior (AP) direction for both inclined conditions. COP velocities are smaller in discrete portions of GI for the UP and DOWN conditions. COM displacement was greater in the ML direction during anticipatory postural adjustments (APA) in the UP condition, and COM moves faster in the ML direction during APA in the UP condition but slower at the end of GI for both the UP and the DOWN conditions. The COP-COM vector showed a greater angle in the DOWN condition. We observed changes for COP and COM in GI in both the UP and the DOWN conditions, with the latter showing changes for a great extent of the task. Both the UP and the DOWN conditions showed increased COM displacement and velocity. The predominant characteristic during GI on inclined surfaces, including APA, appears to be the displacement of the COM.     

Kinematics of knees with osteoarthritis show reduced lateral femoral roll-back and maintain an adducted position. A systematic review of research using medical imaging

BackgroundWhile several studies describe kinematics of healthy and osteoarthritic knees using the accurate imaging and computer modelling now possible, no systematic review exists to synthesise these data.MethodA systematic review extracted quantitative observational, quasi-experimental and experimental studies from PubMed, Scopus, Medline and Web of Science that examined motion of the bony or articular surfaces of the tibiofemoral joint during any functional activity. Studies using surface markers, animals, and in vitro studies were excluded.Results352 studies were screened to include 23 studies. Dynamic kinematics were recorded for gait, step-up, kneeling, squat and lunge and quasi-static squat, knee flexion in side-lying or supine leg-press. Kinematics were described using a diverse range of measures including six degrees of freedom kinematics, contact patterns or the projection of the femoral condylar axis above the tibia. Meta-analysis of data was not possible since no three papers recorded the same activity with the same measures. Visual evaluation of data revealed that knees with osteoarthritis maintained a more adducted position and showed less posterior translation of the lateral femoral condylar axis than healthy knees. Variability in activities and in recording measures produced greater variation in kinematics, than did knee osteoarthritis.ConclusionDifferences in kinematics between osteoarthritic and healthy knees were observed, however, these differences were more subtle than expected. The synthesis and progress of this research could be facilitated by a consensus on reference systems for axes and kinematic reporting.     

The development and concurrent validity of a real-time algorithm for temporal gait analysis using inertial measurement units

The use of inertial measurement units (IMUs) for gait analysis has emerged as a tool for clinical applications. Shank gyroscope signals have been utilized to identify heel-strike and toe-off, which serve as the foundation for calculating temporal parameters of gait such as single and double limb support time. Recent publications have shown that toe-off occurs later than predicted by the dual minima method (DMM), which has been adopted as an IMU-based gait event detection algorithm. In this study, a real-time algorithm, Noise-Zero Crossing (NZC), was developed to accurately compute temporal gait parameters. Our objective was to determine the concurrent validity of temporal gait parameters derived from the NZC algorithm against parameters measured by an instrumented walkway. The accuracy and precision of temporal gait parameters derived using NZC were compared to those derived using the DMM. The results from Bland-Altman Analysis showed that the NZC algorithm had excellent agreement with the instrumented walkway for identifying the temporal gait parameters of Gait Cycle Time (GCT), Single Limb Support (SLS) time, and Double Limb Support (DLS) time. By utilizing the moment of zero shank angular velocity to identify toe-off, the NZC algorithm performed better than the DMM algorithm in measuring SLS and DLS times. Utilizing the NZC algorithm’s gait event detection preserves DLS time, which has significant clinical implications for pathologic gait assessment.     

Feasibility of using combined EMG and kinematic signals for prosthesis control: A simulation study using a virtual reality environment

Transhumeral amputation has a significant effect on a person’s independence and quality of life. Myoelectric prostheses have the potential to restore upper limb function, however their use is currently limited due to lack of intuitive and natural control of multiple degrees of freedom. The goal of this study was to evaluate a novel transhumeral prosthesis controller that uses a combination of kinematic and electromyographic (EMG) signals recorded from the person’s proximal humerus. Specifically, we trained a time-delayed artificial neural network to predict elbow flexion/extension and forearm pronation/supination from six proximal EMG signals, and humeral angular velocity and linear acceleration. We evaluated this scheme with ten able-bodied subjects offline, as well as in a target-reaching task presented in an immersive virtual reality environment. The offline training had a target of 4° for flexion/extension and 8° for pronation/supination, which it easily exceeded (2.7° and 5.5° respectively). During online testing, all subjects completed the target-reaching task with path efficiency of 78% and minimal overshoot (1.5%). Thus, combining kinematic and muscle activity signals from the proximal humerus can provide adequate prosthesis control, and testing in a virtual reality environment can provide meaningful data on controller performance.     

Workflow for analysis of high mass accuracy salivary data set using MaxQuant and ProteinPilot search algorithm

LTQ Orbitrap data analyzed with ProteinPilot can be further improved by MaxQuant raw data processing, which utilizes precursor-level high mass accuracy data for peak processing and MGF creation. In particular, ProteinPilot results from MaxQuant-processed peaklists for Orbitrap data sets resulted in improved spectral utilization due to an improved peaklist quality with higher precision and high precursor mass accuracy (HPMA). The output and postsearch analysis tools of both workflows were utilized for previously unexplored features of a three-dimensional fractionated and hexapeptide library (ProteoMiner) treated whole saliva data set comprising 200 fractions. ProteinPilot's ability to simultaneously predict multiple modifications showed an advantage from ProteoMiner treatment for modified peptide identification. We demonstrate that complementary approaches in the analysis pipeline provide comprehensive results for the whole saliva data set acquired on an LTQ Orbitrap. Overall our results establish a workflow for improved protein identification from high mass accuracy data.     

A method for simultaneous determination of solubility and transfer coefficient of oxygen in aqueous media using off-gas mass spectrometry

A static method was developed that simultaneously determined the solubility of oxygen and the oxygen-transfer coefficient in a stirred bioreactor. It was based on the static method developed by van Sonsbeek et al. to determine the ka in a liquid-impelled loop reactor. Only physical properties of the liquid were used to determine both parameters using a mass spectrometer. Data about the solubility of oxygen in water are available from the literature. Therefore, the solubility of oxygen in water was used to compare our data with published data. Furthermore, the solubility of oxygen in trypticase soy broth was compared to literature data. No significant deviations between our data and literature data could be observed. Our static method and the commonly applied dynamic method to determine the oxygen-transfer coefficient yielded similar results. The effect of temperature on the oxygen-transfer coefficient could be expressed as the activation energy needed for the transition of oxygen from the gas to the water phase. This was verified using the Arrhenius equation. (c) 1994 John Wiley & Sons, Inc.     

A rapid approach for phenotype-screening and database independent detection of cSNP/protein polymorphism using mass accuracy precursor alignment

The dynamics of a proteome can only be addressed with large-scale, high-throughput methods. To cope with the inherent complexity, techniques based on targeted quantification using proteotypic peptides are arising. This is an essential systems biology approach; however, for the exploratory discovery of unexpected markers, nontargeted detection of proteins, and protein modifications is indispensable. We present a rapid label-free shotgun proteomics approach that extracts relevant phenotype-specific peptide product ion spectra in an automated workflow without prior identification. These product ion spectra are subsequently sequenced with database search and de novo prediction algorithms. We analyzed six potato tuber cultivars grown on three plots of two geographically separated fields in Germany. For data mining about 1.5 million spectra from 107 analyses were aligned and statistically examined in approximately 1 day. Several cultivar-specific protein markers were detected. Based on de novo-sequencing a dominant protein polymorphism not detectable in the available EST-databases was assigned exclusively to a specific potato cultivar. The approach is applicable to organisms with unsequenced or incomplete genomes and to the automated extraction of relevant mass spectra that potentially cannot be identified by genome/EST-based search algorithms.     

A high-throughput assay for rapid and simultaneous analysis of perfect markers for important quality and agronomic traits in rice using multiplexed MALDI-TOF mass spectrometry

The application of single nucleotide polymorphisms (SNPs) in plant breeding involves the analysis of a large number of samples, and therefore requires rapid, inexpensive and highly automated multiplex methods to genotype the sequence variants. We have optimized a high-throughput multiplexed SNP assay for eight polymorphisms which explain two agronomic and three grain quality traits in rice. Gene fragments coding for the agronomic traits plant height (semi-dwarf, sd-1 ) and blast disease resistance ( Pi-ta ) and the quality traits amylose content ( waxy ), gelatinization temperature ( alk ) and fragrance ( fgr ) were amplified in a multiplex polymerase chain reaction. A single base extension reaction carried out at the polymorphism responsible for each of these phenotypes within these genes generated extension products which were quantified by a matrix-assisted laser desorption ionization-time of flight system. The assay detects both SNPs and indels and is co-dominant, simultaneously detecting both homozygous and heterozygous samples in a multiplex system. This assay analyses eight functional polymorphisms in one 5 µL reaction, demonstrating the high-throughput and cost-effective capability of this system. At this conservative level of multiplexing, 3072 assays can be performed in a single 384-well microtitre plate, allowing the rapid production of valuable information for selection in rice breeding.     

A proteomic method for the analysis of changes in protein concentrations in response to systemic perturbations using metabolic incorporation of stable isotopes and mass spectrometry

While several techniques exist for assessing quantitative differences among proteomes representing different cell states, methods for assessing how these differences are mediated are largely missing. We present a method that allows one to differentiate between cellular processes, such as protein synthesis, degradation and PTMs which affect protein concentrations. An induced systemic perturbation of a cell culture was coupled to a replacement of the growth medium to one highly enriched in the stable isotope 15N. The relative abundance of the 15N- and 14N-enriched forms of proteins, isolated from cell cultures harvested at time points following the onset of the perturbation, were determined by MS. Alterations in protein synthesis and degradation were quantified by comparing proteins isolated from perturbed and unperturbed cultures, respectively. The method was evaluated by subjecting HeLa cells to heat stress. As expected, a number of known heat shock proteins (Hsp) increased in concentration during heat stress. For Hsp27, increased de novo synthesis accounted for the concentration increase, while for Hsp70, decreased degradation accounted for the increase. A protein that was detected only after prolonged heat stress, vimentin, was not primarily synthesized de novo, but appeared rather as a result of PTM.     

A sensitive multidimensional method for the detection,characterization, and quantification of trace free drug species in antibody-drug conjugate samples using mass spectral detection

Conjugation processes and stability studies associated with the production and shelf life of antibody-drug conjugates (ADCs) can result in free (non-conjugated) drug species. These free drug species can increase the risk to patients and reduce the efficacy of the ADC. Despite stringent purification steps, trace levels of free drug species may be present in formulated ADCs, reducing the therapeutic window. The reduction of sample preparation steps through the incorporation of multidimensional techniques has afforded analysts more efficient methods to assess trace drug species. Multidimensional methods coupling size-exclusion and reversed phase liquid chromatography with ultra-violet detection (SEC-RPLC/UV) have been reported, but offer limited sensitivity and can limit method optimization. The current study addresses these challenges with a multidimensional method that is specific, sensitive, and enables method control in both dimensions via coupling of an on-line solid phase extraction column to RPLC with mass spectral detection (SPE-RPLC/MS). The proposed method was evaluated using an antibody-fluorophore conjugate (AFC) as an ADC surrogate to brentuximab vedotin and its associated parent maleimide-val-cit-DSEA payload and the derived N-acetylcysteine adduct formed during the conjugation process. Assay sensitivity was found to be 2 orders more sensitive using MS detection in comparison to UV-based detection with a nominal limit of quantitation of 0.30 ng/mL (1.5 pg on-column). Free-drug species were present in an unadulterated ADC surrogate sample at concentrations below 7 ng/mL, levels not detectable by UV alone. The proposed SPE-RPLC/MS method provides a high degree of specificity and sensitivity in the assessment of trace free drug species and offers improved control over each dimension, enabling straightforward integration into existing or novel workflows.     

A method for profiling classes of plant hormones and their metabolites using liquid chromatography-electrospray ionization tandem mass spectrometry: an analysis of hormone regulation of thermodormancy of lettuce (Lactuca sativa L.) seeds

A highly selective and sensitive method for the simultaneous analysis of several plant hormones and their metabolites is described. The method combines high-performance liquid chromatography (HPLC) with positive and negative electrospray ionization-tandem mass spectrometry (ESI-MS/MS) to quantify a broad range of chemically and structurally diverse compounds. The addition of deuterium-labeled analogs for these compounds prior to sample extraction permits accurate quantification by multiple reaction monitoring (MRM). Endogenous levels of abscisic acid (ABA), abscisic acid glucose ester (ABA-GE), 7'-hydroxy-abscisic acid (7'-OH-ABA), phaseic acid (PA), dihydrophaseic acid (DPA), indole-3-acetic acid (IAA), indole-3-aspartate (IAAsp), zeatin (Z), zeatin riboside (ZR), isopentenyladenine (2iP), isopentenyladenosine (IPA), and gibberellins (GA)1, GA3, GA4, and GA7 were determined simultaneously in a single run. Detection limits ranged from 0.682 fmol for Z to 1.53 pmol for ABA. The method was applied to the analysis of plant hormones and hormonal metabolites associated with seed dormancy and germination in lettuce (Lactuca sativa L. cv. Grand Rapids), using extracts from only 50 to 100 mg DW of seed. Thermodormancy was induced by incubating seeds at 33 degrees C instead of 23 degrees C. Germinating seeds transiently accumulated high levels of ABA-GE. In contrast, thermodormant seeds transiently accumulated high levels of DPA after 7 days at 33 degrees C. GA1 and GA3 were detected during germination, and levels of GA1 increased during early post-germinative growth. After several days of incubation, thermodormant seeds exhibited a striking transient accumulation of IAA, which did not occur in seeds germinating at 23 degrees C. We conclude that hormone metabolism in thermodormant seeds is surprisingly active and is significantly different from that of germinating seeds.