Walking speed and spatiotemporal step mean measures are reliable during feedback-controlled treadmill walking; however,spatiotemporal step variability is not reliable

The purpose of the study was to compare the effects of a feedback-controlled treadmill (FeedbackTM) to a traditional fixed-speed treadmill (FixedTM) on spatiotemporal gait means, variability, and dynamics. The study also examined inter-session reliability when using the FeedbackTM. Ten young adults walked on the FeedbackTM for a 5-minute familiarization followed by a 16-minute experimental trial. They returned within one week and completed a 5-minute familiarization followed by a 16-minute experimental trial each for FeedbackTM and FixedTM conditions. Mean walking speed and step time, length, width, and speed means and coefficient of variation were calculated from all experimental conditions. Step time, length, width, and speed gait dynamics were analyzed using detrended fluctuation analysis. Mean differences between experimental trials were determined using ANOVAs and reliability between FeedbackTM sessions was determined by intraclass correlation coefficient. No difference was found in mean walking speed nor spatiotemporal variables, with the exception of step width, between the experimental trials. All mean spatiotemporal variables demonstrated good to excellent reliability between sessions, while coefficient of variation was not reliable. Gait dynamics of step time, length, width, and speed were significantly more persistent during the FeedbackTM condition compared to FixedTM, especially step speed. However, gait dynamics demonstrated fair to poor reliability between FeedbackTM sessions. When walking on the FeedbackTM, users maintain a consistent set point, yet the gait dynamics around the mean are different when compared to walking on a FixedTM. In addition, spatiotemporal gait dynamics and variability may not be consistent across separate days when using the FeedbackTM.     

A computational method for reliable gait event detection and abnormality detection for feedback in rehabilitation

In this paper, a gait event detection algorithm is presented that uses computer intelligence (fuzzy logic) to identify seven gait phases in walking gait. Two inertial measurement units and four force-sensitive resistors were used to obtain knee angle and foot pressure patterns, respectively. Fuzzy logic is used to address the complexity in distinguishing gait phases based on discrete events. A novel application of the seven-dimensional vector analysis method to estimate the amount of abnormalities detected was also investigated based on the two gait parameters. Experiments were carried out to validate the application of the two proposed algorithms to provide accurate feedback in rehabilitation. The algorithm responses were tested for two cases, normal and abnormal gait. The large amount of data required for reliable gait-phase detection necessitate the utilisation of computer methods to store and manage the data. Therefore, a database management system and an interactive graphical user interface were developed for the utilisation of the overall system in a clinical environment.     

A simple method for the extraction of soil solution by high speed centrifugation

Summary A simple technique for extracting samples of soil solution by high speed centrifugation is described. The method uses inexpensive commerically available centrifuge tubes requiring only slight modification. Aspects of the method are discussed together with an example of its application.     

A novel method to replicate the kinematics of the carpus using a six degree-of-freedom robot

Understanding the kinematics of the carpus is essential to the understanding and treatment of wrist pathologies. However, many of the previous techniques presented are limited by non-functional motion or the interpolation of points from static images at different postures. We present a method that has the capability of replicating the kinematics of the wrist during activities of daily living using a unique mechanical testing system. To quantify the kinematics of the carpal bones, we used bone pin-mounted markers and optical motion capture methods. In this paper, we present a hammering motion as an example of an activity of daily living. However, the method can be applied to a wide variety of movements. Our method showed good accuracy (1.0–2.6°) of in vivo movement reproduction in our ex vivo model. Most carpal motion during wrist flexion–extension occurs at the radiocarpal level while in ulnar deviation the motion is more equally shared between radiocarpal and midcarpal joints, and in radial deviation the motion happens mainly at the midcarpal joint. For all rotations, there was more rotation of the midcarpal row relative to the lunate than relative to the scaphoid or triquetrum. For the functional motion studied (hammering), there was more midcarpal motion in wrist extension compared to pure wrist extension while radioulnar deviation patterns were similar to those observed in pure wrist radioulnar deviation. Finally, it was found that for the amplitudes studied the amount of carpal rotations was proportional to global wrist rotations.     

A simple method to determine the enantiomeric ratio in enantioselective biocatalysis

The enantiomeric ratio (E) is commonly used to characterize the enantioselectivity in enzyme-catalyzed kinetic resolution. In this paper this parameter is directly derived from the enantiomeric excess of substrate and product. This is formally more correct than using Chen's equation after calculating the degree of conversion from both ee values using the relation of Sih and Wu. New expressions and useful graphs have been generated for reversible and irreversible uni-uni reactions. The theoretical predictions have been verified experimentally for various reactions. Values for E and the thermodynamic equilibrium constant,K_EQ, were obtained for a ( -dehalogenase-catalyzed dehalogenation, a hydrolysis reaction by porcine pancreatic lipase, and for C. Cylindracea lipase-catalyzed esterification and transesterification. In view of the current developments in the field of chiral analysis, this method is an easily available tool in the quantitative treatment of enzyme-catalyzed resolution of enantiomers.     

A novel,simple, and sensitive colorimetric method to determine aromatic amino acid aminotransferase activity using the Salkowski reagent

This study describes the development of a new colorimetric assay to determine aromatic amino acid aminotransferase (ArAT) activity. The assay is based on the transamination of l-tryptophan in the presence of 2-oxoglutarate, which yields indole-3-pyruvate (IPyA). The amount of IPyA formed was quantified by reaction with the Salkowski reagent. Optimized assay conditions are presented for ArAT isozymes isolated from Pseudomonas putida. For comparative purposes, ArAT activity was also determined by high-performance liquid chromatography. ArAT activity staining in polyacrylamide gels with the Salkowski reagent is also presented.     

A simple method to determine urea concentration using intact Helicobacter pylori and BromoCresol Purple as a pH indicator

A modified method for urea quantification, by measuring the ammonia formed by urease, used the urease-positive Helicobacter pylori in place of purified urease with a pH indicator dye, BromoCresol Purple, to provide a color change. The color formed was stable for 20-min and could be read at 588-nm for urea quantification. Using this method, urea standard curves were linear up to 8.3-mM. As there was no need for centrifugation or precipitation, the assay was developed for use with 96-well microplates.     

A simple method to determine bioethanol content in gasoline using two-step extraction and liquid scintillation counting

A simple method for determining bioethanol content in gasoline containing bioethanol (denoted as E-gasoline in this study) is urgently required. Liquid scintillation counting (LSC) was employed based on the principle that ¹⁴C exists in bioethanol but not in synthetic ethanol. Bioethanol was extracted in two steps by water from E-gasoline containing 3% (E3) or 10% (E10) bioethanol. The ¹⁴C radioactivity was measured by LSC and converted to the amount of bioethanol. The bioethanol content in E-gasoline was determined precisely from the partition coefficient in the extraction and the amount of bioethanol in the water phases: 2.98 ± 0.10% for E3 and 10.0 ± 0.1% for E10 (means ± SD; n = 3). It appears that this method can be used to determine bioethanol content in E-gasoline quickly and easily.     

A method to determine the mean pollen dispersal of individual plants growing within a large pollen source

Pollen dispersal has been recently focused on as a major issue in the risk assessment of transgenic crop plants. The shape of the pollen dispersal of individual plants is hard to determine since a very large number of plants must be monitored in order to track rare longdistance dispersal events. Conversely, studies using large plots as a pollen source provide a pollen distribution that depends on the shape of the source plot. We report here on a method based on the use of Fourier transforms by which the pollen dispersal of a single, average individual can be obtained from data using large plots as pollen sources, thus allowing the estimation of the probability of long-distance dispersal for single plants. This method is subsequently implemented on simulated data to test its susceptibility to random noise and edge effects. Its conditions of application and value for use in ecological studies, in particular risk assessment of the deliberate release of transgenic plants, are discussed.     

Open-top static respirometry is a reliable method to determine the routine metabolic rate of barramundi,Lates calcarifer

Closed-system respirometry is a standard technique used to determine aerobic metabolism of aquatic organisms. Open-top systems are rarely used due to concerns of gas exchange across the air–water interface. Here, we evaluated an open-top respirometry system by comparing the mass-specific routine metabolic rate (RMR) of the tropical diadromous finfish barramundi, Lates calcarifer, in both closed-top and open-top respirometers. The RMR of 190?g barramundi was determined across broad temperatures ranging from 18 to 38?°C. There was no significant difference in RMR between barramundi in either closed- or open-top respirometers at any temperature (p?>?0.05). To ensure RMR measurements were not an artifact of the respirometry system, barramundi were reciprocally transplanted into either respective closed-top or open-top respirometer and oxygen consumption re-measured at each temperature treatment. The RMR of transplanted barramundi was found to be virtually identical in either respirometer. RMR increased linearly with increasing temperature; the relationship between RMR and temperature (T; 18–38?°C) can be described as 3.658T?36.294?mg?O₂?kg^?0.8?h^?1. The daily energetic cost of RMR was 1.193T?11.838?kJ?kg^?0.8?day^?1. Q₁₀ for barramundi increased significantly with increasing temperature (p?Q_10(18–28) was the lowest at 1.7 and Q_10(28–38) the highest at 1.9, over the whole experiment temp range Q_10(18–28) was 1.8. The current study demonstrates that open-top respirometry is a reliable and practical alternative to closed-top respirometry for accurate determination of the aerobic metabolism of barramundi and has potential application for a number of different aquatic organisms.     

A simple and rapid method to determine the zygosity of uPA-transgenic SCID mice

Successful transplantation of xenogeneic hepatocytes into uPA-transgenic SCID mice depends on the zygosity of the recipient mice. Normally, the difference between homozygous and heterozygous animals is determined via a quantitative Southern blot. We sequenced a part of the mouse genome that is eliminated upon integration of the transgene in the genome. Based on that sequence we developed a multiplex PCR that allows the unambiguous discrimination of negative, heterozygous, and homozygous uPA-transgenic SCID mice in a single day procedure. The speed of the procedure is an essential quality because transplantation of xenogeneic hepatocytes into uPA-SCID mice should be done as soon as possible after birth.     

A method to minimise error in 2D-DLT reconstruction of non-planar markers filmed with a moving camera

This article describes a method that allows estimating, with the 2D version of the direct linear transformation (DLT), the actual 2D coordinates of a point when the latter is not strictly in the calibration plane. Markers placed in vertical line, above, below and in the centre of a horizontal calibration plane were filmed by a moving camera. Without correction, strong errors (up to 64.5%) were noticed for markers out of the calibration plane. After correction, calculated coordinates were consistent with actual values (error < 0.55%). The method was then applied to slip distance measurement, using a marker fixed on the hoof of a horse trotting on a calibrated track while being followed with a camera. The correction effect represented 6.6% of slip distance. Combined with the 2D-DLT transformation, the proposed corrective method allows an accurate measurement of slip distances, for high-speed outdoor locomotion analysis, using a moving camera.     

一种用于昆虫性信息素成分单不饱和双键定位的简便方法

昆虫利用特定的性信息素成分实现种内信息交流和种间生殖隔离,性信息素成分异构体的细微差异都能导致其生物活性的很大改变。本研究利用二甲基二硫醚（DMDS）甲硫基化反应和气相色谱-电子轰击质谱（GC-EI-MS）的方法对脂肪族单不饱和醇、醛和乙酸酯类昆虫性信息素成分双键在碳链中的位置异构进行系统研究。质谱分析结果表明,含有醇和乙酸酯官能团的单不饱和烯烃甲硫基化衍生物的质谱特征碎片分子离子峰（M+）及2个主要的诊断离子m/z 61+14m ［H-（CH₂）_m-CH=S⁺ CH₃］和77+14n［CH₃S⁺ =CH（CH₂）_nOH］（或119+14n,［CH₃S⁺ =CH（CH₂）_nOOCCH₃］）丰度都很强,可以据此直接推断双键在碳链中的位置。尽管单不饱和醛类化合物DMDS衍生物的特征碎片峰也很明显,但是由于羰基的质量数和2个亚甲基的质量数相同,不能由低分辨质谱的特征碎片峰直接推断双键在碳链中的位置。DMDS甲硫基化反应结合GC-EI-MS分析方法鉴定昆虫性信息素成分单不饱和双键在碳链中的位置异构,该方法简单快捷,为昆虫性信息素的结构鉴定工作奠定了坚实的基础。     

A simple and reliable method to conduct and monitor expression cassette integration into the Escherichia coli chromosome

We report a method for the integration of expression cassettes into the Escherichia coli chromosome using rare and dispensable sugar degradation gene loci as sites for integration. Clones carrying successfully recombined DNA fragments in the chromosome are easily screened using a solid differential medium containing the respective sugar compound. As an example for the heterologous expression of a complex natural product biosynthesis pathway, we show the stepwise chromosomal integration of the zeaxanthin biosynthesis pathway from Pantoea ananatis into E. coli.     

A simple and efficient method to determine the terminal sequences of restriction fragments containing known sequences.

We present an improvement of the inverse PCR method for the determination of end sequences of restriction fragments containing unknown DNA sequences flanked by known segments. In this approach, a short "bridge" DNA is inserted during the self-ligation step of the inverse PCR technique. This bridge DNA acts as primer annealing sites for amplification and subsequent direct sequencing. Successive PCR amplifications enable selective amplification of the unknown sequences from a complex mixture. Unlike previously described methods, our method does not require special materials, such as synthetic adapters or biotinylated primers that must be prepared each time to adapt the target. Furthermore, no complex steps such as dephosphorylation or purification are needed. Our method can save time and reduce the cost of cloning unknown sequences; it is ideal for routine, rapid gene walking. We applied this method to a GC-rich bacterial genome and succeeded in determining the end sequences of a 4.5-kb fragment.     

A novel method to evaluate error in anatomical marker placement using a modified generalized Procrustes analysis

As biomechanical research evolves, a continuing challenge is the standardization of data collection and analysis techniques. In gait analysis, placement of markers to construct an anatomical model has been identified as the single greatest source of error; however, there is currently no standardized approach to quantifying these errors. The current study applies morphometric methods, including a generalized Procrustes analysis (GPA) and a nearest neighbour comparison to quantify discrepancies in marker placement, with the goal of improving reliability in gait analysis. An extensive data-set collected by an Expert (n = 340) was used to evaluate marker placements performed by a Novice (n = 55). Variances identified through principal component analysis were used to create a modified GPA to transform anatomical data, and scaled coordinates from the Novice data-set were then scored against the Expert subset. The results showed quantitative differences in marker placement, suggesting that, although training improved consistency, systematic biases remained.     

A method to determine the mode of binding for GCPII inhibitors using bio-layer interferometry

The rapid dilution of the enzyme-inhibitor complex assay to monitor the recovery of enzyme activity is a well-established assay to determine the reversibility of inhibition. Our laboratory has previously employed this method to ascertain the reversibility of known glutamate carboxypeptidase II (GCPII)-targeting agents. Due to the tedious and time-consuming nature of the assay, we sought to develop a facile method to determine the reversibility of well-characterized GCPII inhibitors using bio-layer interferometry (BLI). The results from the BLI assay are in agreement with the rapid dilution method. Herein, we report for the first time, a rapid, novel real-time BLI method to determine reversibility of inhibition.     

A new method for motion capture of the scapula using an optoelectronic tracking device: a feasibility study

Optoelectronic tracking systems are rarely used in 3D studies examining shoulder movements including the scapula. Among the reasons is the important slippage of skin markers with respect to scapula. Methods using electromagnetic tracking devices are validated and frequently applied. Thus, the aim of this study was to develop a new method for in vivo optoelectronic scapular capture dealing with the accepted accuracy issues of validated methods.

Eleven arm positions in three anatomical planes were examined using five subjects in static mode. The method was based on local optimisation, and recalculation procedures were made using a set of five scapular surface markers.

The scapular rotations derived from the recalculation-based method yielded RMS errors comparable with the frequently used electromagnetic scapular methods (RMS up to 12.6° for 150° arm elevation). The results indicate that the present method can be used under careful considerations for 3D kinematical studies examining different shoulder movements.     

A simple but reliable method for measuring 3D Achilles tendon moment arm geometry from a single,static magnetic resonance scan

Current methods for measuring in vivo 3D muscle-tendon moment arms generally rely on the acquisition of magnetic resonance imaging (MRI) scans at multiple joint angles. However, for patients with musculoskeletal pathologies such as fixed contractures, moving a joint through its full range of motion is not always feasible. The purpose of this research was to develop a simple, but reliable in vivo 3D Achilles tendon moment arm (ATMA) technique from a single static MRI scan. To accomplish this, for nine healthy adults (5 males, 4 females), the geometry of a cylinder was fit to the 3D form of the talus dome, which was used to estimate the talocrural flexion/extension axis, and a fifth-order polynomial fit to the line of action of the Achilles tendon. The single static scan in vivo 3D ATMA estimates were compared to estimates obtained from the same subjects at the same ankle joint angles using a previously validated 3D dynamic MRI based in vivo ATMA measurement technique. The ATMA estimates from the single scan in vivo 3D method (52.5 mm ± 5.6) were in excellent agreement (ICC = 0.912) to the validated in vivo 3D method (51.5 mm ± 5.1). These data show reliable in vivo 3D ATMA can be obtained from a single MRI scan for healthy adult populations. The single scan, in vivo 3D ATMA technique provides researchers with a simple, but reliable method for obtaining subject-specific ATMAs for musculoskeletal modelling purposes.