基于替代数据思想的复杂度归一化方法及其在心电信号分析中的应用

基于替代数据(Surrogate)思想的复杂度归一化方法,克服了一般复杂度对信号采样长度与采样频率的敏感性。文章对在生物医学信号复杂度分析中最有潜在应用价值的近似熵和C0复杂度进行了归一化。应用该方法可以有效地反映人体心脏某些病理状态之间的差别。同时,通过比较各种复杂度指标发现,C0复杂度和近似熵对采样长度的敏感性最弱,适用于短数据量的信号分析。     

EMG of upper trapezius − Electrode sites and association with clavicular kinematics

The upper trapezius (UT) has been widely studied and related to alterations in clavicular kinematics in subject with shoulder disorders. However, the most common electrode site used to capture UT EMG is between C7 and the acromion, placing the electrodes over the acromial fibers rather than clavicular ones. Therefore, this study aimed to investigate the relationship between clavicular movements (elevation and retraction) and UT EMG recorded from three electrode sites (traditional electrode positioning and two different sites proposed for clavicular fibers evaluation). Furthermore, the position associated with the highest EMG during maximal isometric voluntary contractions (MVIC), for each electrode site, was determined for normalization purposes. EMG was simultaneously captured in the three electrode sites of 20 healthy subjects, during MVIC at five different positions and during shoulder elevation and abduction in scapular plane. Clavicular kinematics was recorded using an electromagnetic tracking system during the dynamic contractions. Shoulder abduction with head rotation and lateral flexion elicited the highest EMG amplitude on the three electrode sites and was used to normalize the signals. A cross-correlation analysis showed high correlations between all electrode sites and clavicular movements. However, the traditional electrode site seems to record more informative signals in healthy subjects.     

Obtaining maximum muscle excitation for normalizing shoulder electromyography in dynamic contractions

Muscle specific maximal voluntary isometric contractions (MVIC) are commonly used to elicit reference amplitudes to normalize electromyographic signals (EMG). It has been questioned whether this is appropriate for normalizing EMG from dynamic contractions. This study compares EMG amplitude when shoulder muscle activity from dynamic contractions is normalized to isometric and isokinetic maximal excitation as well as a hybrid approach currently used in our laboratory. Anterior, middle and posterior deltoid, upper and lower trapezius, pectoralis major, latissimus dorsi and infraspinatus were monitored during (1) manually resisted MVICs, and (2) maximum voluntary dynamic concentric contractions (MVDC) on an isokinetic dynamometer. Dynamic contractions were performed (a) at 30°/s about the longitudinal, frontal and sagittal axes of the shoulder, and (b) during manual bi-rotation of a tilted wheel at 120°/s. EMG from the wheel task was normalized to the maximum excitation from (i) the muscle specific MVIC, (ii) from any MVIC (MVIC_ALL), (iii) for any MVDC, (iv) from any exertion (maximum experimental excitation, MEE). Mean EMG from the wheel task was up to 45% greater when normalized to muscle specific isometric contractions (method i) than when normalized to MEE (method iv). Seventy-five percent of MEE’s occurred during MVDCs. This study presents an 20 useful and effective process for obtaining the greatest excitation from the shoulder muscles when normalizing dynamic efforts.     

Tissue proteomics of the low‐molecular weight proteome using an integrated cLC‐ESI‐QTOFMS approach

Analysis of the protein/peptide composition of tissue has provided meaningful insights into tissue biology and even disease mechanisms. However, little has been published regarding top down methods to investigate lower molecular weight (MW) (500–5000 Da) species in tissue. Here, we evaluate a tissue proteomics approach involving tissue homogenization followed by depletion of large proteins and then cLC‐MS (where c stands for capillary) analysis to interrogate the low MW/low abundance tissue proteome. In the development of this method, sheep heart, lung, liver, kidney, and spleen were surveyed to test our ability to observe tissue differences. After categorical tissue differences were demonstrated, a detailed study of this method's reproducibility was undertaken to determine whether or not it is suitable for analyzing more subtle differences in the abundance of small proteins and peptides. Our results suggest that this method should be useful in exploring the low MW proteome of tissues.     

Selection of control genes for quantitative RT-PCR based on microarray data

Use of internal reference gene(s) is necessary for adequate quantification of target gene expression by RT-PCR. Herein, we elaborated a strategy of control gene selection based on microarray data and illustrated it by analyzing endomyocardial biopsies with acute cardiac rejection and infection. Using order statistics and binomial distribution we evaluated the probability of finding low-varying genes by chance. For analysis, the microarray data were divided into two sample subsets. Among the first 10% of genes with the lowest standard deviations, we found 14 genes common to both subsets. After normalization using two selected genes, high correlation was observed between expression of target genes evaluated by microarray and RT-PCR, and in independent dataset by RT-PCR (r = 0.9, p < 0.001). In conclusion, we showed a simple and reliable strategy of selection and validation of control genes for RT-PCR from microarray data that can be easily applied for different experimental designs and tissues.     

Use of a fluorescent internal protein standard to achieve quantitative two-dimensional gel electrophoresis

2-DE is a powerful separation method for complex protein mixtures. However, large intergel variations in spot intensity limit its use for quantitative proteomics studies. To address this issue, we developed a fluorescent internal protein standard for use in 2-DE analysis. Protein samples are spiked with an Alexa-labeled internal standard (ALIS) prior to separation with 2-DE. Due to the high extinction coefficient of the Alexa-fluor, incorporation of 0.1% of total protein is sufficient to allow visualization of the internal standard yet low enough to avoid interference in subsequent quantification and identification steps. Following 2-DE, total proteins are visualized with fluorescent postelectrophoretic stains spectrally separated from ALIS. Four protein stains, Deep Purple, Sulforhodamine G, ruthenium II-tris(bathophenanthroline disulfonate) (RuTBS), and SYPRO Ruby, including improved purification and staining protocols for RuTBS and ten-fold dilutions of SYPRO Ruby were evaluated. All staining protocols were compatible with the ALIS method and had similar LODs (1-4 ng) and dynamic ranges (10(3)). ALIS is a powerful normalization method for quantitative 2-DE which avoids potential problems associated with dual spot migration patterns observed in the DIGE method. Furthermore, ALIS provides significantly improved normality in the distribution of spot abundance-variance compared to normalization through division by the total spot volume.