Identification and expression profiling of flax (Linum usitatissimum L.) polyamine oxidase genes in response to stimuli

Polyamine oxidases (PAOs) are known to be involved in either the terminal catabolism or the back conversion of polyamines, which affect a range of physiological processes, including growth, development, and stress responses. In this study, based on genome-wide analysis, we identified five putative PAO genes (LuPAO1 to LuPAO5) in flax (Linum usitatissimum L.) that contain the amino-oxidase domain and FAD-binding-domain. The expression analysis using quantitative real-time PCR revealed spatial variations in the expression of LuPAOs in different organs. In addition, the expression level of LuPAOs in the flax cell suspension culture was increased by treatment with methyl- jasmonate (MeJA) or pectin, but not with salicylic acid or chitosan. This indicates that LuPAOs might be involved in the MeJA-mediated biological activities. Taken together, our genome-wide analysis of PAO genes and expression profiling of these genes provide the first step toward the functional dissection of LuPAOs

An Engineered Approach to Stem Cell Culture: Automating the Decision Process for Real-Time Adaptive Subculture of Stem Cells

Current cell culture practices are dependent upon human operators and remain laborious and highly subjective, resulting in large variations and inconsistent outcomes, especially when using visual assessments of cell confluency to determine the appropriate time to subculture cells. Although efforts to automate cell culture with robotic systems are underway, the majority of such systems still require human intervention to determine when to subculture. Thus, it is necessary to accurately and objectively determine the appropriate time for cell passaging. Optimal stem cell culturing that maintains cell pluripotency while maximizing cell yields will be especially important for efficient, cost-effective stem cell-based therapies. Toward this goal we developed a real-time computer vision-based system that monitors the degree of cell confluency with a precision of 0.791±0.031 and recall of 0.559±0.043. The system consists of an automated phase-contrast time-lapse microscope and a server. Multiple dishes are sequentially imaged and the data is uploaded to the server that performs computer vision processing, predicts when cells will exceed a pre-defined threshold for optimal cell confluency, and provides a Web-based interface for remote cell culture monitoring. Human operators are also notified via text messaging and e-mail 4 hours prior to reaching this threshold and immediately upon reaching this threshold. This system was successfully used to direct the expansion of a paradigm stem cell population, C2C12 cells. Computer-directed and human-directed control subcultures required 3 serial cultures to achieve the theoretical target cell yield of 50 million C2C12 cells and showed no difference for myogenic and osteogenic differentiation. This automated vision-based system has potential as a tool toward adaptive real-time control of subculturing, cell culture optimization and quality assurance/quality control, and it could be integrated with current and developing robotic cell cultures systems to achieve complete automation

Transtorming Growth Factor β1 Induces Epithelial-to-Mesenchymal Transition of A549 Cells

Idiopathic pulmonary fibrosis (IPF) comprises an aggregate of mesenchymal cells. However, the cellular origin of these mesenchymal phenotypes remains unclear. Transforming growth factor β1 (TGF-β1) has been known as the main cytokine involved in the pathogenesis of IPF. We examined whether the potent fibrogenic cytokine TGF-β1 could induce the epithelial-to-mesenchymal transition (EMT) in the human alveolar epithelial cell line, A549, and determined whether snail expression is associated with the phenotypic changes observed in the A549 cells. EMT was investigated with cells morphology changes under phase-contrast microscopy, western blotting, and indirect immunofluorescence stains. E-cadherin and transcription factor, snail, were also evaluated by measuring mRNA levels using reverse transcriptase-polymerase chain rection (RT-PCR) analysis. The data showed that TGF-β1 induced A549 cells with epithelial cell characteristics to undergo EMT in a concentration-dependent manner. Following TGF-β1 treatment, A549 cells induced EMT characterized by cells morphological changes, loss of epithelial markers E-caherin and cytokeratin, increased stress fiber reorganization by F-actin, and cytokeratin replacement by vimentin. Although IL-1β failed to induce A549 cells to undergo EMT, the combination of TGF-β1 and IL-1β showed synergy effects in cells morphology changes and the expression of mesenchymal markers. The snail expression study using RT-PCR analysis provided that loss of E-cadherin expression was associated with snail expression. Stimulation of A54 cells with TGF-β1 plus IL-1β revealed a higher level of snail expression. Our data showed that EMT of A549 cells might be closely associated with snail expression

The Case On Assessment Of Spilled Oil With Mixed Free Product In Seoul

The purpose of this study is to assess sources of contamination by analyzing the free product and oil compounds in groundwater near the Subway Station, located adjacent to the Yongsan Garrison in Seoul, Republic of Korea. All of the samples collected twice were groundwater and free product in a monitoring well. Analysis items are TPH fingerprinting, pristine/phytane ratio, alkylbenzene pattern, PAH&alkyl PAH, antioxidant, icing inhibitor, PIANO, element(C, N, H), and sulfur. Using ratio of pristine/phytane, we were able to distinguished fuel type between kerosene and JP-8 samples, which was impossible by GC/FID pattern. Alkyl benzene pattern was very effective in distinguishing between JP-8 and kerosene. It is very important that 2,4-dimethyl-6-tert-butyl phenol, an antioxidant used only for JP-8, was detected in free product samples. The concentration of sulfur in kerosene fuel is much lower than that of JP-8, and the total contents of sulfur in environmental samples can be used to differentiate the fuel type of spilled oil between kerosene and JP-8. In conclusion, according to the result of a variety of analytical methods to find the source of spilled fuel, it had been found that the fuel type detected in the tunnel of subway station and monitoring wells outside of Yongsan Garrison and the monitoring wells inside of Yongsan Garrison are the same

Design of User motion intention algorithm for improving accuracy for Active upper limb exercise Robot

In this paper, an algorithm for recognizing the user’s intention by utilizing single channel electromyogram sensor and force sensor is proposed as a method for generating a synchronization signal of an active-type upper limb motion robot. In active-type upper limb motion robot system, a physical human-robot interface device physically connects users with the robot, and the motion of the user is recognized and delivered to the robot by the interface device in order to provide the necessary physical force to the user. By using the force sensor that is built inside the interface device, the intention of the user’s motion is recognized. Although the sensor is capable of measuring the magnitude of force, it is unable to distinguish between the force applied by the exterior and the external force generated by the user. Moreover, in case of the electromyogram signal, it is only expressed when the user actually generates the force, whereas its exact magnitude cannot be measured. Therefore, the suggested algorithm includes a step for each of the following: filtering the signal extracted from the electromyogram sensor, determining the recognition of the user’s motion after the filtering, classifying the signal expressed by the force sensor on the orthogonal coordinate space in accordance with the user’s motion, and outputting the user’s motion intention signal by converting the classified signal into a signal, related with the force and torque. In other words, the algorithm lets the user’s motion intention signal measured by the force sensor to be output only when the user’s motion is recognized as the electromyogram signal. According to the suggested method— which relies on eliminating the signals that are output, except for the motions generated by the user—the reliability of the user motion intention signal delivered to the robot can be enhanced