• Journal of Life Science
• /
• v.28 no.1
• /
• pp.99-104
• /
• 2018

Streptococcus pneumoniae is one of the major pathogens in community-acquired diseases, and it contains several factors that promote its pathogenesis, including pneumolysin (PLY). PLY is a member of the cholesterol-dependent cytolysin family, which attacks cholesterol-containing membranes, thereby forming ring-shaped pores. Thus, it is a major key target for vaccines against pneumococcal disease. We cloned the PLY gene from S. pneumoniae D39 and inserted it into the pQE-30 vector. Recombinant PLY (rPLY) was overexpressed in Escherichia coli M15 and purified by $Ni^{2+}$ affinity chromatography. Similarly, a PLY-EGFP fusion gene was produced by inserting the EGFP gene at the 3' end of the PLY gene in the same vector, and the recombinant protein was purified. Sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) showed that both recombinant proteins were purified. rPLY exhibited significant hemolytic activity against 1% human red blood cells (RBCs). Complete hemolysis was obtained at 500 ng/ml, and 50% hemolysis was found with a 240 ng/ml concentration. In contrast, rPLY-EGFP did not show hemolytic activity. However, rPLY-EGFP did bind the RBC membrane, indicating that rPLY-EGFP lost hemolytic activity via EGFP fusion, while retaining its membrane-binding ability. These data suggest that PLY's C terminus is important for its hemolytic activity. Therefore, these two recombinant proteins can be extremely useful for investigating the toxin mechanism of PLY and cell damage during pneumonia.

• Journal of Mechanical Science and Technology
• /
• v.16 no.3
• /
• pp.292-301
• /
• 2002

Ultrasound waves interact strongly with the orientation and sequence of the plies in a layup when propagating in the thickness direction of composite laminates. Also the layup orientation greatly influences its properties in a composite laminate. If the layup orientation of a ply is misaligned, it could result in the part being rejected and discarded. Now, most researchers cut a small coupon from the waste edge and use a microscope to optically verify the ply sequences on important parts. This may add a substantial cost to the production since the test is both labor intensive and performed after the part is cured. A nondestructive technique would be very beneficial, which could be used to test the part after curing and requires less time than the optical test. Therefore we have developed, reduced, and implemented a novel ply-by-ply vector decomposition model for composite laminates fabricated from unidirectional plies. This model decomposes the transmission of a linearly polarized ultrasound wave into orthogonal components through each ply of a laminate. High probability is found, by comparisons between the model and tests, in characterizing cured layups of the laminates by using the proposed method.

• Proceedings of the KSME Conference
• /
• 2001.06a
• /
• pp.401-406
• /
• 2001

When propagating the thickness direction of composite laminates ultrasound waves interacts strongly with the orientation and sequence of the plies in a layup. Also the layup orientation greatly influences its properties in a composite laminate. If one ply of the layup orientation is misaligned, it could result in the part being rejected and discarded. Now, most researchers cut a small coupon from the waste edge and use a microscope to optically verify the ply sequences on important parts. Those may add a substantial cost to the product since the test is both labor hard and performed after the part is cured. A nondestructive technique would be very beneficial, which could be used to test the part after curing and require less time than the optical test. Therefore we have developed, reduced, and implemented a novel ply-by-ply vector decomposition model for composite lam mates fabricated from unidirectional plies. This model decomposes the transmission of a linearly polarized ultrasound wave into orthogonal components through each ply of a laminate. It is found that a high probability shows between the model and tests developed in characterizing cured layups of the laminates.

• Journal of Mechanical Science and Technology
• /
• v.18 no.1
• /
• pp.65-73
• /
• 2004

Composites are a material class for which nondestructive material property characterization is as important as flaw detection. Laminates of fiber reinforced composites often possess strong in-plane elastic anisotropy attributable to the specific fiber orientation and layup sequence when waves are propagating in the thickness direction of composite laminates. So the layup orientation greatly influences its properties in a composite laminate. It could result in the part being .ejected and discarded if the layup orientation of a ply is misaligned. A nondestructive technique would be very beneficial, which could be used to test the part after curing and requires less time than the optical test. Therefore a ply-by-ply vector decomposition model has been developed, simplified, and implemented for composite laminates fabricated from unidirectional plies. This model decomposes the transmission of a linearly polarized ultrasound wave into orthogonal components through each ply of a laminate. Also in order to develop these methods into practical inspection tools, motorized system have been developed for different measurement modalities for acquiring ultrasonic signals as a function of in-plane angle. It is found that high probability shows between the model and tests developed in characterizing cured layups of the laminates.

• Journal of the Korean Society for Nondestructive Testing
• /
• v.23 no.6
• /
• pp.566-576
• /
• 2003

Carbon-fiber reinforced plastics (CFRP) composite laminates often possess strong in-plane elastic anisotropy attributable to the fiber orientation and layup sequence. The layup orientation thus greatly influences its properties in a composite laminate. It could result in the part being rejected or discarded if the layup orientation of a ply is misaligned. A nondestructive technique would be very beneficial, which could be used to test the part after curing and to require less time than the optical test. In this paper, ultrasonic scanners were set out for different measurement modalities for acquiring ultrasonic signals as a function of in-plane azimuthal angle. The motorized scanner was built first for making transmission measurements using a pair of normal-incidence shear wave transducers. Another scanner was then built fer the acousto-ultrasonic configuration using contact transducers. A ply-by-ply vector decomposition model has been developed, simplified, and implemented for composite laminates fabricated from unidirectional plies. We have compared the test results with model data. It is found that strong agreement are shown between tests and the model developed in characterizing cured layups of the laminates.

• Proceedings of the Korean Society of Precision Engineering Conference
• /
• 2001.04a
• /
• pp.744-747
• /
• 2001

This paper shows shear wave behavior of CFRP composite laminates as a polar grid form to evaluate vibration pattern of ultrasonic transducers, which gives measured modelling fundamental contents of nondestructive evaluation. Polarized direction can be obtained by using a c-scanner and sensitivity of transducers is founded when using through-transmission method of two transducers. And modelling of vector decomposition is presented based on ply-to-ply method to apply practicable nondestructive evaluation of CFRP laminate lay up. This modelling decomposes the transmission of linearly polarized wave into orthogonal components through each ply of a laminate. It is found that a high provable shows between the model and experimental developed in characterizing layup of CFRP composite laminates.

• Proceedings of the Korean Society of Precision Engineering Conference
• /
• 2002.10a
• /
• pp.892-895
• /
• 2002

In this study, to grasp the effect of vibration intensity fur the laminated composite plate, the two-dimension plate was shaken by a harmonic point excitation with the natural frequency using the finite element method. As the result, it shows that the vibration intensity according to the change of angle-ply is various and it flows to the direction of length rather than width in the plate. Also this paper represents those results to the vector flow.

• Structural Engineering and Mechanics
• /
• v.40 no.2
• /
• pp.257-277
• /
• 2011

In this paper the geometrically nonlinear continuum plate finite element model, hitherto not reported in the literature, is developed using the total Lagrange formulation. With the layerwise displacement field of Reddy, nonlinear Green-Lagrange small strain large displacements relations (in the von Karman sense) and linear elastic orthotropic material properties for each lamina, the 3D elasticity equations are reduced to 2D problem and the nonlinear equilibrium integral form is obtained. By performing the linearization on nonlinear integral form and then the discretization on linearized integral form, tangent stiffness matrix is obtained with less manipulation and in more consistent form, compared to the one obtained using laminated element approach. Symmetric tangent stiffness matrixes, together with internal force vector are then utilized in Newton Raphson's method for the numerical solution of nonlinear incremental finite element equilibrium equations. Despite of its complex layer dependent numerical nature, the present model has no shear locking problems, compared to ESL (Equivalent Single Layer) models, or aspect ratio problems, as the 3D finite element may have when analyzing thin plate behavior. The originally coded MATLAB computer program for the finite element solution is used to verify the accuracy of the numerical model, by calculating nonlinear response of plates with different mechanical properties, which are isotropic, orthotropic and anisotropic (cross ply and angle ply), different plate thickness, different boundary conditions and different load direction (unloading/loading). The obtained results are compared with available results from the literature and the linear solutions from the author's previous papers.