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Chang, T. Y.

Publications and source records attributed to Chang, T. Y..

At least 19 records
All sourcesNASA NTRSDOE OSTI
NASA NTRS2001

Distribution and Spectroscopy of Green Fluorescent Protein and Acyl-CoA: Cholesterol Acytransferase in Sf21 Insect Cells

Acyl-CoA: cholesterol acyltransferase (ACAT) is thought to significantly participate in the pathway of cholesterol esterification that underlies the pathology of artherosclerosis. This enzyme is a membrane protein known to be preferentially bound within the endoplasmic reticulum of mammalian cells, from which location it esterifies cholesterol derived from low density lipoprotein. Cultures of insect cells were separately infected with baculovirus containing the gene for green fluroescent protein (GFP) and with baculovirus containing tandem genes for GFP and ACAT. These infected cultures expressed GFP and the fusion protein GCAT, respectively, with maximum expression occurring on the fourth day after infection. Extraction of GFP- and of GCAT-expressing cells with urea and detergent resulted in recovery of fluorescent protein in aqueous solution. Fluorescence spectra at neutral pH were identical for both GFP and GCAT extracts in aqueous solution, indicating unperturbed tertiary structure for the GFP moiety within GCAT. In a cholesterol esterification assay, GCAT demonstrated ACAT activity, but with less efficiency compared to native ACAT. It was hypothesized that the membrane protein ACAT would lead to differences in localization of GCAT compared to GFP within the respective expressing insect cells. The GFP marker directly and also within the fusion protein GCAT was accordingly used as the intracellular probe that was fluorescently analyzed by the new biophotonics technique of hyperspectral imaging. In that technique, fluorescence imaging was obtained from two dimensional arrays of cells, and regions of interest from within those images were then retrospectively analyzed for the emission spectra that comprises the image. Results of hyperspectral imaging of insect cells on day 4 postinfection showed that GCAT was preferentially localized to the cytoplasm of these cells compared to GFP. Furthermore, the emission spectra obtained for the localized GCAT displayed a peak blue shift from 518nm obtained in neutral aqueous solution to 505nm obtained in localized regions within the cells. This blue shift indicates change in the fluorescence coupling of the GFP moiety of GCAT. It is hypothesized that change in tertiary environment of GCAT, coincident with intracellular deposition of GCAT, follows from intracellular trafficking of GCAT leading to membrane interactions with the ACAT moiety, and/or self-assembly of GCAT, that alters the chromophore environment of the GFP moiety of GCAT. These findings introduce a new technique of biophotonic imaging to studies of intracellular protein trafficking and interactions. This technique of hyperspectral imaging could contribute to advancing the emergent field of proteomics. Because of the noninvasive nature of this technique, kinetic processes associated with intracellular protein trafficking, and interactions of proteins within cellular domains, can be considered for investigation within a single cell as well as a cell population.

Richmond, R. C.↗
NASA NTRS2001

Expression of the Acyl-Coenzyme A: Cholesterol Acyltransferase GFP Fusion Protein in Sf21 Insect Cells

The enzyme acyl-coenzyme A:cholesterol acyltransferase (ACAT) is an important contributor to the pathological expression of plaque leading to artherosclerosis n a major health problem. Adequate knowledge of the structure of this protein will enable pharmaceutical companies to design drugs specific to the enzyme. ACAT is a membrane protein located in the endoplasmic reticulum.t The protein has never been purified to homogeneity.T.Y. Chang's laboratory at Dartmouth College provided a 4-kb cDNA clone (K1) coding for a structural gene of the protein. We have modified the gene sequence and inserted the cDNA into the BioGreen His Baculovirus transfer vector. This was successfully expressed in Sf2l insect cells as a GFP-labeled ACAT protein. The advantage to this ACAT-GFP fusion protein (abbreviated GCAT) is that one can easily monitor its expression as a function of GFP excitation at 395 nm and emission at 509 nm. Moreover, the fusion protein GCAT can be detected on Western blots with the use of commercially available GFP antibodies. Antibodies against ACAT are not readily available. The presence of the 6xHis tag in the transfer vector facilitates purification of the recombinant protein since 6xHis fusion proteins bind with high affinity to Ni-NTA agarose. Obtaining highly pure protein in large quantities is essential for subsequent crystallization. The purified GCAT fusion protein can readily be cleaved into distinct GFP and ACAT proteins in the presence of thrombin. Thrombin digests the 6xHis tag linking the two protein sequences. Preliminary experiments have indicated that both GCAT and ACAT are expressed as functional proteins. The ultimate aim is to obtain large quantities of the ACAT protein in pure and functional form appropriate for protein crystal growth. Determining protein structure is the key to the design and development of effective drugs. X-ray analysis requires large homogeneous crystals that are difficult to obtain in the gravity environment of earth. Protein crystals grown in microgravity are often larger and have fewer defects than those grown on earth. The analysis of higher quality space-grown crystals will assist in structure-based drug design. We have successfully grown GCAT-infected Sf21 cells in both adhesion and suspension cultures. Expression levels of GCAT in cell lines such as Sf9 and High Five appear to be reduced. We intend to replicate GCAT expression in all three cell lines using the NASA rotating wall bioreactor which effectively duplicates a microgravity environment. The bioreactor itself could be launched to study the expression of the GFP and GCAT proteins in the actual microgravity environment achieved in orbit.

Mahtani, H. K.↗
NASA NTRS2001

Hyperspectral Imaging and Spectroscopy of Fluorescently Coupled Acyl-CoA: Cholesterol Acyltransferase in Insect Cells

A high-performance hyperspectral imaging module with high throughput of light suitable for low-intensity fluorescence microscopic imaging and subsequent analysis, including single-pixel-defined emission spectroscopy, was tested on Sf21 insect cells expressing green fluorescence associated with recombinant green fluorescent protein linked or not with the membrane protein acyl-CoA:cholesterol acyltransferase. The imager utilized the phenomenon of optical activity as a new technique providing information over a spectral range of 220-1400 nm, and was inserted between the microscope and an 8-bit CCD video-rate camera. The resulting fluorescence image did not introduce observable image aberrations. The images provided parallel acquisition of well resolved concurrent spatial and spectral information such that fluorescence associated with green fluorescent protein alone was demonstrated to be diffuse within the Sf21 insect cell, and that green fluorescence associated with the membrane protein was shown to be specifically concentrated within regions of the cell cytoplasm. Emission spectra analyzed from different regions of the fluorescence image showed blue shift specific for the regions of concentration associated with the membrane protein.

Malak, H.↗
NASA NTRS1991

On the symbolic manipulation and code generation for elasto-plastic material matrices

A computerized procedure for symbolic manipulations and FORTRAN code generation of an elasto-plastic material matrix for finite element applications is presented. Special emphasis is placed on expression simplifications during intermediate derivations, optimal code generation, and interface with the main program. A systematic procedure is outlined to avoid redundant algebraic manipulations. Symbolic expressions of the derived material stiffness matrix are automatically converted to RATFOR code which is then translated into FORTRAN statements through a preprocessor. To minimize the interface problem with the main program, a template file is prepared so that the translated FORTRAN statements can be merged into the file to form a subroutine (or a submodule). Three constitutive models; namely, von Mises plasticity, Drucker-Prager model, and a concrete plasticity model, are used as illustrative examples.

Chang, T. Y.↗
NASA NTRS1990

A mixed element for laminated plates and shells

Formulation and numerical evaluation of a simple shear-flexible four-noded quadrilateral laminated composite plate/shell finite-element is presented. The element developed is based on a generalized mixed variational principle with independently assumed displacement and laminate internal strain fields. For the latter, a layer-number-independent polynomial interpolation is utilized, together with a judicious selection of the in-plane (spanwise) distributions for the strain components. This was facilitated by the use of a set of bubble functions as additional kinematic degrees of freedom, which was shown to be crucial for eliminating the locking phenomenon. For dynamic applications, a simplified lumped mass matrix is employed. Finally, an extensive number of critical test cases are given to access the element's preformance and to illustrate its effectiveness in static as well as vibration problems for anisotropic laminated plates and shells.

Wilt, T. E.↗
NASA NTRS1990

Temporal and spatial variability of the tropical radiation budget from ERBS measurements (March 1985 to February 1986)

The diurnal and regional variations of the top-of-atmosphere radiation budget in the tropics are examined and the relative importance of these variations are assessed. The variations of the radiation components with scene type and the radiative effects of clouds are investigated. The LW emission shows the strongest diurnal variation over clear desert and land scenes, with domain-averaged diurnal ranges of approximately 50 W/sq m and 35 W/sq m, respectively. The LW emission over clear and tropical oceans show a small diurnal range of less than 1 W/sq m. A bias in the ERBE scene ID algorithm against clear tropical scenes at night is evident in the LW diurnal variation results for clear scenes. The effect of tropical water vapor resulting in low LW emission to space is diagnosed as the principal source of uncertainty in scene identification at night. Regional cloud forcing results show a strong dependence of net forcing on cloud type. While high cirrus clouds largely contribute to a small positive net forcing, low stratiform clouds primarily account for a large negative net forcing.

Chang, T. Y.↗
NASA NTRS1988

A mixed formulaton of C(0)-linear triangular plate/shell element - The role of edge shear constraints

A simple and effective linear C(0)-tiangular element for plates and shells is developed on the basis of the Hellinger-Reissner mixed variational principle with independently assumed stress and displacement fields. Two main features are emphasized in this development concerning the assumed transverse shear stress field. First, this assumption results in edge-type penalty constraints in the thin plate/shell regimes. Second, the element is used in the form of a four-triangle, cross-diagonal macroelement mesh. As a result, the element is shown to be free from shear locking in thin plate/shell applications, and it satisfies all the appropriate patch tests required for Kirchhoff plate models. In addition, the element exhibits good overall convergence properties in a variety of test problems for plates and shells. Finally, from the computational standpoint, the element is very efficient since all stiffness component matrices are derived in explicit forms.

Saleeb, A. F.↗
NASA NTRS1987

An efficient quadrilateral element for plate bending analysis

A simple, shear flexible, quadrilateral plate element is developed based on the Hellinger/Reissner mixed variational principle with independently assumed displacement and stress fields. The crucial point of the selection of appropriate stress parameters is emphasized in the formulation. For this purpose, a set of guidelines is formulated based on the follwoing considerations: (1) suppression of all kinematic deformation modes, (2) the element has a favorable value for the constraint index in the thin plate limit, and (3) element properties are frame-invariant. For computer implementation the components of the element stiffness matrix are evaluated analytically using the symbolic manipulation package MACSYMA. The effectiveness and practical usefulness of the proposed element are demonstrated by the numerical results of a variety of problems involving thin and moderately thick plates under different loading and support conditions.

Saleeb, A. F.↗
NASA NTRS1987

A quadrilateral shell element using a mixed formulation

A simple quadrilateral shell element consisting of five nodes, four corner nodes and a central node, is developed for linear elastic analysis of thin as well as moderately thick shells. Based on a modified Hellinger-Reissner principle, finite element equations are derived from the assumed displacement and strain fields. By carefully choosing appropriate strain terms, all kinematic deformation modes are suppressed. Although the present element is similar to a displacement-based degenerated shell, no locking is experienced when it is applied to thin shell problems. Five examples are given to illustrate the analysis capability of the shell element. Numerical results indicate that the element shows fast mesh convergence and gives excellent stress predictions.

Saleeb, A. F.↗
NASA NTRS1986

On the numerical performance of three-dimensional thick shell elements using a hybrid/mixed formulation

Three-dimensional thick shell elements with 8, 16, and 18 nodes are formulated by using the hybrid/mixed method. In bending applications, these elements are free from locking effect and give improved stress predictions. Finite element equations are derived from the Hellinger-Reissner variational principle in which both the displacement and stress fields are approximated by independent interpolation functions. For the assumption of stress parameters, three guidelines are followed: (1) suppression of kinematic deformation modes, (2) invariant element property, and (3) the constraint index exhibited by the element, when applied to constrained-media problems, must be greater than or equal to one. Numerical results are presented to show the element's behavior characteristics regarding sensitivity to locking, distortion effect (patch tests), mesh convergence and the accuracy of stress evaluation.

Graf, W.↗
NASA NTRS1986

On the symbolic manipulation and code generation for elasto-plastic material matrices

A computerized procedure for symbolic manipulations and FORTRAN code generation of elastoplastic material matrix for finite element applications is presented. Special emphasis is placed on expression simplifications during intermediate derivations, optimal code generation, and interface with the main program. A systematic procedure is outlined to avoid redundant algebraic manipulations. Symbolic expressions of the derived material stiffness matrix are automatically converted to RATFOR code which is then translated into FORTRAN statements through a preprocessor. To minimize the interface problem with the main program, a template file is prepared so that the translated FORTRAN statements can be merged into the file to form a subroutine (or a submodule). Three constitutive models; namely, von Mises plasticity, the Drucker-Prager model, and a concrete plasticity model, are used as illustrative examples.

Chang, T. Y.↗
NASA NTRS1985

On numerical integration and computer implementation of viscoplastic models

Due to the stringent design requirement for aerospace or nuclear structural components, considerable research interests have been generated on the development of constitutive models for representing the inelastic behavior of metals at elevated temperatures. In particular, a class of unified theories (or viscoplastic constitutive models) have been proposed to simulate material responses such as cyclic plasticity, rate sensitivity, creep deformations, strain hardening or softening, etc. This approach differs from the conventional creep and plasticity theory in that both the creep and plastic deformations are treated as unified time-dependent quantities. Although most of viscoplastic models give better material behavior representation, the associated constitutive differential equations have stiff regimes which present numerical difficulties in time-dependent analysis. In this connection, appropriate solution algorithm must be developed for viscoplastic analysis via finite element method.

Chang, T. Y.↗
NASA NTRS1984

A computer program for predicting nonlinear uniaxial material responses using viscoplastic models

A computer program was developed for predicting nonlinear uniaxial material responses using viscoplastic constitutive models. Four specific models, i.e., those due to Miller, Walker, Krieg-Swearengen-Rhode, and Robinson, are included. Any other unified model is easily implemented into the program in the form of subroutines. Analysis features include stress-strain cycling, creep response, stress relaxation, thermomechanical fatigue loop, or any combination of these responses. An outline is given on the theoretical background of uniaxial constitutive models, analysis procedure, and numerical integration methods for solving the nonlinear constitutive equations. In addition, a discussion on the computer program implementation is also given. Finally, seven numerical examples are included to demonstrate the versatility of the computer program developed.

Chang, T. Y.↗
NASA NTRS1984

Nonlinear Finite Element Analysis of Shells with Large Aspect Ratio

A higher order degenerated shell element with nine nodes was selected for large deformation and post-buckling analysis of thick or thin shells. Elastic-plastic material properties are also included. The post-buckling analysis algorithm is given. Using a square plate, it was demonstrated that the none-node element does not have shear locking effect even if its aspect ratio was increased to the order 10 to the 8th power. Two sample problems are given to illustrate the analysis capability of the shell element.

Chang, T. Y.↗
NASA NTRS1980

Combustion: Structural interaction in a viscoelastic material

The effect of interaction between combustion processes and structural deformation of solid propellant was considered. The combustion analysis was performed on the basis of deformed crack geometry, which was determined from the structural analysis. On the other hand, input data for the structural analysis, such as pressure distribution along the crack boundary and ablation velocity of the crack, were determined from the combustion analysis. The interaction analysis was conducted by combining two computer codes, a combustion analysis code and a general purpose finite element structural analysis code.

Chang, T. Y.↗
NASA NTRS1967

Long-range interatomic forces.

Quadrupole-quadrupole interaction constant for Se and halogen diatomic molecules, discussing long range interatomic forces determination

Chang, T. Y.↗