• BMB Reports
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• v.40 no.3
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• pp.315-324
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• 2007

The novel $\alpha$-amylase purified from locally isolated strain, Bacillus sp. KR-8104, (KRA) (Enzyme Microb Technol; 2005; 36: 666-671) is active in a wide range of pH. The enzyme maximum activity is at pH 4.0 and it retains 90% of activity at pH 3.5. The irreversible thermoinactivation patterns of KRA and the enzyme activity are not changed in the presence and absence of $Ca^{2+}$ and EDTA. Therefore, KRA acts as a $Ca^{2+}$-independent enzyme. Based on circular dichroism (CD) data from thermal unfolding of the enzyme recorded at 222 nm, addition of $Ca^{2+}$ and EDTA similar to its irreversible thermoinactivation, does not influence the thermal denaturation of the enzyme and its Tm. The amino acid sequence of KRA was obtained from the nucleotide sequencing of PCR products of encoding gene. The deduced amino acid sequence of the enzyme revealed a very high sequence homology to Bacillus amyloliquefaciens (BAA) (85% identity, 90% similarity) and Bacillus licheniformis $\alpha$-amylases (BLA) (81% identity, 88% similarity). To elucidate and understand these characteristics of the $\alpha$-amylase, a model of 3D structure of KRA was constructed using the crystal structure of the mutant of BLA as the platform and refined with a molecular dynamics (MD) simulation program. Interestingly enough, there is only one amino acid substitution for KRA in comparison with BLA and BAA in the region involved in the calcium-binding sites. On the other hand, there are many amino acid differences between BLA and KRA at the interface of A and B domains and around the metal triad and active site area. These alterations could have a role in stabilizing the native structure of the loop in the active site cleft and maintenance and stabilization of the putative metal triad-binding site. The amino acid differences at the active site cleft and around the catalytic residues might affect their pKa values and consequently shift its pH profile. In addition, the intrinsic fluorescence intensity of the enzyme at 350 nm does not show considerable change at pH 3.5-7.0.

• Parasites, Hosts and Diseases
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• v.56 no.4
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• pp.325-334
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• 2018

Toxoplasma gondii is an apicomplexan zoonotic protozoan parasite that infects most species of warm-blooded animals, including humans. The heavy incidence and severe or lethal damage caused by T. gondii infection clearly indicate a need for the development of an effective vaccine. T. gondii GRA8 is a member of the dense granules protein family and is used as a marker of acute infection. In the present study, we evaluated the protective immunity induced by DNA vaccination based on a recombinant eukaryotic plasmid, pDsRed2-GRA8, against acute toxoplasmosis in mice. BALB/c mice were intramuscularly immunized with the pDsRed2-GRA8 plasmid and then challenged by infection with the highly virulent GFP-RH strain of T. gondii. The specific immune responses and protective efficacy against T. gondii of this vaccine were analyzed by measuring cytokine and serum antibody titers, splenocyte proliferation assays, and the survival times of mice after challenge. Our results showed that mice immunized with pDsRed2-GRA8 demonstrated specific humoral and cellular responses, induced higher IgG antibody titers with predominant IgG2a production; increased levels of IL-10, IL-12 (p70), $IFN-{\gamma}$, $TNF-{\alpha}$, and splenocyte proliferation; and prolonged survival times compared to those of control mice. The present study showed that DNA immunization with pDsRed2-GRA8 induced humoral and cellular immune responses, and all immunized mice showed greater Th1-type immune responses and longer survival times than those of control mice. These results indicated that T. gondii GRA8 DNA immunization induces a partial protective effect against acute toxoplasmosis.

• Journal of Life Science
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• v.30 no.10
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• pp.919-929
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• 2020

Aromatic compounds are widely used in the chemical, food, polymer, cosmetic, and pharmaceutical industries and are produced by mainly chemical synthesis using benzene, toluene, and xylene or by plant extraction methods. Due to many rising threats, including the depletion of fossil fuels, global warming, the strengthening of international environmental regulations, and the excessive harvesting of plant resources, the microbial production of aromatic compounds using renewable biomass is regarded as a promising alternative. By integrating metabolic engineering with synthetic and systems biology, artificial biosynthetic pathways have been reconstituted from L-tryptophan biosynthetic pathway in relevant microorganisms, such as Escherichia coli and Corynebacterium glutamicum, enabling the production of a variety of value-added aromatic compounds, such as 5-hydroxytryptophan, serotonin, melatonin, 7-chloro-L-tryptophan, 7-bromo-L-tryptophan, indigo, indirubin, indole-3-acetic acid, violacein, and dexoyviolacein. In this review, we summarize the characteristics, usage, and biosynthetic pathways of these aromatic compounds and highlight the latest metabolic engineering strategies for the microbial production of aromatic compounds and suitable solution strategies to overcome problems in increasing production titers. It is expected that strain development based on systems metabolic engineering and the optimization of media and bioprocesses using renewable biomass will enable the development of commercially viable technologies for the microbial production of many aromatic compounds.

• Fire Science and Engineering
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• v.24 no.6
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• pp.7-12
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• 2010

This paper is aimed the measurement and estimation the thermal interrelation based on leakage current. The leakage models are analyzed by parameters in open and closed (Panel) states and with wood dust and rainwater, charcoal powder in leakage current occurred point. The thermal characteristic of open state that leakage current is inputted in 90 mA is higher in charcoal powder ($105^{\circ}C$) than in wood dust and rainwater ($88.8^{\circ}C$). Charcoal powder that is inputted in 110 mA shows high thermal characteristic ($238.6^{\circ}C$) and the thermal ($238.6^{\circ}C$) is caused by material strain of electrical wire insulation and Panel. The closed state in 90 mA is higher in charcoal powder ($250.6^{\circ}C$) than in wood dust and rainwater ($90.8^{\circ}C$). The open and closed state appear thermal characteristics of two times in charcoal powder than in wood dust and rainwater. The thermal data that are open and close states are used electrical fire investigation and thermal characteristic for Leakage current.

• Journal of the Computational Structural Engineering Institute of Korea
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• v.22 no.5
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• pp.463-474
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• 2009

Double symmetric patch repair of existing structures always causes membrane action only, however, in many cases this technique is not practical. On the other hand, the bending stiffness of the patch and the skin increases as tensile loading is increased and affects the bending deformation significantly in the case of single-sided patch repair. In this study, the p-convergent full layerwise model has been proposed to determine the stress concentration factor in the vicinity of a circular hole as well as across the thickness of plates with single-sided patch repair. In assumed displacement field, the strain-displacement relations and 3-D constitutive equations of a layer are obtained by the combination of 2-D and 3-D hierarchical shape functions. The transfinite mapping technique has been used to represent a circular boundary and Gauss-Lobatto numerical integration is implemented in order to directly obtain stresses occurred at the nodal points of each layer without other extrapolation techniques. The accuracy and simplicity of the present model are verified with comparison of the previous results in literatures using experiment and conventional 3-D finite element. Also, the bending effect has been investigated with various patch types like square, circular and annular shape.

• Journal of the Computational Structural Engineering Institute of Korea
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• v.26 no.6
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• pp.423-430
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• 2013

In this research, a paramteric study to account for the effect of interfacial strength and nanotube agglomeration on the elastoplastic behavior of carbon nanotube reinforced polypropylene composites is performed. At first, the elastoplastic behavior of nanocomposites is predicted from molecular dynamics(MD) simulations. By combining the MD simulation results with the nonlinear micromechanics model based on the Mori-Tanaka model, a two-step domain decomposition method is applied to inversely identify the elastoplastic behavior of adsorption interphase zone inside nanocomposites. In nonlinear micromechanics model, the secant moduli method combined with field fluctuation method is used to predict the elastoplastic behavior of nanocomposites. To account for the imperfect material interface between nanotube and matrix polymer, displacement discontinuity condition is applied to the micromechanics model. Using the elastoplastic behavior of the adsorption interphase zone obtained from the present study, stress-strain relation of nanocomposites at various interfacial bonding condition and local nanotube agglomeration is predicted from nonlinear micromechanics model with and without the adsorption interphase zone. As a result, it has been found that local nanotube agglomeration is the most important design factor to maximize reinforcing effect of nanotube in elastic and plastic behavior.

• Applied Microscopy
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• v.17 no.1
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• pp.47-64
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• 1987

Morphological difference of the renal glomerulus at different age groups have been studied in young (three month-old), adult (twelve month-old) and old (thirty month-old) Fisher strain 344 rats. Pieces of the tissues were taken from renal corticies prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde (0.1 M Millonig's phosphate buffer, pH 7.3), following by post-fixation with 1% osmium tetroxide (0.1 M Millonig's phosphate buffer, pH 7.3) and embedded within Araldite. The ultrathin sections contrasted with uranyl acetate and lead citrate were observed under a JEM 100CX electron microscope. The mean thickness of glomerular basal lamina and Bowman's capsule were determined by measuring the thinnest portion of basal lamina, and by taking the average of 50 readings from electron micrographs at different ages. The numerical changes of the slit pores were compared based upon the numbers over the length of 10um of glomerular basal lamina. The results were as follow: 1. The thickness of glomerular basal lamina is increased during aging; 140.4 nm in young rats, 270.0 nm in adult ones, and 437.8 nm in old ones. 2. The thickness of basal lamina of parietal cells of Bowman's capsule is 187.5 nm in young rats, 914.0 nm in adult ones, and 2850.0 nm in old ones. 3. The numbers of the slit pores of basal lamina are reduced during aging, 30.3 slit pores/$10{\mu}m$ in adult ones, and 24.2 slit pores/$10{\mu}m$ in old ones. 4. Accumulation of dense intracytoplasmic filamentous material in the parietal cells of Bowman's capsule is increased in the vicinity of the basal lamina during aging. The proximal tubule-like epithelial cell in Bowman's capsule is observed at one glomerulus in a young rat. 5. The endothelial cells are edematous and form balloon-like structure protruding into capillary lumen in young and old rats. 6. Cytoplasm of the podocyte shows a variety of alteration during aging, such as swelling of mitochondria and of endoplasmic reticulum, and increase of microtubules, microfilaments, lysosomes and lamellated myelin structures, etc. Accumulation of dense intracytoplasmic material in the foot processes is increased in the vicinity of the basal lamina during aging. The podocytic membrane-like structures are seen in young and o]d rats. 7. The mesangial matrices and mesangial cells are increased during aging, and slight swelling of endoplasmic reticulum and Golgi cisternae in young and old rats.

• Tunnel and Underground Space
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• v.17 no.5
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• pp.350-359
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• 2007

Most large cut slopes of open pit mines, roadways, and railways are steeply inclined and composed with rocks that do not contain soils. However, these rock slopes suffer both weathering and fragmentation. In the case of steep slopes, falling rock and collapse of a slope may often occur due to surface erosion. Cast-in place concrete and rubble work are the most widely used earth structure-based pressure supports that act as restraints against the collapse of the rock slope. In order to overcome the shortcomings of conventional retaining walls, a segmental grid retaining wall is being used with connects precasted segments to construct the wall. In this study, laboratory model test was conducted to estimate deformation behavior of segmental grid retaining wall with configuration of rear strecher, height and inclination of the wall. In order to examine the behavior characteristics of a segmental grid retaining wall, this research analyzes the aspects of spacial displacement through relative displacement according to change in the inclination of the wall. Also, the walls behavior according to the formation and status of the rear stretcher which serves the role of transferring the load from the header and the stretcher which make up the wall, the displacement of backfill materials in the wall, and the location of the maximum load were surveyed and the characteristics of displacement in the segmental grid retaining wall were observed. The test results of the segmental grid retaining wall showed that there was a sudden increase in failure load according to the decrease in the wall's height and the size of the in was greatly decreased. Furthermore, it revealed that with identical inclination and height, the structure of the rear stitcher did not greatly affect the starting point or size of maximum horizontal displacement, but rather had a stronger effect on the inclination of the wall.

• Journal of the Korean Society for Nondestructive Testing
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• v.24 no.3
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• pp.259-267
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• 2004

A finite element method is used to simulate interaction of laser-based ultrasounds with surface breaking tracks in elastic media. The laser line source focused on the surface of semi-infinite medium is modeled as a shear dipole in 2-D plane strain finite elements. The shear dipole-finite clement model is found to give correct directivity patterns for generated longitudinal and shear waves. The interaction of surface waves with surface breaking cracks (2-D machined slot) is considered in two ways. Both the source and receiver are fixed with respect to the cracks in the first case, while the source is moving in another case. It is shown that the crack depth tested in the range of 0.3-5.0mm $({\lambda}_R/d=0.21{\sim}3.45)$ can be measured using the corner reflected waves produced by the fixed laser source. The moving laser source is found to cause a large amplitude change of reflected waves near crack, and the crack whose depth is one order lower than the wavelength ran be detected from this change.

• Parasites, Hosts and Diseases
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• v.35 no.2
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• pp.79-86
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• 1997

Toxoplasmn gonnii, an intracellular protozoan infecting many kinds of eukaryotic cells, has been used to experimentally infect macrophages, epithelial cells, fibroblasts, and various cancer cells, but rarely T and B Iymphocytes or granulocytes. The present study was performed to determine the susceptibility of murine (BALB/c or CBA) splenic T and B llrnphocytes, and granulocytes to infection trio T. gondii RH tachyzoites. The ultrastructure of the infected host cells was observed by TEM, and the degree of intracellular parasite proliferation was quantified using 3H-uracil uptake assay. At 24 hrs post-culture, the host cell cytoplasm was found to contain 1 or 2, or a maximum of 7-8 tachyzoites. Infected T Iymphocytes demonstrated a peripherally displaced nucleus, a parasitophorous vacuole enveloping the parasite, and an increased number of mitochondria. In B Iymphocytes infected with tachyzoites, RER was not well developed compared to uninfected B Iymphocytes. Uninfected granulocytes contained many electron dense granules, but T. gondii-infected granulocytes demonstrated a decreased number of granules. Based on the 3H-uracil uptake assay. the susceptibility of T and B Iymphocytes, and granulocytes, to infection with T. gonnii tachyzoites was fairly high irrespective of cell type and strain of mouse. This strongly suggests deterioration in the functioning of infected host immune cells.