• The Korean Journal of Physiology and Pharmacology
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• 제24권3호
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• pp.267-276
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• 2020

T In the present study, we investigated the effect of oncogenic H-Ras on rat mdr1b expression in NIH3T3 cells. The constitutive expression of H-Ras^V12 was found to downregulate the mdr1b promoter activity and mdr1b mRNA expression. The doxorubicin-induced mdr1b promoter activity of the H-Ras^V12 expressing NIH3T3 cells was markedly lower than that of control NIH3T3 cells. Additionally, there is a positive correlation between the level of H-Ras^V12 expression and a sensitivity to doxorubicin toxicity. To examine the detailed mechanism of H-Ras^V12-mediated down-regulation of mdr1b expression, antioxidant N-acetylcysteine (NAC) and NADPH oxidase inhibitor diphenylene iodonium (DPI) were used. Pretreating cells with either NAC or DPI significantly enhanced the oncogenic H-Ras-mediated down-regulation of mdr1b expression and markedly prevented doxorubicin-induced cell death. Moreover, NAC and DPI treatment led to a decrease in ERK activity, and the ERK inhibitors PD98059 or U0126 enhanced the mdr1b-Luc activity of H-Ras^V12-NIH3T3 and reduced doxorubicin-induced apoptosis. These data suggest that Ras^V12 expression could downregulate mdr1b expression through intracellular reactive oxygen species (ROS) production, and ERK activation induced by ROS, is at least in part, contributed to the downregulation of mdr1b expression.

• Journal of Microbiology and Biotechnology
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• 제16권6호
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• pp.952-960
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• 2006

As a unicellular green alga that possesses many of the metabolic pathways present in higher plants, Chlorelia offers many advantages for expression of heterologous proteins. Since strong and constitutive promoters are necessary for efficient expression in heterologous expression systems, the development of such promoters for use in the Chlorella system was the aim of this study. Proteins encoded by the early genes of algal viruses are expressed before viral replication, probably by the host transcriptional machinery, and the promoters of these genes might be useful for heterologous expression in Chlorella. In this study, putative promoter regions of DNA polymerase, ATP-dependent DNA ligase, and chitinase genes were amplified from eight Korean Chlorella virus isolates by using primer sets designed based on the sequence of the genome of PBCV-1, the prototype of the Phycodnaviridae. These putative promoter regions were found to contain several cis-acting elements for transcription factors, including the TATA, CAAT, NTBBF1, GATA, and CCAAT boxes. The amplified promoter regions were placed into Chlorella transformation vectors containing a green fluorescence protein (GFP) reporter gene and the Sh ble gene for phleomycin resistance. C. vulgaris protoplasts were transformed and then selected with phleomycin. The GFP fluorescence intensities of cells transformed with chitinase, DNA polymerase, and DNA ligase gene promoter-GFP fusion constructs were 101.5, 100.8, and 95.8%, respectively, of that of CaMV 35S-GFP-transformed Chlorella cells. These results demonstrate that these viral promoters are active in transformed Chlorella.

• Microbiology and Biotechnology Letters
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• 제48권3호
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• pp.296-302
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• 2020

Rapamycin, derived from Streptomyces rapamycinicus, is an important bioactive compound having a therapeutic value in managing Parkinson's disease, rheumatoid arthritis, cancer, and AIDS. Because of its pharmaceutical activity, studies over the past decade have focused on the biosynthesis of rapamycin to enhance its yield. In this study, the effect of rapG on rapamycin production was investigated. The rapG expression vector was constructed by utilizing the integration vector pSET152 under the control of the erythromycin resistance gene (ermE^⁎), a strong constitutive promoter. The rapamycin yield of wild type (WT) and WT/rapG overexpression mutant strains, under fermentation conditions, was analyzed by high-performance liquid chromatography (HPLC). Our results revealed that overexpression of rapG increased rapamycin production by approximately 4.9-fold (211.4 mg/l) in MD1 containing 15 g/l of glycerol, compared to that of the WT strain. It was also found that Illicium verum powder (10 g/l), containing shikimic acid, enhanced rapamycin production in both WT and WT/rapG strains. Moreover, the amount of rapamycin produced by the WT/rapG strain was statistically higher than that produced by the WT strain. In conclusion, the addition 15 g/l glycerol and 15 g/l I. verum powder produced the optimal conditions for rapamycin production by WT and WT/rapG strains.

• Computational Structural Engineering
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• 제9권1호
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• pp.151-159
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• 1996

A finite element formulation based on the CFT(Compression Field Theory) concept such as the effect of compression softening in cracked concrete, and macroscopic and rotating crack models etc. was presented for the nonlinear behaviour of structural concrete. In this category, tangential or secant material stiffnesses for cracked concrete were also defined and discussed in view of the iterative solution schemes for nonlinear equations. Considering the computational efficiency and the ability of modelling the post-ultimate behaviour as major concerns, the incremental displacement solution algorithm involving initial material stiffnesses and the relaxation procedure for fast convergence was adopted and formulated in a type of 8-noded quadrilateral isoparametric elements. The analysis program NASCOM(Nonlinear Analysis of structrual Concrete by FEM : Monotonic Loading) developed baed on the CFT constitutive relationships and the incremetal solution strategy described enables the predictions of strength and deformation capacities in a full range. crack patterns and their corresponding widths, and yield extents of reinforcement. As the verfication purpose of NASCOM, the prediction of Cervenka's panel test results including the load resistance and the deformation history was made. A limited number of predictions indicate a good correlation in a general sense.

• Nuclear Engineering and Technology
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• 제52권2호
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• pp.417-428
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• 2020

Investigations of the commercial aircraft impact effect on nuclear island infrastructures have been drawing extensive attention, and this paper aims to perform the safety assessment of Generation III nuclear power plant (NPP) buildings subjected to typical commercial aircrafts crash. At present Part III, the local damage of the rigid components of aircraft, e.g., engine and landing gear, impacting the steel concrete (SC) structures of NPP containment is mainly discussed. Two typical SC target panels with the thicknesses of 40 mm and 100 mm, as well as the steel cylindrical projectile with a mass of 2.15 kg and a diameter of 80 mm are fabricated. By using a large-caliber air gas gun, both the projectile penetration and perforation test are conducted, in which the striking velocities were ranged from 96 m/s to 157 m/s. The bulging velocity and the maximal deflection of rear steel plate, as well as penetration depth of projectile are derived, and the local deformation and failure modes of SC panels are assessed experimentally. Then, the commercial finite element program LS-DYNA is utilized to perform the numerical simulations, by comparisons with the experimental and simulated projectile impact process and SC panel damage, the numerical algorithm, constitutive models and the corresponding parameters are verified. The present work can provide helpful references for the evaluation of the local impact resistance of NPP buildings against the aircraft engine.

• Tunnel and Underground Space
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• 제11권3호
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• pp.225-236
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• 2001

The roughness of rock joint is one of the most important parameters in developing the shear resistance and the tendency of dilation. Due to the damage accumulated with shearing displacement, the roughness angle is lowered continuously. It is known that dilation, shear strength hardening, and softening are directly related to the degradation of asperities. Much effort has been directed to incorporate the complicated damage mechanism of asperities into a constitutive model fur rock joints. This study presents an elasto-plastic formulation of joint behavior including elastic deformability, dilatancy and asperity surface damage. It is postulated that the plastic portion of incremental displacement 7an be decomposed into contributions from both sliding along the asperity surface and damage of asperity. Numerical cyclic shear tests are presented to illustrate th? performance of the derived incremental stress-displacement relation. A laboratory cyclic shear test is also simulated. Numerical examples reveal that the elasto-plastic joints model is promising.

• Journal of Ocean Engineering and Technology
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• 제29권3호
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• pp.241-248
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• 2015

A subsea pipeline for oil/gas transportation or gas injection is subjected to extreme variations in internal pressure and temperature, which can involve a strain rate effect on the pipeline material. This paper describes the flow stress characteristics of a pipeline material called API 5L X52N PSL2, using and experimental approach. High-speed tensile tests were carried out for two metal samples taken from the base and weld parts. The target temperature was 100℃, but two other temperature levels of –20℃and 0℃ were taken into account. Three strain rates were also considered for each temperature level: quasi static, 1/s, and 10/s. Flow stress data were proposed for each temperature level according to these strain rates. The dynamic hardening behaviors of the base and weld metals appeared to be nonlinear on the log-scale strain rate axis. A very high material constant value was required for the Cowper-Symonds constitutive equation to support the experimental results.

• Smart Structures and Systems
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• 제24권1호
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• pp.127-139
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• 2019

Residual drifts after an earthquake can incur huge repair costs and might need to replace the infrastructure because of its non-reparability. Proper functioning of bridges is also essential in the aftermath of an earthquake. In order to mitigate pounding and unseating damage of bridges subjected to earthquakes, a self-adaptive Ni-Ti shape memory alloy (SMA)-cable-based frictional sliding bearing (SMAFSB) is proposed considering self-adaptive centering, high energy dissipation, better fatigue, and corrosion resistance from SMA-cable component. The developed novel bearing is associated with the properties of modularity, replaceability, and earthquake isolation capacity, which could reduce the repair time and increase the resilience of highway bridges. To evaluate the super-elasticity of the SMA-cable, pseudo-static tests and numerical simulation on the SMA-cable specimens with a diameter of 7 mm are conducted and one dimensional (1D) constitutive hysteretic model of the SMAFSB is developed considering the effects of gap, self-centering, and high energy dissipation. Two types of the SMAFSB (i.e., movable and fixed SMAFSBs) are applied to a two-span continuous reinforced concrete (RC) bridge. The seismic vulnerabilities of the RC bridge, utilizing movable SMAFSB with the constant gap size of 60 mm and the fixed SMAFSBs with different gap sizes (e.g., 0, 30, and 60 mm), are assessed at component and system levels, respectively. It can be observed that the fixed SMAFSB with a gap of 30 mm gained the most retrofitting effect among the three cases.

• The Plant Pathology Journal
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• 제37권2호
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• pp.182-193
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• 2021

The transcription factor SHE1 was identified as an interacting partner with the cucumber mosaic virus (CMV) 1a protein in the yeast two-hybrid system, by a pull-down assay, and via bimolecular fluorescent complementation. Using fluorescent-tagged proteins and confocal microscopy, the CMV 1a protein itself was found distributed predominantly between the nucleus and the tonoplast membrane, although it was also found in speckles in the cytoplasm. The SHE1 protein was localized in the nucleus, but in the presence of the CMV 1a protein was partitioned between the nucleus and the tonoplast membrane. SHE1 expression was induced by infection of tobacco with four tested viruses: CMV, tobacco mosaic virus, potato virus X and potato virus Y. Transgenic tobacco expressing the CMV 1a protein showed constitutive expression of SHE1, indicating that the CMV 1a protein may be responsible for its induction. However, previously, such plants also were shown to have less resistance to local and systemic movement of tobacco mosaic virus (TMV) expressing the green fluorescent protein, suggesting that the CMV 1a protein may act to prevent the function of the SHE1 protein. SHE1 is a member of the AP2/ERF class of transcription factors and is conserved in sequence in several Nicotiana species, although two clades of SHE1 could be discerned, including both different Nicotiana species and cultivars of tobacco, varying by the presence of particular insertions or deletions.

• Asian Pacific Journal of Cancer Prevention
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• 제15권10호
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• pp.4353-4358
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• 2014

Background: PI3/AKT and NF-kB signaling pathways are constitutively active in acute myeloid leukemia and cross-talk between the two has been shown in various cancers. However, their role in acute myeloid leukemia has not been completely explored. We therefore used cell penetrating inhibitor peptides to define the contributions of AKT and NF-kB to survival and multi drug resistance (MDR) in HL-60 cells. Materials and Methods: Inhibition of AKT and NF-kB activity by AKT inhibitor peptide and NBD inhibitor peptide, respectively, resulted in decreased expression of mRNA for the MDR1 gene as assessed by real time PCR. In addition, treatment of HL-60 cells with AKT and NBD inhibitor peptides led to inhibition of cell viability and induction of apoptosis in a dose dependent manner as detected by flow cytometer. Results: Finally, co-treatment of HL-60 cells with sub-optimal doses of AKT and NBD inhibitor peptides led to synergistic apoptotic responses in AML cells. Conclusions: These data support a strong biological link between NF-kB and PI3-kinase/AKT pathways in the modulation of antiapoptotic and multi drug resistant effects in AML cells. Synergistic targeting of these pathways using NF-kB and PI3-kinase/AK inhibitor peptides may have a therapeutic potential for AML and possibly other malignancies with constitutive activation of these pathways.