The FBJ murine osteosarcoma viral oncogene homolog B (FosB) gene is located at chromosome 19, and encodes 43 Kda protein. Functionally, the FosB gene is important for differentiation, development, and pathogenesis. Furthermore, the FosB gene is suggested as possible biomarker for tracing disease prognosis. In this study, we constructed plasmid containing a FosB promoter region and evaluate its promoter activity. We analyzed the putative promoter region in FosB genomic DNA using bioinformatics program, and we found important regulatory elements in 1 Kb upstream from transcription start site (TSS). Therefore, we performed polymerase chain reaction (PCR) amplification on region from-1,555 upstream to +73 of the FosB genomic DNA, and PCR product was inserted into TA vector to create the $TA-1^{st}FosBp$ plasmid. We then prepared the primer sets, which contain a restriction enzyme site for Kpn1 and Nhe1, in order to reinsert into the TA vector to prepare $TA-2^{nd}FosBp$ plasmid. It was finally subcloned into pGL3-luc vector after enzyme cutting. To evaluate whether the cloned plasmid is useful in cell based experiment, we performed luciferase assay with pGL3-FosBp-luctransfection. FosB promoter activity was increased compared to empty vector, and this activity was significantly increased by treatment of doxorubicin and taxol. We obtained consistent data on regulation of FosB gene expression after anticancer drug treatment using Western blot analysis. The results suggest that promoter cloning of the human FosB gene is very useful for studying gene expression and analyzing biomarkers.
Luminescent lanthanide complexes have been overviewed for advanced photonics applications. Lanthanide(III) ions ($Ln^{3+}$) were encapsulated by the luminescent ligands such as metalloporphyrins, naphthalenes, anthracene, push-pull diketone derivatives and boron dipyrromethene(bodipy). The energy levels of the luminescent ligands were tailored to maintain the effective energy transfer process from luminescent ligands to $Ln^{3+}$ ions for getting a higher optical amplification gain. Also, key parameters for emission enhancement and efficient energy transfer pathways for the sensitization of $Ln^{3+}$ ions by luminescent ligands were investigated. Furthermore, to enhance the optophysical properties of novel luminescent $Ln^{3+}$ complexes, aryl ether-functionalized dendrons as photon antennas have been incorporated into luminescent $Ln^{3+}$ complexes, yielding novel $Ln^{3+}$-cored dendrimer complex such as metalloporphyrins, naphthalenes, and anthracenes bearing the Fr$\acute{e}$chet aryl-ether dendrons, namely, ($Er^{3+}-[Gn-Pt-Por]_3$ (terpy), $Er^{3+}-[Gn-Naph]_3$(terpy) and $Er^{3+}-[Gn-An]_3$(terpy)). These complexs showed much stronger near-IR emission bands at 1530 nm, originated from the 4f-4f electronic transition of the first excited state ($^4I_{13/2}$) to the ground state ($^4I_{15/2}$) of the partially filled 4f shell. A significant decrease in the fluorescence of metalloporphyrins, naphthalenes and anthracene ligand were accompanied by a strong increase in the near IR emission of the $Ln^{3+}$ ions. The near IR emission intensities of $Ln^{3+}$ ions in the lanthanide(III)-encapsulated dendrimer complexes were dramatically enhanced with increasing the generation number (n) of dendrons, due to the site-isolation and the light-harvesting(LH) effects. Furthermore, it was first attempted to distinguish between the site-isolation and the light-harvesting effects in the present complexes. In this review, synthesis and photophysical studies of inert and stable luminescent $Ln^{3+}$ complexes will be dealt for the advanced photonics applications. Also, the review will include the exploratory investigation of the key parameters for emission enhancement and the effective energy transfer pathways from luminescent ligands to $Ln^{3+}$ ions with $Ln^{3+}$-chelated prototype complexes.
Journal of the Earthquake Engineering Society of Korea
/
v.11
no.1
s.53
/
pp.89-97
/
2007
Parameters including the seismic sources and the elastic wave propagation characteristics were analysed using the observed ground motions from 12 Fukuoka region earthquakes. The Levenberg-Marquardt algorithm was applied to invert all the variables non-linearly and simultaneously with S wave energy in fiequency domain. Average stress drop of 12 events and local attenuation parameter $\kappa$ under seismic stations were estimated to about 79.2-bar and 0.043 respectively. Regional attenuation parameter, Qo and ${\eta}$, were also estimated to be about 248.1 and 0.558 respectively. Low value of Qo seems to caused by inhomogeneous tectonic characteristics between Japan island and southern Korean peninsula. $\kappa$ values are much higher than that characterizing EUS (Eastern United States) region, and nearly similar to that of WUS (Western Waited States) region. If the informations on site specific amplification of all the seismic stations are known, $\kappa$ values can be estimated more precisely. All the values including the seismic sources and the site and crustal scale propagation characteristics can be used as seismic design parameters.
One-dimensional equivalent linear site response analysis is widely used in practice due to its simplicity, requiring only few input parameters, and low computational cost. The main limitation of the procedure is that it is essentially a linear method, in which the time dependent change in the soil properties cannot be modeled and constant values of shear modulus and damping is used throughout the duration of the analysis. Various forms of modified equivalent linear analyses have been developed to enhance the accuracy of the equivalent linear method by incorporating the dependence of the shear strain with the loading frequency. The methods are identical in that it uses the shear strain Fourier spectrum as the backbone of the analysis, but differ in the method in which the strain Fourier spectrum is smoothed. This study used two domestically measured soil profiles to perform a series of nonlinear, equivalent linear, and modified equivalent linear site response analyses to verify the accuracy of two modified procedures. The results of the analyses indicate that the modified equivalent linear analysis can highly overestimate the amplification of the high frequency components of the ground motion. The degree of overestimation is dependent on the characteristics of the input ground motion. Use of a motion rich in high frequency contents can result in unrealistic response.
KSCE Journal of Civil and Environmental Engineering Research
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v.42
no.1
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pp.35-44
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2022
The VS30 map is used as a key variable for site amplification in the ShakeMap, which predicts ground motion at any site. However, no VS30 map considering Korean geology and geomorphology has been developed yet. To develop a proxy-based VS30 map, we used 1,101 VS profiles obtained from a geophysical survey and collected proxy layers of geological and topographical information for the Korean Peninsula. Then, VS30 prediction models were developed using linear regression analysis for each geological age considering the distribution of VS30. As a result, models depending on geomorphology were suggested per each geologic group, including Quaternary, Fill, Ocean, Mesozoic group and Precambrian. Resolution of map is doubled from that of VS30 map by U.S. Geological Survey (USGS). Standard deviation of residual in natural log of proxy-based VS30 map is 0.233, whereas standard deviation of slope-based USGS VS30 map is 0.387. Therefore, the proxy-based VS30 map developed in this study is expected to have less uncertainty and to contribute to predicting more accurately the ground motion amplitude.
We installed temporary strong motion seismometers at the ground surface, 1 m, 2 m, and 9 m at an existing seismic station that houses permanent seismometers installed at 20 m and 100 m, to investigate the influence of installation depth on the recorded ambient and anthropogenic noise level and the characteristics of earthquake signals. Analysis of the ambient noise shows that anthropogenic noise dominates where vibration period T < 1 s at the studied site, whereas wind speed appears to be strongly correlated with the noise level at T > 1 s. Frequency-wavenumber analysis of 2D seismometer array suggests that ambient noise in short periods are predominantly body waves, rather than surface waves. The level of ambient noise was low at 9 m and 20 m, but strong amplification of noise level at T < 0.1 s was observed at the shallow seismometers. Both the active-source test result and the recorded earthquake data demonstrated that the signal level is decreased with the increase of depth. Our result also shows that recorded motions at the ground and 1 m are strongly amplified at 20 Hz (T = 0.05 s), likely due to the resonance of the 3 m thick soil layer. This study demonstrates that analysis of ambient and active-source vibration may help find optimal installation depth of strong motion seismometers. We expect that further research considering various noise environments and geological conditions will be helpful in establishing a guideline for optimal installation of strong motion seismometers.
Background: Interleukin-4 plays an important role in pathogenesis of asthma, especially in developing atopy by means of switching B lymphocytes to produce IgE. It has been shown that there is polymorphism in the Interleukin-4 promoter region, transversion of cytosine to thymine at-598 from translation initiation site of IL-4 gene. There has also been quite a few works to reveal the role of the polymorphism of IL-4 gene in patients with asthma. We performed this investigation to determine the role of the polymorphism in the severity of symptoms of patients with asthma. We also examined the frequency and the type of the polymorphism in asthmatics compared with non-asthmatics as well. Method: The subjects enrolled in this study were 49 asthmatics and 33 non-asthmatics. All the asthmatics were classified as mild and moderate to severe by the NHLBI/WHO Workshop. DNA from both asthmatics and non-asthmatics was extracted, then performed ARMS(Amplification Refractory Mutation System) as well as RFLP using BsmFl restriction enzyme in order to confirm the polymorphism of Il-4 gene. Results: There was no significant difference in the occurrence of polymorphism of the IL-4 promoter sequence between asthm and non-asthma groups(P=0.7). Among those with polymorphisms, the number of C/C type was slightly more than C/T type in both asthmatics and non-asthmatics, 26 vs 21 in asthmatics and 18 vs 15 in non-asthmatics, which was, however, insignificant statistically. No significant relationship between the severity of asthma and the polymorphism was found(P=0.7). Conclusion: There was no significant difference between the severity of asthma and the IL-4 promoter polymorphism(P=0.709). Interestingly, the frequency of the polymorphism in both asthmatics as well as non-asthmatics was found to be even higher than that occurred in Caucasians. However, no significant difference in the frequency of the polymorphism was found in both groups.
Proceedings of the Korean Vacuum Society Conference
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2013.08a
/
pp.88-89
/
2013
A variety of influenza A viruses from animal hosts are continuously prevalent throughout the world which cause human epidemics resulting millions of human infections and enormous industrial and economic damages. Thus, early diagnosis of such pathogen is of paramount importance for biomedical examination and public healthcare screening. To approach this issue, here we propose a fully integrated Rotary genetic analysis system, called Rotary Genetic Analyzer, for on-site detection of influenza A viruses with high speed. The Rotary Genetic Analyzer is made up of four parts including a disposable microchip, a servo motor for precise and high rate spinning of the chip, thermal blocks for temperature control, and a miniaturized optical fluorescence detector as shown Fig. 1. A thermal block made from duralumin is integrated with a film heater at the bottom and a resistance temperature detector (RTD) in the middle. For the efficient performance of RT-PCR, three thermal blocks are placed on the Rotary stage and the temperature of each block is corresponded to the thermal cycling, namely $95^{\circ}C$ (denature), $58^{\circ}C$ (annealing), and $72^{\circ}C$ (extension). Rotary RT-PCR was performed to amplify the target gene which was monitored by an optical fluorescent detector above the extension block. A disposable microdevice (10 cm diameter) consists of a solid-phase extraction based sample pretreatment unit, bead chamber, and 4 ${\mu}L$ of the PCR chamber as shown Fig. 2. The microchip is fabricated using a patterned polycarbonate (PC) sheet with 1 mm thickness and a PC film with 130 ${\mu}m$ thickness, which layers are thermally bonded at $138^{\circ}C$ using acetone vapour. Silicatreated microglass beads with 150~212 ${\mu}L$ diameter are introduced into the sample pretreatment chambers and held in place by weir structure for construction of solid-phase extraction system. Fig. 3 shows strobed images of sequential loading of three samples. Three samples were loaded into the reservoir simultaneously (Fig. 3A), then the influenza A H3N2 viral RNA sample was loaded at 5000 RPM for 10 sec (Fig. 3B). Washing buffer was followed at 5000 RPM for 5 min (Fig. 3C), and angular frequency was decreased to 100 RPM for siphon priming of PCR cocktail to the channel as shown in Figure 3D. Finally the PCR cocktail was loaded to the bead chamber at 2000 RPM for 10 sec, and then RPM was increased up to 5000 RPM for 1 min to obtain the as much as PCR cocktail containing the RNA template (Fig. 3E). In this system, the wastes from RNA samples and washing buffer were transported to the waste chamber, which is fully filled to the chamber with precise optimization. Then, the PCR cocktail was able to transport to the PCR chamber. Fig. 3F shows the final image of the sample pretreatment. PCR cocktail containing RNA template is successfully isolated from waste. To detect the influenza A H3N2 virus, the purified RNA with PCR cocktail in the PCR chamber was amplified by using performed the RNA capture on the proposed microdevice. The fluorescence images were described in Figure 4A at the 0, 40 cycles. The fluorescence signal (40 cycle) was drastically increased confirming the influenza A H3N2 virus. The real-time profiles were successfully obtained using the optical fluorescence detector as shown in Figure 4B. The Rotary PCR and off-chip PCR were compared with same amount of influenza A H3N2 virus. The Ct value of Rotary PCR was smaller than the off-chip PCR without contamination. The whole process of the sample pretreatment and RT-PCR could be accomplished in 30 min on the fully integrated Rotary Genetic Analyzer system. We have demonstrated a fully integrated and portable Rotary Genetic Analyzer for detection of the gene expression of influenza A virus, which has 'Sample-in-answer-out' capability including sample pretreatment, rotary amplification, and optical detection. Target gene amplification was real-time monitored using the integrated Rotary Genetic Analyzer system.
In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.
The horizontal and vertical response spectra using the observed ground motion from the recent 5 macro earthquakes were analysed and then were compared to both the seismic design response spectra(Reg Guide 1.60), applied to the domestic nuclear power plants, and the Korean Standard Design Response Spectrum for general structures and buildings(1997). 74 horizontal and 89 vertical observed ground motions, without considering soil types, were used for normalization with respect to the peak acceleration value of each ground motion. The results showed that the horizontal MPOSD(Mean Plus One Sigma Standard Deviation) response spectra revealed much higher values for the whole frequency bands above 1 Hz than Reg. Guide(1.60). For the vertical response spectra, the results showed slightly higher than just between 7 and 8 Hz frequency band. The results were also compared to the Korean Standard Response Spectrum for the 3 different soil types and showed that the horizontal MPOSD response spectra revealed much higher values for the whole periods below 2 second(0.5 Hz) than those of SE soil type. The vertical response spectra showed similar to the values of the Korean Standard Response Spectrum of SD soil type. These spectral values dependent on frequency could be related to characteristics of the domestic crustal attenuation and the effect of each site amplification. However, through the qualitative improvements and quantitative enhancement of the observed ground motions, the conservation of horizontal seismic design response spectrum should be considered more significantly for the whole frequency bands above the 1 Hz.
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