• Animal Bioscience
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• v.36 no.3
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• pp.429-440
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• 2023

Objective: 12-oxo-5Z,8Z,10E,14Z-eicosatetraenoic acid (12-KETE), a metabolite of arachidonic acid, is a strong candidate signal for placenta separation following calf discharge at delivery. In the present study, the effects of 12-KETE on bovine trophoblast cells were investigated to determine its function in the placentome at delivery. Methods: Bovine trophoblast cells derived from blastocysts were used. They were cocultured with or without fibroblasts derived from bovine placentome and/or bovine uterine epithelial cells. 12-KETE was added to the culture medium. Results: Bovine trophoblast cells contained binucleate cells and strongly expressed caudal type homeobox 2 (CDX-2) genes. Addition of 12-KETE to the trophoblast cell colony without feeder cells or that on a fibroblast monolayer induced rapid exfoliation of the colony. After 12-KETE addition, trophoblast cells emitted strong fluorescence caused by the degradation of dye-quenched collagen, indicating that 12-KETE activated matrix metalloproteinase of the trophoblast cells. Exfoliated cell colonies were stained with YOPRO-1, but not propidium iodide (PI). Moreover, DNA fragmentation and Bcl-2 associated X protein (Bax) gene (apoptosis stimulator) upregulation were observed in exfoliated cells, indicating that 12- KETE induced trophoblast cell apoptosis. These results were consistent with previous in vivo observations; however, even a lower concentration of 12-KETE activated trophoblast protease. Meanwhile, fibroblasts derived from the bovine placentome converted arachidonic acid to 12-KETE. Conclusion: These observations indicate that 12-KETE may serve as a signal for placenta separation at delivery.

• Experimental Neurobiology
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• v.26 no.5
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• pp.241-251
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• 2017

Saturation mutagenesis was performed on a single position in the voltage-sensing domain (VSD) of a genetically encoded voltage indicator (GEVI). The VSD consists of four transmembrane helixes designated S1-S4. The V220 position located near the plasma membrane/extracellular interface had previously been shown to affect the voltage range of the optical signal. Introduction of polar amino acids at this position reduced the voltage-dependent optical signal of the GEVI. Negatively charged amino acids slightly reduced the optical signal by 33 percent while positively charge amino acids at this position reduced the optical signal by 80%. Surprisingly, the range of V220D was similar to that of V220K with shifted optical responses towards negative potentials. In contrast, the V220E mutant mirrored the responses of the V220R mutation suggesting that the length of the side chain plays in role in determining the voltage range of the GEVI. Charged mutations at the 219 position all behaved similarly slightly shifting the optical response to more negative potentials. Charged mutations to the 221 position behaved erratically suggesting interactions with the plasma membrane and/or other amino acids in the VSD. Introduction of bulky amino acids at the V220 position increased the range of the optical response to include hyperpolarizing signals. Combining The V220W mutant with the R217Q mutation resulted in a probe that reduced the depolarizing signal and enhanced the hyperpolarizing signal which may lead to GEVIs that only report neuronal inhibition.

• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.72-76
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• 2017

Detection of exon 19 deletion mutation in the epidermal growth factor receptor (EGFR) gene, which results in increased and sustained phosphorylation of EGFR, is important for diagnosis and treatment guidelines in non-small-cell lung cancer. Here, we have developed a simple and convenient detection system using the interaction between G-quadruplex and fluorophore thioflavin T (ThT) for discriminating EGFR exon 19 deletion mutant DNA from wild type without a label and quencher. In the presence of exon 19 deletion mutant DNA, the probe DNAs annealed to the target sequences were transformed into G-quadruplex structure. Subsequent intercalation of ThT into the G-quadruplex resulted in a light-up fluorescence signal, which reflects the amount of mutant DNA. Due to stark differences in fluorescence intensity between mutant and wild-type DNA, we suggest that the induced G-quadruplex structure in the probe DNA can report the presence of cancer-causing deletion mutant DNAs with high sensitivity.

• Molecules and Cells
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• v.24 no.1
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• pp.76-82
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• 2007

By combining in situ hybridization (ISH) and immunocytochemistry (IC), microscopic topological localization of mRNAs and proteins can be determined. Although this technique can be applied to a variety of tissues, it is particularly important for use on neuronal cells which are morphologically complex and in which specific mRNAs and proteins are located in distinct subcellular domains such as dendrites and dendritic spines. One common technical problem for combined ISH and IC is that the signal for immunocytochemical localization of proteins often becomes much weaker after conducting ISH. In this manuscript, we report a simplified but robust protocol that allows immunocytochemical localization of proteins after ISH. In this protocol, we fix cultured cortical or hippocampal neurons with 4% paraformaldehyde (PFA), rinse briefly in PBS, and then further fix the cells with $-20^{\circ}C$ methanol. Our method has several major advantages over previously described ones in that (1) it is simple, as it is just consecutive routine fixation procedures, (2) it does not require any special alteration to the fixation procedures such as changes in salt concentration, and (3) it can be used with antibodies that are compatible with either methanol (MeOH-) or PFA-fixed target proteins. To our best knowledge, we are the first to employ this fixation method for fluorescence ISH + IC.

• Transactions of the Korean Society of Mechanical Engineers B
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• v.20 no.2
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• pp.710-719
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• 1996

Laser applied combustion diagnostic techniques-laser induced fluorescence (LIF) and coherent anti-Stokes Ramann spectroscopy (CARS)-are demonstrated. The profiles of hydroxyl radical (OH) and temperature in the counterflow burner are measured and compared with the numerical results. OH radical is excited on the Q$_1$(6) line of the $A^2$$\sum^+$$\leftarrow$$X^2{\prod}$(1, 0) band transition (281.1 nm) and LIF signal is measured at the the bands of (0, 0) and (1, 1) transition (306~326 nm). Absolute OH radical is obtained by using the laser absorption technique. The quenching effects are considered. Temperature is measured using broadband CARS system. Two dimensional OH radical profile is also obtained. The profiles of OH radical and temperature are found to agree well with those of numerical calculation.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.13 no.5
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• pp.2291-2297
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• 2012

X-ray fluorescence sprectrum signal include the continuum. XRF analysis the components of material by the amplitude of peaks. XRF remove the noise and background. To remove the noise, we apply the smoothing filter. And background removal methods applied such as SNIP, Morphology, Threshold methods. In this paper, we applied Threshold using DWT. Interval threshold method divide the some blocks in particular levels. We propose the method that is divided the particular level.

• Transactions of the Korean Society of Automotive Engineers
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• v.8 no.5
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• pp.105-113
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• 2000

In order to clarify the final process of oil consumption, the distribution and flow of oil through each ring were visualized by induced fluorescence method. Motoring and firing test were performed in a single cylinder research engine with transparent cylinder liner. The appropriate calibration techniques were used to solve the unstability of induced light intensity as well as to know the relation of the oil film thickness and output signal. Oil behavior was also observed at dynamic state by high speed CCD camera. By analyzing the oil film thickness converted from the photographed image, it was observed that the main route of oil transport through each ring is the end gap under the usual operating condition, low engine speed and low load condition. Oil film thickness is observed to be irregular and tend to move in a body horizontally at a given piston land. And it is also found that oil flows through oil ring gap so quickly that it can be observed in a single cycle, but it flows so slowly through top and 2nd compression rings that it takes quite a long time to detect the flow.

• Bulletin of the Korean Chemical Society
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• v.31 no.11
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• pp.3241-3246
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• 2010

NHERF ($Na^+/H^+$ exchanger regulatory factor) and E3KARP (NHE3 kinase A regulatory protein) play important roles in membrane targeting, trafficking and sorting of ion channels, transmembrane receptors and signaling proteins in many tissues. Each of these proteins contains two PDZ (PSD-95/Dlg-1/ZO-1) domains, which mediate the assembly of transmembrane and cytosolic proteins into functional signal transduction complexes. The interaction between NHERF and E3KARP was investigated by surface plasmon resonance spectroscopy (BIAcore), fluorescence measurement, His-tagged pull-down experiment, and size-exclusion column (SEC) chromatography. BIAcore experiments revealed that NHERF bound to E3KARP with an apparent $K_D$ of 7 nM. Fluorescence emission spectra of the NHERF-E3KARP complex suggested that the tight interaction between these proteins was accompanied by significant conformational changes in one or both. The CD spectra of NHERF and E3KARP show that the conformational changes of these proteins were dependent on pH and temperature. These results implicate that the NHERF-E3KARP complex allows intracellular signaling complexes to form through PDZ-PDZ interactions.

• Journal of the Optical Society of Korea
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• v.13 no.3
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• pp.310-315
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• 2009

We report the fabrication of double clad fiber (DCF) and DCF coupler, suitable for fiber based imaging systems requiring the dual-channel transmission. Unlike the conventional DCF which uses silica for both cladding layers, the proposed DCF uses a low-index polymer for its outer-cladding layer coated over the conventional silica inner-cladding layer. The DCF is drawn with a conventional SMF preform but a low-index polymer coating is used for both jacket and outercladding of the fiber. To achieve the cladding mode coupling, a DCF coupler is fabricated by simply twisting two pieces of the proposed DCF after removing the polymer-coating at contacting regions. A cladding mode coupling ratio of 30% was achieved with a contact length of 16 cm. The proposed DCF and DCF coupler were employed in a composite optical coherence tomography (OCT) and fluorescence spectroscopy (FS) system, and both OCT images and FS signal from a plant tissue are measured simultaneously.

• Bulletin of the Korean Chemical Society
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• v.30 no.6
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• pp.1252-1256
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• 2009

A simple, selective and sensitive fluorescence quenching method was developed to the determination Cr(VI). The method is based on the oxidation of $I^-\;to\;{{I_3}^-}$ by Cr(VI) in sulfuric acid solution followed by immediate formation of ion association compound between I3 − and rhodamine 6G in Tween-80 micellar media at room temperature. The influence of several surfactants on rhodamine 6G fluorescence signal was studied; particular attention was paid in the aggregation behavior of rhodamine 6G–Tween-80 system. The experimental parameters (e.g., type of surfactant, reagent concentration) were studied and the optimal conditions were established. The linear calibration graph was obtained in the range 2.0 - 100.0 ng m$L^{-1}$ Cr(VI). The detection limit of the method was 0.37 ng m$L^{-1}$. The relative standard deviation (R.S.D.) is less than 5% (n = 5). The efficiency of the method for the determination of Cr(VI) in the presence of Cr(III) in the sample was investigated. The method was applied successfully to the determination of Cr(VI) and total Cr in water, and liver tissue samples.