• The Journal of Internal Korean Medicine
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• v.30 no.1
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• pp.36-50
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• 2009

Objectives : This study was aimed to verify the cytoprotective function, antioxidative effect and inflammation genes inhibitory effects of Lycium chinense Milie. Therefore the generation of superoxide anion radical ( $O_2\;^-$), peroxynitrite ($ONOO^-$), nitric oxide (NO) and prostaglandin $E_2$ $(PGE_2)$ was investigated in the renal epithelial cells of mouse. Effects of Lycium chinense Milie on the expression of inflammation-related proteins, $IKK-{\alpha}$. $p-IKK-\alpha\beta$, $p-I{\kappa}B-\alpha$, $NF-{\kappa}B$ (p50, p65), COX-2 and iNOS, were examined by western blotting. Methods : For this study, the fluorescent probes were used, namely dihydrorhodamine 123 (DHR 123), 4.5-diaminofluorescein (DAF-2) and 2',7'-dichlorodihydrofluorescein diacetate (DCFDA). Western blotting was performed using anti-$IKK-\alpha$, anti-phospho $IKK-\alpha\beta$, anti-phospho $I{\kappa}B-\alpha$, anti-$NF-{\kappa}B$ (p50, p65), anti-COX-2 and anti-iNOS, respectively. Results : Lyciutn chinense Milie reduced $H_{2}O_{2}$-induced cell death dose-dependently. It inhibited the generation of $O_2\;^-$, $ONOO^-$, NO and $PGE_2$ in the $H_{2}O_{2}$-treated renal epithelial cells of mouse in vitro. Lycium chinense Milie inhibited the expression of $IKK-\alpha$, $p-IKK-\alpha\beta,\;p-I{\kappa}B-\alpha$, COX-2 and iNOS genes by means of decreasing activation of $NF-{\kappa}B$. Conclusions : According to above results. Lycium chinense Milie recommended to be applied in treatment for the inflammatory process and inflammation-related diseases.

• Journal of Genetic Medicine
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• v.3 no.1
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• pp.5-10
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• 1999

This is a case report of 46,XY female phenotype (46,XY karyotype, no pubic hair, blind vagina and absence of uterus)in an 18-year-old patient. To confirm whether a Y chromosome has a structural abnormality, fluorescent in situ hybridization (FISH) with the chromosome X/Y cocktail probe was simultaneously performed, and the six loci [PABY, RPS4Y(sy16, sy17), ZFY, DYS14] on the short arm, one locus (DYZ3) on the centromere and one locus (DYZ1) on the long arm were amplified by polymerase chain reaction (PCR). The probes used FISH hybridized to centromere of the X chromosome and heterochromatin region (Yq12) of the Y chromosome, and all PCR related Y chromosome showed positive band like normal male. From the results obtained, it seemed that the Y chromosome from the 46,XY female was structurely normal. Especially, the SRY gene has been equated with the mammalian testis-determining factor, and absence or point mutation in the SRY gene causes XY female. To detect the point mutations of SRY sequences, single-strand conformation polymorphism (SSCP) assay was used. Our results confirm that this patient has no mutation in the SRY gene on the Y chromosome.

• Journal of Ginseng Research
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• v.43 no.2
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• pp.326-334
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• 2019

Background: The objective of our study was to analyze the neuroprotective effects of ginsenoside derivatives Rb1, Rb2, Rc, Rd, Rg1, and Rg3 against glutamate-mediated neurotoxicity in HT22 hippocampal mouse neuron cells. Methods: The neuroprotective effect of ginsenosides were evaluated by measuring cell viability. Protein expressions of mitogen-activated protein kinase (MAPK), Bcl2, Bax, and apoptosis-inducing factor (AIF) were determined by Western blot analysis. The occurrence of apoptotic and death cells was determined by flow cytometry. Cellular level of $Ca^{2+}$ and reactive oxygen species (ROS) levels were evaluated by image analysis using the fluorescent probes Fluor-3 and 2',7'-dichlorodihydrofluorescein diacetate, respectively. In vivo efficacy of neuroprotection was evaluated using the Mongolian gerbil of ischemic brain injury model. Result: Reduction of cell viability by glutamate (5 mM) was significantly suppressed by treatment with ginsenoside Rb2. Phosphorylation of MAPKs, Bax, and nuclear AIF was gradually increased by treatment with 5 mM of glutamate and decreased by co-treatment with Rb2. The occurrence of apoptotic cells was decreased by treatment with Rb2 ($25.7{\mu}M$). Cellular $Ca^{2+}$ and ROS levels were decreased in the presence of Rb2, and in vivo data indicated that Rb2 treatment (10 mg/kg) significantly diminished the number of degenerated neurons. Conclusion: Our results suggest that Rb2 possesses neuroprotective properties that suppress glutamate-induced neurotoxicity. The molecular mechanism of Rb2 is by suppressing the MAPKs activity and AIF translocation.

• ALGAE
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• v.35 no.4
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• pp.389-404
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• 2020

Red algal fertilization is unusual and offers a different model to the mechanism of intracellular transport of nuclei and polyspermy blocking. A female carpogonium (egg) undergoes plasmogamy with many spermatia (sperm) simultaneously at the receptive structure, trichogyne, which often contains numerous male nuclei. The pattern of selective transport of a male nucleus to the female nucleus, located in the cell body of the carpogonium, remain largely unknown. We tracked the movement of spermatial nuclei and cell organelles in the trichogyne after plasmogamy using time-lapse videography and fluorescent probes. The fertilization process of Bostrychia moritziana is composed of five distinctive stages: 1) gamete-gamete binding; 2) mitosis in the attached spermatia; 3) formation of a fertilization channel; 4) migration of spermatial nuclei into the trichogyne; and 5) cutting off of the trichogyne cytoplasm from the rest of the cell after karyogamy. Our results showed that actin microfilaments were involved in the above steps of fertilization, microtubules are involved only in spermatial mitosis. Time-lapse videography showed that the first ("primary") nucleus which entered to trichogyne moved quickly to the base of carpogonium and fused with the female nucleus. The transport of the primary male nucleus to the egg nucleus was complete before its second nucleus migrated into the trichogyne. Male nuclei from other spermatia stopped directional movement soon after the first one entered the carpogonial base and oscillated near where they entered trichogyne. The cytoplasm of the trichogyne was cut off at a narrow neck connecting the trichogyne and carpogonial base after gamete nuclear fusion but gamete binding and plasmogamy continued on the trichogyne. Spermatial organelles, including mitochondria, entered the trichogyne together with the nuclei but did not show any directional movement and remained close to where they entered. These results suggest that polyspermy blocking in B. moritziana is achieved by the selective and rapid transport of the first nucleus entered trichogyne and the rupture of the trichogyne after gamete karyogamy.

• The Plant Pathology Journal
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• v.38 no.6
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• pp.665-672
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• 2022

Cymbidium mosaic virus (CymMV) is one of economically important viruses that cause significant losses of orchids in the world. In the present study, a reverse transcription recombinase polymerase amplification (RT-RPA) assay combined with a lateral flow immunostrip (LFI) assay was developed for the detection of CymMV in orchid plants. A pair of primers containing fluorescent probes at each terminus that amplifies highly specifically a part of the coat protein gene of CymMV was determined for RT-RPA assay. The RT-RPA assay involved incubation at an isothermal temperature (39℃) and could be performed rapidly within 30 min. In addition, no cross-reactivity was observed to occur with odontoglossum ringspot virus and cymbidium chlorotic mosaic virus. The RT-RPA with LFI assay (RT-RPA-LFI) for CymMV showed 100 times more sensitivity than conventional reverse transcription polymerase chain reaction (RT-PCR). Furthermore, the RT-PCR-LFI assay demonstrated the simplicity and the rapidity of CymMV detection since the assay did not require any equipment, by comparing results with those of conventional RT-PCR. On-site application of the RT-RPA-LFI assay was validated for the detection of CymMV in field-collected orchids, indicating a simple, rapid, sensitive, and reliable method for detecting CymMV in orchids.

• Journal of Korean Society of Environmental Engineers
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• v.27 no.12
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• pp.1311-1320
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• 2005

The purpose of this research is to survey characteristics of microbial community and the removal efficiency of organic materials for biological activated carbon in water treatment plant. Coal based activated carbon retained more attached bacterial biomass on the surface of the activated carbon than the other activated carbon with operating time and materials. The heterotrophic plate count(HPC), eubacteria(EUB) and 4,6-diamidino-2-phenylindole(DAPI) counts were ranged from $0.95{\times}10^7$ to $52.4{\times}10^7$ CFU/g, from $3.8{\times}10^8$ to $134.2{\times}10^8$ cells/g and from $7.0{\times}10^8$ to $250.2{\times}10^8$ cells/g, respectively. The biomass of EUB and DAPI appeared to be much more $10^2$ than HPC, which were increasing in bed volume of 20,000 at the stage of steady-state. The change of microbial community by analyzing fluorescent in situ hybridization(FISH) method with rRNA-targeted oligonucleotide probes, the dominant group was $\alpha$-proteobacteria($\alpha$ group) and high G+C content bacteria(HGC) the lowest distributing rate before reaching the bed volume of 20,000. After reaching the bed volume of 20,000, $\alpha$ group and other groups of bacteria became decreased, on the other hand, the proportion of both $\beta$-proteobacteria($\beta$ group) and $\gamma$-proteobacteri($\gamma$ group) were increasing. Coconut and wood based activated carbons had similar trend with coal based activated carbon, but the rate of $\alpha$ group on coal based activated carbon had gradually increased. Bacterial production with the operating period appeared highest in coal based activated carbon at the range of $1.2{\sim}3.4\;mg-C/m^3{\cdot}h$ while the coconut and wood based activated carbon were ranged from 1.1 to 2.6 $mg-C/m^3{\cdot}h$ and from 0.7 to 3.5 $mg-C/m^3{\cdot}h$ respectively. The removal efficiency of assimilable organic carbon(AOC) showed to be highly correlated with bacterial production. The correlation coefficient between removal efficiency of AOC and bacterial production were 0.679 at wood based activated carbon, 0.291 at coconut based activated carbon and 0.762 at coal based activated carbon, respectively.

• Clean Technology
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• v.21 no.2
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• pp.102-107
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• 2015

Rhodamine dyes are widely used as fluorescent probes because of their excellent photophysical properties, such as high extinction coefficients, excellent quantum yields, great photostability, relatively long emission wavelengths. A great synthetic effort has been focused on developing efficient and practical procedures to prepare rhodamine derivatives, because for most applications the probe must be covalently linked to another (bio)molecule or surface. Sulforhodamine B is one of the most used rhodamine dyes for this purpose, because it carries two sulfoxy functions which can be easily utilized for binding with other molecules. Recently, we needed an expedient, practical synthesis of sulforhodamine derivatives. We found the existing procedure for obtaining those compounds unsatisfactory, particularly, with the cyclization process of the dihydroxytriarylmethane (1) to produce the corresponding xanthene derivative (2). We report here our findings, which represent modification of the existing literature procedure and provide access to the corresponding xanthene derivative (2) in a high yield. Use of methanol as a co-solvent was found quite effective to prohibit the water molecule produced during the cyclization reaction from retro-cyclizing back to the starting dihydroxytriarylmethane and the yield of the cyclization was increased (up to 84% from less than 20%). The reaction temperature was significantly lowered (80 vs. 135 ℃). Thus, the reaction proceeds in a higher yield and energy-saving manner where the use of reactants and the production of chemical wastes is minimized.

• Journal of Physiology & Pathology in Korean Medicine
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• v.30 no.4
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• pp.242-249
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• 2016

This study was to investigate the anti-inflammatory effects of Prunellae Herba(PH). The generation of superoxide anion radical (․O₂-), nitric oxide (NO), peroxynitrite (ONOO-) and Prostaglandin E₂(PGE₂) were measured in the H₂O₂-Treated renal epithelial cells(LLC-PK1 cell) of mouse. And the effects of Prunellae Spica on the expression of NF-κB (p50, p65), IKK-α, phospho-IκB-α and inflammation-related proteins, COX-2, iNOS, IL-1β and VCAM-1, were examined by western blot. The fluorescent probes, 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) and dihyldrorhodamine 123 (DHR 123) were used to estimate the scavenging effect of Prunellae Spica on ․O₂-, NO, ONOO-. Western blot was conducted to assess the protein expression levels of NF-κB (p50, p65), IKK-α, phospho-IκB-α, inflammation-related proteins, COX-2, iNOS, IL-1β, VCAM-1. PH inhibited H₂O₂-treated cell death dose-dependently. It reduced the generation of ·O₂-, NO, ONOO- and PGE₂ in the H₂O₂-treated renal epitheial cells(LLC-PK₁ cell) of mouse in vitro. PH reduced the expression of NF-κB, IKK-α, phospho-IκB-α, COX-2, iNOS, IL-1β and VCAM-1 genes through means of decreasing activation of NF-κB signaling as well. According to these results, PH has an antioxidative activity and anti-inflammatory effect by regulating the NF-κB pathway. This suggest that PH is expected to be used to regulating inflammatory process and treating inflammation-related disease.

• Parasites, Hosts and Diseases
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• v.51 no.6
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• pp.689-694
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• 2013

Opisthorchis viverrini and Clonorchis sinensis are parasites known to be carcinogenic and causative agents of cholangiocarcinoma in Asia. The standard method for diagnosis for those parasite infections is stool examination to detect parasite eggs. However, the method has low sensitivity, and eggs of O. viverrini and C. sinensis are difficult to distinguish from each other and from those of some other trematodes. Here, we report a multiplex real-time PCR coupled with high resolution melting (HRM) analysis for the differentiation of O. viverrini and C. sinensis eggs in fecal samples. Using 2 pairs of species-specific primers, DNA sequences from a portion of the mitochondrial NADH dehydrogenase subunit 2 (nad 2) gene, were amplified to generate 209 and 165 bp products for O. viverrini and C. sinensis, respectively. The distinct characteristics of HRM patterns were analyzed, and the melting temperatures peaked at $82.4{\pm}0.09^{\circ}C$ and $85.9{\pm}0.08^{\circ}C$ for O. viverrini and C. sinensis, respectively. This technique was able to detect as few as 1 egg of O. viverrini and 2 eggs of C. sinensis in a 150 mg fecal sample, which is equivalent to 7 and 14 eggs per gram of feces, respectively. The method is species-specific, rapid, simple, and does not require fluorescent probes or post-PCR processing for discrimination of eggs of the 2 species. It offers a new tool for differentiation and detection of Asian liver fluke infections in stool specimens.

• Archives of Pharmacal Research
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• v.20 no.1
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• pp.1-6
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• 1997

We determined the microviscosity of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine cerebral cortex and liposomes of total lipids (SPMTL) and phospholipids (SPMPL) extracted from SPMV. Changes in the microviscosity induced by the range and rate of lateral diffusion were measured by the intramolecular excimerization of 1, 3-di(1-pyrenyl)propane (Py-3-Py). The microviscosity values of the direct probe environment in SPMV, SPMTL and SPMPL were 38.17, 31.11 and 27.64 cP, respectively, at$37^{\circ}C$and the activation energies $(E_a)$ of the excimer formation of Py-3-Py in SPMV, SPMTL and SPMPL were 8.236, 7.448 amd 7.025 kcal/mol, respectively. Probe location was measured by polarity and polarizability parameters of the probe Py-3-Py and probe analogues, pyrene, 1-pyrenenonanol and 1-pyrenemethyl-3${\beta}$-hydroxy-22, 23-bisnor-5-cholenate (PMC), incorporated into membranes or solubilized in reference solvents. There existed a good linear relationship between the first absorption peak of the $^1_a$ band and the polarizability parameter $(n^{2}-1)/(2n^{2}+1)$.The calculated refractive index values for SPMV, SPMTL and SPMPL were close to 1.50, which is higher than that of liquid paraffin (n=l.475). The probe location was also determined by using a polarity parameter $(f-1/2f^{I})$. Here f=$({\varepsilon}-1)/(2{\varepsilon}+1)$ is the dielectric constant function and $f^I=(n^2-1)/(2n^2+1)$ is the refractive index function. A correlation existed between the monomer fluorescence intensity ratio and the solvent polarity parameter. The probes incorporated in SPMV, SPMTL, and SPMPL report a polarity value close to that of 1-hexanol $({\varepsilon}=13.29)$. In conclusion, Py-3-Py is located completely inside the membrane, not in the very hydrophobic core, but displaced toward the polar head groups of phospholipid molecules, e.g., central methylene region of aliphatic chains of phospholipid molecules.