• Archives of Pharmacal Research
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• v.27 no.9
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• pp.947-953
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• 2004

The antioxidative and hepatoprotective potentials of two anthraquinones, alaternin (2-hydroxy-emodin) and emodin, to scavenge and/or inhibit hydroxyl radicals generated by the Fenton reaction and to protect tacrine-induced cytotoxicity in human liver derived HepG2 cells were evaluated, respectively. The inhibitory activity on hydroxyl radical generated in a cell-free chemical system (FeSO$_4$/$H_2O$$_2$) was investigated by a fluorescence spectrophotometer using a highly fluorescent probe, 2$^1$,7$^1$-dichlorofluorescein. The hydroxyl radical scavenging activity was determined by electron spin resonance spectroscopy using 5,5-dimethy-1-pyrroline-N-oxide as hydroxyl radicals trapping agents. Tacrine-induced HepG2 cell toxicity was determined by a 3-［4,5-dimethylthiazole-2yl］-2,5-diphenyltertrazolium bromide assay. Although the scavenging activity of alaternin on hydroxyl radical was similar to that of emodin in dose-dependent pat-terns, the inhibitory activity exhibited by the former on hydroxyl radical generation was stron-ger than that of the latter, with $IC_{50}$/ values of 3.05$\pm$0.26 $\mu$M and 13.29$\pm$3.20 $\mu$M, respectively. In addition, the two anthraquinones, alaternin and emodin showed their hepatoprotective activ-ities on tacrine-induced cytotoxicity, and the EC$_{50}$ values were 4.02 11M and 2.37 $\mu$M, respec-tively. Silymarin, an antihepatotoxic agent used as a positive control exhibited the EC$_{50}$ value of 2.00 $\mu$M. These results demonstrated that both alaternin and emodin had the simultaneous antioxidant and hepatoprotective activities.ies.

• Korean Journal of Medicinal Crop Science
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• v.20 no.5
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• pp.387-392
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• 2012

This study was carried out to identify Korean ginseng cultivars using peptide nucleic acid (PNA) microarray. Sixty-seven probes were designed based on nucleotide variation to distinguish Korean ginseng cultivars of Panax ginseng. Among those PNA probes, three (PGB74, PGB110 and PGB130) have been developed to distinguish five Korean ginseng cultivars. Five Korean ginseng cultivars were denoted as barcode numbers depending on their fluorescent signal patterns of each cultivar using three probe sets in the PNA microarray. Five Korean ginseng cultivars, Chunpoong, Yunpoong, Gopoong, Gumpoong and Sunpoong, were simply denoted as '111', '222', '211', '221' and '122', respectively. This is the first report of PNA microarray which provided an objective and reliable method for the authentication of Korean ginseng cultivars. Also, the PNA microarray will be useful for management system and pure guarantee in ginseng seed.

• Journal of Periodontal and Implant Science
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• v.34 no.2
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• pp.357-366
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• 2004

Chronic exposure to high levels of manganese leads a pronounce and debilitating disorder known as manganism. Research on the toxic manifestation of manganese have focused primarily on its neurological effects because exposure to high levels of the metal produces a distinct and irreversible extrapyramidal dysfunction resembling the dystonic movements associated with Parkinson's physiological and biochemical systems in the body. The purpose of this study was to evaluate the effect of manganeses on primary rat calvarial cell growth and toxicity. The experimental groups were in concentration of 0, 10, 30, 60, 100, 300 ${\mu}M$. Cell activity was assessed at day 1 and day 3 using a fluorescent molecular probe. Cell proliferation was evaluated at day 1 and day 3 by MTT assay. The amount of total protein synthesis was measured at day 3 and day 7. The results were as follows: The proliferation of primary rat calvarial cells were inhibited by $MnCl_2$ in the concentration exceeding $100{\mu}M$. The primary rat calvarial cells treated with $MnCl_2$ showed similar protein synthesis to the control group except in 100 ${\mu}M$. These result suggest that manganese suppress the viability and protein synthesis of primary rat calvarial cells in concentration exceeding $100{\mu}M$.

• BMB Reports
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• v.33 no.3
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• pp.279-284
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• 2000

Using fluorescence probes, 2-(9-anthroyloxy) stearic acid (2- AS) and 12-(9-anthroyloxy) stearic acid (12-AS), we determined the differential effects of local anesthetics (tetracaine-HCI, bupivacaine-HCI, lidocaine-HCI, prilocaine-HCI and procaine-HCI) on the differential rotational rate between the surface (in carbon number 2 and its surroundings including the head group) and the hydrocarbon interior (in carbon number 12 and its surroundings) of the outer monolayer of the total phospholipid fraction liposome that is extracted from synaptosomal plasma membrane vesicles. The anisotropy (r) values for the hydrocarbon interior and the surface region of the liposome outer monolayer were$0.051{\pm}0.001$ and $0.096{\pm}0.001,$ respectively. This means that the rate of rotational mobility in the hydrocarbon interior is faster than that of the surface region. Local anesthetics in a dosedependent manner decreased the anisotropy of 12-AS in the hydrocarbon interior of the liposome outer monolayer, but increased the anisotropy of 2-AS in the surface region of the monolayer. These results indicate that local anesthetics have significant disordering effects on the hydrocarbon interior, but have significant ordering effects on the surface region of the liposome outer monolayer.

• Journal of Genetic Medicine
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• v.14 no.2
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• pp.62-66
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• 2017

Interchromosomal insertion of Y chromosome heterochromatin in an autosome was identified in a fetus and a family. A fetal karyotype was analyzed as 46,XX,dup(7)(?q22q21.1) in a referred amniocentesis at 16 weeks of gestation for advanced maternal age. In the familial karyotype analyses for identification of der(7), the mother, the first daughter and the maternal grandmother showed the same der(7) as the fetus's. CBG-banding was positive at 7q22 region of der(7) that indicated inserted material was originated from heterochromatin. The origin of heterochromatic insertion region in der(7) of the fetus and the mother was found in Yq12 region by fluorescent in situ hybridization with a DYZ1 probe. In the specific analysis of Y chromosomal heterochromatic region of ins(7;Y) of the mother, 15 sequence tagged sites from Yp11.3 region including SRY to Yq11.223 region was not detected. Final karyotypes of the mother, the first daughter and the maternal grandmother were reported as 46,XX,der(7)ins(7;Y)(q21.3;q12q12). All female carriers of ins(7;Y) in the family showed normal phenotype and the mother and the maternal grandmother were fertile. A healthy girl was born at term. We report a rare case of familial interchromosomal insertion of Y chromosome heterochromatin detected only in female family members with normal phenotype that was diagnosed prenatally.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.7
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• pp.949-956
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• 2002

There are several physiological and pharmacological evidences indicating that opening of voltage dependent $Ca^{2+}$ channels play a critical role in induction of acrosome reaction in mammalian sperm. We determined the intracellular free $Ca^{2+}$ concentration in ejaculated goat sperm using a fluorescent, $Ca^{2+}$-specific probe, Fura2/AM, after the suspension of sperm in KRB medium, capable of sustaining capacitation and the acrosome reaction. We used nifedipine, D-600 and diltiazem, the $Ca^{2+}$ channel antagonists belonging to the classes of dihydropyridines, phenylalkylamines and benzothiazepines, to investigate the possibility that L-type voltage gated $Ca^{2+}$ channels play a role in the progesterone-stimulated exocytotic response. Progesterone promoted a rise in intracellular $Ca^{2+}$ in goat sperm and addition of nifedipine (100 nM) just prior to progesterone induction, significantly inhibited both intracellular $Ca^{2+}$ rise and exocytosis suggesting that $Ca^{2+}$ channels are involved in the process. However, the intracellular $Ca^{2+}$ increase during the process of capacitation was not affected with the addition of nifedipine suggesting a role of focal channel for $Ca^{2+}$ during capacitation. Studies using monensin and nigericin, two monovalent cation ionophores showed that an influx of $Na^+$ also may play a role in the opening of $Ca^{2+}$ channels. These results strongly suggests that the entry of $Ca^{2+}$ channels with characteristics similar to those of L-type, voltage-sensitive $Ca^{2+}$ channels found in cardiac and skeletal muscle, is a crucial step in the sequence of events leading to progesterone induced acrosome reaction in goat sperm.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.73-73
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• 2002

Nuclear transfer (NT) techniques have advanced in the last years, and cloned animals have been produced by using somatic cells in several species including pig. However, it is difficult that the nuclear transfer porcine embryos development to blastocyst stage overcoming the cell block in vitro. Abnormal segregation of chromosomes in nuclear transferred embryos on genome activation stage bring about embryo degeneration, abnormal blastocyst, delayed and low embryo development. Thus, we are evaluated that the correlations of the frequency of embryo developmental rates and chromosome aberration in NT and In viかo fertilization (IVF) derived embryo. We are used for ear-skin-fibroblast cell in NT. If only karyotyping of embryonic cells are chromosomally abnormal, they may difficultly remain undetected. Then, we evaluate the chromosome aberrations, fluorescent in situ hybridization (FISH) with porcine chromosome 1 submetacentric specific DNA probe were excuted. In normal diploid cell nucleus, two hybridization signal was detected. In contrast, abnormal cell figured one or three over signals. The developmental rates of NT and IVF embryos were 55% vs 63%, 32% vs 33% and 13% vs 17% in 2 cell, 8 cell and blastocyst, respectively. When looking at the types of chromosome aberration, the detection of aneuploidy at Day 3 on the embryo culture. The percentage of chromosome aneuploidy of NT and IVF at 4-cell stage 40.0%, 31.3%, respectively. This result indicate that chromosomal abnormalities are associated with low developmental rate in porcine NT embryo. It is also suggest that abnormal porcine embryos produced by NT associated with lower implantation rate, increase abortion rate and production of abnormal fetuses.

• Journal of Photoscience
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• v.6 no.4
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• pp.187-191
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• 1999

Using fluorescent probe fura-2 acetoxymethyl ester, we studied effects of N-Methyl-D-aspartate (NMDA) on free intracellular $Ca^{2+}$ concentration ([$Ca^{2+}$]$_{i}$) and interaction of ethanol with NMDA-mediated response in freshly dissociated brain cells from newborn rats. Twenty five micromolar NMDA significantly increased ($Ca^{2+}$), and this increasing effect could be prevented or reversed by the NMDA antagonists $Mg^{2}$(1.0 mM) and 2-amino-5-phosphonovalerate (AP5, 100 ${\mu}$M). Ethanol at concentrations from 2.5 to 100 mM inhibited NMDA-mediated calcium current in a concentration-dependent manner. Maximal inhibition of NMDA-mediated calcium current by ethanol was 82% at 50 mM. The ethanol inhibition at 100 mM was not significantly different from the inhibition at 50 mM.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.9
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• pp.1285-1292
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• 2016

Changes in surface hydrophobicity of soy protein isolate (SPI), which plays an important role in the functional characteristics of protein, were measured according to various SPI concentrations, pH levels, electrolytes concentrations, and alginate molecular weights by using 1-anilino-8-naphthalene sulfonic acid as a fluorescent probe. SPI surface hydrophobicity decreased as SPI concentrations increased. SPI surface hydrophobicity reached a maximum at pH 7.0. SPI surface hydrophobicity rapidly increased as the NaCl concentration of SPI solution increased up to 100 mM, and showed no large increases above 100 mM. However, SPI surface hydrophobicity radically decreased until the $CaCl_2$ concentration reached 50 mM and revealed no large variations above 50 mM. A similar trend was exhibited in the case of $MgCl_2$. As both the concentration and molecular weight of sodium alginate increased, SPI surface hydrophobicity decreased. The increasing rate of SPI surface hydrophobicity decreased as the molecular weight of sodium alginate increased.

• Proceedings of the Microbiological Society of Korea Conference
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• 2007.05a
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• pp.83-85
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• 2007

Deep-sea bacteria are adapted to extreme environments, such as high pressures and cold temperatures. We have isolated many piezophiles which grow well even under high pressures from deep-sea sediment. Shewanella violacea DSS12 and Moritella japonica DSK1 have the ability to grow at up to 70 MPa, and those bacteria have unique mechanisms of gene expression in response to high pressure conditions. The combination of gene expression systems in piezophiles, like the high pressure-dependent promoters and GFP reporter gene, may reveal highly fluorescent cells when exposed to high hydrostatic pressure conditions. It is predicted that a novel bio-sensing system can be made to probe high pressure environments using living bacteria. First, gene transformation into our piezophiles, strains DSS12 and DSK1, were examined. Eschericha coli S17-1 was used for bacterial conjugation with those piezophiles. As a result, the broad host range vector, pKT231, and the shuttle vector, pTH10, were successfully introduced to DSS12 and DSK1, respectively. Next, The pressure regulated promoters from DSS12 and DSK1 were cloned into proper vectors and combined with GFP as a reporter gene downstream of each promoter. The transformants of DSK1 and DSS12 with the recombinant pTH10 and pKT231 plasmid, which has cadA and glnA promoters (each of them is a pressure regulated promoter from DSK1 and DSS12, respectively) and GFP, were grown under high pressure and gene expression of GFP promoted by 50 MPa pressure was confirmed. This is a critical point to create a pressure-sensing bacteria, as the "High Pressure Glow Cells", which will indicate the level of environmental pressure using fluorescence of GFP as a reporter gene.