• Toxicological Research
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• v.16 no.2
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• pp.125-131
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• 2000

The ethanol extract of Rhus verniciflua Stokes (RVS), the Korean Lacquer tree, was subsequentely isolated and fractioned into two portions using distilled water (SED) and 99% ethanol (SEE) as elution buffers through silica gel column (4x28 em, 22 $\AA$. 28~200 mesh). To know the antioxidative effect of the RVS extracts, primary hepatocytes were exposed to hydroxyl radical generated by 20 mU/$m\ell$ glucose oxidase with SED or SEE for 4 hr. The addition of 100$\mu\textrm{g}$/$m\ell$ SED in culture medium showed good protection from glucose oxidase (GO)-mediated cytotoxicity of hepatocytes, showing approximately equivalent to control. When the hepatocytes were incubated with 100 $\mu\textrm{g}$/$m\ell$ SED or SEE only for 4 hr. the activities of cell-associated superoxide dismutase (SOD) and catalase were elevated up to 1.22 fold and 1.4 fold, respectively, compared to control. Further increase, 1.88fold in SOD activity or 1.64fold in catalase activity, was also observed when the hepatocytes were incubated with 100 units/$m\ell$ of commercial SOD or catalase for 4 hr. Moreover. the glucose oxidase-mediated cytotoxicity in cultured hepatocytes was generally reduced upon addition of lysate obtained from SED or SEE-stimulated hepatocytes in a dose-dependent manner. From these results, we suggest that, in cultured hepatocytes, RVS ethanol extract can efficiently reduce cytotoxicity induced by glucose oxidase and may increase the activity of cell-associated SOD and/or catalase, thereby preventing and/or scavenging superoxides and hydroxyl radicals in this experiment.

• Journal of Oral Medicine and Pain
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• v.43 no.1
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• pp.1-7
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• 2018

Purpose: The purpose of the study was to investigate the influences of hyaluronic acid on the candidacidal activities of lysozyme, the peroxidase system, and the glucose oxidase-mediated peroxidase (GO-PO) system at different concentrations of antimicrobial enzymes. Methods: Hyaluronic acid was used at a final concentration of 0.5 mg/mL. Hen egg-white lysozyme (HEWL) was used at concentrations ranging from 10 to $100{\mu}g/mL$. The peroxidase system included bovine lactoperoxidase (bLPO), potassium thiocyanate (KSCN, 1 mM), and hydrogen peroxide ($100{\mu}M$). The GO-PO system included bLPO, KSCN (1 mM), glucose oxidase (10 units/mL), and glucose ($30{\mu}g/mL$). The final concentration of bLPO in the peroxidase and GO-PO systems ranged from 12.5 to $100{\mu}g/mL$. Candida albicans strains ATCC 10231, 11006, and 18804 were utilized. Candidacidal activities of antimicrobials and the influence of hyaluronic acid on their candidacidal activities were determined based on colony forming units. Results: Candidacidal activities of the peroxidase and GO-PO systems increased with increasing concentrations of bLPO. This tendency was the same in the presence or absence of hyaluronic acid. Candidacidal activity of HEWL was not significantly concentration-dependent. Candidacidal activities of the GO-PO system were higher than those of the corresponding peroxidase system. Candidacidal activity was inhibited in the presence of hyaluronic acid in the following order: HEWL, the peroxidase system, and the GO-PO system. Conclusions: Hyaluronic acid inhibited the candidacidal activities of HEWL, the peroxidase system, and the GO-PO system. The GO-PO system exhibited better candidacidal activity than HEWL and the peroxidase system both in the presence and absence of hyaluronic acid.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.2
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• pp.343-347
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• 2002

In order to darify the neuroprotective effect of Gamibojungikki-tang(GBJIKT) water extract on cultured mouse spinal sensory neuron damaged by glucose Oxidase (GO), NR (Neutral Red) assay and LDH (Lactate Dehydrogenase) activity assay were carried out after the cultured mouse spinal sensory neuron were preincubated with various concentrations of GBJIKT water extract for 3 hours prior to exposure of GO. Cell viability of cultured mouse spinal sensory neurons exposed to various concentrations of GO for 8 hours was decreased in a dose-dependent manner. NR/sub 50/ values were 50 mU/ml GO. Cultured mouse spinal sensory neurons in the medium containing various concentration of GO for 8 hours showed increasing of LDH activity. We knew that GO was toxic on cultured spinal sensory neurons. Pretreatment of GBJIKT water extract for 3 hours following GO prevented the GO-induced neurotoxicity such as increasing of LDH activity. These results suggest that GO shows toxic effect on cultured spinal sensory neurons and GBJIKT water extract is highly effective in proecting the neurotoxicity induced by GO.

• Korean Journal of Clinical Laboratory Science
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• v.43 no.2
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• pp.75-81
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• 2011

To clarify the oxidative stress of reactive oxygen species (ROS) and the effect of Chrysanthemum morifolium L. (CM) flower extract on the cultured neuroglial cells (C6 glioma) damaged by ROS, cell adhesion effect was measured by colorimetric assay after cultured C6 glioma cells were treated with various concentrations of glucose oxidase (GO) for 5 hours. For the antioxidative effect of CM flower extract, cell adhesion activity (CAA), superoxide dismutase (SOD)-like activity and lactate dehydrogenase (LDH) activity were assessed against GO-induced cytotoxicity on same cultures. In this study, GO remarkably decreased CAA dose-dependently, and the $XTT_{90}$ and $XTT_{50}$ values were measured at 15 mU/mL and 50 mU/mL following the treatment of C6 glioma cells with 5~60 mU/mL of GO. The CM flower extract significantly increased cell adhesion activity damaged by GO-induced cytotoxicity, and it also showed the SOD-like activity and the decrease of LDH activity. From these results, it is suggested that GO was cytotoxic on cultured C6 glioma cells, and CM flower extract showed antioxidative effects as shown by the increased CAA, SOD-like activity and the decrease of LDH activity on GO-induced cytotoxicity on the same cultures.

• Journal of Sensor Science and Technology
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• v.23 no.5
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• pp.295-298
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• 2014

A novel approach to fabricating high-performance glucose sensors is reported, which is based on the process of self-assembled monolayers (SAMs). In this study, we have particularly used ${\alpha}$-lipoic acid (LA) SAMs for the glucose sensors. To our best knowledge, this study is the first one to use LA as SAMs for this purpose. N-hydroxysuccinimide (NHS) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) were deliberately attached at the same time on the LA SAM. Then, glucose oxidase ($GO_X$) and horseradish peroxidase (HRP) were sequentially immobilized. Thus, the HRP/$GO_X$/NHS-EDC/LA-SAM/Au/Cr/glass working electrode was developed. The glucose-sensing capability of the fabricated sensor was systematically measured by the use of cyclic voltammetry in the range of 1-30 mM glucose in phosphate-buffered saline. The result showed a good sensitivity, that is, as high as $27.5{\mu}A/(mM{\cdot}cm^2)$. This result conspicuously demonstrates that LA can be one of promising substances for use as SAMs for accurately monitoring trace levels of glucose concentration in human blood.

• Bulletin of the Korean Chemical Society
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• v.18 no.5
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• pp.515-519
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• 1997

In a competitive enzyme immunoassay, the performance was tested with different analyte-enzyme conjugates (signal generators) in their binding constants to antibody. Analyte (progesterone)-enzyme (glucose oxidase; GO) conjugates were chemically synthesized and purified by using a gel column with an immobilized antibody to progesterone. In an elution range from the column, four peaks were detected by measuring total enzyme activities. Results from further analysis indicated that the first peak contained mainly unreacted GO while the next three peaks conjugated GO with progesterone. These three conjugate preparations were compared in dose-response curves along with the unpurified mixture. The purified conjugates showed higher detection capabilities than did the mixture. Especially, the preparation in the second peak next to the free GO peak improved the detection limit five times. This performance was comparable to that of a progesterone-horseradish peroxidase conjugate that has been identified to have one progesterone ligand.

• Journal of Sasang Constitutional Medicine
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• v.15 no.2
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• pp.137-150
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• 2003

The purpose of this study is to examine the toxic effects caused by glucose oxidase (GO) and the effects of Hyungbangjihwangtang (HJT) water extracts on the treatment of the toxic effects. For this purpose, experiments with the cultured hippocampal cells from new born mice were done. The results of these experiments were as follows. 1. GO decreased the survival rate of the cultured cells on MTT assay and NR assay. 2. HJT has the efficacy of decreasing the lipid peroxidation. 3. HJT has the efficacy of increasing the amount of neurofilaments. 4. HJT has the efficacy of decreasing the activation of protein kinase C(PKC). From the above results, it is concluded that HJT has marked efficacy as a treatment for the damages caused by the GO-mediated oxidative stress. Further clinical study of this pharmacological effects of H]T should be completed.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.2
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• pp.499-502
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• 2003

Neurotoxicity of reactive oxygen species(ROS) and neuroprotective effect of Aconiti Radix(AR) against ROS-induced cytotoxicity were determined on cultured mouse cerebral neurons by MTT assay after cerebral neurons were cultured for 5 hours in various concentrations of GO. GO was toxic in a dose-dependent manner on cultured cerebral neurons after cerebral neurons were incubated for 5 hours in media containing 5～40mU/ml GO. While, cultures were pretreated with 180 μg/ml AR for 2 hours increased remarkably cell viability. From these results, it is suggested that GO has toxic effect on cultured mouse cerebral neurons by the decrease of cell viability. And also, herb extract such as AKR is very effective in the protection pf neurotoxicity induced by GO.

• Advances in Traditional Medicine
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• v.1 no.2
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• pp.59-65
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• 2000

To clarify the toxic effect of oxidative stress, hydrogen peroxide $(H_{2}O_{2})-induced$ neurotoxicity was examined in cultured newborn mouse spinal motor neurons after spinal motor neurons were grown in the media containing various concentrations of glucose oxidase (GO). And also, the protective effect of Rhizoma gastrodiae extract against GO-induced neurotoxicity was evaluated. Cytotoxicity was expressed as a cell viability by 3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) assay. In this study, exposure of motor neurons to GO-induced cell death significantly, in a dose- and time-dependent manners in spinal motor neuron cultures. The decrease in cell viability of motor neurons damaged by GO was proventioned by Rhizoma gastrodiae extract. These results suggest that the neuroprotective effect of Rhizoma gastrodiae extract on GO-induced neurotoxicity may result from a attenuation of $H_{2}O_{2}-induced$ oxidative stress.

• Toxicological Research
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• v.14 no.2
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• pp.171-176
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• 1998

The biological effects of the water extracts of Rhus Verniciflua Stokes (RVS) were evaluated by protection against hydroxyl radicals. Antioxidative activities were measured using both 1,1-diphenyl-2-picrylhydrazyl (DPPH) and thiocyanate method. Also we used the Glucose oxidase (GO) 20 mU/$\textrm{m}{\ell}$ hydroxyl radical generating system in mouse forebrain cell culture. Water was used for ex-traction from RVS as a solvent which has high polarity especially. In DPPH method, the antioxidative activities of the crude water extract were stronger than any other extracts of low polar-solvents. In the antioxidative effects of mouse forebrain culture using 20 mU/$\textrm{m}{\ell}$ GO, cell viabilities were evaluated 65.6%, 68.8% at 1 $\mu\textrm{g}$. 5 $\mu\textrm{g}$ addition of crude water extracts (30 mg/$\textrm{m}{\ell}$) respectively. 10 $\mu\textrm{g}$ addition of crude water extracts had more than 86.1% cell viabilities, P<0.0l significantly, compared with the group treated with GO alone. In comparison with the antioxidative activities of several commercial antioxidants (ascorbic acid, $\alpha$-tocopherol, catalase, serum), 273 $\mu\textrm{g}$/$\textrm{m}{\ell}$ addition of crude water extracts (300 $\mu\textrm{g}$/$\textrm{m}{\ell}$) showed equivalent antioxidative effect to 25 uM ascorbic acid.