• BMB Reports
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• v.48 no.6
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• pp.313-318
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• 2015

Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that degrade the extracellular matrix (ECM) and regulate the extracellular microenvironment. Despite the significant role that MMP activity plays in cell-cell and cell-ECM interactions, migration, and differentiation, analyses of MMPs in vitro and in vivo have relied upon their abundance using conventional immunoassays, rather than their enzymatic activities. To resolve this issue, diverse nanoprobes have emerged and proven useful as effective activity-based detection tools. Here, we review the recent advances in luminescent nanoprobes and their applications in in vitro diagnosis and in vivo imaging of MMP activity. Nanoprobes with the purpose of sensing MMP activity consist of recognition and detection units, which include MMP-specific substrates and luminescent (fluorescent or bioluminescent) nanoparticles, respectively. With further research into improvement of the optical performance, it is anticipated that luminescent nanoprobes will have great potential for the study of the functional roles of proteases in cancer biology and nanomedicine. [BMB Reports 2015; 48(6): 313-318]

• Applied Microscopy
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• v.17 no.2
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• pp.55-71
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• 1987

To investigate the effects of lead on the electron transport system and ultrastructure of mouse kidney mitochondria, various lead acetate concentrations were treated in vitro and respiration rate, enzyme activities were measured. Ultrastructural changes at state IV respiration were also observed. To compare with in vivo experiments, mouse were injected intraperitoneally of 100 mg lead acetate per kg body weight and state IV respiration rate and enzyme activities were measured. Ultrastructure of renal proximal tubular cells were also observed. In in vitro treatement, decreased state IV respiration, decreased enzyme activities, ruptured membranes and inhibition of condensed to orthodox transformation were observed. In in vivo treatment, decreased state IV respiration and decreased enzyme activities were observed after 24 hrs of i.p. injection. Cytochrome c oxidase activity showed twice the inhibition compared to NADH-CoQ reductase activity at 24 hrs. Continuous decreased state IV respiration was observed after 48 and 72 hrs of injection, however, the enzyme activities were increased to control level. Lead-protein complex which probably inhibits the toxic effects of lead appeared. To conclude, dominant effect of lead on the electron transport system appeared at cytochrome c oxidase activity, and the increased enzyme activities may be a result of appearance of lead-protein complex.

• Korean Journal of Microbiology
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• v.50 no.3
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• pp.239-244
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• 2014

ErmSF is one of the four antibiotic resistance factor proteins expressed by Streptomyces fradiae, antibiotic tylosin producer, which renders $MLS_B$ (macrolide-lincosamide-streptogramin B) antibiotic resistance through dimethylating A2058 of 23S rRNA, thereby reducing the affinity of antibiotic to ribosome. Unlike other Erm proteins, ErmSF harbors long N-terminal end region. To investigate its role in enzyme activity, mutant ErmSF deleted of 1-38 amino acids was overexpressed and activity in vivo and in vitro was observed. In vitro enzymatic assay showed that mutant protein exhibited reduced activity by 20% compared to the wild type enzyme. Due to the reduced activity of the mutant protein, cells expressing mutant protein showed weaker resistance to erythromycin than cells with wild type enzyme. Presumably, the decrease in enzyme activity was caused by the hindrance in substrate binding and (or) product release, not by defect in the methyl group transfer occurred in active site.

• Journal of Aquaculture
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• v.16 no.3
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• pp.196-201
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• 2003

Sessile organisms such as the oyster Crassostrea gigas have been given much attention as a potential biomonitoring indicator to assess the impact of toxicants on aquatic organism. In this study, we exposed cells isolated from gill of oyster (Crassostrea gigas) to hydrogen peroxide in vitro. In addition oysters were in vivo exposed to pyrene and benzo(a)pyrene at various concentrations for 2 weeks. Comet assay was used to detect DNA single strand breaks and to investigate the application of this technique as a tool for aquatic biomonitoring. Hydrogen peroxide increased DNA single strand break with increasing concentration after 30 minutes exposure in vitro. Pyrene and benzo(a)pyrene caused DNA damage only at very high concentration (100 $\mu\textrm{g}$/L or 1000 $\mu\textrm{g}$/L) at two week exposure in vivo. DNA damage was relatively higher at hemocyte than at gill. It suggested that metabolized PAHs are transferred to hemolymph from digestive gland which have a relatively high enzyme activity, and attacked the DNA of hemocyte, while gill accumulated PAHs without degrading them to their metabolites due to low enzyme activity at gill. Both in vitro and in vivo exposure experiments showed that the comet assay is an effective tool on screening whether the organism are exposed to genotoxic contaminants.

• Journal of Environmental Science International
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• v.11 no.4
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• pp.391-397
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• 2002

This study was carried out with the inhibition of the cholinesterase activity by carbamate insecticides in the chicken in vivo and in vitro. The optimum pH of cholinesterase was 8.0. The cholinesterase activity used the acetylcholin as substrate in plasma was 24.6 $\mu$mol/min/g protein. After oral administration with 0.32 mg/kg of BPMC as carbamate pesticide, the cholinesterase activity was inhibited to 60% of control after 15min in vivo. Then the recovery of cholinesterase activity followed to 97% of control after 12hr. I$_{50}$, such as concentration required for 50% inhibition of enzyme activity, of phenyl N-methylcarbamate were 329 $\mu\textrm{g}$/$\ell$ of XMC, 214 $\mu\textrm{g}$/$\ell$ of metolcarb, 111 $\mu\textrm{g}$/$\ell$ of BPMC, 107 $\mu\textrm{g}$/$\ell$ of propoxur and 104 $\mu\textrm{g}$/$\ell$ of isoprocarb. I$_{50}$ of aromatic N-methylcarbamate were 280 $\mu\textrm{g}$/$\ell$ of carbaryl and 114 $\mu\textrm{g}$/$\ell$ carbofuran.ran.

• Korean Journal of Pharmacognosy
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• v.30 no.2
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• pp.145-150
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• 1999

To explain the theory of KIMI which is the theory of therapeutics in oriental medicine, monoamine oxidase(MAO) activities were measured in the brain and liver of mice which were orally administered oriental medicinal herbs which were classified into cold and hot drugs. Rheum palmatum, Anemarrhena asphodeloides, Gardenia jasminoides, Scutellaria baicalensis and Coptis japonica were considered as the cold drugs and Zingiber officinale, Aconitum carmichaeli, Asiasarum sieboldi, Evodia officinalis and Cinnamomum cassia were included in the hot drugs. The effects of cold and hot drugs on in vitro enzyme activities were measured and compared with the in vivo effects. Serotonin is important neurotransmetter involved in the control of body temperature. The MAO plays a central role in the metabolism of many neurotransmetter monoamines including serotonin. MAO is a flavoprotein found exclusively in the mitochondrial outer membrane, occuring in the MAO-A and MAO-B subtypes. MAO-A deaminates serotonin and noradrenaline, whereas MAO-B prefers phenylethylamine and benzylamine as substrates. Coptis japonica and Aconitum carmichaeli elevated the in vivo MAO activities and especialy, in vivo MAO-B activities were significantly increased. In vitro MAO-A activities were increased by hot drugs, whereas the in vitro MAO-B activities were inhibited. Cold drugs inhibited both enzyme activities in vitro.

• Journal of Life Science
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• v.11 no.6
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• pp.574-581
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• 2001

This study was performed to evaluate the inhibition effects of fermented soybean on lipid perosidation and antioxidative relative enzyme activity. in vivo. Fermented soybean was induced the high SOD activity, while significantly inhibited on the peroxide value of linoleic acid and lipid perxidation from rat microsome induced by Fe$^{2+}$ ascorbate system, Sprague-Dawley(SD) male rats were fed basic diet, and experimental diets group added 200 or 500 mg/kg fermented soybean for 2 weeks. The effect of fermented soybean is also significantly increased catalase and glutathione peroxidase activities, while significantly inhibited the lipid peroxidation of rat liver microsome in a dose dependent manner. Therefore, these results suggest that fermented soybean has antioxidative activity which is related enzyme to prevention of oxidative stress.s.

• Journal of the East Asian Society of Dietary Life
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• v.4 no.2
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• pp.59-67
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• 1994

The present study was conducted to evaluate the hepatoprotective effect of puerariae Radix methanol extract on benzo(a) pyrene(B(a)P) - induced liver injuries in rats. In vitro experiment, primary cultured hepatocytes (5X105 cells/$m\ell$) were cultured for 20~24 hours after adding puerariae Radix mehtanol extract(32$\mu\textrm{g}$/$m\ell$) and B(a)P(50 uM). In vivo experiment, Puerariae Radix methanol extract(0.25 g/kg/day, per os) was administered for 7 days and B(a)P(0.1 mg/kg/day, intraperitoneally) was given after the last administration of extract. And then the hepatoprotective effect of Puerariae Radix methanol extract was investigated biochemically through in vitro and in vivo experiments. Namely, activities of enzymes (GOT, GPT and LDH) were measured and 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide(MTT) assay were carried out in vitro cell culture study and GOT, GPT, LDH and ALP activities and HDL-cholesterol, total cholesterol and triglyceride contents were performed in vivo study. In vitro experiment, as a result of enzyme activity measurement(GOT, GPT and LDH) and MTT assay, GOT,GPT and LDH activities changed by B(a)P were recovered to normal levels and hepatocytes impaired by B(a)P were recovered to normal. In vivo experiment, Puerariae Radix methanol extract significantly decreased the enzyme activities(GOT, GPT, ALP and LDH in serum and GPT and ALP in tissue) and lipid contents in comparison to B(a)P-treated group.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.3
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• pp.1163-1169
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• 2014

Astragalus, a commonly used traditional Chinese medicine, has exhibited antitumor actions in patients. In this study, in vitro and in vivo antitumor effects of astragalus and synergistic antitumor efficacy in combination with pterostilbene were investigated. Melanoma cells were treated with pterostilbene (Pt), graduated doses of astragalus injection (AI), or these in combination. Cell viability was measured using a MTT assay. Released nucleosomes and caspase activity were measured using enzyme-linked immunosorbent assay. Growth inhibition in vitro and in vivo was also assessed. Analysis of variance and t tests were used for statistical analysis. Significant reduction (p<0.05) in cellular proliferation were observed with AI and AI-Pt in a time- and concentration-dependent manner. Apoptosis and caspase-3/7 activity were significantly increased by AI and AI-Pt treatment (p<0.05). In vivo, AI inhibited melanoma tumor growth, with inhibition rates ranging from 36.5 to 62.3%, by inducing apoptosis via up-regulation Bax expression and the Bax/Bcl-2 ratio and down-regulating Bcl-2 expression. AI significantly inhibits the growth of melanoma in vitro and in vivo by inducing apoptosis. These data suggest that combined treatment of astragalus with pterostilbene enhances antitumor efficacy.

• Journal of Life Science
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• v.11 no.2
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• pp.94-98
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• 2001

Type I signal peptidase cleaves the signal sequence from the amino terminus of membrane and secreted proteins afters these protein insert across the membrane. This enzyme serves as a potential target for the development of novel antibacterial agents due to its unique physiological and biochemical properties. Despite considerable research, the signal peptidase assay still remains improvement to provide further understanding of the mechanism and high-throughput inhibitor screening of this enzyme. In this paper, three known signal peptidase assays are tested with an E. coli D276A mutant signal peptidase to distinguish the sensitivity of each assays. In vitro assay using the procoat synthesized by in vitro transcription translation shows that the D276A signal peptidase I was inactive while in vivo processing of pro-OmpA expressed in the temperature-sensitive E. coli strain IT41 as well as in vitro assay using pro-OmpA nuclease A substrate show that D276A signal peptidase I has activity like wild-type signal peptidase. These results suggest that in vitro assay using the pro-OmpA nuclease A and in vivo pro-OmpA processing assay are more sensitive monitors than in vitro assay using the pro-coat. In conculsion, caution should be used when interpreting the in vitro results using the procoat.