• Journal of Microbiology and Biotechnology
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• v.21 no.7
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• pp.675-678
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• 2011

DNA methylation in Streptomyces griseus IFO 13350 was analyzed by high-performance liquid chromatographic analysis and bisulfite-based analysis to reveal two methylation sites, 5'-$GC^{5m}$ CGGC-3' and 5'-$GAG^{5m}$ CTC-3'. The methylation was reconstituted in Escherichia coli by simultaneous expression of S. griseus SGR4675 and S. achromogenes M.SacI. The E. coli cells produced plasmids that mimicked the methylation profile of S. griseus DNA, which was readily introduced into S. griseus. The results of this study raise the possibility of a promising approach to establish efficient transformation in several streptomycetes.

• The Korea Journal of Herbology
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• v.25 no.4
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• pp.55-59
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• 2010

Objective : This study was conducted to investigate the antioxidant activity and inhibitory effect on oxidative DNA damage of Nelumbinis Semen Extracts Methods : Nelumbins semen were extracted with hot-water and ethylacetate (EtOAC). The 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radical scavenging assay and $Fe^{2+}$ chelating assay were performed for antioxidative effect and ${\phi}X$-174 RF I DNA cleavage assay and intracellular DNA damage assay were used for inhibitory effect on intracellular DNA damage. Results : In DPPH, Hydroxyl radical scavenging activity and $Fe^{2+}$ chelating activity of EtOAC extracts were 96.22%, 53.53%, 64.72%, while those of hot-water extracts were 20.86%, 10.72%, 29.74% at $200{\mu}g/m{\ell}$, respectively. In ${\phi}X$-174 RF I plasmid DNA cleavage assay, the protective effects of EtOAC and hot-water extracts against oxidative DNA damage were 76% and 6% at $200{\mu}g/m{\ell}$, respectively. Conclusion : These results indicated that the seed extracts of Nelumbo nucifera can be used as a natural antioxidants, which effectively inhibits the oxidative DNA damage.

• The Korea Journal of Herbology
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• v.33 no.5
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• pp.105-110
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• 2018

Objectives : We tried to observe the fluorescence anisotropy and intensity of ethidium ion in the intercalating binding interaction between DNA and ethidium ions in the presence of berberine, and then tried to explain the effect of berberine on the intercalating interaction of ethidium ion with DNA. Methods : DNA(calf thymus DNA), berberine and ethidium bromide(EtBr) were purchased from Sigma-Aldrich Co. Proper amount of each compound was dissolved in 20 mM sodium phosphate buffer(pH 7.0) containing 100 mM of NaCl to prepare stock solutions. Collections of the fluorescence anisotropy and intensity data were performed on JASCO FP-8300 spectrofluorometer equipped with a polarizer and a Peltier temperature controller. The excitation of ethidium ion was done at 550 nm and the emission data were collected at 600 nm. For Stern-Volmer plot, the fluorescence data were collected at $18^{\circ}C$ and $30^{\circ}C$. Results : According to the results of this research, the weak competitive binding pattern between ethidium ion and berberine appeared in binding with DNA at low ratio of DNA to ethidium ion. But at high ratio of DNA to ethidium ion, this weak competition disappeared. Instead, berberine might bind to DNA by intercalating way. In other words, berberine could de-intercalate ethidium ion from DNA at low concentration of DNA relative to ethidium ion, but could not at high concentration of DNA relative to ethidium ion. In addition, the mechanism of fluorescence quenching of ethidium ion could also proceed differently, depending on the ratio of the amount of DNA to that of ethidium ion. Conclusions : The effect of berberine on the DNA-ethidium ion intercalating interaction could work differently, depending on the relative ratio of the amount of DNA to that of ethidium ion. This study also showed that fluorescence anisotropy analysis is very useful method to obtain detailed information for investigation of the complex binding interactions. In order to fully understand the mechanism of action of the pharmacological effect by berberine, studies on the effect of berberine on the action of proteins such as various enzymes closely related to berberine-induced medicinal effects should be continued.

• Journal of Plant Biology
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• v.34 no.4
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• pp.267-273
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• 1991

Polyamine levels in the male and female cells as well as DNA methyltransferase activity in the female cells during gametogenesis of Chlamydomonas reinhardtii indicated that both spermidine and spermine levels were decreased while DNA methyltransferase activity was markedly increased about 12 hours after the onset of gametogenesis. In vitro, putrescine and spermine at 1 mM inhibited methylation of chloroplasts DNA isolated from vegetative female cells by 35% and 65%, respectively. Spermine was found to be more inhibitory than putrescine at all concentrations tested. The pattern of the inhibition by polyamines appeared different from that caused by cations. The results obtained in this work suggest that the polyamine inhibition of DNA methylation is due to an action of polyamines on the enzyme involved instead of on the DNA itself.

• Reproductive and Developmental Biology
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• v.28 no.4
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• pp.211-219
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• 2004

The effects of adenosine 5'-triphosphate (ATP) and ATP analogs, P/sub 2y/ purinoceptor agonists, on growth of normal mouse mammary epithelial cells (NMuMG) were examined. Cells were plated onto 24 well plates in DMEM supplemented with 10 % fetal calf serum. After serum starvation for 24 hours, ATP, P/sub 2y/ purinoceptor agonists (AdoPP[NH]P, ATP-α-S, ATP-γ-S, β, γ-me-ATP and 2me-S-ATP), P/sub 2u/ purinoceptor agonist (UTP) and P/sub 2y/ purinoceptor antagonists (Reactive Blue 2, more selective to P/sub 2y/ receptor than PPADS; PPADS) were added. DNA synthesis was estimated as incorporation of 3H-thymidine into DNA (1 hour pulse with 1 μ Ci/ml, 18～19 hours after treatment). ATP, Adopp[NH]P, ATP-α-S or ATP-γ-S, significantly increased DNA synthesis at 1, 10 and 100 μM concentrations with dose-dependency (P<0.05), and the maximum responses of ATP and ATP analogs were shown at 100 μM concentration (P<0.05). The potency order of DNA synthesis was ATP≥ATP- γ -S>Adopp [NH]P>ATP-α-S. β, γ -me-ATP, 2me-S-ATP and UTP did not increase DNA synthesis. In autoradiographic analysis of percentage of S-phase cells, similar results were observed to those of DNA synthesis. Addition of 1, 10 or 100 μM Reactive Blue 2 or PPADS significantly decreased ATP (100 μM)-induced DNA synthesis, however, PPADS was less effective than Reactive Blue 2. In Elvax 40P implant experiment, ATP directly stimulated mammary endbud growth in situ suggesting the physiological regulator of ATP in mammary growth. ATP 100 μM rapidly increased MAPK activity, reaching a maximum at 5 min and then gradually decreasing to the base level in 30 min. ATP analogs, Adopp[NH]P and ATP-γ-S also increased MAPK activity, however, β, γ-me-ATP and 2me-S-ATP did not. The inhibitor of the upstream MAPK kinase (MEK), PD 98059 (25 μM), effectively reduced ATP (100 μM) or EGF(10 ng/ml, as positive control)-induced MAPK activity and DNA synthesis (P<0.05). These results indicate that ATP-induced DNA synthesis was prevented from the direct inhibition of MAPK kinase pathway. Overall results support the hypothesis that the stimulatory effects of normal mouse mammary epithelial growth by addition of ATP or ATP analogs are mediated through mammary tissue specific P/sub 2y/ purinoceptor subtype, and MAPK activation is necessary for the ATP-induced cell growth.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.5
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• pp.545-551
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• 2017

Curcuma longa L. (CL) is widely used as a spice and coloring agent in several foods, such as curry and mustard, as well as cosmetics and drugs. In this study, we investigated the protective effects of CL extracted with various solvents [methanol (MC), ethanol (EC), acetone (AC)] on $H_2O_2-induced$ DNA damage in human leukocytes along with total polyphenol contents (TPC) and antioxidant properties. The antioxidant effects of CL were determined by measuring 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity (RSA) and superoxide dismutase (SOD)-like activity. The preventive effect of CL on oxidative stress-induced DNA damage and DNA repair capacities were assessed using comet assay. MC showed the highest TPC (11.17 g gallic acid equivalents/100 g) and antioxidant properties among the solvent extracts. The $SC_{50}$ for DPPH RSA was MC: 35.0 > AC: 45.8 > EC: $57.8{\mu}g/mL$ and SOD-like activity was MC: 46.6 > EC: 141.5 > AC: $296.4{\mu}g/mL$. In the comet assay, the $ED_{50}$ value of MC showed the highest inhibition ($86.7{\mu}g/mL$) of $H_2O_2-induced$ DNA damage, followed by AC ($110.0{\mu}g/mL$) > EC ($115.8{\mu}g/mL$). Analysis of the percentage of damaged cells showed that repair capacity significantly decreased at 4, 8, and 12 h from $H_2O_2-induced$ oxidative stress in each extract. After 12 h, level of DNA damage recovery was similar to the negative control level. These results suggest that CL has potential antioxidant activity and a protective effect against oxidation-induced DNA damage, and the methanol extract of CL was the most effective.

• The Korean Journal of Nuclear Medicine Technology
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• v.18 no.1
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• pp.153-157
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• 2014

Purpose: SLE (systemic lupus erythematosus) is an inflammatory autoimmune disease, characterized by various autoantibody. The detection of Anti double-stranded DNA (Anti-ds DNA) is important in the diagnostics of SLE, and include the American College of Rheumatology (ACR) diagnostic criteria for SLE. Also SLE disease activity and correlativity with the level Anti-ds DNA antibody have been reported and Anti-ds DNA antibody quantitative test is very useful for tracing before and after SLE treatment. When These Anti-ds DNA antibody test (Farr assay: $^{125}I$ labeled ds-DNA and bound Anti-ds DNA antibodies complex in serum is precipitated by ammonium sulfate and used to centrifugation, measured it) inhaled supernatant after centrifugation, a lipemic specimen does not facilitate the formation of precipitate and also occurs situation was inhaled with precipitate. To solve these problems, The Influence of the degree of lipemic specimen was evaluated. Materials and Methods: September 2012 to February 2013, We selected lipemic samples (n=81) of specimen commissioned by Anti-ds DNA antibody test. Lipemic samples were done pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) used a micro-centrifuge (Eppendorf Model 5415D). At the same time lipemic specimen and pre-treatment samples were performed Anti-ds DNA antibody test (Anti-ds DNA kit, Trinity Biotech, Ireland). Statistical analysis were analyzed Pearson's correlation coefficients and regression and paired t-test, and Difference (%). Results: Experimental group 1 (Lipemic Specimen Anti-ds DNA Ab concentration ${\leq}7IU/mL$) at y=0.368X+4.732, $R^2=0.023$, Pearson's correlation coefficient was 0.154, paired t-test (P=0.003), Difference (%) mean 65.7 and showed a statistically significant difference. Experimental group 2 (Lipemic Specimen Anti-ds DNA Ab concentration ${\geq}8IU/mL$) at y=0.983X+0.298, $R^2=0.994$, Pearson's correlation coefficient showed 0.997, paired t-test (P=0.181), Difference (%) mean -5.53 made no statistically significant difference. Conclusion: Lipemic sample of low Anti-ds DNA Ab concentration (2.5-7 IU/mL) and the result is obtained pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) were made a significant difference statistically. Anti-ds DNA is one of the primary auto-antibodies present in patients with SLE, and remain an important diagnostic test for SLE. Therefore, we recommend preprocessing (high-speed centrifugation: 14,000 rpm 5 mins) in order to exclude the influence of lipemic specimen.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.4
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• pp.618-623
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• 2014

Sarijang, a soy sauce made from fermented black soybean (Rhynchosia nulubilis), sulfur fed duck, dried bark of Ulmus davidiana, Allium sativum, and bamboo salt, has been demonstrated to exert anti-inflammatory and anti-tumor activities. However, the antioxidant properties of Sarijang have not yet been elucidated. In this study, the antioxidant effects of Sarijang were investigated by determining total phenolic content (TPC), DPPH radical scavenging activity (DPPH RSA), total radical trapping antioxidant potential (TRAP), oxygen radical absorbance capacity (ORAC), and cellular antioxidant capacity (CAC). The inhibitory effects of Sarijang on oxidative stress-induced DNA damage (200 ${\mu}M$ $H_2O_2$, 250 ${\mu}M$ Fe-NTA, and 200 ${\mu}M$ HNE) in human leukocytes were evaluated by comet assay. The TPC of Sarijang was $1.04{\pm}0.01$ mg GAE/mL. DPPH RSA, TRAP, and ORAC values of Sarijang increased in a dose-dependent manner. The $IC_{50}$ for DPPH RSA of Sarijang was $11.2{\pm}0.3$ mg/mL, whereas $IC_{50}$ of TRAP was $209.5{\pm}2.0$ mg/mL. 2,2'-Azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress and oxidative stress-induced DNA damage in HepG2 cells were effectively abrogated by all tested concentrations of Sarijang (1~100 ${\mu}g/mL$). These results suggest that Sarijang has antioxidative activity and protective effects against oxidative DNA damage.

• Bulletin of the Korean Chemical Society
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• v.27 no.10
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• pp.1655-1658
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• 2006

The solution of $[Ru(phen)_2(dppz)]^{2+}$ and SDS has high Resonance Light Scattering (RLS) signals due to $[Ru(phen)_2(dppz)]^{2+}$ assemble on the surface of the SDS micelle. Because of the high affinity ($KB\geq10^6\;L\;mol^{-1}$) between $[Ru(phen)_2(dppz)]^{2+}$ and DNA, the adding of DNA in the solution of $[Ru(phen)_2(dppz)]^{2+}$-SDS makes the dissociation of $[Ru(phen)_2(dppz)]^{2+}$-SDS, and results in decreasing of the RLS signals and increasing of the absorbance. Based on this, a novel method is proposed for DNA assay. Under optimum condition, good linear relationship was obtained within the concentration range of 0.018-1.26 $\mu g\;mL^{-1}$, the linear equation is $I_{RLS}$ = 504.8-348.8 c (c: $\mu g\;mL^{-1}$) and the correlation coefficient (r) is 0.9992. The detect limit for calf thymus DNA is 8.6 ng $mL^{-1}$. The proposed method was successful applied to determine the extracted colibacillus plasmid DNA.

• Bulletin of the Korean Chemical Society
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• v.30 no.9
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• pp.1951-1955
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• 2009

A Thermodynamic study on the interaction of bovine calf thymus DNA with new designed Pd(II) complex (Ethylendiamine- 8-hydroxyquinolinato Palladium(II) chloride) was studied by using isothermal titration calorimetry (ITC) at 27 ${^{\circ}C}$ in Tris buffer solution at pH = 7.5. The enthalpies of Pd(II) complex + DNA interaction are reported and analysed in terms of the new solvation theory. It was indicated that there are three identical and non-cooperative sites for Pd(II) complex. The binding of a Pd(II) complex is endothermic with association equilibrium constants of 428.03 m$M^{-1}$ at 27 ${^{\circ}C}$. The binding of Pd(II) complex can cause some changes in the stability of the DNA at low and high Pd(II) complex concentrations. Our results suggested that this complex might interact with DNA as an intercalator, thus interfering with DNA replication and cell proliferation.