• Korean Journal of Microbiology
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• v.34 no.1_2
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• pp.51-57
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• 1998

A study was carried out to understand some parameters affecting on RAPD analysis with Lactobacillus species. From the results, we found that appearance of specific DNA bands were very influenced by the concentration of $MgCl_2$ but it was overcome by applying enough amount of Taq DNA polymerase. Other parameters such as concentrations of template DNA, random primers and Taq DNA polymerase have enhanced the production of specific DNA bands by increasing their concentration applied. However, we noticed that G/C contents of random primers did not show any correlations with number of specific RAPD bands generated but the RAPD results were heavily influenced by the characteristics of the random primers, that is, the sequences of the oli.

• Environmental Mutagens and Carcinogens
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• v.10 no.2
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• pp.85-92
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• 1990

The effects of aphidicolin (APC), 2`,3`-dideoxythymidine 5`-triphosphate (ddTTP), and novobiocin (NOV) on the frequencies of chromosome aberrations induced by ethyl methanesulfonate (EMS) or bleomycin (BLM) were examined in synchronized Chinese hamster ovary (CHO)-K$_1$ cells. The cells were synchronized by the thymidine double block method. APC, ddTTP and NOV alone did not affect the frequencies of chromosome aberrations. The cells in late G$_1$ and early S phases were sensitive to the induction of chromosome aberrations by EMS, wherase cells in G$_2$ phase were most sensitive to chromosme aberration by BLM.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1326-1334
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• 2008

The prime-boost vaccination with DNA vaccine and recombinant viral vector has emerged as an effective prophylactic strategy to control infectious diseases. Here, we compared the protective immunities induced by multiple alternating immunizations with DNA vaccine (pCIgB) and replication-incompetent adenovirus (Ad-gB) expressing glycoprotein gB of pseudorabies virus (PrV). The platform of pCIgB-prime and Ad-gB-boost induced the most effective immune responses and provided protection against virulent PrV infection. However, priming with pCIgB prior to vaccinating animals by the DNA vaccine-prime and Ad-boost protocol provided neither effective immune responses nor protection against PrV. Similarly, boosting with Ad-gB following immunization with DNA vaccine-prime and Ad-boost showed no significant responses. Moreover, whereas the administration of Ad-gB for primary immunization induced Th2-type-biased immunity, priming with pCIgB induced Th1-type-biased immunity, as judged by the production of PrV-specific IgG isotypes and cytokine IFN-$\gamma$. These results indicate that the order and injection frequency of vaccine vehicles used for heterologous prime-boost vaccination affect the magnitude and nature of the immunity. Therefore, our demonstration implies that the prime-boost protocol should be carefully considered and selected to induce the desired immune responses.

• Reproductive and Developmental Biology
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• v.29 no.1
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• pp.9-13
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• 2005

This study was performed to analyze the sequence variation in mtDNA D-loop and their effects on milk and milk fat production in Holstein cows. The analyzed sequences were compared with previously published sequences from other cattle breeds (GenBank J01394). PCR was performed to amplify a total of 964 bp between nucleotide 15758 and 383 within D-loop region of mtDNA using specific primers. Thirty five polymorphic sites by nucleotide substitution were found in mtDNA. The frequencies of positions at 106, 169, 16057, 16231 and 16255 nt with high levels of sequence polymorphism were 0.090, 0.555, 0.055, 0.090 and 0.050, respectively. The substitution effect at 169 nt was found significant on milk production, and substitution effect at 16118, 16139 and 16302 nt was highly significant (p<0.1) on milk fat production. Polymorphism of mtDNA sequence in D-loop region might be useful for the analysis of cytoplasmic genetic variation and associations with the other economically important traits and maternal lineage analysis in Holstein cows.

• Clinical and Experimental Pediatrics
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• v.48 no.6
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• pp.634-639
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• 2005

Purpose : An understanding of the immunological process is required if primary prevention of atopic diseases is to be developed in early childhood. But, it is too hard to distinguish atopy from nonatopy under the age of two clinically, because the expression of phenotype and cytokines is vague in early childhood. We evaluated DNA methylation changes at Th2 interleukin-4 gene in peripheral blood from atopic children. Methods : We selected 15 allergic children(mild : eight, moderate to severe : seven) and seven normal controls by using family allergy scores and clinical histories. We measured Total IgE and Der f II specific IgE levels and cultured peripheral blood mononuclear cells with Der f II stimulation and extracted DNA from Der f II specific T cells. We examined the change of CpG methylation in DNA from atopic and nonatopic children. Results : In T cells from normal children, IL-4 DNA were predominantly methylated; otherwise, CpG demethylation occurred in Der f II specific T cells from allergic children. Conclusion : IL-4 DNA methylation changes occurred in T genes from allergic children and DNA methylation assay in early childhood.

• Korean Journal of Plant Resources
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• v.29 no.2
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• pp.163-170
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• 2016

Antioxidant activity and inhibitory effect on oxidative DNA damage of ethyl acetate fractions extracted from Cone of Red Pine (Pinus densiflora) were investigated to find utilization of Cone, by-product of Red Pine, thrown out after berry shatter, as a new natural plant resource. Cone from P. densiflora was extracted with methanol (MeOH) and separated to petroleum ether, ethyl acetate and water fraction. Among them, ethyl acetate fraction was used. The antioxidant activity was conducted by the 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, 2, 2'-Azino-bis (3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS) radical scavenging assay, Fe²⁺ chelating assay and reducing power assay. The inhibitory effect on oxidative DNA damage was determined by DNA cleavage assay using φX-174 RF I plasmid. The results of DPPH and ABTS radical scavenging activity at 200 ㎍/㎖ of extracts were 86.50% and 95.80% respectively, which were similar figures compared with L-ascorbic acid as control. Fe²⁺ chelating activity was 77.96% and reducing power was 0.77 at 200 ㎍/㎖. Total phenolic component was 27.29±0.3 ㎎/g and Vitamin C content was 1.84±0.1 ㎎/g. Also ethyl acetate fraction from Cone has inhibitory effect, using φX-174 RF I plasmid on DNA cleavage assay. In conclusion, Cone, by-product of P. densiflora, showed high antioxidant activity and inhibitory effect on oxidative DNA damage. Therefore this study suggests Cone, useless by-product, can be developed as a new natural plant resource with lots of utilization such as an effective antioxidant, natural medicine, food, cosmetics and so on.

• Journal of Animal Science and Technology
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• v.46 no.5
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• pp.729-734
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• 2004

This study was performed to analyze the sequence variations of mtDNA D-loop and their effects on milk in Hanwoo(Korean cattle). The resulting sequences were compared with previously published sequences for other cattle breeds (GenBank JOI394). The Polymerase Chain Reaction was performed to amplify a total of 964 bp between nucleotide 15758 and 383 within D-loop region of mtDNA using specific primers. Twenty polymorphic sites by nucleotide substitution were found in mtDNA D-loop region of Hanwoo. The frequencies of positions at 8, 169, 16042, 16051, 16057, 16093, 16119, 16122, 16209, 16255 and 16302 nt with high levels of sequence polymorphism were 0.150, 0.950, 0.085, 0.138, 0.106, 0.085, 0.138, 0.212, 0.085, 0.148 and 0.180, respectively. The substitution effect at 16119(p<0.1) and 16185(p< 0.05) nt was found significant on milk production. Polymorphism of mtDNA sequence in D-Ioop region could be useful for the analysis of cytoplasmic genetic variation and associations with the other economically important traits and maternal lineage analysis in Hanwoo.

• Korean Journal of Pharmacognosy
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• v.30 no.1
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• pp.1-6
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• 1999

To evaluate cytotoxic effect and topoisomerase I inhibition activity of Caesalpinia sappan L., both water and methanol extracts were examined using in vitro assay. The cytotoxic effect of Caesalpinia sappan L. examined using MTT and SRB assay and $IC_{50}$ values were measured against U937, HL60, HepG2, SNU-1, SNU-16 cancer cell lines. Among them the representative cytotoxic results are shown as follows; water extract (U937=13.39 ${\mu}g/ml$, HL60=8.65 ${\mu}g/ml$, HepG2=38.48 ${\mu}g/ml$, SNU-1=7.72 ${\mu}g/ml$, SNU-16=25.49 ${\mu}g/ml$), methanol extract (U937=13.35 ${\mu}g/ml$, HL60=9.43 ${\mu}g/ml$, HepG2=25.67 ${\mu}g/ml$, SNU-1=8.37 ${\mu}g/ml$, SNU-16=28.64 ${\mu}g/ml$). The inhibitory concentration of DNA topoisomerase I activity against water extract was 100 ${\mu}g/ml$ and the inhibitory concentration of DNA topoisomerase I against methanol extract was 400 ${\mu}g/ml$.

• 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.

• Biomolecules & Therapeutics
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• v.19 no.2
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• pp.261-266
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• 2011

As recent reports suggest that nanoparticles may penetrate into cell membrane and effect DNA condition, it is necessary to assay possible cytotoxic and genotoxic risk. Three different sizes of magnetic nanoparticle silica (MNP@$SiO_2$) (50, 100 and 200 nm diameter) were tested for cytotoxicity and DNA damage using L5178Y cell. MNP@$SiO_2$ had constant physicochemical characteristics confirmed by transmission electron microscope, electron spin resonance spectrometer and inductively coupled plasma-atomic emission spectrometer for 48 h. Treatment of MNP@$SiO_2$ induced dose and time dependent cytotoxicity. At 6 h, 50, 100 or 200 nm MNP@$SiO_2$ decreased significantly cell viability over the concentration of 125 ${\mu}g/ml$ compared to vehicle control (p<0.05 or p<0.01). Moreover, at 24 h, 50 or 100 nm MNP@$SiO_2$ decreased significantly cell viability over the concentration of 125 ${\mu}g/ml$(p<0.01). And treatment of 200 nm MNP@$SiO_2$ decreased significantly cell viability at the concentration of 62.5 ${\mu}g/ml$ (p<0.05) and of 125, 250, 500 ${\mu}g/ml$ (p<0.01, respectively). Furthermore, at 48 h, 50, 100 or 200 nm MNP@$SiO_2$ decreased significantly cell viability at the concentration of 62.5 ${\mu}g/ml$ (p<0.05) and of 125, 250, 500 ${\mu}g/ml$ (p<0.01, respectively). Cellular location detected by confocal microscope represented they were existed in cytoplasm, mainly around cell membrane at 2 h after treatment of MNP@$SiO_2$. Treatment of 50 nm MNP@$SiO_2$ significantly increased DNA damage at middle and high dose (p<0.01), and treatment of 100 nm or 200 nm significantly increased DNA damage in all dose compared to control (p<0.01). Taken together, treatment of MNP@$SiO_2$ induced cytotoxicity and enhanced DNA damage in L5178Y cell.