• Title/Summary/Keyword: DnaB

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Antioxidative Activity of the Extract from the Inner Shell of Chestnut

  • SON Kyung Hun;YANG He Eun;LEE Seung Chul;CHUNG Ji Hun;JO Byoung Kee;KIM Hyun Pyo;HEO Moon Young
    • Biomolecules & Therapeutics
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    • v.13 no.3
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    • pp.150-155
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    • 2005
  • The ethanolic extract of chestnut (Castanea crenata S. et Z., Fagaceae) inner shell (CISE) and one of its components, ellagic acid (EA), were evaluated for their protective effects against 1, 1-diphenyl-2-picryl hydrazine (DPPH) free radical generation and hydrogen peroxide-induced oxidative DNA damage in a mammalian cell line. CISE and EA were shown to possess the free radical scavenging effect against DPPH radical generation, significantly. They were also found to strongly inhibit hydrogen peroxide-induced DNA damage from Chinese hamster lung (CHL) cell, assessed by single cell gel electrophoresis assay and 8-hydroxy -2'-deoxy guanosine (8-OH-2'dG) assay. Furthermore, topical application of CISE [$12.5\%$(w/w) cream] and ellagic acid [$1.0\%$(w/w) cream] for 14 days potently inhibited malondialdehyde (MDA) formation of mouse dorsal skin (a marker of lipid peroxidation) induced by ultraviolet B exposure. Therefore, CISE and its component, ellagic acid, may be the useful natural antioxidants by scavenging free radicals, inhibition of lipid peroxidation and protecting oxidative DNA damage when topically applied.

Quantitative Assay for the Binding of Jun-Fos Dimer and Activator Protein-1 Site

  • Lee, Sang-Kyou;Park, Se-Yeon;Jun, Gyo;Hahm, Eun-Ryeong;Lee, Dug-Keun;Yang, Chul-Hak
    • BMB Reports
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    • v.32 no.6
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    • pp.594-598
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    • 1999
  • The Jun and Fos families of eukaryotic transcription factors form heterodimers capable of binding to their cognate DNA enhancer elements. We are interested in searching for inhibitors or antagonists of the binding of the Jun-Fos heterodimer to the activator protein-1 (AP-1) site. The basic-region leucine zipper (bZIP) domain of c-Fos was expressed as a fusion protein with glutathione S-transferase, and allowed to form a heterodimer with the bZIP domain of c-Jun. The heterodimer was bound to glutathione-agarose, to which were added radiolabeled AP-1 nucleotides. After thorough washing, the gel-bound radioactivity was counted. The assay is faster than the coventional electrophoretic mobility shift assay because the gel electrophoresis step and the autoradiography step are eliminated. Moreover, the assay is very sensitive, allowing the detection of picomolar quantities of nucleotides, and is not affected by up to 50% dimethylsulfoxide, a solvent for hydrophobic inhibitors. Curcumin and dihydroguaiaretic acid, recently known inhibitors of Jun-Fos-DNA complex formation, were applied to this Jun-GST-fused Fos system and revealed to decrease the dimer-DNA binding.

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Isolation of Novel Non-Toxic Bacillus thuringiensis from Soil Samples in Korea (한국 토양으로부터 새로운 무독성 Bacillus thuringiensis 균주의 분리)

  • 노종열;박현우;김호산;진병래;강석권
    • Korean journal of applied entomology
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    • v.34 no.4
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    • pp.373-377
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    • 1995
  • Four Bacillus thuringiensis isolates obtained from soil samples in Korea produce parasporal inclusions non-toxic to 10 insect species of three orders, Lepidopera, Diptera and Coleoptera. These four isolates are named NTB-1, NTB-2, NTB-3 and NTB-4, respectively. The morphology of parasporal inclusions of four isolates observed by phase contrast- and scanning electron microscope was all ovoid. Characterization of four non-toxic B. thuringiensis isolates was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and restriction endonuclease analysis. The results showed that parasporal inclusion proteins and total plasmid DNA profiles of four isolates are different from other known non-toxic B. thuringiensis strains', suggesting that four isolates are novel.

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Combined Genotoxic Effects of Aflatoxin B1, Ochratoxin A and Zearalenone in Rat Bone Marrow and Blood Leukocytes

  • Tigran, Harutyunyan;Anna, Karapetyan;Galina, Hovhannisyan;Rouben, Aroutiounian
    • Korean Journal of Environmental Biology
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    • v.31 no.3
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    • pp.189-191
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    • 2013
  • Mycotoxins such as aflatoxin B1 (AFB1), ochratoxin A (OTA) and zearalenone (ZEA) are widespread contaminants of food and feedstuffs. It is very likely, that humans and animals are always exposed to mixtures of mycotoxins rather than to individual compounds. Therefore, risk assessments should consider mixture toxicity data. In the present study the combination of AFB1, OTA and ZEA was tested for genotoxicity in rat bone marrow and blood leukocytes after 15, 30 and 60 days treatment. The level of DNA damage was determined by the comet assay. The tail intensity and Olive tail moment in leukocytes and bone marrow cells were significantly higher than in controls. At the same time, the level of DNA damage in bone marrow cells was higher than in leukocytes. The data suggests that prolonged exposure to mycotoxins combination through food consumption can induce DNA damage contributing to the harmful effects in vivo.

Synthesis and Characterization of Oligonucleotides Containing Site-Specific Bulky $N^2$-Aralkylated Guanines and $N^6$-Aralkylated Adenines

  • Moon, Ki-Young;Kim, Yeong-Shik
    • Archives of Pharmacal Research
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    • v.23 no.2
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    • pp.139-146
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    • 2000
  • 7- Bromomethylbenz[a]anthracene is a known mutagen and carcinogen. The two major DNA adducts produced by this carcinogen, i.e., $N^2$-(benz[a]anthracen-7-yl methyl)-2'-deoxyguanosine (2, b[a]$a^2$G) and $N^6$-(benz[a]anthracen-7-ylmethyl)-2'-deoxyadenosine (4, b[a]$a^6$/A), as wel 1 as the simpler benzylated analogs,$N^2$-benzyl-2'deoxyguanosine (1, $bn^2$G) and $N^6$-benzyl-2'-deoxyadenosine (3, $bn^6$/A), were prepared by direct aralkylation of 2'-deoxyguanosine and 2'-deoxyadenosine. To determine the site-specific mutagenicity of these bulky exocyclic amino-substituted adducts, the suitably protected nucleosides were incorporated into 16-base oligodeoxyribonucleotides in place of a normal guanine or adenine residues which respectively are part of the ATG initiation codon for the lac Z' \alpha-complementation gene by using an in situ activation approach and automated phosphite triester synthetic methods. The base composition and the incorporation of the bulky adducts into synthetic oligonucleotides were characterized after purification of the modified oligonucleotides by enzymatic digestion and HPLC analysis.

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Genetic Variation in the Asian Shore Crab Hemigrapsus sanguineus in Korean Coastal Waters as Inferred from Mitochondrial DNA Sequences

  • Hong, Sung-Eic;Kim, Jin-Koo;Yu, Jeong-Nam;Kim, Keun-Yong;Lee, Chung-Il;Hong, Kwan-Eui;Park, Kie-Young;Yoon, Moon-Geun
    • Fisheries and Aquatic Sciences
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    • v.15 no.1
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    • pp.49-56
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    • 2012
  • Genetic variation in the Asian shore crab Hemigrapsus sanguineus was determined from partial mitochondrial DNA (mtDNA) sequences of the cytochrome b (Cytb) gene. Samples included 143 crabs from six localities along three coastlines in South Korea. A nucleotide sequence analysis revealed 38 variable sites in a 470-bp sequence, which defined 37 haplotypes. The haplotypes were not associated geographically and had a shallow genealogy. Pairwise $F_{ST}$ tests and a two-dimensional scaling analysis revealed no significant genetic differentiation among most of the populations. The low pairwise comparison values, but significant genetic differentiation of a northeastern population from all other populations, might have been influenced by a restriction in gene flow caused by hydrographic conditions such as ocean boundaries. The high haplotype diversity, low nucleotide diversity, and time since H. sanguineus expansion in Korean coastal waters indicate rapid population growth and a recent, sudden expansion in the Late Pleistocene.

Protective Effects of Probiotic Lactobacillus rhamnosus IMC501 in Mice Treated with PhIP

  • Dominici, Luca;Villarini, Milena;Trotta, Francesca;Federici, Ermanno;Cenci, Giovanni;Moretti, Massimo
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.371-378
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    • 2014
  • The aim of the present study was to investigate the antigenotoxic properties of the probiotic Lactobacillus rhamnosus IMC501; DNA damage was induced by one representative food mutagen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). Mice were treated orally with suspensions of lactobacilli for 10 days before administration of food mutagen. During the treatment, the abundance of lactobacilli in feces, as assessed by qPCR analysis, increased, whereas ${\beta}$-glucuronidase and N-acetyl-${\beta}$-glucosaminidase activities decreased. The extent of DNA damage was measured in colon and liver cells by comet assay. In colonocytes, diet supplementation with IMC501 resulted in a significant inhibition of DNA damage induced by PhIP. The results obtained in this in vitro study suggest that Lactobacillus rhamnosus IMC501 used as a dietary supplement can provide a useful integration of antimutagen food components of the normal diet, which are generally lower than the protective level.

Molecular Cloning and Characterization of the Gene encoding $\beta$-N-acetylhlucosaminidases Homologue from Bombyx mandarina (멧누에(Bombyx mandarina)로부터 $\beta$-N-Acetyglunosamicidase를 코딩하는 cDNA의 분리 및 염기서열 결정)

  • 구태원;황재삼;성규병;윤은영;방혜선;권오유
    • Journal of Sericultural and Entomological Science
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    • v.41 no.3
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    • pp.147-153
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    • 1999
  • Chitinolytic enzymes such as ${\beta}$-N-acetylglucosaminidase are major hydrolases involved in insect molting. We have isolated, sequenced a CDNA encoding ${\beta}$-N-acetylglucosaminidase from the silkworm, Bombyx mandarina, and compared its sequence with genes encoding chitinolytic enyzmes from other sources. The insert DNA in the clone is 3,284 nucleotides long with an open reading frame of 1,788 nucleotides that encodes a protein of 596 amino acids with a molecular weight of 68.2 kDa. There is a 3’-untranslated region composed with 1.479 nucleotides and are several potential polyadenylation signals. The predicted amino acid sequence apparently contains a leader peptide of 23 amino acids. A search of the amino acids sequence databases for sequences similarities to other ${\beta}$-N-acetylglucosaminidases or ${\beta}$-N-acetylhexosaminidases. The highest similarity matched with the enzyme from B. mori, which has a sequence identity of 95%. On the other hand, the identity between the B. mandarina enzyme and those from M. sexta and human are 70% and 24%, respectively.

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Therapeutic Efficacy of Prednisolone Withdrawal Followed by Recombinant ${\alpha}$ Interferon in Children with Chronic Hepatitis B (소아 만성 B형 간염 환자에서 스테로이드 이탈 요법 후 인터페론 병용 투여의 치료 효과)

  • Ryu, Na-Eun;Kim, Byung-Ju;Ma, Jae-Sook;Hwang, Tai-Ju
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.2 no.2
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    • pp.169-177
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    • 1999
  • Purpose: To evaluate the efficacy of interferon alpha therapy with or without prednisolone in children with chronic hepatitis B. Methods: Twenty-eight children (22 boys, 6 girls, mean age 130 months) had seropositive results for HBsAg, HBeAg and HBV DNA; 11 had chronic persistent hepatitis and 17 had chronic active hepatitis. The patients were divided into two groups depending upon their inflammatory activity on liver biopsy, pretreatment serum ALT levels and HBV DNA levels. Fourteen children (group 1: chronic active hepatitis, ALT ${\geq}$ 100 IU/L and HBV DNA ${\leq}$ 100 pg/$300\;{\mu}L$) received interferon alpha 2a 5 $MU/m^2$ of body surface three times weekly for 6 months. Fourteen children (group 2: chronic persistent hepatitis or chronic active hepatitis with ALT < 100 IU/L or HBV DNA > 100 pg/$300\;{\mu}L$) received prednisolone in decreasing daily doses of 60 mg/$m^2$, 40 mg/$m^2$, and 20 mg/$m^2$, each for 2 weeks, followed after 2 weeks by interferon alpha 2a on the same schedule. At the end of therapy, 3 end points were analyzed: HBeAg seroconversion, serum ALT normalization rate and clearance of serum HBV DNA. Results: At the end of treatment, HBe antigen-to antibody seroconversion was higher but not more significant in group 1 than group 2 (71.4% vs. 50.0%). Only one patient in group 2 who lost HBeAg, also cleared HBsAg. ALT normalization was similar in both groups (64.3% in group 1 vs. 55.6% in group 2). Clearance of serum HBV DNA was observed in 78.6% of patients in group 1 and 64.3% in group 2, but no significant differences. Complete response was similarly achieved in both groups (57.1% in group 1 vs. 50.0% in group 2). Interferon alpha therapy with prednisolone priming was well tolerated and all children finished therapy. Conclusion: The combined therapy with prednisolone followed by interferon alpha may be safe and effective in inducing a serological and biochemical remission of the disease in approximately 50% of children with chronic hepatitis B and with a high level of viral replication and less active disease. However, a controlled study should be performed to confirm these results.

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Molecular cloning of casein gane which is expressed in mammary glands (유선조직에서 특이적으로 발현되는 카제인 유전자의 클로닝(I))

  • Choe, In-Ho;Bae, Bong-Jin;Lee, Chang-Su
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.21 no.1
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    • pp.53-66
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    • 1995
  • The gene for ${\gamma}$-casein, a milk protein, is a member of a family of casein gene which is expressed in mammary glands of the animal during the late gestation and lactation periods binder the influence of various hormones. In order to elucidate tile mechanisms b)'which hormones regulate the coordinate induction of milk protein genes, the mouse ${\gamma}$-casein gene was isolated and characterized. The ${\gamma}$-casein gene was screened from a mouse genomic library constructed in bacteriophage EMBL3 with the ${\gamma}$-casein CDNA used as probe and one clone was obtained. The ${\gamma}$-casein CDNA as probe was partially sequenced and contained ATG start codon and 5'-noncoding region. The cloned genomic DNA was digested with Sal I restriction enzyme, by which the insert DNA can be isolated from EMBL3 vector. Three DNA bands were observed and the size of DNAs was approximately 28kb, 14kb and 9Kb, respectively Accordingly the size of the insert DNA was calculated with approximately 23Kb. The result of Southern blot analysis, however, showed that the cloned genomic DNA was not hybridized with the synthetic oligonucleotides (40 mer) of cDNA 5'-end region, but it was hybridized with the y -casein CDNA. This means that tile cloned y -casein gene may not contain its promoter region. The ${\gamma}$ -casein genomic DNA containing the promoter region has been screening from mouse genomic library with oligonucleotides of CDNA 5'-end region as probe, and twenty-nine clones was obtained.

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