• Title/Summary/Keyword: mDNA

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THE EFFECT OF SODIUM FLUORIDE AND SODIUM ORTHOVANADATE ON OSTEOBLASTIC CELL LINE MC3T3-E1 CELLS (Sodium fluoride와 Sodium orthovanadate가 조골세포주 MC3T3-E1에 미치는 영향에 관한 연구)

  • Kim, Won-Jin;Chung, Kyu-Rhim
    • The korean journal of orthodontics
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    • v.21 no.1 s.33
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    • pp.97-111
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    • 1991
  • It is the aim of this study to investigate the effects of sodium fluoride and sodium orthovanadate upon the proliferation and activity of the osteoblast (MC3T3-E1 cells). MC3T3-E1 cells were cultured in $\alpha-MEM$ containing $10\%$ FBS and various concentration of sodium fluoride and sodium orthovanadate was appended to serum free media. DNA synthesis was examined through the $[^3H]$ thymidine incorporation into DNA. Collagen synthesis was examined through the $[^3H]$ proline incorporation into collagenase digestible protein and noncollagen protein. The following results were drawn; 1. Sodium fluoride stimulated the DNA synthesis of osteoblast significantly in dose-dependent manner within the concentration from $2{\mu}M$ to $10{\mu}M$ (P < 0.005). 2. Sodium orthovanadate stimulated the DNA synthesis of osteoblast significantly in dose-dependent manner within the concentration from $2{\mu}M\;to\;8{\mu}M$, however showed diminution at $10{\mu}M$ (P < 0.001). 3. Sodium fluoride and sodium orthovanadate stimulated the percent collagen synthesis of osteoblast significantly in dose-dependent manner within the concentration from $5{\mu}M$ to $10{\mu}M$ (P < 0.001). 4. Sodium fluoride and sodium orthovanadate stimulated the noncollagen synthesis of osteoblast significantly in dose-dependent manner within the concentration from $5{\mu}M\;to\;10{\mu}M$ (P < 0.001). In conclusion, sodium fluoride and sodium orthovanadate stimulate the proliferation and activity of osteoblast by stimulation of DNA synthesis and collagen and noncollagen synthesis in osteoblast.

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Antioxidative and Antigenotoxic Activity of Extracts from Rhododendron mucromulatum Turcz. Flowers (진달래 꽃 추출물의 항산화 및 항유전독성 활성)

  • Lee, Bo-Bae;Chun, Ji-Hae;Lee, Suck-Hee;Park, Hae-Ryong;Kim, Jung-Mi;Park, Eun-Ju;Lee, Seung-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.12
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    • pp.1628-1632
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    • 2007
  • The present study describes the preliminary evaluation of antioxidant activities and antigenotoxic effect of Rhododendron mucromulatum Turcz. flowers (RMF). The samples were prepared by extracting RMF with four different solvents (methanol, ethanol, acetone, and water), and antioxidant properties were evaluated by determining total phenolic contents (TPC), DPPH radical scavenging activity (RSA), and reducing power (RP). Water extract showed the highest total phenol content (328.1 mg/g gallic acid equivalents). Acetone extract showed the most potent RSA and RP. The $IC_{50}$ for RSA and RP in the acetone extract were 78 mg/mL and 454 mg/mL, respectively. The 200 mM $H_2O_2$ induced DNA damage, measured by Comet assay, was inhibited with water, methanol and acetone extract in dose dependent manner in human leukocytes. The inhibition rates were 42, 62, and 52% at the concentrations of 50 mg/mL of water, methanol and acetone extracts, respectively. These results suggest that R. mucromulatum Turcz. has significant antioxidative activity and protective effect against oxidative DNA damage.

Mechanism of DNA Cleavage Induced by Fe2+ Autoxidation

  • Kim, Jong-Moon;Kim, Seog-K.
    • Bulletin of the Korean Chemical Society
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    • v.32 no.3
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    • pp.964-972
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    • 2011
  • This work investigated the difference between $Fe^{2+}$ autoxidation-induced and Fenton-type cleavage of pBR322 plasmid DNA. $^{\cdot}OH$ generation reactions in the absence and presence of $H_2O_2$ under various conditions were also investigated. Although both the $Fe^{2+}$ autoxidation and Fenton-type reactions showed DNA cleavage and $^{\cdot}OH$ generation, there were significant differences in their efficiencies and reaction rates. The rate and efficiency of the cleavage reaction were higher in the absence of 1.0 mM of $H_2O_2$ than in its presence in 20 mM phosphate buffer. In contrast, the $^{\cdot}OH$ generation reaction was more prominent in the presence of $H_2O_2$ and showed a pH-independent, fast initial reaction rate, but the rate was decreased in the absence of $H_2O_2$ at across the entire tested pH range. Studies using radical scavengers on DNA cleavage and $^{\cdot}OH$ generation reactions in both the absence and presence of $H_2O_2$ confirmed that both reactions spontaneously involved the active oxygen species $^{\cdot}OH$, ${O_2}^{\cdot-}$, $^1O_2$ and $H_2O_2$, indicating that a similar process may participate in both reactions. Based on the above observations, a new mechanism for the $Fe^{2+}$ autoxidation-induced DNA cleavage reaction is proposed.

Light Regulation of rbcL Transcript and Protein-binding Region on rbcL Promoter in Maize

  • Lee, Jae-Seon;Sim, Woong-Seop
    • Journal of Plant Biology
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    • v.39 no.4
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    • pp.279-286
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    • 1996
  • To know the changes of rbcL mRNA level by illumination, Northern hybridization analysis was performed with maize (Zea mays L.cv. Golden X Bantam). The average level of rbcL. mRNA in the light-grown shoots was 3.1 times higher than that of the dark-grown shoots after 6 to 10 growth days. The maximum difference of rbcL mRNA level between the dark-grown and the light-grown shoots was 5.1 folds. These results indicate that accumulation of rbcL mRNAin maize shoots is induced by light. Since the transcriptional DNA binding proteins and their cognate promoter elements, we carried out gel-retardation assays to elucidate the specific binding proteins on the rbcL promoter. It was found that plastid proteins of light-grown shoots bound to the R2 DNA fragment (-33 to -229) and R3 DNA fragment (-230 to -418 from ATG) of the rbcL promoter. From the results of competitive binding assays and heat or protease treatments, it was demonstrated that the bindings were sequence-specific DNA-protein interactions. Therefore, it could be concluded that the rbcL promoter region has at least two specific recognition sites for plastid proteins.

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Virological Characteristics in Chronic Hepatitis B Patients with Concurrent HBsAg and anti-HBs Positivity

  • Lee, Hyeok-Jae;Lee, Min-Hyeok
    • Biomedical Science Letters
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    • v.18 no.3
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    • pp.283-290
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    • 2012
  • In this study, we investigated the virological characteristics, HBV DNA levels and presence of mutations of "a" determinant in the HBsAg S gene in chronic hepatitis B patients with coexisting HBsAg and anti-HBs. The 18 patients who were diagnosed with chronic hepatitis B were both positive for HBsAg and anti-HBs. HBV Among them, 15 patients were DNA positive. The median of HBV DNA levels in serum was $2.18{\times}10^7$ copies/mL with the HBsAg+/anti-HBsAb+ patients. Also, 4 of 8 HBeAg negative patients had HBV DNA levels higher than $10^4$ copies/mL and the median of HBV DNA levels was $2.03{\times}10^6$ copies/mL, which were significantly high. These results showed that viral replication still existed in most of the patients of the concomitant HBsAg and anti-HBs, and even in the some HBeAg negative patients. Furthermore, mutation within the "a" determinant of HBV were found in 7 of 15 patients. The most frequent changes were located at positions aa126. In addition, one mutation observed for HBsAg only positive.

Temperature dependent hydrogen exchange study of DNA duplexes containing binding sites for Arabidopsis TCP transcription factors

  • Kim, Hee-Eun;Choi, Yong-Geun;Lee, Ae-Ree;Seo, Yeo-Jin;Kwon, Mun-Young;Lee, Joon-Hwa
    • Journal of the Korean Magnetic Resonance Society
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    • v.18 no.2
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    • pp.52-57
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    • 2014
  • The TCP domain is a DNA-binding domain present in plant transcription factors and plays important roles in various biological functions. The hydrogen exchange rate constants of the imino protons were determined for the three DNA duplexes containing the DNA-binding sites for the TCP11, TCP15, and TCP20 transcription factors using NMR spectroscopy. The M11 duplex displays unique hydrogen exchange property of the five base pairs in the first binding site (5'-GTGGG-3'). However, the M15 and M20 duplexes lead to clear changes in thermal stabilities of these five base pairs. The unique dynamic features of the five base pairs in the first binding site might play crucial roles in the sequence-specific DNA binding of the class I TCP transcription factors.

Analysis of Double Stranded DNA-dependent Activities of Deinococcus radiodurans RecA Protein

  • Kim, Jong-Il
    • Journal of Microbiology
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    • v.44 no.5
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    • pp.508-514
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    • 2006
  • In this study, the double-stranded DNA-dependent activities of Deinococcus radiodurans RecA protein (Dr RecA) were characterized. The interactions of the Dr RecA protein with double-stranded DNA were determined, especially dsDNA-dependent ATP hydrolysis by the Dr RecA protein and the DNA strand exchange reaction, in which multiple branch points exist on a single RecA protein-DNA complex. A nucleotide cofactor (ATP or dATP ) was required for the Dr RecA protein binding to duplex DNA. In the presence of dATP, the nucleation step in the binding process occurred more rapidly than in the presence of ATP. Salts inhibited the binding of the Dr RecA protein to double-stranded DNA. Double-stranded DNA-dependent ATPase activities showed a different sensitivity to anion species. Glutamate had only a minimal effect on the double-stranded DNA-dependent ATPase activities, up to a concentration of 0.7 M. In the competition experiment for Dr RecA protein binding, the Dr RecA protein manifested a higher affinity to double-stranded DNA than was observed for single-stranded DNA.

Evaluation of two DNA extraction methods on exhumed bone samples: Ultrafiltration versus column affinity (유골에서 DNA 추출법 비교 연구: Ultrafiltration과 Column affinity)

  • Kim, Soonhee;Hong, Seungbeom;Kemp, Brian M.;Park, Kiwon;Han, Myunsoo
    • Analytical Science and Technology
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    • v.21 no.4
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    • pp.338-343
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    • 2008
  • Extraction of DNA from skeletal material is of great importance in the identification of human remains, but is particularly difficult because the high amount of microbial DNA was often co-extracted with human bone DNA. We found that a phenol/chloroform extraction, followed by ultrafiltration, and cleanup by via the $QIAquick^{(R)}$ PCR purification kit yields higher amounts of human genomic DNA compared with extraction by the column affinity $method^{(R)}$ alone. Ultrafiltration extraction of human DNA from ten exhumed bone samples yielded $0.041-1.120ng/{\mu}L$ DNA (mean = $0.498ng/{\mu}L$ DNA), and purification using the column affinity resulted in $0.016-0.064ng/{\mu}L$ DNA (mean = $0.034ng/{\mu}L$ DNA). Although the STR genotyping by the column affinity method was partially successful, all DNA samples by the ultrafiltration method produced full profiles from the multiplex PCR. The efficiency of STR genotyping was in accordance with the amounts of the human DNA extracted.

Molecular Biological Identification of Bacteria in Middle Ear Effusion Using 16S rDNA Multiplex PCR (중이 삼출액 미생물의 16S rDNA 복합중합효소연쇄반응을 이용한 분자생물학적인 진단)

  • 이정구;이인숙;박지연;정상운;오충훈
    • Korean Journal of Microbiology
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    • v.39 no.1
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    • pp.36-39
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    • 2003
  • The rapid and reliable 16S rDNA multiplex polymerase chain reaction (PCR) assay was established to characterize bacterial etiologies of middle ear effusion. These etiologies included Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumonia, which were detected in middle-ear effusion (MEE) samples taken from patient with otitis media. A total of 39 MEE samples were aspirated from 26 patients. DNA was extracted from MEE samples, and PCR was done with DNA extracts by using the common primers, which is localized at C4 region in the 16S rDNA gene of all bacterial species, and species-specific primers: (i) Haemophilus-specific primer, (ii) Moraxella- specific primer, and (iii) Streptococcus-specific primer. Among 39 samples tested, 24 (61.5%) were positive for H. influenzae, 10 (25.6%) were positive for M. catarrhalis, 3(7.7%) were positive for S. pneumonia, and 11 (28%) were negative for 165 rDNA multiplex PCR reaction. Nine samples (28.6%) exhibited a mixed infection and were positive for both H. infuenzae and M. catarrhalis. We suggested that 16S rDNA multiplex PCR is a useful method to identify rapidly for rapid identification of the pathogenic bacteria and characterization of bacterial etiologies of middle ear effusion.

Research on the detection of LCN DNA from traces on firearms (총기 흔적흔에서의 low copy number(LCN) DNA 검출에 관한 연구)

  • Jeon, Chung-Hyun;Park, Sung-Woo
    • Analytical Science and Technology
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    • v.24 no.1
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    • pp.51-59
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    • 2011
  • Genetic Identification has become an important forensic investigation method which discerns identity through analysis of physical samples discovered in various crime scenes. Recently more samples are being requested to undergo A-STR analysis of low copy number (LCN) DNA, which is known as touch evidence-type sample and left on various objects such as a pen briefly used by the criminal, the gear of the car used for driving, the handle, and various buttons inside a car. This research attempted to extract the LCN DNA of the touch evidencetype left on crushed fingerprints on firearms, etc. and examine the genotyping success rate. Four types of firearms (M16, K1A, COLT 45 Pistol, M29 Revolver) were fired individually and physical samples were gathered from four parts of each firearm. Subsequently, in order to extract the LCN DNA, Microkit and $Prepfiler^{TM}$ were used to compare and analyze the quantity of DNA extracted and the genotyping success rate. Analysis results showed that the quantity of DNA extracted by $Prepfiler^{TM}$ was on average 1.7 times higher than that of Microkit, and in genotype analysis success rate $Prepfiler^{TM}$ also demonstrated 24.9% on average in contrast to 0% for Microkit. In regards to the grip part of the K1A, $Prepfiler^{TM}$'s success rate was as high as 50.6%.