• Title/Summary/Keyword: inducibility

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EFFECTS OF CHITOSAN ON HUMAN OSTEOBLASTS (키토산이 조골세포에 미치는 영향)

  • Kim, Ki-Hong;Park, Young-Ju;Park, Jun-Woo;Lee, Yong-Chan;Cho, Byoung-Ouck;Ahn, Byoung-Keun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.28 no.4
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    • pp.280-285
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    • 2002
  • As the result of the study concerning "bone inducibility of chitosan", 1. "BMP-2" was observed mainly through the test when the "osteoblast" is exposed to the "chitosan". The expression of BMP-2 was 542.63 times compared to control after 2 hours exposure and it was maintained 16.60 times till 24 hours. 2. The expression of BMP-4 was decreased compared to control during exposure. 3. The expression of BMP-7 revealed two peaks during exposure. 4. The expression of osteocalcin was increased in early phase, and then decreased. Although it is not clear whether the "chitosan" is clinically effective material as a "bone induction material", we could say that it has a function for bone induction. Further detailed study will be required.

Expression of Metallothionein mRNA in Cadmium Treated Leydig Cells (테스토스테론생성 레이디히세포(Leydig)에서의 메탈로치오닌 유전자 발현특성연구)

  • Park Kwangsik
    • Environmental Analysis Health and Toxicology
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    • v.19 no.3
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    • pp.261-269
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    • 2004
  • Although the biological functions of metallothioneins (MTs) are still being investigated, they have been suggested to be involved in detoxification of heavy metals, scavenging of free radicals, and protection against alkylating agents. MTs have been reported to be induced in most of animal tissues by heavy metals such as zinc, copper, mercury and cadmium, and the proteins have binding affinities to the metals. However, the presence or induction of MTs was reported not to be clear in leydig cells, which produce testosterone for the maturation of spermatozoa in male testes. In this study, we investigated the inducibility of metallothionein isomers by cadmium in cultured mouse leydig cells. Total RNA was extracted from the near confluent grown leydig cells and RT-PCR was Performed using the Primers which were synthesized on the basis of MT-1, 2, 3 and 4 cDNA from GenBank database. As results, MT-1 and MT-2 mRNA were found to be expressed in cadmium non-treated control cells and MT 1 mRNA expression was dose-dependent when leydig cells were treated with cadmium chloride. But MT-3 which is known to be brain specific and MT-4 which is another isoform of metallothionein, were not expressed. Other genes induced or depressed in cadmium treated leydig cells were also identified by microarray techniques.

The RecA-like protein of Schizosoccharomvces pombe: its cellular level is induced by DNA-damaging agents (DNA 상해요인에 의한 Schizosaccharomyces pombe RecA 유사 단백질의 유도생성)

  • 이정섭;박상대
    • The Korean Journal of Zoology
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    • v.37 no.2
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    • pp.232-239
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    • 1994
  • RecA protein plans a central role in homologous recombination and DNA repair in Escherichia cofi (E. colD. The function 8nd structure of this protein are universal in prokarvotes and also conserved in eukaryotes such as yeast. The RecA-like protein with 74 lInDa in size has already been identified and purified from a fission yeast Schizosaccharomyces pombe (5. pommel (Lee, 19911. From this study it was revealed that the RecA-like protein of 5. pombe was highly inducible to various DNA damaging agents and inhibitors of nucleotide pool svnthesizins enzymes. The cellular level of the 5. pombe RecA-like protein wi,u markedly increased, upto 5- to 10-fold, by treatment with various DNA-damains agents including ultraviolet (UV) light, methyl methanesulfonate WS),4-nitroquinoline-1-oxide (4-NQO), and mitomycin-C (MMC), similar to E. cofi RecA protein. Interestingly, the protein level was also increased by inhibitors of nucleotide pool forming enzlwnes such as methotrexate (MTX) and hvdroxvurea (HU). The most effective doses for the inducibility of 4-NQO, MMS, W, MMC, MTX, and HU were 0.2 Ug/ml, 30 mM, 200 J/ma, 0.4 $\mus/ml,$ 1 Ug/ml, and 100 mM, respectively. The range of effective duration time for the inducibilitv of RecA-like protein was from 270 to 450 mins. These results suggest that the 5. pombe RecA-like protein also platys an imortant role in cellular responses to DNA damage as in E. coli system.

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Molecular Characterization of an Apple cDNA Encoding Cinnamyl Alcohol Dehydrogenase

  • Kim, Sung-Hyun;Lee, Jae-Rin;Shin, Yong-Uk;An, Gyn-Heung;Kim, Seong-Ryong
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.475-481
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    • 1999
  • The study of lignin, a major component of secondary cell wall, has been partly focused on its removal from the woody part in the kraft pulping industry. Cinnamyl alcohol dehydrogenase (CAD; EC 1.1.l95) catalyzes the synthesis of cinnamyl alcohols from corresponding cinnamaldehydes. A cDNA clone, MdCADl, encoding putative CAD from apples (Malus domestica Borkh. cv Fuji) was characterized in this study. The clone contains an open reading frame of 325 amino acid residues, which shows a greater than 80% identity with Eucalyptus CADl. MdCADl mRNA was detectable in vegetative tissues and was strongly expressed in the fruit. The expression pattern of MdCADl mRNA in the fruit peel after light exposure was also examined. The mRNA was rapidly increased until 1 day after light exposure and remained stable thereafter, suggesting that MdCADl is light inducible. The inducibility of the MdCADl gene was examined using several environmental stresses. Mechanical wounding of leaves increased the MdCADl mRNA level and the induction was further increased by salicylic acid. Southern blot hybridization showed that there is either one or a few copies of CAD genes in apples. To our knowledge, it is believed that MdCADl is the first CAD clone expressed predominantly in fruit.

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Genotoxic Evaluation of Surfactin C in Chinese Hamster Lung Cell Line

  • Lim, Jong-Hwan;Song, In-Bae;Park, Byung-Kwon;Kim, Myoung-Seok;Hwang, Youn-Hwan;Yun, Hyo-In
    • Toxicological Research
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    • v.25 no.1
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    • pp.47-50
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    • 2009
  • To investigate the mutation inducibility of surfactin C, we performed the chromosome aberration assay with Chinese hamster lung cells in vitro. The colorimetric MTT screening assay was carried out to determine the cytotoxicity index ($IC_{50}$) of surfactin C. The $IC_{50}$ value was $125{\mu}g/ml$. For the chromosome aberration test of surfactin C, the maximum concentration was employed as $125{\mu}g/ml$, followed by 62.5 and $31.25{\mu}g/ml$ for the lower concentrations, with or without metabolic activation (S9). Cyclophosphamide and mitomycin C were used as positive controls in the presence and absence of S9 metabolic activation, respectively. These results showed that surfactin C was not capable of inducing chromosome aberration, as measured by the chromosome aberration test using Chinese hamster lung cell line. There is no evidence for surfactin C to have a genotoxic potential.

Isolation and Characterization of a Salt Inducible Promoter from Chlorella vulgaris PKVL7422

  • Min-Jeong Kim;Su-Hyun Kim;Najib Abdellaoui;Tae-Jin Choi
    • Journal of Microbiology and Biotechnology
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    • v.33 no.7
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    • pp.955-963
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    • 2023
  • Chlorella is a eukaryotic organism that can be used as an industrial host to produce recombinant proteins. In this study, a salt-inducible promoter (SIP) was isolated from the freshwater species Chlorella vulgaris PKVL7422 from the screening of genes that were upregulated after salt treatment. Several cis-acting elements, including stress response elements, were identified in the isolated SIP. Moreover, the Gaussia luciferase gene was cloned after the SIP and transformed into C. vulgaris to test the inducibility of this promoter. Reexamination of transcriptome of C. vulgaris revealed that genes involved in the synthesis of methyl jasmonic acid (MeJA), gibberellin (GA), and abscisic acid (ABA) were upregulated when C. vulgaris was treated with salt. Furthermore, the expression level of recombinant luciferase increased when the transformed C. vulgaris was treated with salt and MeJA, GA, and ABA. This study represents the first report of the C. vulgaris SIP and highlights how transformed microalgae could be used for robust expression of recombinant proteins.

Characterization of hrp2 + Gene Related to SNF2 Family in Schizosaccharomyces pombe (Schizosaccharomyces pombe에서 SNF2에 속하는 hrp2+ 유전자의 특성 연구)

  • Park, In-Soon
    • Environmental Mutagens and Carcinogens
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    • v.22 no.3
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    • pp.137-141
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    • 2002
  • The SNF2/SW12 family comprises proteins from a variety of species with in vivo functions, such as transcriptional regulation, maintenance of chromosome stability during mitosis, and various types of DNA repair. This study was shown the characterization of hrp2+ gene which was isolated by PCR amplification using the conserved domain of SNF2 motifs. Sequence analysis of hrp2+ gene showed striking evolutionary conservation among the SNF2 family of proteins. The transcript of hrp2+ gene was found to be a 4.7 kb as identified by Northern hybridization. In addition, to determine the transcription initiation site of hrp2+ gene, primer extension analysis was performed. This result showed the band of 64 bp. The transcriptional start point was mapped to a position of 47 base pair from the first ATG codon of translational initiation codon. In order to investigate the inducibility of hrp2+ gene, transcript levels were examined after treating the cells to various DNA damaging agents. The transcripts of hrp2+ were induced by UV-irradiation. But the transcripts were not induced by treatment of 0.25% Methylmethane sulfonate (MMS). These results implied that the effects of damaging agents are complex and different regulatory pathways exist for the induction of this gene.

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Altered expression of mud loach (Misgurnus mizolepis; Cypriniformes) hepcidin mRNA during experimental challenge with non-pathogenic or pathogenic bacterial species

  • Lee, Sang-Yoon;Kim, Dong-Soo;Nam, Yoon-Kwon
    • Journal of fish pathology
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    • v.24 no.3
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    • pp.279-287
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    • 2011
  • Transcriptional response patterns of mud loach (Misgurnus mizolepis; Cypriniformes) hepcidin, a potential ortholog to human hamp1, in response to experimental challenges with non-pathogenic and pathogenic bacterial species were analyzed based on the semi-quantitative reverse transcription-PCR assay. Mud loach hepcidin transcripts were much more preferentially induced by pathogenic bacterial species (Edwardsiella tarda and Vibrio anguillarum) causing apparent pathological symptoms than by non-pathogenic species (Escherichia coli and Bacillus thuringiensis) displaying neither clinical signs nor mortality. However in overall, the induced amounts of hepcidin transcripts were positively related with the number of bacterial cells delivered in both pathogenic and non-pathogenic bacterial species. Inducibility of hepcidin transcripts were variable among three tissues examined (liver, kidney and spleen) in which kidney and spleen were more responsive to the bacterial challenge than liver. Time course expression patterns of hepcidin mRNAs after challenge were different between groups challenged with pathogenic and non-pathogenic species, although the overall pattern of hepcidin expression was in accordance with that generally observed in battery genes appeared during early phase of inflammation. Fish challenged with E. coli (non-pathogenic) showed the significant induction of hepcidin transcripts within 24 hr post injection (hpi) but the level was rapidly declined to the basal level either at 48 or 96 hpi. On the other hand, hepcidin transcript levels in E. tarda (pathogenic)-challenged fish were continuously elevated until 48 hpi, then downregulated at 96 hpi, although the level at 96 hpi was still significantly higher than control level observed in non-challenged fish. This expression pattern was consistent in all the three tissues examined. Taken together, our data indicate that hepcidin is tightly in relation with pathological and/or inflammation status during bacterial challenge, consequently providing useful basis to extend knowledge on the host defensive roles of hepcidin under infectious conditions in bony fish.

Freeze Tolerance Enhanced by Antifreeze Protein in Plant

  • Hwang, Cheol-Ho;Park, Hyun-Woo;Min, Sung-Ran;Liu, Jang-Ryol
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.4
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    • pp.339-343
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    • 2000
  • When plants are exposed to subfreezing temperatures ice crystals are forming within extracelluar space in leaves. The growth of ice crystal is closely related to the degree of freezing injury. It was shown that an antifreeze protein binds to an ice nucleator through hydrogen bonds to prevent growth of ice crystal and also reduces freezing damage. The antifreeze proteins in plants are similar to PR proteins but only the PR proteins induced upon cold acclimation were shown to have dual functions in antifreezing as well as antifungal activities. Three of the genes encoded for CLP, GLP, and TLP were isolated from barley and Kentucky bluegrass based on amino acid sequence revealed after purification and low temperature-inducibility as shown in analysis of the protein. The deduced amino acid of the genes cloned showed a signal for secretion into extracellular space where the antifreezing activity sup-posed to work. The western analysis using the antisera raised against the antifreeze proteins showed a positive correlation between the amount of the protein and the level of freeze tolerance among different cultivars of barely. Besides it was revealed that TLP is responsible for a freeze tolerance induced by a treatment of trinexapac ethyl in Kentucky bluegrass. Analysis of an overwintering wild rice, Oryza rufipogon also showed that an acquisition of freeze tolerance relied on accumulation of the protein similar to CLP. The more direct evidence for the role of CLP in freeze tolerance was made with the analysis of the transgenic tobacco showing extracellular accumulation of CLP and enhanced freeze tolerance measured by amount of ion leakage and rate of photosynthetic electron transport upon freezing. These antifreeze proteins genes will be good candidates for transformation into crops such as lettuce and strawberry to develop into the new crops capable of freeze-storage and such as rose and grape to enhance a freeze tolerance for a safe survival during winter.

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Molecular Analysis of the Salmonella Typhimurium tdc Operon Regulation

  • Kim, Min-Jeong;Lim, Sang-Yong;Ryu, Sang-Ryeol
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1024-1032
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    • 2008
  • Efficient expression of the Salmonella Typhimurium tdc ABCDEG operon involved in the degradation of L-serine and L-threonine requires TdcA, the transcriptional activator of the tdc operon. We found that the tdcA gene was transiently activated when the bacterial growth condition was changed from aerobic to anaerobic, but this was not observed if Salmonella was grown anaerobically from the beginning of the culture. Expression kinetics of six tdc genes after anaerobic shock demonstrated by a real-time PCR assay showed that the tdc CDEG genes were not induced in the tdcA mutant but tdcB maintained its inducibility by anaerobic shock even in the absence of tdcA, suggesting that an additional unknown transcriptional regulation may be working for the tdcB expression. We also investigated the effects of nucleoid-associated proteins by primer extension analysis and found that H-NS repressed tdcA under anaerobic shock conditions, and fis mutation delayed the peak expression time of the tdc operon. DNA microarray analysis of genes regulated by TdcA revealed that the genes involved in N-acetylmannosamine, maltose, and propanediol utilization were significantly induced in a tdcA mutant. These findings suggest that Tdc enzymes may playa pivotal role in energy metabolism under a sudden change of oxygen tension.