• Korean Journal of Weed Science
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• v.10 no.4
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• pp.328-337
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• 1990

The incividual and combined effects of the chloroacetanilide herbicide pretilachlor and of the safener fenclorim on the growth and selected physiological processes of rice (Oryza sativa L., var 'Lemont')were evaluated under greenhouse and laboratory conditions. Fenclorim applied at rates ranging from 50 to 300 g a.i./ha antagonized the injurious effects caused by 150 to 900 g a.i./ha of pretilachlor on 15-day old wet-sown rice grown under greenhouse conditions. When used rates of 150 g/ha or higher, fenclorim reversed completely the effects of all doses of pretilachlor on rice. When the two compounds were given simultaneously, fenclorim enhanced the uptake of $^{14}C$pretilachlor into rice leaf mesophyll protoplasts measured for 1 hr, indicating that competition for uptake at the protoplast level is not involved in the protective action of this safener. The safener-induced stimulation of pretilachlor uptake was particularly evident when fenclorim was used at concentrations of 10, 20 and $40{\mu}M$. Following 4 hr of incubation, individual treatments with pretilachlor inhibited the in vitro incorporation of radiolabeled precursors into proteins, DNA, and lipids of rice leaf protoplasts only when used at the high concentration of $100{\mu}M$M. Individual treatments with high concentrations (10 or $100{\mu}M$) of the safener fenclorim inhibited the incorporation of radiolabeled precursors into proteins and lipids of rice protoplasts, but had no DNA synthesis. The combined effects of pretilachlor and fenclorim on the incorporation of radiolabeled precursors into these macromolecules of isolated rice mesophyll protoplasts appeared to be additive or slightly synergistic rather than antagonistic. Fenclorim at $1{\mu}M$ antagonized the effects of pretilachlor on total lipids of rice leaf protoplasts. In addition, individual and combined treat-menu with pretilachlor and fenclorim influenced the incoroporation of$^{14}C$acetate into polar lipids, triglycerides and steryl esters of rice leaf protoplas causing a redistribution of carbon in these lipid fractions. However, these effects were not large enough to explain the herbicidal activity of pretilachlor or to account for the protective action of the safener fenclorim. Overall, the uesults of the present study idnicate that the safener fenclorim does not seem to protect rice against pretilachlor injury by antagonizing its effects on protein, DNA, or lipid syntheses.

• Molecular & Cellular Toxicology
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• v.1 no.2
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• pp.92-98
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• 2005

DNA microarray analysis of gene expression in toxicogenomics typically requires relatively large amounts of total RNA. This limits the use of DNA microarray when the sample available is small. To confront this limitation, different methods of linear RNA amplification that generate antisense RNA (aRNA) have been optimized for microarray use. The target preparation strategy using amplified RNA in DNA microarray protocol can be divided into direct-incorporation labeling which resulted in cDNA targets (Cy-dye labeled cDNA from aRNA) and indirect-labeling which resulted in cRNA targets (i.e. Cy-dye labeled aRNA), respectively. However, despite the common use of amplified targets (cDNA or cRNA) from aRNAs, no systemic assessment for the use of amplified targets and bias in terms of hybridization performance has been reported. In this investigation, we have compared the hybridization performance of cRNA targets with cDNA targets from aRNA on a 10 K cDNA microarrays. Under optimized hybridization conditions, we found that 43% of outliers from cDNA technique and 86% from the outlier genes were reproducibly detected by both targets hybridization onto cDNA microarray. This suggests that the cRNA labeling method may have a reduced capacity for detecting the differential gene expression when compared to the cDNA target preparation. However, further validation of this discordant result should be pursued to determine which techniques possesses better accuracy in identifying truly differential genes.

• 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.

• Archives of Pharmacal Research
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• v.16 no.4
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• pp.259-264
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• 1993

Three cell lines, CHO, L929 and B16 which are non-tumorigenic and cancer cells, respectively, were first tested for their survival in the presence of radioprotective ginseng protein fraction(GPF0. The influence of three radioprotectors-CPF, cysteamine, and 1-Methyl-2-bis[(2-methylthio)vinyl] quinolinium iodide (MVQI) on DNA repair capacity of UV damaged cells survival test, the GPF showed higher cytotoxicity in L929 and B16 than in CHO cells. However, the degree of cell killing was also investigated by measuring $^3H$-thymidine incorporation of PUVA treated cells. In cell survival test, the GPF showed higher cytotoxicity in L929 and B16 than in CHO cells. However, the degree of cell killing was not high enough to consider it as an antitumorigenic agent. Variable results were obtained in the effects on DNA repair capacity depending on the protectors and cell lines used. In pretreatment, the presence of GPF and MVOI brought about a sinificant increase in the capacity in both CHO and B16 cells. However, in L929, the enhancing effect was not shown. In all three cell lines, cysteamine showed lower repair capacity than control, suggesting the primary damage reduction in stronger enhancing effects in L929 and B16 cells, while it was weaker in CHO cells. Here also cystemine hsowed a very little or no increase in the capacity in all three cell lines. These results demonstrate that GPF has mild cytotoxicity in tumorignic cells and that GPF and MVQI enhance DNA repair capacity of UV damaged cells, whether they are tumorigenic or not. On the other hand, cysteamine shows only damage reduction effect. Celles of different genetic origin seem to give different responses to the modifier and different modifiers may possibly work by different mechanisms.

• BMB Reports
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• v.36 no.1
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• pp.12-19
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• 2003

Fragmentation of purine imidazole ring and production of formamidopyrimidines in deoxynucleosides (Fapy lesions) occurs upon DNA oxidation as well as upon spontaneous or alkali-triggered rearrangement of certain alkylated bases. Many chemotherapeutic agents such as cyclophosphamide or thiotepa produce such lesions in DNA. Unsubstituted FapyA and FapyG, formed upon DNA oxidation cause moderate inhibition of DNA synthesis, which is DNA polymerase and sequence dependent. Fapy-7MeG, a methylated counterpart of FapyG-, a efficiently inhibits DNA replication in vitro and in E.coli, however its mutagenic potency is low. This is probably due to preferential incorporation of cytosine opposite Fapy-7MeG and preferential extension of Fapy-7MeG:C pair. In contrast, FapyA and Fapy-7MeA possess miscoding potential. Both lesions in SOS induced E.coli preferentially mispair with cytosine giving rise to A$\rightarrow$G transitions. Fapy lesions substituted with longer chain alkyl groups also show simult aneous lethal and mutagenic properties. Fapy lesions are actively eliminated from DNA by repair glycosylases specific for oxidized purines and pyrimidines both in bacteria and eukaryotic cells. Bacterial enzymes include E.coli formamidopyrimidine-DNA-glycosylase (Fpg protein), endonuclease III (Nth protein) and endonuclease VIII (Nei protein).

• Rapid Communication in Photoscience
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• v.3 no.4
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• pp.76-78
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• 2014

Fluorescent nucleic acids were prepared utilizing the polymerase extension (PEX) reaction to incorporate fluorescent molecules. 2'-Deoxyuridine triphosphate (dUTP) derivatives possessing pyrene molecules as fluorophores were synthesized using the aqueous-phase Sonogashira coupling between 5-Iodo-dUTP and acetylene-linked pyrene molecules. The incorporation of the pyrene (Py)-labeled deoxyuridine triphosphates (PyU) into DNA by polymerase was evaluated by polyacrylamide gel electrophoresis, demonstrating that the PyU can work as a good substrate for the PEX reaction. The fluorescent properties of the functionalized DNA prepared by the PEX reaction were characterized by steady-state fluorescence measurements. The Py-conjugated DNA showed typical emission spectra of the pyrene, and the DNA with two pyrene molecules connected to each other by a diethylene glycol linker exhibited a broadened emission attributed to the electronic interaction between the Py molecules.

• The Journal of Korean Medicine
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• v.20 no.3 s.39
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• pp.54-65
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• 1999

Objectives: Hepatoma is a very serious disease in Korea and vvorldwiclc. Hepatitis B vims (HBV) has proved the most significant cause of hepatoma. We canied out this study to investigate the effect of Acanthopanax sessilifloms on inhibiting cell proliferation and DNA synthesis in HepG2.2.15 cell line and on inhibiting phosphorilation of oncogene (MAP kinase) in NIT/3T3-HBx ceIl. Methods: To investigate the anti-cancer effect of Acanthopanax sessiliflorus, we did the CellTiter 96 Aqueous Non-radioactive Cell Proliferation assay (Promega); MTS/PMS assay, [$^3H$]-thymicline incorporation assay, and we measured the gene expression through westem blotting. Results: Acanthopanax sessiliflorus showed an inhibiting effect on the increase of HepG2.2.15 in the NTS/PMS assay. It also showed an inhibiting effect on DNA synthesis of HepG2.2.15 in the [$^3H$]-thymidine incorporation assay. Acanthopanax sessiliflorus showed an inhibiting effect of phosphorilation of MAP kinase in HBV - X genes. too. Conclusions: The results suggested that this herb had an anti cancer effect. We may discover an effective anti-cancer herb medicine through further studies on this herb medicine.

• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.881-884
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• 1999

The gene encoding the RNA-dependent RNA polymerase of the hepatitis C virus was cloned and expressed with a C-terminal hexahistidine tag. The protein was purified from Escherichia coli to near homogeneity and characterized in vitro. When the 21 amino acids from the C-terminus of the protein were deleted, an inclusion body was not formed and a better purification yield was achieved. However, the activity of the purified enzyme decreased compared to that of the full length protein. The purified enzyme did exhibit ribonucleotide-incorporation activity on an in vitro transcribed RNA containing the 3' end of the HCV genome. It also possessed ribonucleotide incorporation activity, to a lesser extent, on in vitro transcribed foreign RNA templates when RNA or DNA primers were present. The activity was higher with DNA primers than with RNA primers. Accordingly, this assay system will facilitate the screening of inhibitors for hepatitis C virus replication.

• The Korean Journal of Zoology
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• v.20 no.4
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• pp.159-168
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• 1977

In order to evaluate the mutagenic potentential in mammalian system for those pesticides which were proved to be mutagenic in Salmonella microsome assay system, we have studied drug-resistant mutagenesis in cultured L5178Y cells and unscheduled DNA synthesis in human lymphocytes in vitro. We have tested five pesticides: insecticides DDVP and trichlorfon, fungicide TMTD and herbicides MO and NIP. Of these pesticides, TMTD induced weak mutation to MTX-resistance in L5178Y cells in vitro and gave positive responses in DNA repair assay system. Therefore, its potential genetic risks in human beings should be re-evaluated. DDVP and trichlorfon gave negative response in L5178Y mutagenesis test system but stimulated incorporation of $^{3}H$-TdR in DNA repar assay. MO and NIP gave also negative responses both in L5178Y mutagenesis test systemand in DNA repair assay system.

• Journal of Microbiology and Biotechnology
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• v.16 no.6
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• pp.933-938
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• 2006

Decaprenyl diphosphate synthase (DPS) is the key enzyme for the production of coenzyme $Q_{10}$ ($CoQ_{10}$). A dps gene from Sinorhizobium meliioti KCCM 11232 (IFO 14782) was isolated by PCR and then cloned in Escherichia coli. DNA sequencing analysis revealed an open reading frame of 1,017 bp encoding a 338-amino-acid protein. The protein was identical at the 98% level to the putative octaprenyl diphosphate synthase (IspB) of S. meliloti 1021. The deduced amino acid sequence included the DDxxD domains conserved in the majority of the prenyl diphosphate synthases. Heterologous expression in E. coli BL21 (DE3) was carried out, and the $CoQ_{10}$ produced was then analyzed by HPLC. E. coli BL21 (DE3) harboring the dps gene from S. melioti produced CoQ$_{10}$ in addition to endogenous coenzyme Q$_8$ (CoQ$_8$), whereas wild-type E. coli BL21 (DE3) host did not have the ability of producing CoQ$_{10}$. The results suggest that the putative dps from S. meliloti KCTC 2353 encoded the DPS.