• Title/Summary/Keyword: [$^3$H]thymidine incorporation

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Comparison of Ag-NORs stain and [3H]thymidine incorporation in antiproliferative effect of caffeine on NIH3T3 cells (Ag-NORs 염색법과 [3H]thymidine incorporation법에 의한 caffeine의 NIH3T3 세포증식 억제효과 비교측정)

  • Kim, Sung-ho;Lee, Cha-soo
    • Korean Journal of Veterinary Research
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    • v.30 no.4
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    • pp.459-464
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    • 1990
  • Inhibitory effect of caffeine on NIH3T3 cell proliferation was studied by using [$^3H$]thymidine incorporation and a modified one-step silver staining technique. The latter technique shows argyrophilic nucleolar organizer region-associated proteins (Ag-NORs), which are seen in nuclei as black dots. The result was compared with the counts of [$^3H$] thymidine incorporation. The results were summarized as follows; 1. The Ag-NORs numbers of NIH3T3 cells were $6.81{\pm}1.38$ at 24 hrs, $7.13{\pm}1.26$ at 48 hrs, $8.50{\pm}2.04$ at 72 hrs after incubation. 2. The numbers of Ag-NORs were significantly decrease in caffeine treated groups (p<0.01). 3. The counts of [$^3H$] thymidine incorporation were similar to the result of using Ag-NORs staining technique. It is concluded that Ag-NOR is a rapid, simple and compatible method to quantitate cell proliferation as compared with [$^3H$]thymidine incorporation.

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Comparison of Methods for the Measurement of Bacterial Production in a Lake Ecosystem (호수 생태계에서 세균 생산량 측정 방법의 비교)

  • 김명운;강찬수;김상종
    • Korean Journal of Microbiology
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    • v.28 no.4
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    • pp.318-323
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    • 1990
  • The bacterial secondary production was measured at 6 sites of Lake Soyang in October, 1989 by $^{3}$H-thymidine incorporation rate. Verfication for the method of bacterial secondary production measurement showed that $^{3}$H-thymidine incorporated into DNA, RNA and protein by average percentage of 38.45, 42.27 and 20.07%, respectively. THe more increased incoporated $^{3}$H-thymidine, the more increasde DNA fraction, but protein fraction was generally low. Incorporation of rate of /usp 3/H-thymidine. $^{3}$H-leucine into protein correlated with protein fraction of incorporated $^{3}$H-thymidine. Conversion factors were calculated as follows; $1.83*10 ^{20}$ cells/moles of thymidine incorporated/hr and 1.69*10$^{22}$ cells/moles of leucine incorporated/hr.

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Effects of Glutamine, Glycine and Nucleosides/Nucleotide Mixture on Intestinal Mucosal Growth in Rats (흰쥐의 소장 점막 세포의 성장에 미치는 Glutamine, Glycine과 Nucleosides/Nucleotide 혼합물의 효과)

  • 이선영;오현인
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.1
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    • pp.130-136
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    • 1997
  • Total parenteral nutritional effect was induced by surgical creation of Thiry-Vella fistula(TVFs) in rats. Glutamine, glycine or nucleosides/nucleotide mixture in solution was injected into the loops for 2, 4, 6, 8 days. Control animals received a 0.9% saline solution. Results include weight gain, total protein, DNA, [$^3$H] thymidine incorporation into DNA, morphometry of the intestine in both TVFs and intestine in continuity. Perfusion of nucleosides/nucleotide mixture into the bypassed loops caused an increase in total protein, DNA content, villous height, villous surface area in loops. The injection of glycine into loops caused an increase in [$^3$H] thymidine incorporation but the mean values of the protein and DNA contents were not significantly different from those in group Cont and group Nuc. Overall values for group Gln were slightly higher than those of the control but the differences were not statistically significant. This study suggests that this animal model may be useful for studying the effect of dietary factors on intestinal growth and maturation, separating the direct effect of diet from systemic effect on the intestine.

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CHEMOSENSITIVITY OF CANCER CELLS TO ANTICANCER DRUGS USING DYE EXCLUSION ASSAY, [3H] THYMIDINE INCORPORATION, AND CLONOGENIC ASSAY (두경부악성종양세포주의 항암제감수성 시험에 관한 실험적 연구)

  • Jin, Woo-Jeong
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.15 no.1
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    • pp.35-48
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    • 1993
  • The in vitro predictive tests in cancer chemotherapy of cancer cell lines to anticancer drugs were determined using novel dye exclusion assay [NDEA], [3H] thymidine incorporation, and clonogenic assay [CA>. Antitumor effect of Bleomycin, Cis-platin, Vinblastine, Methotrexate to HEp-2, B16 cell lines using rapid assays was compared with [CA> in this study. In dye exclusion assay of B l6 cell line, cancer cells were sensitive to Bleomycin at all concentrations, to Vinblastine at the level of peak plasma concentration [PPC], ${\times}1/10$ [PPC](P<0.05). And Bleomycin revealed relatively good cytotoxicity than that of CDDP and vinblastine at ${\times}10$[PPC], (P<0.05). HEp-2 cells were resistive to methotrexate at the level of ${\times}100$[PPC] (P<0.05) In [3H] thymidine incorporation assay, B 16 cells were sensitive to Bleomycin, CDDP, Vinblastine at the level of [PPC], ${\times}10$ [PPC](P<0.01). Dose-dependent drugs of bleomycin, CDDP were more sensitive than Vinblastine at high concentration (P<0.05). In clonogenic assay, HEp-2 cell line was sensitive to three drugs of all concentrations except ${\times}10$ [PPC] of CDDP. B 16 cell line was sensitive to all drugs(P<0,01). In comparison of chemosensitivity tests among three assays, the results were correlated(${\gamma}=0.99$, P<0.05).

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Interaction of $17{\beta}-Estradiol$ with EGF and IGF-I on Proliferation and $P_i$ Uptake in Primary Cultured Rabbit Renal Proximal Tubular Cells

  • Han, Ho-Jae;Lee, Yeun-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.4
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    • pp.493-501
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    • 1998
  • The most significant direct role of estrogen in vivo is its ability to elicit receptor-mediated cellular proliferation in mammalian target tissues. However, the mechanism by which exogenously added estrogen causes the neoplastic transformation of renal cortical cells is yet to be uncovered. The present study was designed to evaluate interaction of $17{\beta}-estradiol\;(E_2)$ with epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I) on proliferation and $P_i$ uptake in primary cultured rabbit renal proximal tubular cells in phenol red-free, hormonally defined-medium. $[^3H]-thymidine$ incorporation increased markedly by about 133% and 141% more in the presence of $10^{-9}\;and\;10^{-6}\;M\;E_2$, respectively, than that of control. Cell count was 162% and 143% greater in the presence of $10^{-9}\;and\;10^{-6}\;M\;E_2$ , respectively, compared with control. Among all time points examined, there was an increase in $[^3H]-thymidine$ incorporation in the presence of $10^{-9}\;M\;E_2$ at day 9 or 13, respectively. However, $E_2$ ($10^{-9}\;M$) significantly drove up cell count to 160% of that of control at day 13, while it had a slight but statistically insignificant effect at day 9. $E_2-induced$ stimulation of $[^3H]-thymidine$ incorporation was completely reversed by $E_2$ antagonists (progesterone or tamoxifen). $E_2$ ($10^{-9}\;M$) or EGF ($10^{-8}\;M$) significantly stimulated $[^3H]-thymidine$ incorporation by 144% and 154% of control. $E_2$ plus EGF was synergistic on $[^3H]-thymidine$ incorporation (204% of control), while $E_2$ plus IGF-I showed a slight but no significant synergistic effect. Cell number also displayed similar pattern. $E_2$ ($10^{-9}\;M$) significantly stimulated $P_i$ uptake to 134% of control. $E_2$-induced stimulation of $P_i$ uptake was partially reversed by $E_2$ antagonists. EGF or IGF-I ($10^{-8}\;M$) significantly also increased $P_i$ uptake to 132% or 129% of control. $E_2$ plus EGF had synergistic effect on $P_i$ uptake, while $E_2$ plus IGF-I did not. In conclusion, $E_2$ may act not only directly interaction with its receptors but also indirectly as a modulator of EGF in proliferation and $P_i$ uptake of primary cultured rabbit renal proximal tubular cells.

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Antiproliferative Effects of Panu ginseng Glycosides on DNA Synthesis in Cultured Mouse Fibroblasts (인삼에서 추출한 글리코시드가 배양한 마우스 섬유아세포의 핵산합성에 미치는 영향)

  • Byun, Boo-Hyeong;Shin, In-Cheol;Lee, Lil-Ha;Joe, Cheol-O
    • Journal of Ginseng Research
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    • v.19 no.2
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    • pp.114-116
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    • 1995
  • Panax ginseng ginsenosides were examined for their affects on the DNA synthesis. The DNA 1 synthesis was measured by the thymidine incorporation into NIH3T3 cells. The ginsenoside, panaxytriol, $Rh_1$ and $Rh_2$ showed reduced [$^{3}H$]-thymidine incorporation. However, other ginsenosides of $Rh_1$, $Rh_2$ and $Rh_3$ did not inhibit DNA synthesis. Among the various ginsenosides, ginsenoside $Rh_2$ was found to be the most inhibitory on DNA synthesis. We suggest $Rh_2$ as one of the potential choice of antiproliferative drugs.

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THE EFFECTS OF MECHANICAL FORCE ON CULTURED PERIODONTAL LIGAMENT CELLS IN VITRO (물리적 외력이 배양중인 치주인대세포에 미치는 영향)

  • Kim, Hyun-young;Cha, Kyung-Suk
    • The korean journal of orthodontics
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    • v.24 no.2
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    • pp.295-301
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    • 1994
  • The movement of teeth during orthodontic treatment requires bone remodeling process in periodontal tissue. To find out the changes occuring in the cell itself, mechanical force was applied to the cultured periodontal ligament cells. Following results were obtained from measuring the changes in cyclic AMP and $PGE_2$, $^3H$-thymidine incorporation amount in time lapse after application of mechanical force. 1. When mechanical force was applied to cultured PDL cells, the amount of cAMP in cells were increased significantly after 15 min. of force application, but were decreased gradually as time lapsed. 2. When mechanical force was applied to cultured PDL cells, the amount of PGE2 were increased at 20,40,60 min. and was significantly increased at 20 min. 3. When mechanical force was applied to cultured PDL cells, the amount of $^3H$-thymidine incorporation was some increased, but was not statistically significant.

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Cytological Studies of Diapause in the Silkworm, Bombyx mori L. (가잠의 휴면성에 관한 세포학적 연구)

  • Park, Kwang E.
    • Journal of Sericultural and Entomological Science
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    • v.18 no.2
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    • pp.1-60
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    • 1976
  • In present study, radioautography and electron-microscopy were applied by the author to the elucidation of the mechanism of diapause in Bombyx mori L. (1). Patterns of nucleic acid and protein syntheses during embryonic development of silkworms, incubated at high and at low temperatures, were demonstrated by means of radioautography with labelled precursors of nucleic acid and protein. On the third day after blastokinesis the embryo incubated at high temperature generally incorporated much of the $^3$H-glycine into the brain and the suboesophageal ganglion, and some into other regions. When incubated at low temperature, no difference in the pattern between the brain, the suboesophageal ganglion and other parts was observed. Radioautography with $^3$H-thymidine revealed no significant difference in DNA synthesis in embryos incubated at high and low temperatures. In diapausing eggs twenty days after deposition, only a few cells of the mesoderm incorporated the labelled material into their nuclei, but in the hibernated eggs all the nuclei of the mesoderm had thymidine incorporation. After blastokinesis only the anterior portion of the embryo around the brain and the suboesophageal ganglion had thymidine incorporation; this was not observed in the posterior portion.

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Alkoxybenzylcyanoguanidine Analogs as a Novel Class of Inhibitors for Restenosis

  • Lee, Sun-Kyung;Yi, Kyu-Yang;Hwang, Sun-Kyung;Suh, Jee-Hee;Lee, Byung-Ho;Yoo, Sung-Eun
    • Bulletin of the Korean Chemical Society
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    • v.25 no.7
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    • pp.1003-1008
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    • 2004
  • A novel class of alkoxybenzylcyanoguanidine analogs as the inhibitors of restenosis was discovered, which showed the inhibitory effects on angiotensin II-induced cell proliferation, determined by $[^3H]$thymidine incorporation method. The compound, N'-(4-nitrophenyl)guanidine analog 19, showed 62% inhibition of $[^3H]$thymidine incorporation at 1 ${\mu}M$ concentration. In addition, the compound 19 inhibited intimal thickening dose-dependently after balloon injury, which suggests the therapeutic potential for restenosis.

Activity of the Extracts from Polyporus umbellatus as a Apoptosis Inducer (저령의 암세포증식 억제효능)

  • 신유진;김봉선;서용준;조덕제;최원일;이용규
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.6
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    • pp.1074-1077
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    • 2004
  • The effect of gradient eluted fractions from Et$_2$O extracts of Polyporus umbellatus screlotium was investigated on the viability of leukemia cell lines, K-562, L-1210, HL-60 and U-937 cells. Among those fractions, fraction 2 showed mild cytotoxic effect on L-1210 and HL-60 cells. Fraction 3 showed cytotoxic effect on 4 cell lines, and cytotoxic effect was the most potent on L-1210. The hallmark of apoptosis, DNA fragmentation, also appeared by fraction 3 on L-1210 after 48 hr treatment. Furthermore, this fraction was shown to be able to induce cell death on L-1210 cells by the inhibition of DNA synthesis in [$^3$H]thymidine incorporation test. From these results, P. umbellatus involves a potent chemical component that inhibits the viability of leukemia cell lines, L-1210. Further studies about the components of fraction 3 to function as a apoptosis inducer are necessary.