• Title/Summary/Keyword: thymidine

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In Vitro Cytotoxicity of Pt(II) Complexes Containing Ethylenediamine in Rabbit Kidney Proximal Tubular and Human Renal Cortical Cells (에틸렌디아민을 배위자로 한 백금(II)착체의 토끼 및 인체 신장세포에 대한 in vitro 독성)

  • Rho, Young-Soo;Lee, Kyung-Tae;Jung, Jee-Chang;Chang, Sung-Goo
    • YAKHAK HOEJI
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    • v.40 no.2
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    • pp.218-224
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    • 1996
  • This laboratory has recently reported the synthesis and in vitro antitumor activity of PT(II) complexes containing ethylenediamine and diphosphine. In view of the reports of others, cisplatin is toxic to the kidney since the kidney's vulnerability to PT(II) complexes may originate in its ability to accumulate and retain platinum to a greater degree than other organs. The in vitro cytotoxicity of these synthetic PT(II) complexes on the primary cultured proximal tubular cells of rabbit kidney and renal cortical cells of human kidney was investigated. Three endpoints for cytotoxicity tests were evaluated:3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), $^3H$-thymidine uptake and the glucose consumption tests. The rank order of sensitivity exhibited $^3H$-thymidine uptake>MTT>glucose consumption test. The agents with diphosphine leaving group were significantly less cytotoxic than cisplatin. Moreover, 1,2-bis(diphenylphosphino)ethane (DPPE) exhibited less cytotoxicity than 1.3-bis (diphenylphosphino)propane (DPPP) against on rabbit and human cultured kidney cells. Based on these results, the decreased nephrotoxicity of these new complexes over cisplatin appeared to be partially attributable to a leaving group of DPPP and DPPE. This novel class of platinum compound represents a valuable lead in the development of a "third-generation" agent.

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Lipocortin 1 Mediates the Suppressive Effects of Dexamethasone on ConA-induced Proliferative Response and Nitric Oxide Production in Rat Splenic Leukocytes

  • Jang, Yeon-Jin;Park, Hyoung-Sup;Kang, Soon-A;Yang, Sus-Jung;Na, Doe-Sun
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.1
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    • pp.91-96
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    • 1997
  • Lipocortin 1 has been proposed as a putative mediator of anti-inflammatory actions of glucocorticoids. We investigated the role of lipocortin 1 in the effect of dexamethasone using rat splenic leukocytes. Concanavalin A(ConA; 1 ${\mu}g/ml$) increased the leukocyte proliferation and nitric oxide(NO) generation, which were measured as $[^3H]-thymidine$ uptake by the cells and nitrite accumulation in the culture media, respectively. Dexamethasone suppressed ConA-induced cell proliferation, in a concentration-dependent manner with $EC_{50}$ around 50nM. The addition of anti-lipocortin l(Anti-LCl) reversed dexamethasone effects: 0.24, 1.2, 6 ${\mu}g/ml$ of Anti-LC1 reversed dexamethasone(50 nM)-induced suppression of thymidine uptake by $9{\pm}3%$, $16{\pm}3%$, $36{\pm}5%$, respectively; 0.24, 1.2, and 6 ${\mu}g/ml$ of Anti-LCI reversed dexa-methasone-induced decrease of nitrite concentration by $49{\pm}16%$, $61{\pm}20%$, $77{\pm}19%$, respectively. The present data indicate that lipocortin 1 mediates, at least in part, glucocorticoids-induced suppression of leukocyte proliferation and blockade of NO generation.

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The Effect of Epidermal Growth Factor on Cell Proliferation and Its Related Signal Pathways in Pig Hepatocytes

  • Kim Dong-Il;Han Ho-Jae;Park Soo-Hyun
    • Biomedical Science Letters
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    • v.12 no.3
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    • pp.249-254
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    • 2006
  • It has been reported that liver is a very important organ to xenotransplantation. Pig is known to be a most suitable species in transplantation of human organs. However, the physiological function of pig hepatocytes is not clear elucidated. Epidermal growth factor (EGF) is known to be a mitogen in various cell systems. Thus, we examined the effect of EGF on cell proliferation and its related signal cascades in primary cultured pig hepatocytes. EGF stimulates cell proliferation in a dose (>1ng/ml) dependent manner. EGF-induced increase of $[^3H]-thymidine$ incorporation was blocked by AG 1478 ($10^{-6}M$, an EGF receptor antagonist) genistein and herbymycin A (tyrosine kinase inhibitors, $10^{-6}M$), suggesting the role of activation and tyrosine phosphorylation of EGF receptor. In addition, EGF-induced increase of $[^3H]-thymidine$ incorporation was prevented by neomycin $(10^{-4}M)$, U73122 $(10^{-5}M)$ (phospholipase C [PLC] inhibitors), staurosporine ($(10^{-8}M)$, or bisindolylmaleimide I $(10^{-6}M)$ (protein kinase C [PKC] inhibitors), suggesting the role of PLC and PKC. Moreover, EGF-induced increase of $[^3H]-thymidine$ incorporation was blocked by PD 98059 (a p44/42 mitogen activated protein kinase [MAPK] inhibitor), SB 203580 (a p38 MAPK inhibitor), and SP 600125 (a JNK inhibitor). EGF increased the translocation of PKC from cytosol to membrane fraction and activated p42/44 MAPK, p38 MAPK and JNK. In conclusion, EGF stimulates cell proliferation via PKC and MAPK in cultured pig hepatocytes.

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A Study on the Relationship between the Uptake of $^{67}Ga-citrate$ and the Incorporation Rate of $^3H-thymidine$ and Metabolism of Protein in the Rat Livers Treated with $CCl_4$ (사염화탄소투여(四鹽化炭素投與)후 백서간세포(白鼠肝細胞)에서 $^{67}Ga$섭취율(攝取率)과 $^3H-thymidine$ 결합율(結合率) 및 단백대사(蛋白代射)와의 관계(關係)에 관(關)한 연구(硏究))

  • Hong, Seong-Woon
    • The Korean Journal of Nuclear Medicine
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    • v.19 no.1
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    • pp.83-93
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    • 1985
  • The ability of $^{67}Ga$, administered carrier free as the citrate complex, to localize in human and animal tumors to an extent sufficient to permit visualization of the lesion by scanning is well established. However, neither the mechanism of $^{67}Ga$ uptake by tumors or inflammatory cells nor its relationship to cell type or to the biochemical status of the cell is yet understood. Author investigated the uptake of $^{67}Ga-citrate$ using subcellular tissue fractionation of rat livers treated with $CCl_4$ associated with the $^3H-thymidine$ incorporation rate to detect subcellular localization of $^{67}Ga$ and it's relationship in DNA synthesis. Large amounts of $^{67}Ga$ associated with the soluble portion of tissue homogenate rather than with isolated cell organelles and not related nuclei residue in the regenerating period after hepatocellular injury caused by $CCl_4$. The elevated uptake of $^{67}Ga$ in the livers of $CCl_4$ treated rats was also inhibited when protein synthesis was stopped by cyclohexamide. Thus protein and the soluble portion of issue homogenates seems to play an important role in the elevated uptake of $^{67}Ga$ in liver injury induced by $CCl_4$ treated rats.

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Constructions of a Transfer Vector Containing the gX Signal Sequence of Pseudorabies Virus and a Recombinant Baculovirus

  • Lee, Hyung-Hoan;Kang, Hyun;Kim, Jung-Woo;Hong, Seung-Kuk;Kang, Bong-Joo;Song, Jae-Young
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.541-547
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    • 1999
  • Constructions of a transfer vector and a recombinant baculovirus using the thymidine kinase gene of the Herpes simplex virus type 1 strain F (HSV -1) were carried out. Newly cloned transfer vector, pHcgXIIIB, was constructed by insertion of the glycoprotein gX gene signal peptide sequence of Pseudorabies virus into the baculovirus vector pHcEV-IV. The gX sequence was inserted just downstream from the promoter for the polyhedrin gene of the Hyphantria cunea nuclear polyhedrosis virus (HcNPV). HSV-1 thymidine kinase(tk) gene (1.131 kb) was used as a candidate gene for transferring into the baculovirus expression system. The tk gene was inserted into a BamHI site downstream from the gX sequence-promoter for the polyhedrin gene in the pHcgXIIIB transfer vector and was transferred into the infectious lacZ-HcNPV expression vector. Recombinant virus was isolated and was named gX-TK-HcNPV. The recombinant virus produced a 45 kDa gX-TK fusion protein in Spodoptera frugiperda cells, which was confirmed by Western blot analysis. Microscopic examination of gX-TK-HcNPV-infected cells revealed normal multiplication. Fluorescent antibody staining indicated that the gX-TK fusion protein was present in the cytoplasm. These results indicated that the transfer vector successfully transferred the gX-tk gene into the baculovirus expression system.

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A STUDY ON THE CYTOTOXICITY OF THE ORTHODONTIC BONDING MATERIALS (교정용 접착제의 세포독성에 관한 실험적 연구)

  • Sa, Myung-Hee;Yang, Won Sik
    • The korean journal of orthodontics
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    • v.22 no.1
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    • pp.147-158
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    • 1992
  • This study was aimed to compare the relative cytotoxicity of the five common orthodontic bonding materials (Concise. Mon-lok, Ortho-One, Super C, Transbond) using cell culture technique. DNA synthesis of the fibroblasts was assessed by $^3H$-thymidine uptake to evaluate the effect of the bonding materials on the growth of the cells. The human gingival fibroblasts were explanted from the buccal gingiva of 10 year-old girl and cultured in $\alpha$-MEM/10% FBS/1% antibiotics medium, $37^{\circ}C$, 5% $CO_2$ incuvator. The gingival fibroblasts were tested with the medium into which the bonding materials had been soaked for 1 week. Or the bonding materials were placed on the cells immediately or 2 weeks after polymerization. After 22 hours, $^3H$-thymidine was added into the microtest wells and after 24 hours, the uptake of $^3H$-thymidine was determined by liquid scintilation counter. The results of this study were as follows. 1. DNA synthesis was significantly decreased with Super C and Transbond than Ortho-One, when treated with medium into which the bonding materials had been soaked for 1 week. 2. DNA synthesis was significantly decreased with Concise, Super C and Transbond than control, when treated immediately after polymerization. 3. DNA syntehsis was significantly decreased with Concise, Super C and Transbond than Ortho-One, when immediately after polymerization. 4. There was no significant difference in DNA synthesis between the bonding materials, when treated 2 weeks after polymerization.

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Cloning, Sequencing and Expression in Escherichia coli of Herpes simplex virus Type-1 Thymidine Kinase Gene

  • Lee, Hyung-Hoan;Kim, Jung-Woo;Kang, Hyun;Cha, Sung-Chul
    • The Journal of Korean Society of Virology
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    • v.28 no.3
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    • pp.215-224
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    • 1998
  • Cloning, sequencing and expressing in E. coli of the thymidine kinase (TK) gene of Herpes simplex virus type-1 (HSV-1) strain F was investigated. The TK gene, located in the BamHI 3.74 kb DNA fragment of the plasmid pHLA-12, was amplified by polymerase chain reaction (PCR). The 1,131 kb PCR product was cloned into the BamHI and EcoRI sites of pBacPAK9 plasmid and then named pBac-TK recombinant. The TK gene was subcloned into the BamHI and BglII sites of pQE-30, and named pQE-TK recombinant. The nucleotide sequence of the 1,131 kb TK gene was determined, and the GC content was 65.13%. There were deduced 367 amino acid residues with a total molecular weight of 43 kDa. The weight was confirmed by the protein produced by E. coli M15/pQE-TK on the SDS-PAGE and Western blot. The production of the TK protein in the IPTG induced cells was measured over 4 h. At the end of 1, 2 and 3 h the level increased by 146, 204 and 242%, respectively. The amount of the protein at the highest fraction purified with Ni-NTA resin chromatography was $0.68\;{\mu}g$ per ml. The soluble state TK protein was present in the cytoplasm. In these results the F strain was different in base sequence and amino acid sequence from that of the CL101 strain, which caused difference in their strains.

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Effects of Gamgung-tang on Proliferation and cAMP Accumulation of Thyroid Cells (감궁탕이 갑상선세포의 증식과 cAMP 축적에 미치는 영향)

  • Kim, Mee-Kyung;Shon, Yun-Hee;Nam, Kyung-Soo;Son, Ok-Lye;Kim, Cheorl-Ho;Jeon, Byung-Hun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.3
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    • pp.623-627
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    • 2005
  • Abnormal thyroid cell proliferation has a very important role in hypothyroidism. Thyroid stimulating hormone (TSH) stimulates proliferation and maintains differentiated function in thyroid follicular cells. A functioning rat thyroid cell line (FRTL) was used to study the effect of Gamgung-tang (GGT, Glycyrrhizae Radix, black beans, Angelicae Radix and Cnidii Rhizoma) on proliferation and cAMP accumulation of thyrocytes. Proliferation of cell was assessed by DNA synthesis and incorporation of $[^3H]thymidine$ into DNA. The concentration of cAMP was measured simultaneously with growth assessment. Extract of GGT ($0.15{\sim}0.9\;mg/ml$ increased DNA synthesis in a dose-dependent manner. GGT at 0.6 (p<0.05) and 0.9 mg/ml (p<0.01) significantly increased $[^3H]thymidine$ incorporation. A comparable effect was observed with TSH. GGT also enhanced cAMP accumulation. These results indicate that GGT increases the proliferation of thyrocytes and may be considered a promising agent for the treatment of autoimmune thyroid disease.